PLENARY SESSION ABSTRACTS Theme: IMMUNITY AND ...

PLENARY SESSION ABSTRACTSTheme: IMMUNITY AND AUTOIMMUNITY

State-of-the-Art AddressWhat’s new in autoimmune blistering diseases?

D. F. MURRELLDepartment of Dermatology, St George Hospital, andUNSW Faculty of Medicine, Sydney, New South Wales,Australia

There are several blistering diseases which occur natu-rally in other species as well as in humans; for example,pemphigus occurs naturally in dogs and horses and theinherited blistering disease, epidermolysis bullosa, alsooccurs in dogs. Several new validated scoring systemsto measure the severity of autoimmune blistering dis-ease (AIBD) have been developed which assist indemonstrating efficacy of new treatments, such as thePemphigus Disease Area Index (PDAI) for pemphigusand Bullous Pemphigoid Disease Area Index (BPDAI)for pemphigoid. Pemphigus is due to autoantibodies todesmogleins 1 and 3 in human pemphigus foliaceus andvulgaris and desmocollin1 in canine pemphigus foli-aceus, generated by the late onset activation of theadaptive immune response. Paraneoplastic pemphigus,in which there are also antibodies to plakin proteins inthe skin and mucosae, also has occurred in canines.The B-cell CD-20 inhibitor, Rituximab, with short-course steroids has been shown to be more effectiveand safer than traditional high-dose steroids alone in alarge human pemphigus RCT.Pemphigus also involves the innate immune response,with induction of interleukin (IL)-1 and IL-6. A canineversion of the PDAI has been developed and used in apivotal study of the first Bruton Tyrosine Kinase Inhi-bitor (BTKI), which can inhibit both the innate andadaptive immune response, to be used in an AIBD, indogs, as solo open-label treatment. Two BTKIs givenorally once daily demonstrated excellent efficacy andsafety, as proof-of-concept for trials in patients withpemphigus. Open-label Phase 2 studies in pemphigushave shown rapid responses with rilzabrutinib and thephase 3 PEGASUS trial is recruiting. CAR-T cell trialsin pemphigus are also ongoing. Bullous pemphigoid(BP) is increasing in incidence, particularly in patientsover 70 years. Case control studies have shown thatprior neurological damage, particularly multiple sclero-sis, and also dementia and Parkinson disease, increasethe risk. Multiple medications increase the risk of trig-gering BP, particularly gliptins used to treat diabetesmellitus. Eotaxin 1, IL-17 and IL-4/13 are pathwayswhich are increased in BP and trials of new therapiestargeting these pathways are underway. Epidermolysisbullosa acquisita (EBA) develops due to autoantibodiesto collagen VII, the anchoring fibril protein present inskin and stratified mucosae. Inflammatory forms ofEBA have been reproduced in mice and respond tocomplement inhibitors and inhibition of the Fli-1 path-way, which is increased in the EBA mouse and inhereditary dystrophic EB, a disease with gene defects incollagen VII.Source of funding: Self-funded.

Conflict of interest: None declared.

Supporting ReviewEpithelial, immune cell and microbial cross-

talk in homeostasis and atopic dermatitis

T. KOBAYASHILaboratory for Innate Immune Systems, RIKEN Centerfor Integrative Medical Sciences (IMS), Yokohama,Japan

Skin is a complex and dynamic ecosystem, wherein theepithelial cells, immune cells and microbiota engage inactive dialogues and maintain barrier integrity andfunctional immunity. Alterations of the peaceful coexis-tence with the resident microbiota, referred to as dys-biosis, lead to dysregulation of host immunity. It hasbeen long debated whether the dysbiosis in the skin ofatopic dermatitis is merely a consequence of chronicskin inflammation or whether it is actively involved indriving skin inflammation. Microbiome analysis by 16SrRNA sequencing in humans and dogs with atopic der-matitis showed the shifts in microbial diversity repre-sented by increased proportion of Staphylococcus spp.Direct evidence has been found in mice deficient in adisintegrin and metalloproteinase 17 (ADAM17), whichspontaneously developed eczematous dermatitis withdysbiosis that was predominated by S. aureus andCorynebacterium spp. Targeting the dysbiotic flora withan antibiotic cocktail reversed dysbiosis and extin-guished eczematous inflammation, suggesting crucialroles of dysbiotic flora during atopic inflammation.Symbiotic relationships between the host and micro-biota must be constitutively maintained. Detailed mech-anisms on how host immunity regulates commensalbacteria in the steady state are now being revealed.Skin harbours a myriad of tissue-resident immune cellswhich include both innate and adaptive immune cells.Recent studies have highlighted a fundamental role ofinnate lymphoid cells (ILCs) in the maintenance of bar-rier functions and tissue homeostasis. A study in micerevealed that ILCs and T lymphocytes differentially reg-ulate skin microbiota. In particular, epidermal ILCs reg-ulate production of antimicrobial lipids from sebaceousglands to maintain homeostatic balance of skin micro-biota. Cross-talk between epithelial cells and immunecells determines immunological tones in the barrier tis-sue. ILCs directly respond to tissue-derived signals andplay an essential role in barrier immunity. Epithelial cellsproduce alarmins such as thymic stromal lymphopoietin(TSLP), interleukin (IL)-33 and IL-25, all of which acti-vate group 2 ILCs (ILC2s), which produce type 2 cytoki-nes such as IL-5 and IL-13, and boost type 2 immunereactions against parasitic infections, allergens, mites andtoxins. Dysregulation of epithelial–ILC cross-talk resultsin allergic inflammation. Single cell RNA-sequencinganalysis uncovered unique characteristics of skin ILCsand the regulatory mechanisms in the context of atopicinflammation. Understanding active interactions betweenthe host epithelial and resident immune cells and micro-biota provides a foundation of novel therapeutic strate-gies of inflammatory skin diseases.Source of funding: Self-funded.


© 2020 The Authors. Veterinary Dermatology © 2020 ESVD and ACVD,

Veterinary Dermatology, 31 (Suppl. 1), 6–109

Theme: ALLERGY AND ATOPIC DERMATITIS

State-of-the-Art AddressA world of inflammation: ecological solutions

for people, place and planet

S. L. PRESCOTTSchool of Paediatrics and Child Health, University ofWestern Australia, Perth, Western Australia, Australia

The ecology of the early environment determines life-long health, including microbial diversity, nutrition,nature, social interactions and the wider “exposome”.Almost all of these factors impacting immunity hadhave implications for all aspects of health and reliance.In particular, the emergence of “microbiome science”provides new evidence of vital relationships betweenbiodiversity and health at every level. Allergy was thefirst discipline to link adverse changes in early life ecol-ogy with the epidemic of immune disease – with muchwider implications for other systems. New perspectivesof ecological interdependence connect personal andplanetary health; the human health crisis cannot be sep-arated from the social, political and economic “ecosys-tems” otherwise driving “dysbiosis” (life in distress) atevery level. Changes in macroscale ecology – of foodsystems, lifestyle behaviours, socioeconomic disadvan-tage and environmental degradation – all impact themicrobial systems sitting at the foundations of allecosystems. In particular, changes in the function andcomposition of the human-associated microbiome hasbeen implicated in the mounting global burden of non-communicable diseases (NCDs), exacerbating inflam-mation and metabolic dysregulation through multiplepathways along lifespans. This underscores the need forecological approaches aimed at restoring symbiosis, bal-ance and mutualism. Although there is promise withsupplement-based strategies (e.g. probiotics, prebiotics),it is essential to focus on upstream factors implicated indysbiosis, including the health of wider environments,lifestyle, nature relatedness, and the social policies andpractices which can facilitate or inhibit “dysbioticdrift”. This underscores the necessity for ambitiousintegrative approaches which not only define theseinterconnections, but also capitalize on them to createnovel, collaborative and mutualistic solutions to ourvast interdependent global challenges.Source of funding: Self-funded.


Supporting Review

Advances in our understanding of canine atopic

dermatitis

R. MARSELLADepartment of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA

Much progress has been made in our understandingcanine atopic dermatitis (cAD), a clinical syndromewith intricate pathogenesis. Canine Atopic dermatitisresults from the complex interaction between geneticand environmental factors. It is becoming clear thatcAD is not a single disease but rather a clinical

syndrome, and that different genetic alterations play arole in certain breeds and geographical locations andnot in others. Disease development is modulated by theinteraction between skin barrier alterations, lifestyleconditions, microbiome and immunological responses.As is the case in people, dogs living on a farm withincreased exposure to the outdoors and other dogs havedecreased risk of development of disease. This likelyresults from a modulation of immunological responseby increased exposure to beneficial bacteria andincreased biodiversity. Skin barrier impairment plays arole in promoting cutaneous dysbiosis, increased aller-gen penetration and excessive T helper 2 (Th2)response. It is under investigation whether primarydefects exist in dogs, as reported in people, or whetheralterations are purely secondary to inflammation. Doc-umented abnormalities range from ultrastructural (e.g.disorganized lipid lamellae) to chemical (e.g. decreasedceramide levels) and functional (e.g. increased transepi-dermal water loss). These alterations are present in clin-ically normal atopic skin and are intensified in lesionalatopic skin. It is important to stress that clinically nor-mal atopic skin is not the same as normal skin and thatlow-grade inflammation is present in atopic skin, thusmaking possible that these changes are secondary.Future studies should include control groups withinflammatory diseases that are not AD (besides healthycontrols) to distinguish between changes that are simplyconsequences of inflammation and those that are speci-fic signatures of atopic disease. Decreased transepithe-lial electrical resistance of monolayers of atopickeratinocytes in culture suggests a role of tight junctionsor other proteins involved in cell–cell connection. Pro-teins important for keratinization such as filaggrin(s)have been investigated. In genetic studies, it was foundthat filaggrin single nucleotide polymorphism (SNP)may play a role in some breeds and not others. Ofinterest is a SNP in the gene encoding for thymic stro-mal lymphopoietin (TSLP) receptor, which wasdetected in all tested breeds. TSLP is an important linkbetween skin barrier damage and modulation of Th2response. Interestingly, staphylococcal wall componentscan induce transcription of TSLP. Documented defectsof extrusion of antimicrobial peptides potentially canexplain reduced killing activity in atopic patients andthe important role of Staphylococcus in promotingflares of AD. Decreased biodiversity and increasedstaphylococcal colonization is present in atopic skinand aggravated during active flares. A dysregulatedimmune response in cAD includes increased Th2, Th17and CD4+ CD25+ regulatory T cells. Interleukin (IL)-31 has been identified as an important mediator of pru-ritus and as a possible biomarker for disease severity.As results have varied among studies, more work isneeded to evaluate IL-31 expression in various pheno-types of cAD. In summary, our understanding hasexpanded from viewing cAD as a histamine-drivenType I hypersensitivity to a very complex multifacetedsyndrome.Source of funding: Self-funded.


Abstracts 7



Theme: ECOSYSTEM AND INFECTIOUS DISEASE

State-of-the-Art AddressMeticillin-resistant Staphylococcuspseudintermedius – this is your life

D.J. TROTT*, J.M. NORRIS†, J. KOVAC* and S.ABRAHAM‡

*Australian Centre for Antimicrobial ResistanceEcology, School of Animal and Veterinary Sciences, TheUniversity of Adelaide, South Australia, Australia;†School of Veterinary Science, University of Sydney,New South Wales, Australia; ‡Antimicrobial Resistanceand Infectious Diseases Laboratory, School ofVeterinary Life Sciences, Murdoch University, Murdoch,Western Australia, Australia

In the year 2020, meticillin-resistant Staphylococcuspseudintermedius (MRSP) is officially 21 years of age.Although first reported in Spain in 1995, it was notuntil 1999 that a MRSP strain, isolated from a dog inthe United States, was confirmed to contain the mecAgene. At such an auspicious coming of age, it is timelyto look back on the key developments in our under-standing of MRSP evolution as well as crystal-ball-gazeinto the future now MRSP has become endemic inmost countries. This review will cover the key aspectsof MRSP’s “birth” and “adolescence” (i.e. moleculargenetics, clonal diversity and epidemiological spread)and summarise some of the possible reasons for its“global success”. Key “newsworthy” events, cautionarytales and successful treatment of problem casesthroughout MRSP’s “life” will be explored, includingdevelopment of resistance to critically important andtopically applied antimicrobial agents. Given that Syd-ney NSW was to be the location for the World Veteri-nary Dermatology Conference, a uniquely Australianview on MRSP also will be included. It will concludewith a summary of what we have learned thus far andwhat technological developments such as Next-Genera-tion Sequencing (NGS) and rapid susceptibility testingare available to help us devise the treatment and con-trol plans of the future.The first bacterial genome (Haemophilus influenzae) wassequenced in 1995. In 2005, the estimated cost for gen-erating a bacterial genome sequence was approximatelyAU$50,000. Since then, the availability of rapid andaccurate NGS platforms such as Illumina MiSeq andan array of free, online bioinformatics tools has almostmade whole genome sequencing cheap enough torequest as a diagnostic test on a single isolate. IlluminaMiSeq generates large numbers of short DNA readsideal for quickly and accurately interrogating the gen-ome for multilocus sequence types, antimicrobial resis-tance and virulence genes. Additionally, screeningmultiple genomes for single nucleotide polymorphismsand accessory genes provides unprecedented power forunderstanding clonality, epidemiological spread and theinfluence of genetic recombination on MRSP popula-tion structure. More expensive deep sequencing, how-ever, is still required to close genomes and tounderstand the genomic context of identified mobilegenetic elements. The first reported fully annotatedMRSP genome sequence, from a ST71 strain isolatedfrom a canine bite wound in Denmark, was announced

in 2013. In little more than seven years, recent papershave now compared NGS data for large numbers(>100) of MRSP isolates. Whilst the population struc-ture of MRSP is highly clonal, many MRSP cloneshave acquired important genes such as those encodingantimicrobial resistance through multiple, independent,horizontal gene transfer events. Furthermore, strainswithin the epidemiological successful clones ST71 andST68 contain unique prophage-mediated resistance andvirulence genes that may aid in fitness and clonalspread. In all ST71 strains, prophages inserted intogenes affecting natural competency, prevent acquisitionof horizontal gene flow by genetic transformation andmay further drive clonal spread. These results, onlymade possible through breakthroughs in the analysis ofthe pangenome, are shedding new light on the reasonsfor MRSP’s “global success”.Source of funding: Research on Australian isolates of

MRSP was supported by ARC Linkage Grant

LP130100736 with Zoetis and Luoda Pharma as the

main Linkage Partners.

Conflicts of interest: DJT has received research funding

from Luoda Pharma, Neoculi, Zoetis, Bayer, Boehringer

Ingelheim, Virbac and Elanco.

Supporting Review

Using a One Health approach to chart a path

out of the Pandemic Era

J. MAZETDepartment of Epidemiology and Disease Ecology,School of Veterinary Medicine, University of California-Davis, Davis, CA, USA

Threats posed by pandemics and epidemics are nowclear to every household around the world. A singlelethal microbe can emerge suddenly and spread rapidlyto every community without regard to national bordersor to social and economic standing. The SARS, Ebolaand Zika outbreaks did little to prepare us for theCOVID-19 pandemic sweeping the world, which servesas a clarion call that we are vulnerable to emergingviral threats. Since the mid-20th Century, new anddeadly diseases have emerged at an alarming rate inanimals and people, and the threats from this vast poolof unknown viruses are accelerating exponentially, dri-ven by our expanding population and an increasinglyinterconnected world. After a decade of employing aOne Health approach to viral detection, discovery andcharacterization, we can reasonably estimate that thereare >500,000 zoonotic viruses, about which we knownothing or very little, that have the potential to spillover from evolutionary host species to vulnerable onesand cause disease. We know how to identify nearly allof the viral diversity in every species and can begin torank the risks of these viruses for interspecies transmis-sion. Understanding the disease threats and developingmitigation strategies to prevent infection and spread ofviruses are key to preventing future devastating pan-demics. The Global Virome Project is a strategicresponse to the growing need to better predict, preventand respond to future viral pandemic threats, and to

8 Abstracts



protect us all from their worst consequences. By bring-ing health professionals and scientists from many disci-plines together to share protocols and data under oneumbrella, a global consortium can be established toimplement a joint coordinated effort, whereby countries

will gain the benefits of shared global and regionalvirus identification and disease mitigation strategies,while maintaining autonomy to respond to local needs.Source of funding: Self-funded.


Abstracts 9



Theme: SKIN BIOLOGY AND GENETICS

State-of-the-Art AddressGrainy head-like genes are key epidermal

regulators across 360 Myr of evolution

Z. DENG*, S. M. JANE*, M. CANGKRAMA† andM. R. CARPINELLI**Department of Medicine, Central Clinical School,Monash University, Melbourne, Victoria, Australia;†Institute of Molecular Health Sciences, ETH Z€urich,Switzerland

There has been strong selective pressure to maintain afunctional skin barrier since terrestrial animals evolved360 Million years ago. In order to sustain terrestrial liv-ing habits, these animals acquired a unique integumen-tary system with a keratinized, stratified, squamousepithelium surface barrier. The barrier performs inside-out roles safeguarding against dehydration, and out-side-in roles preventing entry of microbes and toxins.The skin barrier centres on the stratum corneum layerof the epidermis, and consists of cornified envelopescemented by the intercorneocyte lipid matrix in abricks-and-mortar-like scenario. Multiple componentsof the barrier undergo cross-linking by transglutami-nase (TGM) enzymes, while keratins provide additionalmechanical strength. Cellular tight junctions also arecrucial for barrier integrity. Our understanding of themolecular basis of barrier formation, maintenance andrepair is still evolving. Previous studies have identifiedhighly conserved grainy head-like (GRHL) transcrip-tion factors that regulate the formation and mainte-nance of the integument in diverse species across700 Myr of evolution. Grhl3 is essential for formationof the skin barrier during embryonic development andnull mice die soon after birth from excessive transepi-dermal water loss. The Grhl3 null epidermis is abnor-mally thick and shows failed terminal differentiation ofkeratinocytes accompanied by excess proliferation.Tgm1, encoding a transglutaminase, is a direct target ofGrhl3 in epidermis, and loss of the cross-linking func-tion of this enzyme underpins the barrier defect. Sur-prisingly, postnatal inactivation of the Grhl3 gene inconditional knock-out mice showed that it was dispens-able for maintenance of the skin barrier, once formed.This was due to compensatory upregulation of Grhl1,which activates another transglutaminase, Tgm5, thusmediating adequate cross-linking. Concomitant loss ofGrhl1 and Grhl3 from postnatal mouse skin led to com-plete regression of the formed skin barrier and was notcompatible with life. These findings identify the GRHLfactors as potential therapeutic targets in human dis-eases in which barrier formation and function is com-promised. In addition to its barrier function, Grhl3 alsoplays key roles in wound repair and as an epidermaltumour suppressor. In its former role, Grhl3 activatesthe planar cell polarity signalling pathway to mediateepidermal wound-healing by providing directional

migration cues. Grhl3 null mouse embryos fail to healwounds and keratinocytes with reduced Grhl3 expres-sion display chaotic migratory patterns. In squamousepithelium from any tissue source, Grhl3 regulates thebalance between proliferation and differentiation, andits loss induces squamous cell carcinoma. In the skin,this is mediated through increased levels of miR21,which reduces the expression of Grhl3 and its direct tar-get, PTEN leading to activation of the PI3K-AKT sig-nalling pathway. Taken together, these data positionthe GRHL family as master regulators of epidermalhomeostasis across a vast gulf of evolutionary history.Source of funding: Australian National Health and Medi-

cal Research Council.


Supporting Review

An approach to cornification and hereditary

cornification disorders in dogs

E. MAULDINDepartment of Pathobiology, School of VeterinaryMedicine University of Pennsylvania, Philadelphia, PA,USA

The orderly formation of the stratum corneum (SC) isvital to the health of the skin and body. The SC pro-vides the ultimate barrier between the vertebrate bodyand the ambient environment. It serves to restrict waterflow in and out of the body to maintain hydration andprevent ingress of pathogens and toxins. The barrier iscomprised of terminally differentiated keratinocytes(corneocytes) that are sandwiched between lipid bilay-ers. Primary disorders of cornification arise from Men-delian defects in the structural proteins, lipids andenzymes that normally orchestrate formation of theskin barrier. Tremendous innovations in moleculartechniques have led to the identification of many disor-ders in both pure-bred and mixed breed dogs over thepast decade. These advances have far outpaced the gen-eration of therapeutics. Disorders of cornification aremanaged rather than cured. Awareness of the lesionpattern, character and age of onset will help avoid pit-falls in diagnosis. Disorders of cornification can be gen-eralized (e.g. autosomal recessive ichthyosis) orlocalized to paw pads (e.g. palmar plantar kerato-derma) or nasal planum (e.g. hereditary nasal paraker-atosis). Some disorders mimic those in humans whileothers are unique to the dog. This review will focus onthe epidermal structure and function in the process ofcornification and specific defects that have been identi-fied in canine species.Source of funding: Self-funded.


10 Abstracts



Theme: DIAGNOSIS AND THERAPY

State-of-the-Art Address

Advances in diagnosis and therapy of equine

Culicoides hypersensitivity

E. MARTIDepartment of Clinical Research and Veterinary PublicHealth, Vetsuisse Faculty, University of Bern, Bern,Switzerland

Culicoides hypersensitivity (CH) is an allergic,immunoglobulin (Ig)E-mediated, Th2-type dermatitis ofhorses caused by bites of insects of the genus Culi-coides. CH occurs worldwide except in Iceland. Theprevalence of CH is particularly high in horses born inIceland and exported to Culicoides-rich environmentsas adults. The performance of in vitro diagnostic testsand the efficacy of allergen-specific immunotherapy(ASIT) is questionable. This probably results from theuse of crude whole-body extracts from laboratory-bredCulicoides species. For these reasons, salivary glandallergens from Culicoides have been identified andalmost 30 produced as pure recombinant (r-) proteins.These r-Culicoides allergens bind IgE from sera of CH-affected horses with varying frequencies. Proteinmicroarrays represent efficient tools for determinationof the IgE sensitization pattern to allergens also inhorses. Testing of sera from horses living in variousEuropean countries has resulted in the identification ofthe most relevant r-Culicoides allergens for CH. A com-bination of seven major r-Culicoides allergens allows aspecific (95%) and sensitive (90%) serological diagnosisof CH.A preventive ASIT is under development with the aimof decreasing the incidence of CH in horses exportedfrom Iceland. An immunization protocol was estab-lished in Culicoides-unexposed horses living in Iceland.Immunization into the submandibular lymph node witha combination of r-Culicoides allergens, using Alumand MPLA as adjuvants, is able to induce an IgGresponse without induction of IgE. The induced anti-bodies are able to block binding of IgE to these sameallergens, an important feature of successful ASIT. Fur-thermore, immunization with Alum/MPLA led to ahigher production of interferon (IFN)-( and interleukin(IL)-10 than with Alum only, indicating that this com-bination of adjuvants is best suited for ASIT in horses.The efficacy of a preventive ASIT against CH will beevaluated in a cohort of Icelandic horses exported fromIceland to Switzerland. Therapeutic monoclonal anti-bodies (mAb) targeting key cytokines or their receptorare gaining in importance for treatment of human andalso canine allergy. This approach, however, may notbe available for equine patients due to the high amountof mAbs required. A new approach was thus devel-oped: By contrast to passive immunization with mAb,horses were actively immunized against selected cytoki-nes, linked to virus-like particles to enable antibodyproduction against a self-antigen. The first cytokine tar-geted using this approach was IL-5, important for thedifferentiation, migration, activation and survival ofeosinophils, which play an important role in the patho-genesis of CH. This therapeutic vaccine induced anti-IL-5 antibodies, was safe, reduced eosinophil countsand significantly improved clinical signs of CH in com-parison to the placebo group. IL-31 is involved in theinduction of pruritus and strong pruritus is a hallmark

of CH. The same VLP approach was used for inductionof anti-IL31 antibodies in CH-affected horses andresulted in a decrease of CH lesions scores. Theserecent advances should lead to the development of bet-ter in vitro diagnostic tests and new treatment optionsfor CH, and are worthwhile studying for other equineallergic diseases.Source of funding: EM is funded by the Swiss National

Science Foundation, the Morris Animal Foundation and

Stiftung Forschung f€ur das Pferd.Conflict of interest: None declared.

Supporting Review

Bacteriophage therapy

R. A. SQUIRESDiscipline of Veterinary Science, James Cook University,Townsville, Queensland, Australia

Bacteriophages (or just phages) are naturally occurringviruses that infect and kill bacteria. They are remark-ably numerous, diverse and easily found, for examplein sewage and dog faeces. Each phage typically has anarrow host range, infecting only certain strains of aparticular bacterial species. Yet a large majority of bac-terial species studied to date play host to bacterio-phages, explaining the remarkable diversity. Phageswere discovered just over 100 years ago and were firstused to treat bacterial infections in domestic animalsand people in the 1920s. Phages grown in simple brothcultures of host bacteria were used successfully to treatsalmonellosis in chickens, bacillary dysentery in chil-dren, and cutaneous furuncles and carbuncles in adulthumans. Phages have been safely administered orally,topically, intravenously, directly into infected lesionsand via inhaled aerosol. In many parts of the world,interest in bacteriophage therapy waned after penicillinbecame available. Interest continued in France andparts of Eastern Europe. In Georgia, phages continueto be used to treat human bacterial infections, forexample diabetic foot ulcers. During the 1980s, veteri-narian H. Williams Smith and colleagues investigatedphage therapy in the UK. They sought to prevent andtreat severe Escherichia coli diarrhoea in calves, pigletsand lambs. In 1987, they showed that severe, experi-mental E. coli diarrhoea in calves could be cured by asingle dose of 100,000 phage particles and could be pre-vented by doses as low as 100 particles. They obtainedtheir phages from ordinary sewage. The worseningantimicrobial resistance crisis has recently led to resur-gent interest in bacteriophage therapy. Importantly,phages can kill their host bacteria regardless of whetheror not the host is antimicrobial drug-resistant. This isbecause drug resistance usually has nothing to do withphage resistance. There recently has been a markedincrease in the number and quality of research publica-tions dealing with phage therapy, mostly for use inhuman medicine but also in veterinary science. A fewpromising publications have described successful, topi-cal phage therapy of Pseudomonas aeruginosa otitisexterna (OE) in dogs. Consequently, a veterinary phageproduct for treatment of Pseudomonas OE in dogs(containing six different bacteriophage strains) waslicensed and became commercially available in Europe.There have been some proof-of-concept studies. A New

Abstracts 11



Zealand-based research group showed that bacterio-phages readily found in sewage could kill a large pro-portion of different uropathogenic canine and felineE. coli strains. A very recent publication described theisolation and characterisation of phages with preferen-tial activity against meticillin-resistant Staphylococcuspseudintermedius strains, mainly from Europe andNorth America. The anti-staphylococcal phages wereisolated from canine faeces. Despite the fact that bacte-riophages have been used to treat bacterial infections

for nearly 100 years, surprisingly little has been pub-lished about their use in companion animals. What hasbeen published looks promising. Bacterial skin diseasesof dogs (including otitis externa) have been studied themost, although only to a small extent at this stage.Phage therapy represents a promising and potentiallyfruitful area of study for companion animal researchersin the future.Source of funding: Self-funded.


12 Abstracts



Theme: ALOPECIA

State-of-the-Art AddressPathogenesis and treatment of alopecia in

humans

R. SINCLAIRSinclair Dermatology, Melbourne, Victoria, Australia

Hair is a defining characteristic of mammals. The hairfollicle is a complete mini-organ that lends itself as amodel for investigation of a variety of complex biologi-cal phenomena including stem cell biology, organregeneration and cloning. The hair follicle has a com-plex immunology with constitutive expression of humanleucocyte antigen G (HLA-G), yet low or absentexpression of class 1-HLA. This is the basis of hair fol-licle relative immune privilege. Loss of this immuneprivilege is thought to underlie alopecia areata (AA).AA severity varies from a single small patch to com-plete loss of scalp hair, body hair, eyelashes and eye-brows. Approximately 40% of all affected individualsonly ever get one patch and will achieve a spontaneous,complete, durable remission within six months. AA thatpersists beyond 12 months is defined as chronic; with-out treatment, 55% of individuals with chronic AA willhave persistent multifocal relapsing and remitting dis-ease and 45% will ultimately develop alopecia totalis oruniversalis. The unpredictable course contributes to thepsychological distress attributable to AA. Numeroustopical, intralesional and systemic agents are currentlyused to treat AA; however, Janus kinase-signal trans-ducer and activator of transcription (JAK/STAT) inhi-bitors are emerging as a safe, effective treatment formoderate to severe chronic AA. Other common causesof alopecia in humans are androgenetic alopecia(AGA) and cicatricial alopecia. AGA is a disorder per-petuated by androgens that occurs only in geneticallypredisposed individuals. The pivotal role of androgensand more specifically 5a reductase, the enzyme thatconverts testosterone into dihydrotestosterone (DHT),has been known since the mid-1970s. However, thegenetic and epigenetic bases of AGA are still not fullyelucidated. In addition the paradoxical site specificityof the androgen effect on hair growth remains anenigma. The search for a cure for baldness has led tosignificant recent advances in fundamental hair science.These include the description of scalp follicular units,the hierarchy of androgen sensitivity within follicularunits and the central role of the arrector pili muscle inmaintenance of follicular unit integrity. Current treat-ments for AGA include finasteride, dutasteride, minoxi-dil and prostaglandin analogues. Emerging treatmentsinclude low-level laser, platelet-rich plasma injectionsand microneedling. The cicatricial alopecias include apoorly understood group of hair disorders linked by acommon final pathway of permanent hair follicle lossand replacement by fibrous tissue in involved areas.There are several well-recognized secondary causes ofhair loss including trauma, infiltration (e.g. metastatictissue) and infection. Primary cicatricial alopecias resultfrom pathology centred on the hair follicle itself, ratherthan the scalp, and include lichen planopilaris, discoidlupus erythematosus, frontal fibrosing alopecia, dissect-ing cellulitis and folliculitis decalvans. Progress in ourunderstanding and treatment of these disorders hasbeen slow. Recently sunscreen use has been implicatedin the development of frontal fibrosing alopecia.

Improved understanding of the general clinical and his-tological presentations in the early, mid- and late stagesof these disorders will facilitate their understanding.Management strategies for these conditions are limitedyet a number of biologics therapy show promise in themanagement of cicatricial alopecia.Source of funding: Self-funded.


Supporting Review

Molecular mechanisms of noninflammatory

alopecia in dogs

M. M. WELLE*, S. R €UFENACHT†, T. LEEB‡ andD. J. WIENER§

*Institute for Animal Pathology, Vetsuisse Faculty,University of Bern, Dermfocus,Bern, Switzerland; †DermaVet, Tierklinik Aarau West,Oberentfelden, Switzerland; ‡Institute of Genetics,Vetsuisse Faculty, University of Bern, Bern, Switzerland;§Department of Veterinary Pathobiology, College ofVeterinary Medicine & Biomedical Science, Texas A&MUniversity, TX, USA

Lifelong recurrent hair growth is needed to renew theprotective covering of the body and to enable adaptionof coat quality and colour to environmental changes.An intact hair coat is provided by hair follicle morpho-genesis during embryonic development and subse-quently maintained postnatally by the hair cycle,during which hair follicles undergo recurrent phases ofgrowth (anagen), regression (catagen) and relative pro-liferative quiescence (telogen). The hair shaft is shed ina phase called exogen. At the end of telogen, new ana-gen is induced by the activation of stem cells, located inthe secondary hair germ and in the bulge. If anageninduction does not occur, the telogen hair follicle willeventually lose the club hair and a hair follicle in thequiescent kenogen stage remains until appropriate sig-nals initiate a new anagen phase. Hair follicle morpho-genesis during embryogenesis and the maintenance ofthe hair cycle during postnatal life are tightly regulatedand highly conserved processes, involving numeroussignals derived from epithelial, neuroendocrine andmesenchymal cells. These signals are assembled in thehair follicle stem cells and the interplay of all factorsbelonging to several signalling pathways finally resultin stem cell quiescence or activation. Once activated,stem cells give rise to rapidly proliferating transientamplifying cells, which terminally differentiate into thedifferent hair follicle layers and the layers of the hairshaft. In dogs noninflammatory alopecia can be subdi-vided into four categories: (i) Inherited alopecia withan impaired hair follicle morphogenesis affecting eitherhair follicle induction, organogenesis or cytodifferentia-tion during embryonic development; (ii) Alopecia withimpaired hair follicle or hair shaft regeneration due toa genetic predisposition and postnatal clinical manifes-tation; (iii) Acquired alopecia associated with animpaired hair cycle and eventual subsequent follicularatrophy (hair cycle arrest); and (iv) Acquired alopeciaassociated with factors such as trauma, ischaemia andpoor nutrition. Some of the inherited congenital alope-cias represent monogenic traits that are caused, forexample, by variations in the FOXI3 or SGK3 genes. In

Abstracts 13



genetically predisposed noninflammatory alopecias, themode of inheritance appears to be more complex. Tran-scriptome analyses in these disorders reveals differentderegulated genes encoding for follicular stem cells,proteins involved in Wingless-related integration site,Sonic hedgehog and bone morphogenic processing sig-nalling, as well as proteins important for Vitamin Dmetabolism and steroidogenesis. In the presentation

and the supporting review the following topics will beaddressed: (i) molecular regulation of the hair cycle; (ii)genetic variants and associated molecular mechanismsunderlying inherited alopecias; and (iii) deregulation ofgenes and the associated consequences in alopeciadeveloping postnatally.Source of funding: Self-funded.


14 Abstracts



SUPPORTING ORIGINAL STUDY ABSTRACTS

SOS-1

Narrow-band ultraviolet B therapy attenuates

immunological responses in hapten-induced

contact dermatitis in beagles

S. ONISHI-SAKAMOTO*, K. MAKISHI*, K.TAKAMI†, R. ASAHINA‡, S. MAEDA§, M.NAGATA¶, P. F. MOORE**, K. IDE* and K.NISHIFUJI**Laboratory of Veterinary Internal Medicine, TokyoUniversity of Agriculture and Technology, Tokyo, Japan;†Ushio Inc., Tokyo, Japan; ‡Graduate School ofMedicine, Kyoto University, Kyoto, Japan; §Faculty ofApplied Biological Sciences, Gifu University, Gifu,Japan; ¶Synergy Animal Hospital, Saitama, Japan;**School of Veterinary Medicine, University ofCalifornia Davis, Davis, CA, USA

In human medicine, narrow-band ultraviolet B (NB-UVB) phototherapy is known as a safe and effectivetherapy in various inflammatory skin diseases such asatopic dermatitis (AD) and early phase epidermotropicT-cell lymphoma. Phototherapy causes cell death ofimmune cells in the skin while minimising tissue dam-age. To obtain efficacious NB-UVB therapy, minimalerythema dose (MED), which is a minimal dose ofUVB irradiation that causes gross erythema on humanskin, is applied to the lesional skin. However, the effectof NB-UVB on the cutaneous immune cells in dogs hasnot yet been analysed. The purpose of this study was toinvestigate the effect of NB-UVB therapy on hapten-induced contact dermatitis in dogs. To determine MEDof NB-UVB in the individual dog, seven healthy bea-gles were treated with NB-UVB doses of 300, 560, 750,920 and 1,000 mJ/cm2. The average of the NB-UVBMEDs in six of seven dogs was 785 � 88 mJ/cm2

(range: 560–1,000 mJ/cm2), although one dog did notexhibit gross erythema with 1,000 mJ/cm2 of NB-UVB.To determine whether NB-UVB MED affects theimmunological responses in inflammatory skin of dogs,2,4-dinitrochlorobenzene (DNCB) was applied topicallyto the skin of six healthy beagles. DNCB is known tocause type 1 helper T cell (Th1)- and/or cytotoxic T cell(Tc)-induced skin inflammation. Treated skin then wasirradiated by NB-UVB MEDs. The skin samples weresubjected to immunohistochemical analysis as well asquantitative reverse transcription-PCR for cytokinetranscription analysis. As a result, numbers of dermalCD4+ and CD8+ positive cells in irradiated skin(185.0 � 58.3 cells/mm2 and 508.7 � 217.2 cells/mm2,respectively) were significantly lower than those in non-irradiated skin (287.2 � 84.5 cells/mm2 and844.7 � 286.9 cells/mm2; Wilcoxon signed rank test,P < 0.05). TdT-mediated dUTP nick-end labellingrevealed that dermal apoptotic cells were significantlyincreased in irradiated skin (47.6 � 12.8 cells/mm2)compared to those in nonirradiated skin(26.1 � 6.8 cells/mm2; Wilcoxon signed rank test,P < 0.05). Frequency of apoptotic keratinocytes in theirradiated skin was <0.5 cells/mm2 of basement mem-branes. qRT-PCR revealed that transcription of genesencoding interferon c- and Tumour necrosis factor-a,which are Th1 cytokines, were significantly lower inirradiated skin than in nonirradiated skin (Wilcoxonsigned rank test, P < 0.05). These findings indicate that

NB-UVB MED attenuates Th1 or Tc cell-mediatedskin inflammation, minimising epidermal damage in acanine model of hapten-induced contact dermatitis. Ourfindings implicate that NB-UVB phototherapy can be asafe adjunct therapy for canine skin diseases associatedwith Th1 or Tc cells, such as the chronic phase ofcanine AD or canine epitheliotropic lymphoma.Source of funding: Self-funded.

Conflict of interest: KT is an employee of Ushio Inc.

SOS-2

Cytokine expression in feline allergic

dermatitis and feline asthma

C. E. OLDER*, A. B. DIESEL¶, J. C. HESELTINE¶,A. FRIEDECK¶, C. HEDKE§, S. PARDIKE§, K.BREITREITER†, M. A. ROSSI‡, J. MESSAMORE§,G. F. BAMMERT§, A. J. GONZALES§ and A.RODRIGUES HOFFMANN**Department of Veterinary Pathobiology, College ofVeterinary Medicine & Biomedical Sciences, TexasA&M University, College Station, TX, USA;¶Department of Small Animal Clinical Sciences, Collegeof Veterinary Medicine & Biomedical Sciences, TexasA&M University, College Station, TX, USA;§Veterinary Medicine Research and Development, GlobalTherapeutics Research, Zoetis Inc, Kalamazoo, MI,USA; †South Austin Cat Hospital, Austin, TX, USA;‡Coastal Veterinary Dermatology & Ear Clinic,Houston, TX, USA

The pathogenesis of feline allergic dermatitis is unclear,with several differences from allergic dermatitis in dogsand humans. Few treatment options exist, and thoseavailable may not be effective and may have adverseeffects. To further understand the pathogenesis of felineallergies, we performed a survey of various cytokineswithin cats affected with allergic dermatitis and nonaf-fected cats. We hypothesized that cats with allergic der-matitis would have higher cutaneous expression ofinterleukin (IL)-31 and IL-31 receptor alpha (IL-31RA)than nonaffected controls, given their role in canineand human atopic dermatitis (AD). To measure cyto-kine levels, formalin-fixed, paraffin-embedded skinbiopsies from cats with and cats without allergic der-matitis were obtained from two service archives. Usingquantitative PCR, mRNA expression was evaluated forIL-31 and IL-31RA. Using RNAScope, mRNA expres-sion of IL-5, IL-31, IL-31RA, IL-33 and Oncostatin Mreceptor (OSMR)-beta, part of the heterodimeric recep-tor for IL-31, also was evaluated in tissues. Addition-ally, IL-31 protein levels were evaluated viaimmunohistochemistry (IHC). IL-31 and IL-31RA werelow or not detected with qPCR in most samples. RNA-scope corroborated qPCR results indicating extremelylow levels of mRNA expression for IL-31 and IL31RA.IL-5 and IL-33 also were expressed in low quantities.By contrast, OSMR-beta was highly expressed andincreased in the skin of allergic cats versus healthy cats(P < 0.05). IHC revealed that IL-31 protein wasexpressed likewise in allergic and healthy cats. Both IL-31 protein and mRNA were predominantly expressedby follicular epithelial cells and keratinocytes. Weaker

Abstracts 15



expression also could be observed in muscle, sebaceousand apocrine glands, and occasionally in infiltratinginflammatory cells. We also investigated circulatinglevels of cytokines. In addition to healthy cats andthose affected by allergic dermatitis, patients with felineasthma – an incompletely understood allergic conditionthought to be due to a Th2 immune response – weresampled. We hypothesized that cats with either cuta-neous or respiratory allergic disease would demonstratehigher levels of Th2 cytokines. For systemic measure-ment of cytokine levels, serum was collected from a dif-ferent cohort of cats including 17 healthy cats, 18 catswith allergic dermatitis and 18 cats with asthma. Sam-ples were subjected to a Luminex panel of 19 cytokinescovering both Th1- and Th2-associated molecules andan IL-31 ELISA. None of the cytokines were differentbetween healthy cats and those affected by allergic der-matitis and/or asthma. Interestingly, IL-31 was detectedin a greater proportion of samples from cats with aller-gic dermatitis (10/18) than from healthy (4/17) andasthmatic cats (5/18).These results suggest that feline patients suffer fromallergic diseases that may exhibit different pathomecha-nisms from allergic diseases affecting other species,which typically exhibit a Th2-skewed immune system.Although IL-31 is shown to be relevant in human andcanine AD, the results of this study did not reveal dra-matic differences in IL-31 levels in tissue or serum,instead showing an upregulation of the IL-31 co-recep-tor OSMR-beta. These findings implicate aspects of theIL-31 pathway that may be dysregulated and suggestthat the co-receptor OSMR-beta has an important rolein the pathogenesis of feline allergic skin disease.Source of funding: Zoetis Inc.

Conflicts of interest: CEO, ABD, JCH and ARH

received partial salaries from Zoetis Inc. CH, SP, JM,

GFB and AJG are employees of Zoetis Inc.

SOS-3

Structural and molecular comparative analysis

of normal and atopic canine skin

D. COMBARROS*,†, D. GOUDOUNECHE‡, M. C.CADIERGUES*,† and M. SIMON§

*Small Animal Clinic, Universit�e de Toulouse, Toulouse,France; †UDEAR, Universit�e de Toulouse, Toulouse,France; ‡Centre de Microscopie Electronique Appliqu�ee�a la Biologie, Facult�e de M�edecine Rangueil, Universit�ede Toulouse, Toulouse, France; §UDEAR U1056,Universit�e de Toulouse, Toulouse, France

The pathogenesis of atopic dermatitis (AD) is complexand not fully understood. Similar to people, dogsdevelop spontaneous AD, suggesting this species couldbe a useful model of study. Skin barrier defects, suchas filaggrin (FLG) gene loss-of-function mutations andFLG decreased expression, are well-known in humansand are considered to play a key role in the pathogene-sis of AD. However, little information exists concerningthe expression of skin barrier-related proteins in dogsowing to the low number of studies and of well-

characterized dog-specific antibodies. The objective ofthis study was to comparatively analyse, at both struc-tural and molecular levels, canine healthy and atopicskin. Six healthy and six spontaneously atopic labora-tory beagles housed in identical environmental condi-tions (CanCog Technologies; Ontario, Canada) wereincluded in the study after validation of the experimen-tal protocol by CanCog ethics committee (referenceVRI185-18245-CO), and in accordance with the princi-ples of the Animal for Research Act of Ontario and theguidelines of Canadian Council on Animal Care. Dogswere clinically characterized [general examination,Canine Atopic Dermatitis Extent and Severity Index,4th iteration (CADESI-04) evaluation and transepider-mal water loss (TEWL) measurement] and skin biopsieswere performed in healthy, lesional and non-lesionalskin. Samples were analysed using haematoxylin &eosin staining and transmission electron microscopy(TEM). The expressions of FLG, corneodesmosin(CDSN), claudin-1 and involucrin were studied, usingpreviously validated antibodies, by indirect immunoflu-orescence and Western blotting, as well as by quantita-tive reverse transcription PCR. TEWL was significantlyhigher in atopic dogs when compared to controls, sug-gesting skin barrier impairment. Blinded evaluation ofH&E stained skin sections did not allow discriminationbetween the healthy and non-lesional atopic skin. TEMexamination showed abnormal morphology of the stra-tum corneum (SC) in the non-lesional atopic skin,including thicker corneocytes (P = 0.02) with very irreg-ular morphology and a sometimes vesicular aspect. TheSC compactum was either absent or of increased thick-ness, when compared to the healthy skin. Thesechanges were even more pronounced in lesional skin.No significant differences were observed betweengroups in the number nor the size of keratohyalin gran-ules (containing FLG and loricrin), nor in the numberof corneodesmosomes (containing CDSN). Blindedqualitative evaluation of immunofluorescence reactivityby two investigators failed to show any significant dif-ferences between groups for any of the proteins studied.This result was confirmed by an objective evaluation ofgrey levels using IMAGEJ software. Intensity of Westernblotting bands was normalized to actin and statisticalcomparison between groups further confirmed theabsence of differences in protein expression (FLG,CDSN, claudin-1 and involucrin). Expression also wassimilar at the mRNA level. As in humans, epidermaldifferentiation seemed to be impaired in canine AD, asshown by both abnormalities of the SC structure andhigh water permeability. However, the expression ofsome major epidermal proteins was not found to bealtered. Further studies including a larger number ofanimals and use of additional validated antibodiesdirected against epidermal proteins are necessary todetermine the cause of these abnormalities in the SC.Source of funding: The project was partially financed by

the BQR 2019 research grant of the National Veterinary

School of Toulouse (ENVT).


16 Abstracts



SOS-4

Survey of cytokines on the ocular surface of

atopic dogs by multiplex analysis using two

sampling methods: a pilot study

C. PRESSANTI*,†, E. RAVAILHE*, J. BRUN*, N.AMALRIC‡, L. A. LECRU*, J. KONDRAJTEVA*,F. MOOG*, D. COMBARROS*,†, J. Y. DOUET*,§

and M. C. CADIERGUES*,†

*Small Animal Clinic, Universit�e de Toulouse, Toulouse,France; †UDEAR, Universit�e de Toulouse, Toulouse,France; ‡Synelvia, Lab�ege, France; §IHAP, Universit�e deToulouse, Toulouse, France

Conjunctivitis in atopic dogs has been described and islikely underdiagnosed in practice. Consistent conjuncti-val infiltration by eosinophils in atopic dogs, as com-pared with healthy controls, was reported in a previousstudy of conjunctival biopsies as a possible hallmark ofan allergic reaction. Elevated levels of inflammatorycytokines have been reported in tears from ocular aller-gic disease states in humans. The purpose of thisprospective study (Ethics Committee approval no.SSA_2019_005) was to assay various cytokines in tearsand conjunctivae from atopic and normal dogs, and tocompare canine atopic dermatitis-associated conjunc-tivitis with controls. Ocular surfaces from 10 atopicand 10 normal dogs were sampled bilaterally using twodifferent methods. Tear samples were obtained with asterile swab previously moistened with saline solution(method A). Conjunctival impressions were obtainedwith a validated device (Eyeprim, Opia Technologies;Paris, France) (method B). For each sample, the con-centrations of a panel of cytokines, comprising inter-leukin (IL)-2, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18,tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, granulocyte-macrophage colony-stimulatingfactor (GM-CSF), IFN-gamma–induced protein 10 (IP-10), monocyte chemoattractant protein 1 (MCP-1) andkeratinocyte-derived chemokine (KC)-like, were mea-sured in duplicate by multiplex analyses (#CCYTMG-90K-PX13, Merck Millipore; Darmstadt, Germany).Canine Atopic Dermatitis Extent and Severity Index,4th iteration (CADESI-04), pruritus (PS) and conjuncti-val (CS) scores were determined. Cytokine levels werecompared by Wilcoxon–Mann–Whitney U-test; correla-tions with CS, PS and CADESI-04 were determined bySpearman rank correlation test. Median age in normaldogs was 5.3 years (range 1–11) and in atopic dogs 4.4years (range 1–10). In atopic dogs, the median CS was1.5/12 (range 0–7), the median CADESI-04 was 43.5(range 39–57) and the median PS was 10.2/16 (range2.8–14.7). Among the measured biomarkers, only GM-CSF, KC-like and IL-8 were above the limit of quan-tification in most samples. Absolute amounts of eachbiomarker were always higher in samples obtained withmethod A than with method B. GM-CSF amountswere lower in atopic dogs (method A, P = 0.02; methodB, P=0.0005). KC levels were higher in atopic dogs, yetthe differences were not significant. IL-8 amounts werehigher in atopic dogs (method A: 378 and 1504 pg/mLin normal and atopic dogs, respectively, P = 0.0003;method B: 57 and 382 pg/mL in normal and atopicdogs, respectively, P = 0.006). In atopic dogs, IL-8 con-centrations obtained with method B were positively cor-related with CS (P = 0.008) and PS (P = 0.036).Regardless of the method, these preliminary results sug-gest an overexpression of IL-8 in conjunctivae and tears

of atopic dogs despite subtle conjunctival clinical signs.As IL-8 is commonly found in many inflammatory con-ditions, further studies are needed to determine itsspecificity in atopic conjunctivitis. Although higher con-centrations of markers were obtained with method A,method B gave similar results. The Eyeprim device iseasy to use and allows specific sampling from the con-junctiva. Canine tears have been shown to containcytokines which are not necessarily linked to theinflammatory status of the conjunctiva. The Eyeprimdevice could be more discriminant than swabbing toevaluate conjunctival markers and would limit tear col-lection and be more representative of ocular status.Source of funding: Self-funded.


SOS-5

Interleukin-31 and IL-31 receptor expression

in acute experimental canine atopic dermatitis

skin lesions

C. TAMAMOTO-MOCHIZUKI and T. OLIVRYDepartment of Clinical Sciences, College of VeterinaryMedicine, NC State University, Raleigh, NC, USA

Interleukin (IL)-31 is one of the earliest and mostimportant pruritogenic cytokines in canine atopic der-matitis (AD). To optimize the IL-31 signalling blockadetherapy in AD, an understanding of the chronologicalchanges in IL-31 and its receptor (IL-31RA) in canineskin lesions is needed. This study aimed to, after anallergen challenge, (i) assess the chronology of theexpression of IL-31 during experimental acute AD skinlesions and compare it with the serum IL-31 level, and(ii) determine the identity of cells producing IL-31 andexpressing the IL-31RA. Skin and blood samples wereobtained from four Maltese-beagle atopic dogs at 0(baseline), 24, 48 and 96 h after epicutaneous housedust mite (HDM) application. Immunofluorescence wasperformed with a Zoetis-developed mouse anti-canine-IL-31 monoclonal antibody and goat anti-human-IL-31RA polyclonal antibodies. Each section also wasdouble-stained for IL-31/CD3, IL-31/CD4 and IL-31RA/b3-tubulin (a neuronal marker). The IL-31-posi-tive round/dendritic cells were subjectively scored fortheir distribution (0, none; 1, one focal area; 2, multifo-cal areas; 3, diffuse) and count (0, none; 1, low; 2, med-ium; 3, high) to obtain an expression score (distributionscore x cell count score). The serum IL-31 level wasmeasured using an ultrasensitive signal molecule array.A Spearman rank correlation test was used to investi-gate the correlation between the IL-31 expression scoreand the serum IL-31 level. All of the procedures wereapproved by our university’s Institutional Animal Careand Use Committee. On the one hand, the peak IL-31expression scores for each dog occurred at 24 h and/or48 h after HDM provocation; it started to decrease at96 h, except in Dog 3 whose score continued toincrease throughout the study period. On the other, theserum IL-31 levels for each dog increased throughoutthe study, except, again, for Dog 3 whose serum IL-31level was unchanged. Overall, there was only a weaknegative correlation between the IL-31 expression scoreon the skin and the serum IL-31 level (r = -0.25;P = 0.35). The majority of IL-31-positive cells in thedermis co-expressed CD3 (range 91–100%) and CD4

Abstracts 17



(range 63–100%), indicating that they were likely Th2cells. Unexpectedly, sebaceous glands were stronglystained with anti-IL-31 antibodies; the extinction of thispositivity by pre-incubating the antibodies with recom-binant canine IL-31 further supported the presence ofIL-31 in sebaceous glands. The IL-31RA immunofluo-rescence staining showed an epidermal intercellular pat-tern at all time points, suggesting its expression bykeratinocytes. Some of the dermal b3-tubulin-positivelinear structures were double-positive for IL-31RA,indicating that it was constitutively expressed on dermalnerves. Herein, we showed, in experimental acutecanine AD skin lesions, the early and transient secre-tion of IL-31 by helper T cells – likely Th2 – that doesnot parallel the serum IL-31 level. The detection of IL-31RA in keratinocytes and peripheral nerves in canineskin indicates the potential roles of IL-31 as a pro-inflammatory factor involving keratinocytes and dermalnerves in the transmission of itch in atopic dogs.Source of funding: Zoetis.

Conflict of interest: The PhD programme of CTM is

sponsored by Zoetis.

SOS-6

Association of prick-and-patch test in the

evaluation of adverse food reaction in dogs

with allergic dermatitis

J. POSSEBOM, A. CRUZ, V. GMYTERCO and M.FARIASDepartment of Veterinary Medicine, School of LifeSciences, Pontif�ıcia Universidade Cat�olica do Paran�a,Curitiba, PR, Brazil

Adverse food reaction (AFR) in dogs is assessedthrough an elimination diet for eight weeks, followedby food provocation, which often has poor complianceby owners and variable results in diagnosis. Previousstudies have shown that the patch test with foodextracts was adequate to assist formulations of elimina-tion diets in patients with suspected AFR. Associationof prick-and-patch tests assist diagnosis of AFR in chil-dren with atopic dermatitis (AD); however, little isknown about these tests in dogs. The aim of this studywas to evaluate the effectiveness of prick-and-patch testin guiding elimination diet protocols and in the foodprovocation in dogs with AD that have an AFR. Thestudy was approved by the Ethics Committee ofPUCPR (01252). Thirty-eight dogs with AD wereselected on which to perform prick tests with foodextracts (bovine protein, chicken, pork, fish, egg, milk,soy, wheat, rice, potato and manioc) and with extractsof dust mite and pollen allergens (Veterinary Allergen-ics; Rio de Janeiro, Brazil). The patch test was per-formed using the same food sources, placed inAllergochambers containers (Neoflex; S~ao Paulo, Bra-zil) and then kept for 48 h in contact with the skin,and the presence of erythematous papules, spots or pla-ques were considered positive. Patients with positivereactions to at least one food extract in the prick testand/or patch test were submitted to an elimination dietand evaluated on day (D)0, D30 and D60. In patientsthat had improvement, a provocation test was per-formed with a food that had positive and a negativereaction, at D75 and D90, respectively. The pruritusVisual Analog Scale (pVAS) and Canine Atopic

Dermatitis Extent and Severity Index, 4th iteration(CADESI-04) scores were evaluated every 30 days andanalysed by ANOVA of repeated measurements(P < 0.05). Sensitivity (SE), specificity (SP), positivepredictive value (PPV), negative predictive value (NPV)and accuracy (AC) and Kappa values between prick-and-patch test were calculated. Twenty-one (55%) ofthe 38 dogs had a positive reaction to at least one testand of these, 14 (66.7%) presented sensitivity to envi-ronmental allergens. A significant average improvementin pVAS and CADESI-04 scores was obtained by 17(81%) of the 21 dogs after 30 (P<0.01) and 60 (P<0.01)days of dietary exclusion, and all 17 dogs presented sig-nificant increased scores after food allergen re-exposure.There was no difference in pVAS and CADESI-04scores between D30 and D60. The combination of testsshowed values of SE, SP, VPP, VPN and AC of 80%,66.7%, 66.7%, 80% and 73%, respectively. There waslow kappa agreement (j = –0.17) between the two aller-gic tests. It is concluded that most dogs with chronicpruritus associated with AD are polysensitized to foodand environmental allergens. The lack of agreementbetween the tests suggests the need to perform both.These tests can represent an important method forestablishing dietary protocols and provocative testing,allowing an early diagnosis of AFR, with a 30 dayelimination diet.Source of funding: Coordenac�~ao de Aperfeic�oamento de

Pessoal de N�ıvel Superior (CAPES).Conflict of interest: None declared.

SOS-7

Intradermal immunotherapy with

Actinomycetales preparations as treatment for

feline hypersensitivity dermatitis: a preliminary

randomized, placebo-controlled, double-blinded

study

D. SANTORO, L. ARCHER and L. FAGMANDepartment of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA

Feline hypersensitivity dermatitis (FHD) is an extre-mely common skin disease affecting ≤33% cats seen bydermatology specialty practices. Causes of FHD includeallergic dermatitis such as fleabite hypersensitivity,cutaneous adverse food reaction, urticaria, angioedemaand feline atopic syndrome. Although very common,few therapeutic options are available for cats withFHD. Very recently, the use of a single intradermalinjection of heat-killed Actinomycetales (Mycobacteriumvaccae, Gordonia bronchialis, Rhodococcus coprophilusand Tsukamurella inchonensis) as bacterialimmunomodulators has shown potential beneficialeffects on canine allergies. To date, no studies havebeen published on the use of such therapy for FHD.Thus, the purpose of this double-blinded, randomized,placebo-controlled clinical trial was to evaluate the clin-ical effects of heat-killed Actinomycetales[G. bronchialis (GB) and R. coprophilus (RC) alone orin combination) in FHD. Privately owned cats with adiagnosis of FHD were included in this study. Catswere randomly allocated in one of three treatmentgroups (GB, RC and GB/RC combination) or placebo.

18 Abstracts



Five intradermal injections [day (D)0, D20, D40, D90and D180) were performed over a one year period.Because of the duration of the study, rescue antipruriticmediations were allowed, if necessary, and frequencyand doses recorded. At each visit (D0, D20, D40, D60,D90, D180 and D360), the severity of the clinical signs(lesional score and pruritus), global owner assessmentscore, use of rescue medications, clinical adverse effects,skin hydration and cutaneous pH were assessed. AFriedman’s test and a Kruskal-Wallis test followed byDunn’s multiple comparison test were performed toanalyse the behaviour of each variable; P ≤ 0.05 wasconsidered statistically significant. A total of 17 cats(GB: n = 6; RC: n = 3; GB/RC: n = 4; placebo: n = 4)were enrolled in this study. All cats except two (Sia-mese) were domestic short hair cats. The average (�SD) age at the time of enrolment was 5.7 � 3.5 yearswith no significant difference among groups (P > 0.6).The median weight at enrolment was 4.8 kg with nosignificant difference among groups (P > 0.2) or overtime (P > 0.1). Compared to placebo, GB significantlyimproved the clinical score from D90 (P < 0.05). In theGB group, an over-time clinical score reduction wasseen from D60 (P ≤ 0.03). A complete remission of theclinical signs was seen in 30–67% of cats in all treat-ment groups at various time points. A significant reduc-tion in the pruritus score was only achieved by RC onD360 (P = 0.04) when compared to D0. Significant dif-ferences in the other parameters were not seen. Adverseeffects were not seen in any cat. In conclusion, the useof intradermal injections of GB is an effective and well-tolerated treatment for FHD. Because of the low costand the lack of adverse effects, GB – alone or in com-bination with other treatments – could offer a beneficialtreatment option for affected cats. A larger study isneeded to confirm these data and to evaluate theimmunological changes occurring in the treated cats.Source of Funding: This study was funded by the Univer-

sity of Florida Research Foundation.


SOS-8

Investigation of the skin microbiota in equine

pastern dermatitis

S. KAISER-THOM*, M. HILTY† and V. GERBER**Swiss Institute of Equine Medicine (ISME), VetsuisseFaculty, University of Bern, Bern, Switzerland; †Institutefor Infectious Diseases, University of Bern, Bern,Switzerland

Equine pastern dermatitis (EPD) is a multifactorial syn-drome that has variable degrees of severity. Despite thewidespread use of antibiotics in horses with this condi-tion, the role of bacteria has not been adequately inves-tigated. This study explored the bacterial skinmicrobiota of affected (EPD) and control pasterns, andinvestigated associations with the clinical presentationand previous treatments. The study protocol wasapproved by the veterinary ethical committees of all 26cantons in Switzerland. The study design was case con-trol. All legs of each horse were examined with a stan-dardized procedure. One affected and one control-suitable pastern in the same horse was a prerequisitefor participation. Eighty horses fulfilled the inclusioncriterion. Affected pasterns were classified into mild,

exudative or proliferative according to clinical signs.All previous treatments of EPD of the last six monthswere recorded and categorized as disinfectant, antibioticor no antibacterial. Skin swabs were obtained from themost severely affected pastern and one control pastern.DNA was extracted from the swabs and the bacterial16S rRNA gene region was amplified. Samples weresubmitted for paired-end sequencing on an IlluminaMiSeq platform (Illumina Inc.; San Diego, CA, USA).Alpha diversity (i.e. within-sample diversity) was signifi-cantly reduced in affected pasterns in comparison totheir respective controls as determined by a univariateWilcoxon signed rank test (P = 0.001, r = 0.54). A mul-tilevel mixed effects model showed that the Shannondiversity index was significantly associated with theform of EPD (P = 0.001) and not with the type of pre-vious treatment (P = 0.16). Analyses of beta-diversity(i.e. between-samples dissimilarity) showed a disorder-ing of the microbiota in affected pasterns. A Procrustesanalysis revealed that proliferative EPD induced thelargest alteration in microbiota, followed by exudativeEPD and mild EPD, with mean residual values of 0.2,0.18 and 0.14, respectively. Much of this divergencewas driven by staphylococci bacteria, with affected pas-terns having a significantly higher relative abundancethan their controls (univariate Wilcoxon signed ranktest, P = 0.011, r = 0.37). On average, the relativeabundance of staphylococci increased by 7.8% inaffected pasterns (SD: 18.1). However, the intraindivid-ual increase varied greatly between horses, being partic-ularly large in exudative EPD lesions and in pasternsthat had previously received antibacterial treatment. Inconclusion, our study highlights the influence that EPDhas on the pastern bacterial skin microbiota, confirm-ing that bacteria are an important factor in the courseof this multifactorial disease. In addition, althoughantibacterial treatment may not be the dominant factorfor the overall reduction of bacterial diversity in EPD,it might be a predisposing factor for the growth ofstaphylococci as opportunistic pathogens.Source of funding: Self-funded.


SOS-9

Whole genome sequencing and de novoassembly of Staphylococcus pseudintermedius:a novel pangenome approach to unravel

pathogenesis of canine pyoderma

L. FERRER*, R. FONTICOBA*, D. P�EREZ†, J.VI ~NES†, S. MADRO ~NERO†, A. CUSCO†, L.MIGURA‡ and O. FRANCINO†

*Department de Medicina I Cirurgia Animals,Universitat Aut�onoma de Barcelona, Spain; †ServeiVeterinari de Gen�etica Molecular, Universitat Aut�onomade Barcelona, Spain; ‡Centre de Recerca en SanitatAnimal, IRTA, Bellaterra, Barcelona, Spain

The majority of canine pyodermas are caused by Sta-phylococcus pseudintermedius, which is part of the nor-mal skin microbiota of the dog. The mechanisms thatlead a commensal bacterium to transform itself into apathogen have so far remained elusive to research.However, understanding the whole pathogenic processis of outmost importance to develop new treatment andprevention strategies. Whole genome sequencing (WGS)

Abstracts 19



is regarded currently as the most comprehensive way ofobtaining relevant genomic information of micro-organisms. We hypothesized that comparative analysisof the complete genome of S. pseudintermedius strainsisolated from healthy dogs and dogs with pyodermacould help to understand the pathogenesis of pyo-derma. The aims of this study were (i) to visualize andunderstand the pangenome of S. pseudintermedius (coregenome and accessory genome) and (ii) to find differ-ences between strains isolated from healthy and affecteddogs. Sequencing was performed using Oxford Nano-pore MinION, a new technology that delivers longerreads (up to tens or hundreds of kilobases) in a hand-held device. The long reads obtained allow de novoassembly of bacterial genomes, and its portability andreal-time results are essential to ultimately develop aclinical metagenomic approach to canine pyoderma.Five strains of S. pseudintermedius isolated from theabdominal skin of five healthy dogs and 27 strains iso-lated from lesional skin of 27 dogs with pyoderma wereanalysed. DNA was extracted using the DNA Micro-prep kit (ZymoBIOMICS). DNA quality was assessedwith a Qubit fluorimeter. The libraries for Nanoporesequencing were prepared by transposase fragmentationusing the Rapid Barcoding kit and sequenced in aMinION FLO-MIN106 v9.4.1 flow cell. The fast5 readswere basecalled with Guppy, assembled with Flye, andcorrected with racon and medaka. The pangenome wasanalysed and visualized with Anvi’o 6.1. Nanoporesequencing was effective and successful and allowed theassembly of the complete genome of S. pseudinter-medius (2.69 Mb; range 2.55–2.89 Mb) in a single con-tig. The mean genome size of S. pseudintermediusstrains isolated from healthy dogs (2.55 Mb) was smal-ler than the mean size of strains isolated from lesionsof dogs with pyoderma (2.71 Mb). The Anvi’o platformallowed an accurate visualization of the core and acces-sory genomes, and the detection of differences betweencommensal and pathogenic strains. The genome ofS. pseudintermedius is an example of an open pangen-ome. The core genome represents 45% of the total andis present in all of the strains (housekeeping genes).The accessory genome (55%) is more prominent in thepathogenic strains and comprises antimicrobial resis-tance genes, virulence factors and bacteriophages. Theanalysis of the pangenome of S. pseudintermedius isvery promising as a tool to understand the pathogenesisof canine pyoderma and the transformation of a com-mensal into a pathogen. Recent studies have demon-strated the aetiological diversity and complexity ofcanine pyoderma. Traditional culture techniques anddetermination of antimicrobial sensitivity have limita-tions facing this complexity. The analysis of the pan-genome provides more accurate and clinically relevantinformation.Source of funding: Research Project RTI2018-101991-B-

I00 (“From whole genome sequencing to clinical metage-

nomics: investigations on the pathogenesis of Staphylo-coccus pseudintermedius pyoderma in the dog”), Spanish

Ministry of Science, Innovation and Universities.

Conflict of interest: LF has received unrelated honoraria

for lecturing from Zoetis, Bayer, LETI and Affinity Pet-

care.

SOS-10

The efficacy of olanexidine gluconate in canine

superficial pyoderma: in vitro and in vivoanalysis

Y.H. HSIAO, K. SHIMOIKE and K. IYORIVet Derm Tokyo, Kanagawa, Japan

Superficial pyoderma caused by meticillin-resistant Sta-phylococcus pseudintermedius (MRSP) is an emergingproblem and it often leads to limited options for sys-temic antimicrobial therapy. By contrast, topicalantiseptic treatments such as chlorhexidine shampoo orsolution have been reported to be effective treatmentoptions for MRSP skin infections in dogs. Recently, anovel biguanide of olanexidine gluconate (Olanedine,Otsuka Pharmaceutical Co., Ltd; Tokyo, Japan) waslaunched as a broad spectrum bactericidal solution fordisinfecting human skin. However, olanexidine glu-conate has not been evaluated sufficiently in caninesuperficial pyoderma. The aim of the study was toinvestigate the clinical utility of olanexidine gluconatecompared with topical chlorhexidine and determine itsminimum inhibitory concentration (MIC) in S. pseudin-termedius. In an in vivo study, 33 client-owned dogswith superficial pyoderma caused by S. pseudinter-medius infection were recruited. Species identificationand meticillin resistance were detected using a multiplexPCR method for identifying the thermonuclease andmecA genes. Dogs were allocated randomly to olanexi-dine or chlorhexidine treatment groups. In the olanexi-dine group (n = 15), the dogs received a gauze-dippingtreatment of 1.5% olanexidine gluconate solution onthe lesion sites once daily. In the chlorhexidine group(n = 17), dogs received 3% chlorhexidine gluconateshampoo (Douxo, CEVA; Lenexa, KS, USA) once aweek according to the product instruction. All dogswere not allowed antibiotics or anti-inflammation medi-cations either systematically or topically. Clinical scoresof superficial pyoderma were evaluated day (D)0 andD14 following the guidelines of the Japanese Society ofAntimicrobials for Animals. All investigators wereblinded to treatment allocation. All patients were ableto finish the trial, and MRSP was identified in sevendogs in the olanexidine group and 10 dogs in thechlorhexidine group. At D14, clinical scores were signif-icantly reduced compared to D0 in both groups(P < 0.05). The improvement rate of clinical score atD14 was 46% and 43% in the olanexidine andchlorhexidine groups, respectively; there was no signifi-cant difference in the improvement of clinical scoresbetween the groups (P > 0.05). Two dogs developedmild scaling after one week of olanexidine applicationand they recovered spontaneously after the trial. Wefurther performed an in vitro study to determine theMIC both in MRSP and meticillin-susceptible S. pseud-intermedius (MSSP) isolated from dogs with superficialpyoderma. Fifty-seven strains of S. pseudintermedius(29 MRSP and 28 MSSP strains) were included in thein vitro study. In the broth microdilution method,MICs of olanexidine gluconate were 1.46 lg/mL inMRSP and 0.73 lg/mL in MSSP. Therefore, 1.5%olanexidine gluconate solution contains 1027.4- to2054.8-fold higher concentration than the determinedMIC. From these findings, 1.5% olanexidine gluconatesolution could be an effective and tolerable topical ther-apy option for dogs with superficial pyoderma even inMRSP infections.

20 Abstracts



Source of funding: Self-funded.


SOS-11

Advantages of restricting antimicrobial use

with antibiograms for reducing antimicrobial

resistance rate in dogs with Staphylococcuspseudintermedius infections

K. IYORI*, T. SHISHIKURA†, K. SHIMOIKE*, K.MINOSHIMA* and Y. TOYODA**Vet Derm Tokyo, Kanagawa, Japan; †Daktari AnimalHospital Tokyo Medical Center, Tokyo, Japan

The antibiotic resistance of Staphylococcus pseudinter-medius (SP) and meticillin-resistant SP (MRSP) isincreasing dramatically in dogs worldwide. An antibi-ogram is a periodic summary of the test results ofantimicrobial susceptibility of specific micro-organismsto batteries of antimicrobial drugs. Antibiograms areused to select appropriate empirical antimicrobial treat-ments and monitor the trends in antimicrobial resis-tance. This study evaluated whether the restricted useof antimicrobials with antibiograms can reduce theantimicrobial resistance of SP strains isolated fromdogs with pyoderma and otitis externa (OE). The rela-tionship between hospital size and MRSP isolation ratein Japan was analysed. A total of 95 private clinics and2,294 dogs with SP infections between 2017 and 2018were selected. SP and meticillin resistance were detectedby a multiplex PCR strategy for identifying the ther-monuclease gene and the oxacillin susceptibility test,respectively, according to the Clinical and LaboratoryStandards Institute (CLSI) Guideline. MRSP wasdetected in 987 dogs (43.0%). The isolation rate ofMRSP in large clinics (n = 32) with >10 veterinarypractitioners was 51.2% (411/803), which was signifi-cantly higher (P < 0.01) than that in small clinics(n = 36) with <2 veterinary practitioners (34.45%, 154/447) and medium-sized clinics (n = 37) with 3–9 veteri-nary practitioners (40.4%, 422/1,044). An antibiogramsubsequently was constructed based on the periodicalantimicrobial susceptibility data for 12 antibiotics,including oxacillin (MPIPC), clavulanic acid-amoxicillin(AMPC/CVA), cefalexin (CEX), cefpodoxime (CPDX),enrofloxacin (ERFX), gentamycin (GM), trimethoprim-sulfamethoxazole (ST), clindamycin (CLDM), doxycy-cline (DOXY), minocycline (MINO), chloramphenicol(CP) and fosfomycin (FOM), from one large veterinaryclinic in Tokyo, with 11 veterinary practitioners, from2017 to 2018. The antimicrobial susceptibility wasdetected by disk diffusion susceptibility tests in accor-dance with the CLSI Guideline. Fifty-three SP strainswere collected from dogs with pyoderma (n = 30) andOE (n = 24). The resulting antibiogram depicted lowsusceptibility rates to MPIPC (38%), CEX (49%),CPDX (34%), ERFX (19%), GM (42%), ST (32%),CLDM (17%), DOXY (15%) and MINO (23%), andmoderate susceptibility rates to AMPC/CVA (66%),CP (51%) and FOM (58%). Based on these results, thehospital introduced strict antimicrobial selection criteriafrom 2018 for treating dogs with pyoderma and OE,which included: (i) fluoroquinolones, first and thirdgeneration cephalosporins for systemic treatment wererestricted, (ii) CP and FOM could be used for empiricaltreatment, and (iii) ST, CLDM, DOXY and MINO

should be used according to the results of susceptibilitytests. Following the restricted use of antimicrobials, anantibiogram was reconstructed using the susceptibilitytest data between January and December 2019. Fifty-three SP strains were isolated from dogs with pyoderma(n = 30) and OE (n = 22). The susceptibility rates ofMPIPC (60%), CEX (66%) and MINO (56%) in 2019were significantly higher than those between 2017 and2018 (P < 0.05). The other drugs showed higher suscep-tibility rates than those observed in 2017, with theexception of ST (24%). These findings indicated thatthe restricted use of antimicrobials for over a year,based on antibiograms, reduced the rate of antibioticresistance of SP strains isolated from dogs with pyo-derma and OE.Source of funding: Self-funded.

Conflict of interests: None declared.

SOS-12

Persistent activation of the mammalian target

of rapamycin signalling pathway in cutaneous

squamous cell carcinomas in cats

B. L. SANZ RESSEL*,†, A. R. MASSONE‡ and C. G.BARBEITO*,†

*Laboratorio de Histolog�ıa y Embriolog�ıa Descriptiva,Experimental y Comparada (LHYEDEC), Facultad deCiencias Veterinarias (FCV), Universidad Nacional deLa Plata (UNLP), La Plata, Buenos Aires, Argentina;†FCV, Consejo Nacional de Investigaciones Cient�ıficas yT�ecnicas (CONICET), UNLP, La Plata, Buenos Aires,Argentina; ‡Laboratorio de Patolog�ıa EspecialVeterinaria Dr. Bernardo Epstein, FCV, UNLP, LaPlata, Buenos Aires, Argentina

Cutaneous squamous cell carcinoma (CSCC) representsthe most common malignant tumour of the feline skin.Research aimed at clarifying how the deregulated activ-ity of signalling pathways contributes to CSCC progres-sion can help to identify suitable molecular targets forthe development of novel therapeutic approaches. Inthis regard, recent evidence indicates that the develop-ment and progression of human CSCC involve the dys-regulated activation of the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) signalling pathway, which is a key regulator ofcell growth and cell fate. Thus, the present study inves-tigated the immunohistochemical expression pattern ofrelevant signalling proteins involved in the PI3K/Akt/mTOR signalling pathway including, pEGFRTyr1068,pAktSer473, pS6Ser235/236 combined with Ki-67, and thetumour suppressor protein PTEN. These signalling pro-teins were investigated in 45 feline CSCC samples usinga tissue microarray. In normal feline skin controls, allproteins showed a low expression in the epidermis,except pEGFRTyr1068. In CSCC, pEGFRTyr1068 immun-odetection showed that the persistent activation of thisreceptor was a frequent event in 31 of 45 samples,showing membrane and cytoplasmic immunoreactivityin the epidermal cells of the basal and spinous strata.Likewise, pAktSer473 was overexpressed in 30 of 45CSCC samples, showing cytoplasmic and nuclearimmunoreactivity in the basal and suprabasal cellstrata. Forty-one of 45 CSCC samples also showed ahigh epidermal expression for pS6Ser235/236, with manyof the cells having cytoplasmic staining located in the

Abstracts 21



basal and parabasal cells strata. Indeed, the doubleimmunostaining of pS6Ser235/236 combined with the pro-liferation marker Ki-67 showed the co-expression ofthese molecules in some basal and parabasal neoplasticcells of most CSCC samples, a finding that was notobserved in normal feline skin. We also have identifieda group of CSCC feline patients displaying active Aktand S6 in the absence of EGFR activation. Expressionof PTEN characterized by a cytoplasmic immunoreac-tivity in the basal and spinous cell strata was reducedor absent in 15 of 45 CSCC samples. These findingsshow that the persistent activation of the PI3K/Akt/mTOR signalling pathway represents one of the keyevents during CSCC progression in cats, pointing tothis signalling pathway as a potential therapeutic targetin CSCC feline patients. These results also demonstratethat the activation of the PI3K/Akt/mTOR signallingpathway is independent from EGFR activation in somefeline CSCC, suggesting that therapeutic approachesaimed at modulating the growth and survival oftumour cells by the use of EGFR inhibitors would beineffective as sole therapeutic agents in some CSCCfeline patients.Source of funding: Self-funded.


SOS-13

A heterozygous novel non-sense variant in the

COL5A1 gene in two likely related Bombay

cats with Ehlers–Danlos syndrome

J. SCHISSLER*, F. LEUTHARD†,‡, V.JAGANNATHAN†,‡ and T. LEEB†,‡

*Department of Clinical Sciences, Colorado StateUniversity College of Veterinary Medicine andBiomedical Sciences, Fort Collins, CO, USA; †Instituteof Genetics, Vetsuisse Faculty, University of Bern, Bern,Switzerland; ‡DermFocus, University of Bern, Bern,Switzerland

Ehlers–Danlos syndrome (EDS) in humans is classifiedinto 13 subtypes involving not only cutaneous asthenia,but also musculoskeletal, vascular, neurological andmast cell activation disorders. EDS is a result of colla-gen and/or extracellular matrix changes and its preva-lence in people is estimated at 1:2,500–5,000individuals. The most common form, classical EDS, istypically caused by COL5A1 gene variants resulting inabnormal collagen type V. In cats, presumptive EDSwith cutaneous laxity has been rarely described, withvariable concurrent vasculopathy, joint laxity and epi-lepsy. An 8-month-old female spayed Bombay cat(Patient A) presented for lumbar self-trauma, periauralpruritus, and four wounds of the neck and lumbarregion since adoption from a shelter four months previ-ously. Histopathological evaluation indicated severeleucocytoclastic vasculitis and superficial collagen calci-fication. Videos demonstrated cutaneous trunci spasmswith frantic licking and biting of the dorsum. Examfindings included a skin extensibility index of 19%,focal alopecia and papulocrustous dermatitis of thebase of the neck/top of head, and a 7.5 cm full-thick-ness lumbar ulcer. Three months after presentation,Patient A experienced multiple partial and grand malseizures necessitating phenobarbital therapy. A secondfemale spayed Bombay cat diagnosed with EDS

(Patient B) was reported to the author. The veterinar-ian and owner were contacted and permitted release ofPatient B’s medical record for study. It was discoveredthat Patient B was obtained from the same shelterapproximately one year before Patient A and bothpatients originated from Rock Springs, Wyoming,USA. Patient B’s record described 13 documentedwounds of the ear, neck, elbow or shoulder over a per-iod of 28 months, starting at approximately 5 monthsof age. Ulcers were reported to occur after play or epi-sodes of scratching. Wounds continued despite reardeclaw. Additionally, a Grade 3 patellar luxation wasobserved at approximately 18 months of age. Ethylene-diaminetetraacetic acid blood samples were obtainedand genome sequencing was performed in bothpatients. A non-sense variant in the COL5A1 gene wasidentified, XM_023242950.1:c.3514A>T orXP_023098718.1:p.Lys1172*. Sanger sequencing con-firmed that both patients were heterozygous at thisnon-sense variant, indicating a common ancestor of thetwo cats and an autosomal dominant mode of inheri-tance. The finding of a COL5A1 variant with autoso-mal dominant inheritance as well as the array ofclinical signs displayed suggest that cats may be a suit-able clinical model of Classical EDS in humans.Source of funding: Self-funded.


SOS-14

Onset and duration of action of lokivetmab at

2 mg/kg in a canine model of IL-31-induced

pruritus

A. GONZALES, T. FLECK, L. NORRIS, S.MAHABIR, S. DUNHAM and O. MARTINONVeterinary Medicine Research & Development, Zoetis,Inc., Kalamazoo, MI, USA

Lokivetmab (Cytopoint) is a caninized monoclonalantibody that specifically targets canine interleukin(IL)-31 and is approved for dogs with allergic and ato-pic dermatitis. Lokivetmab functions to block IL-31-mediated cell signalling, providing relief from condi-tions associated with allergic dermatitis such as pruritusand skin inflammation. Two studies utilized an estab-lished, well-characterized canine model of IL-31-induced pruritus in which pruritic behaviours such aslicking, scratching, head-shaking and body rubbing areobserved by video surveillance (in real time) and scoredover a 2 h period following injection of purified canineIL-31. In both studies, 24 laboratory beagles ranging inage from 1.5 to 4.7 years of age (n = 6/group) wereevaluated for pruritus. Categorical presence/absence(yes/no) of pruritic behaviour displayed by each animalwas continuously measured via real-time video surveil-lance in discrete 1 min periods for 2 h and hypotheseswere tested at the two-sided 10% significance level. Thefirst study objective was to evaluate the duration ofantipruritic activity of multiple dose levels of lokivet-mab. Subcutaneous administration of placebo or loki-vetmab (0.125, 0.5 or 2.0 mg/kg) occurred on study day(D)0. Pruritic activity of dogs then was evaluated in themorning on D14, D28, D42 and D56. On those days,dogs were given a 1.75 µg/kg intravenous bolus ofcanine IL-31 approximately 15–20 min before thescheduled 2 h observation period. The duration of

22 Abstracts



action was defined by a statistically significant reduc-tion in pruritic behaviour relative to placebo. Studyresults showed that a significant reduction in prurituswas observed up to D14, D28 and D42 for doses of0.125, 0.5 and 2.0 mg/kg, respectively. Statistical analy-sis of the data with mean pruritic scores and corre-sponding P-values relative to placebo for D 14 (91versus 52, P < 0.001), D28 (81 versus 55, P = 0.0218)and D42 (89 versus 60, P = 0.0257) support theantipruritic properties of lokivetmab. In order to evalu-ate the onset of action of lokivetmab, laboratory bea-gles were given 2.0 mg/kg lokivetmab s.c., then IL-31was administered 2.5 h post-lokivetmab administration,and dogs were evaluated for pruritus between 3 and5 h post-treatment. These results demonstrate that asingle subcutaneous 2 mg/kg dose of lokivetmab signifi-cantly reduces pruritic activity in a canine model of IL-31-induced pruritus when efficacy is evaluated 3–4 h(P < 0.0001, mean pruritic score for placebo = 37 ver-sus 10), 4–5 h (P < 0.0001, 52 versus 20) or 3–5 h(P < 0.0001, 90 versus 29) post-antibody administrationand the duration of antipruritic activity was sustainedfor ≥42 days.Source of funding: Zoetis Inc., Parsippany, NJ, USA.

Conflict of interest: All authors received reimbursements,

fees, funding or salary from Zoetis Inc.

SOS-15

Detection and inhibition of IgE antibodies

reactive with cross-reactive carbohydrate

determinants in an ELISA for allergen-specific

IgE in horses

K. ENCK*, K. LEE*, B. MCKINNEY*, K.BLANKENSHIP* and C. MONTESANO†

*Stallergenes Greer, Lenoir, NC, USA; †Montesano &Tallarico, D.V.M., LLP, New York, NY, USA

Recent findings demonstrate that IgE antibodies speci-fic for cross-reactive carbohydrate determinants (CCD)

are present in the serum of allergen-sensitized humans,dogs and cats, and these CCD-specific antibodies mightyield false positive results in serological tests for detec-tion of allergen-specific immunoglobulin (Ig)E. Theobjective of this study was to determine whether or notsimilar CCD-reactive antibodies occur in horses, and toinvestigate the prevalence of CCD-reactive IgE antibod-ies in equine sera using a previously defined inhibitor(BROM-CCD) and a monoclonal cocktail basedenzyme-linked immunosorbent assay designed to detectallergen-specific IgE in horses. Sera from 28 horses,presumed to be allergic, were evaluated with and with-out BROM-CCD using a panel of 72 different allergensthat included 15 grasses, 17 trees, nine weeds, fivemites, 12 fungi, 12 insects and two environmental aller-gens. Without BROM-CCD, 22 horses reacted to atleast one of the allergens in the panel. Of these reactivehorses, 15 horses were reactive to ≥10 allergens, andnine of these had demonstrable IgE to >25 of the aller-gens. BROM-CCD had little or no effect on the mitereactivity in any of the positive samples; however, sub-stantial inhibition for pollen allergens (trees, grassesand weeds) was demonstrable. Complete inhibition ofsignal was evident in only two of the reactive samples.However, reduction in signal of ≥20% was evident in20 of the reactive samples and reduction >50% wasobserved in eight of the reactive horses. Forty percentof the signals evident in 18 of the samples were inhib-ited by CCD-BROM. These results demonstrate thatCCD-reactive IgE antibodies are evident in horses andthat BROM-CCD is effective in reducing reactions withthese irrelevant carbohydrates. Thus, inhibition ofCCD reactivity will likely yield a more accurate in vitroallergen reactivity profile to be used in the selection ofallergens for an immunotherapeutic regime.Source of funding: Funding for this study was provided

by Stallergenes Greer.

Conflict of Interest: KE, KL, BMcK and KB are employ-

ees at Stallergenes Greer.

Abstracts 23



FREE COMMUNICATION ABSTRACTS

ALOPECIA

FC-1

Comparison of skin thickness and hair follicle

density in horses with pituitary pars intermedia

dysfunction (PPID) and aged non-PPID

horses

A. PETERSEN*, S. FRENCH†, A. HASBACH* andH. SCHOTT II‡

*Department of Small Animal Clinical Sciences,Veterinary Medical Center, College of VeterinaryMedicine, Michigan State University, East Lansing, MI,USA; †Veterinary Diagnostic Laboratory, College ofVeterinary Medicine, Michigan State University, EastLansing MI, USA; ‡Department of Large AnimalClinical Sciences, Veterinary Medical Center, College ofVeterinary Medicine, Michigan State University, EastLansing, MI, USA

There is a paucity of reports describing histopathologi-cal characterization of equine skin. A recent studyreported that horses with PPID have thinner neck skinas compared to aged horses without PPID. Followingeuthanasia (late autumn), we collected skin biopsiesfrom nine different locations in PPID horses (n = 9)and aged non-PPID horses (n = 5). Areas includedproximal and distal dorsal forelimb, mane, tail, fore-head, side of neck, jugular groove, ventral abdomenand rump. Biopsies were processed routinely; sagittaland transverse (at the level of the sebaceous duct paral-lel to the skin surface) plane sections were examined.Skin thickness was measured and the number of hairfollicles were counted in a 2 x 2 mm square superim-posed on photomicrographs of x4 microscopic magnifi-cation of haematoxylin & eosin stained sides. Nosignificant differences in skin thickness or hair follicledensity were found between PPID samples and agednon-PPID horses. Consequently, data for both groupswere combined for comparison of skin thickness andhair follicle density between the nine body regions.Mane and tail skin were significantly thicker than theother seven sites. In addition, hair follicle density wasgreater in the face than other body sites. In conclusion,we could not replicate a previous finding of thinnerneck skin of PPID horses versus aged non-PPIDhorses. Nevertheless, we were able to show that skinthickness is greater in mane and tail as compared tothe other sites. Additionally, density of hair follicles isgreater in the face than other body sites.

Funded by a grant from ACVD.

Conflicts of interest: None declared.

FC-2

Clinical features of hair cycle arrest: a

retrospective study of 63 cases in Japan

K. MORI, Y. OSHIMA-TERADA and N.MURAYAMADermatology Services for Dogs and Cats, Tokyo, Japan

Hair cycle arrest (HCA) is a common disease yet thereare few retrospective studies. This study aimed to assessthe clinical features of and therapeutic outcomes forHCA in Japan. Sixty-three dogs with HCA wererecruited based on the clinical, endocrinological andhistopathological features. The therapeutic evaluationincluding four sites (neck, dorsum, ventral region andcaudal portion) before and after a therapy used a hairregrowth scale (0, 0–25%; 1, 25–50%; 2, 50–75%; 3,75–100%) based on clinical records and digital images.Treatment success was defined as improvement of morethan four scores before and after a therapy. Toy poo-dles (30) and Pomeranians (22) were significantly over-represented (P < 0.01). Forty-seven dogs were assessedfor therapeutic outcomes and 48.9% of these showedtreatment success. Oral tocopherol nicotinate (JuvelaN, Eisai; Tokyo, Japan) with L-cysteine (HYTHIOL,Hisamitsu Pharmaceutical; Tokyo, Japan), oral osater-one acetate (UROACE, Aska Animal Health; Tokyo,Japan), oral essential fatty acids (Antinol, V and P;Tokyo, Japan) and a prescription diet (Derm Defence,Hill’s-Colgate; Tokyo, Japan) were used to treat; suc-cess rates were 39.5% (17/43), 27.3% (6/22), 18.2% (2/11) and 37.5% (3/8), respectively. The median time totreatment success was 148 days (63–1,239 days). Theduration-by-treatment success rates for <3 months, 3–6months, 6–9 months, 9–12 months and >12 monthswere 13.0% (3/23), 47.8% (11/23), 8.7% (2/23), 4.3%(1/23) and 26.1% (6/23), respectively. This study indi-cated that oral tocopherol nicotinate and L-cysteine wasthe most effective therapy, with 60.8% treatment suc-cess by six months and 26.1% with hair regrowth after12 months.Source of funding: Self-funded.


24 Abstracts



NEOPLASTIC SKIN DISEASES

FC-3

Clinical, cytological, histopathological and

immunohistochemical findings of cutaneous

non-epitheliotropic lymphoma in a cat: case

report

M. PONTES*, A. CASTRO†, T. SILVA†, H.PESSOA†, F. TOLEDO‡, D. SOUZA§ and F.SUEIRO¶

*Department of Veterinary Medicine, UNIBRA, PE,Brazil; †Autonomous Veterinary, PE, Brazil;‡Department of Veterinary Clinical and Surgical Studies,Federal University of Minas, Gerais, MG, Brazil;§Department of Veterinary Medicine, UNIBRA, PE,Brazil; ¶Faculty of Agricultural and Veterinary Sciences,UNESP, SP, Brazil

Cutaneous manifestation of primary lymphoma of theskin in cats is a rare condition. This report describes acase of primary non-epitheliotropic cutaneous lym-phoma in an 8-year-old mixed breed cat presenting withmultifocal and nodular skin lesions not adhered to deeptissues. Some nodules were erythematous and hairless.Cytological findings from fine needle aspirates revealeda high population of atypical lymphoid cells character-ized by scant to moderate amounts of basophilic cyto-plasm, round and eccentric nucleus, displaying intenseanisocytosis and anisokaryosis, little mitosis and abun-dant lymphoglandular bodies in the background. Histo-logically, tumours were replacing subcutaneous tissueand were composed by a substantial monomorphicpopulation of large lymphocytes arranged in mantle orsolid pattern. The cells had scant to moderate amountsof eosinophilic cytoplasm and rounded cleaved nucleiwith prominent small central nucleoli. The cells hadintense anisocytosis and anisokaryosis. There generallywere eight to nine mitosis per high power field (x400).Epitheliotropism was not seen and the neoplasia wasrestricted to the subcutaneous tissue. The neoplasticcells immunoexpressed CD79a and MUM1 and did notexpress CD3, Tryptase, PAX5 or C-Kit. There was pos-itive proliferation marker in approximately 70% ofneoplastic cells. The diagnosis of non-epitheliotropiclymphoma of large cells was made. Diagnosis of pri-mary cutaneous non-epitheliotropic lymphoma may bebased on cytological and histological findings, yetmolecular diagnosis is the gold standard for treatment,although prognosis was very poor for this case.Source of funding: Self-funded.


FC-4

The characterization of the cellular immune

environment in equine genital papillomas and

squamous cell carcinomas

I. PORCELLATO*, M. ORLANDI*, P. MODESTO‡,V. CIMADOMO‡, C. BRACHELENTE*, L.MECHELLI*, B. PASSERI†, K. CAPPELLI*, S.MECOCCI*, R. GIALLETTI*, A. GHELARDI§ andE. RAZZUOLI‡

*Department of Veterinary Medicine, Universit�a degliStudi di Perugia, Perugia, Italia; †Department ofMedical Veterinary Sciences, University of Parma,Parma, Italy; §Azienda Usl Toscana Nord-Ovest, UOCOstetricia e Ginecologia, Ospedale Apuane, Massa,Italy; ‡Istituto Zooprofilattico Sperimentale delPiemonte, Liguria e Valle d’Aosta, Genova, Italy

Equine genital squamous cell carcinomas (SCCs) arecommon skin tumours, often caused by Equus caballuspapillomavirus 2 (EcPV-2) infection, characterized byhigh local invasiveness and a 20% risk of metastasis tolymph nodes. Growing evidence, both in humans andanimals, indicates that presence and type of immuneinfiltrate within the tumour microenvironment can dee-ply influence the behaviour of cancer. As no data onthe immune environment of genital equine SCCs havebeen reported, our aim was to characterize the cellularimmune infiltrate in this tumour. Twenty-five formalin-fixed, paraffin-embedded (FFPE) cases of equine geni-tal SCCs were retrospectively retrieved from ourarchives and submitted for histological evaluation ofinflammatory cells. Real-time PCR was performed onFFPE samples to assess the presence of EcPV-2 withinthe lesions. Immunohistochemical analysis for CD3,CD20, MUM1, MAC387, IBA1 and FoxP3 was per-formed. Results confirmed the presence of EcPV-2 inall tested cases and showed in most of the cases a mas-sive plasmacytic immune infiltrate (MUM1+), withfewer B (CD20+) and T (CD3+) lymphocytes. Numer-ous histiocytes (IBA1+), mostly located within thetumour stroma and also infiltrating nests of neoplasticcells, were present with fewer macrophages and neu-trophils expressing calprotectin (MAC387+). Neu-trophils were abundant in ulcerated lesions, whereas insome cases eosinophils were predominant. The immuneinfiltrate observed in equine genital SCCs shows somesimilarities with human HPV-induced cervical cancer.Moreover, the presence of plasma cells and B cells,IBA1+ and FoxP3+ cells, may suggest the presence ofa tumour-promoting and actively immunosuppressiveenvironment in these tumours.Source of funding: Ministry of Health (Ministero della

Salute).


Abstracts 25



PARASITES AND PARASITOLOGY

FC-5

Severe eosinophilic dermatitis in Jersey cows

infested with Leptotrombidium spp. (Acari:

Trombiculidae)

M. C. CADIERGUES*,†, J. P. MOULIN‡, M. N.LUCAS§, P. DORCHIES* and F. SCHELCHER§,¶

*Small Animal Clinic, Universit�e de Toulouse, ENVT,Toulouse, France; †UDEAR, Universit�e de Toulouse,INSERM, ENVT, Toulouse, France; ‡Cliniquev�et�erinaire, Saint-C�er�e, France; §Large Animal Clinic,Universit�e de Toulouse, ENVT, Toulouse, France;¶IHAP, Universit�e de Toulouse, INRA, ENVT,Toulouse, France

A serious outbreak of pruritic dermatitis was investi-gated in mid-October in an organic farming dairy herdwith 21 Jerseys, three Bretonne pie noir and fourcrossed Abondance–Montb�eliarde cows in south westFrance. Lactating cows (17) had permanent grazingaccess except for indoor milking times. Five 2-year-oldand six 1-year-old heifers had been permanently kept intwo different meadows, with no contact between themor with adults, since the beginning of the summer. Noantiparasitic treatment had been administered for theprevious five months. Severe and extensive lesions ofalopecia, lichenification and crusting were observed onthe jaws, dewlap, face, perineum, above the udder anddistal limbs, associated with moderate pruritus. OnlyJersey cows were affected (10 of 13 lactating cows andall three 2-year-old heifers) with various degrees oflesions severity. Heifers displayed more intense lesionsthan adults. Microscopic examination of deep skinscrapes showed numerous specimens of Trombiculidaelarvae identified as Leptotrombidium spp. and no otherectoparasites. Histopathological examination of skinbiopsy specimens included severe hyperplastic, superfi-cial and deep perivascular, oedematous, eosinophilicdermatitis. Complete blood counts revealed mild tomoderate eosinophilia. The final clinicopathologicaldiagnosis was eosinophilic dermatitis suspected to be aconsequence of Trombiculidae bites. The lesions com-pletely self-resolved by the end of December. This casereport emphasizes the need to include Trombiculidae inthe differential diagnosis of bovine pruritic dermatoses.Finding lesions only in Jersey cows remained unex-plained and could be hypothetically consistent withindividual genetic susceptibility to develop hypersensi-tivity reactions, or other predisposing breed factors(e.g. feeding habits or skin characteristics).Source of funding: Self-funded.


Published in Veterinary Dermatology, https://doi.org/10.1111.vde.12895

FC-6

Development of a quantitative PCR to

evaluate accurately the treatment response of

dogs with demodicosis

S. MART�INEZ*,†, N. SASTRE‡, O. FRANCINO‡, L.MAT�E*, S. SANCHEZ BRUNI*, M. J. DEL SOLE*,†

and L. FERRER§

*Centro de Investigaci�on Veterinaria Tandil(CIVETAN), CIC-CONICET-UNCPBA, Tandil,Buenos Aires, Argentina; †Hospital Escuela de Peque~nosAnimales (HEPA), Facultad de Ciencias Veterinarias dela Universidad Nacional del Centro de la Provincia deBuenos Aires (FCV-UNCPBA), Tandil, Buenos Aires,Argentina; ‡Servei Veterinari de Gen�etica Molecular,Universitat Aut�onoma de Barcelona, Bellaterra,Barcelona, Spain; §Departament de Medicina i CirurgiaAnimals, Universitat Aut�onoma de Barcelona, Bellaterra,Barcelona, Spain

Canine demodicosis is the result of an overpopulationof Demodex mites, normal inhabitants of most skin ofmammals. There is no reliable method to quantify thenumber of Demodex mites present in the skin to pro-vide accurate information about treatment response.The objective of this study was to develop a moleculartechnique to quantify the Demodex load in canine skinin order to evaluate the treatment response in dogs withdemodicosis. For this purpose, a real-time quantitative(q)PCR was developed using primers that amplified afragment of the D. canis 18S rRNA gene. Ten-fold dilu-tions of the DNA extracted from isolated mites wereused to elaborate the standard curve. In addition, 44skin biopsies (4 mm) obtained from nine canine cadav-ers and 18 skin biopsies from seven dogs with demodi-cosis in day (D)0 and D14, D35 and D56 post-treatment (Animal Welfare Committee, FCV-UNCPBA) were used to assess the number of mites inthe skin of healthy dogs and of dogs with demodicosis.The skin biopsies of healthy dogs were negative in 38/44 samples and positive (>one mite) in the rest (6/44).However, in skin biopsies of diseased dogs the initialDemodex load was mostly >100 mites, decreasing to50–100, 10–50 and 0–10 mites at D14, D35 and D56post-treatment, respectively. The homology between theqPCR product and the 18S rRNA gene was confirmedby DNA sequencing (GenBank KC010485 – D. canis).This simple technique could be a useful tool to evaluateaccurately the response to treatment of dogs withdemodicosis.Source of funding: PICT 2016-0608, ANPCyT, Argen-

tina (SSB) and HEPA, FCV, UNCPBA (MJDS).


26 Abstracts



https://doi.org/10.1111.vde.12895

https://doi.org/10.1111.vde.12895

FC-7

A case report of Demodex cyonis in a toy

poodle

K. MORI*, T. MORITA†, Y. OSHIMA-TERADA*, I.TODA‡, H. KONDO§ and N. MURAYAMA**Dermatology Services for Dogs and Cats, Tokyo,Japan; †School of Veterinary Medicine, NipponVeterinary and Life Science University, Tokyo, Japan;‡Toda Animal Hospital, Tokyo, Japan; §NihonUniversity College of Bioresource Sciences, Kanagawa,Japan

Several species of Demodex mites parasitizing dogs,D. canis, D. injai, D. cornei and D. cyonis have beendescribed morphologically. Demodex cyonis has beenreported but without details of the clinical course. Thisreport describes the clinical course in a toy poodle withD. cyonis. A 15-year-old, intact female toy poodle pre-sented with pruritus, erythema and seborrhoea on thedorsum one year previously. The dog had been treatedwith oral oclacitinib (Apoquel, Zoetis Japan; Tokyo,Japan) at 0.5 mg/kg twice daily for one year. Hairexamination and skin scraping showed short- and wide-

bodied Demodex mites. The histopathological findingson the dorsum revealed orthokeratotic hyperkeratosison the epidermis and the hair follicles, sebaceous glandhyperplasia and apocrine gland dilution. The hair folli-cles contained Demodex mites. However, there were noapparent inflammatory cells in the epidermis, dermisand hair follicles. Microscopic evaluation for the Demo-dex mites by transmission electron microscopy? identi-fied the typical opisthosomal organ of male D. cyonis.The diagnosis based on the clinical and examinationfindings was canine demodicosis caused by D. cyonis.Oral sarolaner (Simparica, Zoetis Japan) at 10 mg totaldose was administered, and the administration of oclac-itinib was stopped. One month later, pruritus disap-peared and Demodex mites were not detected by hairexamination or skin scraping. Pruritus immediatelyimproved with administration of oral sarolaner andthus D. cyonis could be associated with pruritus.Although this and previous reports suggest that D. cy-onis might favour sebum, more cases will be needed toassess predisposing factors.Source of funding: Self-funded.


Abstracts 27



OTITIS

FC-8

Partial traction/avulsion followed by systemic

and topical glucocorticoid therapy for the

management of feline aural inflammatory

polyps

J. BACHTEL*, J. SCHISSLER† and R. ROSYCHUK†

*Veterinary Referral Center of Colorado, Englewood,CO, USA; †Department of Clinical Sciences, College ofVeterinary Medicine and Biomedical Sciences, ColoradoState University, Fort Collins, CO, USA

Traction/avulsion therapy for aural polyps in cats has areported 33–85.7% recurrence, likely due to re-expan-sion of the residual polyp in the middle ear. Post-proce-dural glucocorticoid therapy may facilitate residualpolyp regression. The efficacy and recurrence of partialtraction/avulsion followed by systemic and topical glu-cocorticoid therapy for polyps perforating the tympanicmembrane (TM) and systemic and topical glucocorti-coid therapy for nonperforating polyps were evaluatedfor 25 aural polyps in 22 cats. Three of 22 cats hadbilateral polyps, each with a perforating and nonperfo-rating polyp. Five cats had only a nonperforatingpolyp. All but one (16/25) perforating polyps had allpolypoid material removed from the horizontal canalby traction/avulsion (biopsy forceps) to the level of orjust within the TM. Other than for biopsy, cats withnonperforating polyps did not have polyp materialremoved from the middle ear. Post procedure, 21/22cats (24/25 polyp ears) were treated with oral pred-nisolone at a mean dose of 2.37 mg/kg (range of 0.91-3.2 mg/kg) and tapered over 6–18 weeks (mean 8.72weeks). One cat received subcutaneous dexamethasone.Polyp ears received topical glucocorticoid therapythroughout systemic glucocorticoid treatment. Follow-up ranged from 95 days to 10.4 years (mean 3.7 years).No recurrence of perforating polyps and no recurrence/progression was noted for the nonperforating polyps.Based on these results, partial avulsion/traction fol-lowed by an eight-week course of prednisolone startingat 2–3 mg/kg/day with concurrent topical corticosteroidtherapy is as effective as aggressive surgical removal(ventral bulla osteotomy, ear canal ablation, middle earcurettage).Source of funding: Self-funded.

Conflict of Interest: None declared.

FC-9

Efficacy of antibiotic treatment in

Pseudomonas aeruginosa otitis in dogs:

prospective study in 12 dogs

V. BRUET*, T. BR�EMENT*, M. FUSELIER† and P.BOURDEAU**Dermatology, Parasitology, Mycology Service, Oniris,Nantes, France; †Diagnostic Imaging Service, Oniris,Nantes, France

Recent literature on medical management of Pseu-domonas otitis and the impact of different approaches(antibacterial, glucocorticoid) on healing are lacking. Inthis prospective study, the aim was to evaluate the

efficacy of a treatment focused on a unique antibacte-rial approach in a two month protocol. Bacterial isola-tion and antibiogram were performed at day (D)0 andD60. Magnetic resonance imaging also was evaluated.The protocol was to flush the external ear and, in thecase of media otitis, the middle ear; no glucocorticoidswere used. Medical treatment for otitis externa (OE)included local instillation of gentamycin (G4, Virbac;Carros, France) used off label and EpiOtic (Virbac)every day. For otitis media, treatment included mar-bofloxacin (Marbocyl, V�etoquinol) topically (off label)and orally and topical EpiOtic. Meloxicam (Metacam,Boehringer Ingelheim) was used to control pain. Twelvedogs representing seven breeds were treated; most werecocker spaniels (four) and German shepherd dogs(three). The median age was 6.4 years. Otitis was uni-lateral in seven dogs and bilateral in five. There werenine OE cases and three individuals with otitis media.Pseudomonas aeruginosa isolates were sensitive to gen-tamycin in 5/12 dogs and sensitive to marbofloxacin in7/12. After two months, six dogs were free of Pseu-domonas (four without otitis, two with other organ-isms), while two dogs still had Pseudomonas otitis.Three dogs were excluded owing to excessive pain (two)or deviation from the protocol (one); one dog was lostto follow-up. This study shows the improvementobtained with a unique antibacterial approach.Source of funding: Study funded by Virbac Corporate,

France.


FC-10

Bacterial chronic otitis: correlation between

cytological inflammatory patterns, clinical

aspect and bacteriology in 64 dogs

V. BRUET, T. BREMENT, M. RIVALAIN and P.BOURDEAUDermatology, Parasitology, Mycology Service, ONIRIS,Nantes, France

The objective of this study was to evaluate cytologicalpatterns in chronic bacterial otitis to characterize theirclinical or bacteriological implications. One hundredand four ears (64 dogs) with chronic (minimum twoweeks) bacterial otitis externa (bacteria observed thenidentified bacteriologically) were included. Primary fac-tors, chronicity, previous topical antibacterial treat-ments, pain, clinical scoring (OTIS-3) and classification[erythematous (Er), ceruminous (Cer), erythemato-ceru-minous (ErCer), suppurative (Sup)] were collected. Oti-tis was classified on inflammatory pattern fromcytological results: bacterial overgrowth (no inflamma-tory cells) (BOG) (43 ears, 41.4%), suppurative (>90%neutrophils) (Pyo) (55 ears, 52.9%), pyogranulomatous(>50% neutrophils and macrophages) (PyoG) (six ears,5.8%). Chi-squared test or Fisher’s exact test were used(P < 0.05). No correlation was observed between thecytological pattern and chronicity (over two months:76% BOG, 85% Pyo, 100% PyoG), pain, OTIS-3 orthe presence of Malassezia. There was a statistical cor-relation between cytological pattern and: primary fac-tors (BOG observed in 89.3% of allergic otitis)

28 Abstracts



(P = 0.02); number of previous antibacterial treat-ments [0 (58% BOG, 31% Pyo, 17% PyoG) versus 1, 2or 3 (P = 4.6 9 10-2)]; and clinical type of otitis [BOGwith ErCer (61.9%), Pyo and PyoG with Sup (59.2%and 66.7%, respectively)] (P = 5 9 10-3). There alsowas a statistical correlation with bacteria isolated: Sta-phylococcus and Corynebacterium with BOG (53.8%and 50%, respectively), Pseudomonas with Pyo/PyoG(80%) (P = 0.04); abundance of Malassezia [moderate,high level with BOG (84.6%)] (P = 2.4 9 10–3). Theseresults highlight the importance of cytological inflam-matory patterns for describing otitis and possible rela-tionships with bacterial causal agent.Source of funding: Study funded by Virbac Corporate,

France.


FC-11

Clinical and microbiological performances and

effects on lipid and cytokine production of a

cerumenolytic ear cleaner in canine

erythemato-ceruminous otitis

F. MOOG*, J. MIVIELLE*, J. BRUN*, N.AMALRIC†, L. A. LECRU*, C. PRESSANTI*,‡, J.KONDRATJEVA*, D. COMBARROS*,‡, O.FANTINI§ and M. C. CADIERGUES*,‡

*Small Animal Clinic, Universit�e de Toulouse, ENVT,Toulouse, France; †Synelvia, Lab�ege, France; ‡UDEAR,Universit�e de Toulouse, INSERM, ENVT, Toulouse,France; §Vetoquinol, Paris, France

Erythemato-ceruminous otitis (ECO) is the most com-mon type of otitis in dogs and generally is associatedwith bacterial and/or fungal infections. Performance ofthe ear cleaner Sonotix (Vetoquinol; Lure, France) wasassessed over two weeks in canine ECO, associated witha mild or moderate secondary infection, in a prospec-tive open-label study (Ethics Committee approval no.SSA_2019_003). Forty ear canals with ECO not receiv-ing any type of aural treatment or cleaning for sevendays were included. Pruritus (PS), OTIS3 and 0–4 scalecytological (CYTO) scores were assessed on Day (D)0,D7 and D14. Aural surface concentrations of a panelof 13 cytokines and lipid profiles of the exudate weremeasured on D0 and D14. Ear cleaning was performeddaily if the secretion score (SEC) was 3/3, every secondday if 2/3 and every third day if 1/3 from D0 to D12or D13. PS, OTIS3, SEC and CYTO were significantlylower on D7 compared to baseline (–40%, –31%, –36%, –34%, respectively; P < 0.001). PS, OTIS3, SECand CYTO further decreased on D14 (–60%, –53%, –61%, -73%, respectively; P<0.0001) and Interleukin 8and chemokine (KC)-like amounts also were reducedcompared to baseline (–62%, P < 0.05; –36%, P = 0.3,respectively). The lipid profile also was modified withreduced free lipids and increased bound lipids. The useof Sonotix at a frequency based on the amount of auralsecretions significantly decreased the infectious andinflammatory components of ECO and could providean additional tool for managing this type of otitis.Source of funding: Vetoquinol, Lure, France.

Conflicts of interest: MCC has previously been a consul-

tant for Vetoquinol. The residency programme of LAL

was supported by Vetoquinol. NA is employed by

Synelvia. OF is employed by Vetoquinol. FM, JM, JB,

CP, JK and DC declare no conflicts of interest.

FC-12

Florfenicol in vitro antibacterial activity and

genetic resistance of Staphylococcuspseudintermedius strains selected from dogs

with otitis

A. CORONA, A. VERCELLI, L. GUIDI andL. CORNEGLIANIClinica Veterinaria Citt�a Di Torino, Italy

Florfenicol (FFC) is a broad-spectrum fluorinatedantibiotic, derived from chloramphenicol (Cf) andactive against Gram-positive and -negative bacteria.The aims of this study were to describe the in vitroantibacterial activity of FFC against Staphylococcuspseudintermedius (SP) strains and to evaluate the pres-ence of resistance genes. Six hundred SP strains fromear canals of dogs with otitis were identified using API(bioM�erieux; Marcy-l’�Etoile, France). In vitro suscepti-bility was tested with the “minimum inhibitory concen-tration (MIC) Test Strip” method (Liofilchem; Rosetodegli Abruzzi, Italy) with values between 0.016 and256 µg/mL. After heat treatment, extracted bacterialDNA was resuspended in Tris-HCl-EDTA (TLE) buf-fer and stored at -20°C. Real-time PCR was performedwith Step One (Life Technologies Corporation, USA)for identification of the cfr (chloramphenicol/florfenicolresistance) and fexA genes. The reaction included aFast SYBR Green Master Mix with primers cfr-fw/cf-rv and fexA-fw/fexA-rv. Melting temperature analysesand relative curves were performed. The amplificationproducts were tested using a 2% agarose gel elec-trophoretic run containing ethidium bromide, to showthe bands of at 746 and 1,272 bp corresponding to cfrand fexA, respectively. Of the SP strains, 99.3% had aMIC to florfenicol from 0.064 to 4 µg/mL and 0.7%showed a MIC of ≥24 µg/mL. The PCR for cfr andflexA genes were positive for 100% of four strains withhigh MIC (≥24 µg/mL) and corresponded to 0.7% ofall SP tested, demonstrating an effective inhibitingaction against SP. However, as resistance genes can beacquired during FFC-based treatments, FFCs shouldbe used with caution.Source of funding: Self-funded.


FC-13

Evaluating factors associated with Malasseziapachydermatis presence in canine otitis externa

A. FLORES*, C. ROMERO† and L. G. B. G�OMEZ‡

*University of the State of Mexico (UAEM),Amecameca, Mexico; †DERMAVET, Iztapalapa,Mexico City, Mexico; ‡University Center-AmecamecaUnit of the Autonomous University of the State ofMexico UAEM, Amecameca, Mexico

Factors such as breed, age and air conditioning expo-sure influence the development of yeast otitis. The dis-ease often is diagnosed by cytological evaluation yetPCR is useful to detect Malassezia pachydermatis. This

Abstracts 29



study aimed at evaluating the presence of yeast in theexternal ear canal using PCR and cytological methodsin order to establish the correlation between yeast andfactors associated with canine otitis. A comparativecross-sectional study approved by the ethics committeeof the UAEM was conducted (registration no. 228/CUAMECAMECA / UAEMEX / 2019). Sixty dogswere examined. NL1 and NL4 primers were used in thePCR test. Additionally, a 600 bp fragment wassequenced. BLAST was used to compare the obtainedDNA sequence and reported sequences. We conducteda phylogenetic analysis using MEGA X 10.0 software.We used a chi-squared test (P < 0.05) to assess thefactors associated with the presence of M. pachyderma-tis detected by PCR and cytological evaluation. ByPCR we determined that the associated factors are pre-vious otitis, itching and air conditioning exposure.Cytological results corresponded with predisposing fac-tors of itching, ear shape, hair inside the ear and skinhealth. The obtained sequence was combined at 99.9%with M. pachydermatis. The phylogenetic analysisshowed that this sequence is close to sequences reportedin Colombia and Kuwait corresponding to a zoonoticstring endangering owners and veterinarians. Knowingthe predisposing factors for otitis externa is a tool toobtaining the diagnosis.Source of funding: Self-funded.


FC-14

Cytological and microbial culture from 60

normal feline ears

L. J. VOGELNEST*,†, A. NESSIEM† and J. M.NORRIS†

*Small Animal Specialist Hospital, North Ryde, NSW,Australia; †School of Veterinary Science, University ofSydney, NSW, Australia

This prospective study (University of Sydney ethicsN2016/1088) evaluated cytological findings and concur-rent microbial culture from 60 normal ears (30 cats).Sterile swabs inserted 1–2 cm into external canals wererotated for 1–3 s. An initial swab placed into transportmedium was inoculated onto sheep blood agar andSabouraud’s dextrose agar within 10 h. Plates wereincubated aerobically at 37C for 48 h and bacterial iso-lates identified using matrix-assisted laser desorption/ionization-time of flight. A second swab was rolledonto a glass slide (2 x 2 cm area) and stained withmodified Wright’s stain. Keratinocytes were identifiedunder x40 magnification, and nine oil-immersion fields(OIF; x100 magnification) with keratinocytes were eval-uated for microbes. Bacteria were cultured from 50 ears(83%) with 63 species isolated. Staphylococcus (25%),Bacillus (22%) and Lactobacillus (6%) were the mostfrequent genera, and S. felis, S. epidermidis and Micro-coccus luteus the most frequent isolates; Pseudomonas

spp. was isolated from one ear. Overall, cocci werefound from 40 ears (67%) and rods/coccobacilli from33 ears (55%). Malassezia were not found. There waspoor correlation of culture and cytological results, withmicrobes not apparent in 43 ears (72%) and 95% ofOIF. Cocci were detected in four ears (7%; median 0,range 1–3/OIF), rods in 12 ears (20%; median 0, range1–14/OIF), and yeast in five ears (8%; median 0, range1–2/OIF). By contrast with previous studies, bacterialrods were present in normal feline ear microbiota andmicrobes were sparse.Source of funding: Self-funded.


FC-15

Cytological and microbial culture from 60

normal canine ears

L. J. VOGELNEST*,†, A. P. KING† and J. M.NORRIS†

*Small Animal Specialist Hospital, North Ryde NSWAustralia; †School of Veterinary Science, University ofSydney, NSW Australia

No studies on cytological findings from normal canineears evaluated with oil-immersion microscopy havebeen published. This prospective study (University ofSydney ethics N2016/1089) evaluated cytological resultsand concurrent microbial culture from 60 normal ears(30 dogs). Two sterile swabs were collected; the firstplaced into transport medium and inoculated ontosheep blood agar and Sabouraud’s dextrose agar within10 h, with plates incubated aerobically at 37(C for48 h. Bacterial isolates were identified using matrix-assisted laser desorption/ionization-time of flight. Thesecond swab was rolled onto a glass slide (2 x 2 cmarea) which was stained with modified Wright’s stain.Keratinocytes were identified under x40 magnification,and nine oil-immersion fields (OIF; x100 magnification)containing keratinocytes evaluated for microbes. Bacte-ria were cultured from 48 ears (80%) with 93 speciesisolated. Staphylococcus (62%), Bacillus (33%) andMicrococcus (30%) were the most frequent genera, andS. pseudintermedius, Micrococcus luteus and S. hoministhe most frequent isolates. Cocci were isolated from 44ears (73%), rods/coccobacilli from 37 ears (61%),Malassezia from 29 ears (48%), Pseudomonas spp. fromsix ears and Aspergillus flavus from one ear. Cytologicalresults revealed cocci in 58 ears (97%; median 1, range1–8/OIF), rods in 56 ears (93%; median 1, range 1-6/OIF) and yeast in 40 ears (67%; median 5, range 1 to>100/OIF). Five dogs with >100 yeasts in at least oneOIF were excluded from median counts. By contrast toother studies, bacterial rods were identified as part ofnormal canine ear microbiota and yeasts were plentifulin some normal ears.Source of funding: Self-funded.


30 Abstracts



SKIN BIOLOGY AND ECOLOGY

FC-16

Carriage of Malassezia yeasts on healthy dogs

in Malaysia

A. L. BOOPet First Veterinary Centre, Kuala Lumpur, Malaysia

Malassezia spp. yeasts are both commensal organismsand opportunistic pathogens of canine skin. Althoughcanine Malassezia dermatitis is a common diagnosis inMalaysia, little is known about their presence on theskin of healthy dogs there. This study was performedto evaluate the prevalence of carriage and quantifica-tion of Malassezia spp. yeasts in healthy dogs inMalaysia. Thirty-eight clinically healthy dogs admittedfor neutering in a small animal veterinary clinic wereenrolled in the study. Swabs were collected from bilat-eral ear canals and tape-strip samples were obtainedfrom interdigital skin of both forelimbs. Cytologicalexamination was performed and results were expressedas the mean number of yeasts per x1,000 microscopicfield per dog. Fifty percent of the otic samples (n = 74)and 68% of the samples from the interdigital skin(n = 38) showed presence of Malassezia spp. yeasts oncytological examination. The mean number of Malasse-zia spp. yeasts was 1.86/oil-immersion field (high-powerfield, HPF) and 1.19/HPF for the otic samples andinterdigital skin, respectively. There was no statisticallysignificant difference in prevalence and number ofMalassezia spp. yeasts between dogs of different ages,sex, breed and area of sampling (P > 0.05). Theseresults are in accordance with previous findings in theliterature which suggested that a clinical diagnosis ofMalassezia dermatitis may be established if >2/HPFMalassezia spp. yeasts are found on cytological analysiscoupled with compatible skin lesions.Source of funding: Self-funded.


FC-17

Comparative study of microbubble wash

system and regular shampooing in the effect on

detergency and skin barrier function of dogs

K. SHIMADA* and T. IWASAKI†

*Azabujuban Dog and Cat Clinic, Tokyo, Japan; †VetDerm Osaka, Osaka, Japan

Microbubble wash system (MWS) is a new technologyto wash animal skin by using microbubbles withoutscrubbing. The purpose of this study was to evaluatethe effects of MWS on the canine skin detergency andbarrier function in comparison with regular shampoo-ing. B-WASH-ST-W (Thales Inc.; Tokyo, Japan) wasused as a microbubble generator in conjunction withbath salts and enzymes. A shampoo containing nomoisturizing components was used as control. Sham-pooing and MWS were performed on each side of thelateral thorax of five mixed-breed dogs with healthyskin. The detergency was measured from the amount ofsebum extracted from the hair coat (per g) with chloro-form and methanol solutions (2:1) at time (T, min) 0,T5, T10, T15 and T20 of washing. Skin barrier functionwas evaluated by measuring transepidermal water loss(TEWL) with a closed chamber evaporimeter (CC-01,Kao Corporation; Tokyo, Japan) at T0, T10 and T20of washing. The detergency of MWS at T15 of washingwas equivalent to that of shampooing at T5 (P = 0.855,paired Student’s t-test). At T10 and T20, the TEWLafter the shampooing treatment was significantly higher(21.51 � 1.36 g/m2/h and 26.13 � 2.42 g/m2/h) com-pared to after the MWS (17.40 � 3.63 g/m2/h and16.62 � 2.43 g/m2/h) (P < 0.05, one-factor ANOVA).These observations suggest that MWS can clean theskin surface and hair coat of dogs without scrubbingand impairs the skin barrier function less than sham-pooing.Source of funding: Self-funded.


Abstracts 31



IMMUNE-MEDIATED DISEASES

FC-18

Oclacitinib in the treatment of pemphigus

foliaceous in dogs

A. M. CORDERO*, C. LOPEZ-MARQUEZ*, G.SHEINBERG† and C. ROMERO§

*Vetderm, Guadalajara, Mexico; †Centro VeterinarioMexico, Mexico City, Mexico; §Dermavet, Mexico City,Mexico

Pemphigus foliaceous (PF) is the most commonautoimmune skin disease in dogs. Its treatment requireslong-term management with immunosuppressive drugs,often needed life-long. The objective of this study wasto evaluate the therapeutic effect of Oclacitinib (Apo-quel, Zoetis; New Jersey, NY, USA) in the treatmentof PF in dogs. Eight dogs diagnosed clinically, cytologi-cally and histologically for PF were divided into twogroups. Owners signed a consent form. Group 1 wasinduced with immunosuppressive doses of prednisoneand/or azathioprine in addition to oclacitinib at 1 mg/

kg twice daily. Group 2 was induced with oclacitinib ata mean dose of 1 mg/kg twice daily. Mean age of dogswas 6.5 years. Patients were evaluated at four visits onday (D)1, D14, D30 and D60 using a validated lesionscore system. Patients in Group 2 had a decrease ofclinical score of 65% and 84.8% at visits 3 and 4,respectively. Group 1 had a 15% decrease in clinicalscore at visit 4. One patient in Group 1 discontinuedfollow-up at the third visit. One dog in Group 2 devel-oped a fibrosarcoma and another was diagnosed withcutaneous T-cell lymphoma; both were removed fromthe study at visits 3 and 4. Oclacitinib seems to be analternative for maintenance and induction in the treat-ment of PF in dogs. Further studies are required toevaluate high dose and long-term use of oclacitinib indogs and different treatment combinations to achieveclinical control or remission for PF.Source of funding: Self-funded.


32 Abstracts



FOLLICULAR/CORNIFICATION FUNCTION AND PATHOLOGY

FC-19

Effects on clinical signs, lipids and hydration

factors of combined applications of shampoo

and mousse containing ophytrium and

seboliance in seborrhoeic dogs

J. KONDRATJEVA*, J. BRUN*, N. AMALRIC†, F.MOOG*, D. COMBARROS*,‡, C. PRESSANTI*,‡, C.ZEMIRLINE§, N. MAUBERT§, M. GATELLET¶ andM. C. CADIERGUES*,‡,¶

*Small Animal Clinic, Universit�e de Toulouse, ENVT,Toulouse, France; †Synelvia, Lab�ege, France; ‡UDEAR,Universit�e de Toulouse, INSERM, ENVT, Toulouse,France; §Ceva Sant�e Animale, Louvern�e, France; ¶CevaSant�e Animale, Libourne, France

Rapid improvement of skin appearance in case of seb-orrhoeic disorders is important to bring comfort to thedog and encourage owner compliance. The objective ofthis study was to evaluate the tolerance, performanceand effect on hair lipids and skin hydration of a sham-poo and mousse containing plant extracts (ophytriumand seboliance, DOUXO S3 SEB, Ceva Sant�e Animale;Libourne, France) in seborrhoeic dogs. Eighteen dogswith idiopathic chronic seborrhoea oleosa were enrolledin a prospective controlled study. Six control dogs werewashed with plain water on day (D)0. Twelve dogswere shampooed on D0 followed by eight mousseapplications every 48–72 h from D2 to D18. Clinicalscore (CS), natural moisturizing factors (NMF) andhair lipids (HL) were assessed on D0, D0 + 4h, D7,D14 and D24. At baseline, CS, NMF and HL were notsignificantly different between groups and remainedstable throughout the study in the control group. A sta-tistically significant reduction in CS from D0 to D7 (–53%) was observed in the test group reaching 91% onD24 (P < 0.05). No adverse effects were noted. NMFand HL levels decreased in the test group on D0+4h (–73%, P < 0.0001 and –50%, P = 0.14, respectively) andreturned to baseline values on D14 and D7, respec-tively. The combination of one shampoo and subse-quent mousse applications every 48-72 h containingophytrium and seboliance can be recommended as aconvenient and safe protocol to quickly improve haircoat quality in seborrhoeic dogs without dehydratingthe skin.Source of funding: Ceva Sant�e Animale, Libourne,

France.

Conflicts of interest: MCC has previously been consul-

tant for Ceva Sant�e Animale. NA is employed by Synel-

via. CZ, NM, MG are employed by Ceva Sant�eAnimale. JK, JB, FM, DC and CP declare no competing

interests.

FC-20

Performance of combined shampoo and mousse

applications containing ophytrium and

seboliance in dogs with scaling disorders: a

European field trial

M. GATELLET*, S. �AHMAN†, V. BRUET‡, P. M.CADOT§, R. S. MUELLER¶, C. NOLI††, T.NUTTALL‡‡, E. OLLIVIER*, T. BLONDEL* and N.SAVELLI§§

*Ceva Sant�e Animale, Libourne, France; †VetaDermVeterin€arklinik, Lomma, Sweden; ‡SkinVetDermatologie V�et�erinaire, Nantes, France; §Cliniquev�et�erinaire SeineVet, Rouen & Boos, France; ¶LudwigMaximilian University, Munich, Germany; ††ServiziDermatologici Veterinari, Peveragno, Italy; ‡‡The Royal(Dick) School of Veterinary Studies and The RoslinInstitute, Roslin, Scotland; §§Nad�ege Savelli EIRL,Villeneuve Loubet, France

Besides treating the underlying disease, it is essential toimprove skin appearance and odour in dogs with dryor greasy scaling. The objective of this study was toevaluate the performance of the combined applicationof a shampoo and mousse containing the plant extractsophytrium and seboliance (DOUXO S3 SEB Shampooand Mousse, Ceva Sant�e Animale; Libourne, France)in dogs with scaling disorders. Nineteen dogs with dryor greasy scaling were included in this prospective, mul-ticentric, open study. Dogs were shampooed on day(D)0 and received eight mousse applications every 48-72 h. Skin Seborrhoeic Index (SSI) was assessed by adermatologist on D0, D7 and D21. This includedodour, erythema, scaling, greasiness, extent and sec-ondary signs, each scored on a 0–3 scale. On D21, own-ers and veterinarians scored the protocol performanceand satisfaction. Mean SSI decreased significantly after21 days (P < 0.05) with a mean 33.9% improvement.On D21, the mean odour score decreased by 54.5%,extent by 46.1%, greasiness by 40.3% and scaling by33.3%. Most owners (84.2%) were satisfied or very sat-isfied with the performance of the protocol, emphasiz-ing the good appearance of the skin in 89.4% of cases.Veterinarians assessed the protocol as satisfactory,good or excellent in 84.2% of cases. One shampoo andeight mousse applications of the products containingophytrium and seboliance improved dry or greasy scal-ing in the study dogs. There was a noticeable improve-ment in some parameters important for pet owners,such as odour, resulting in high owner satisfactionlevels.Source of funding: Ceva Sant�e Animale, Libourne,

France.


fees, funding or salary from Ceva Sant�e Animale.

Abstracts 33



ALLERGIC DISEASES: PATHOBIOLOGY AND DIAGNOSIS

FC-21

Sensitization to the crude Dermatophagoidesfarinae extracts Der f 2 and Zen 1 in dogs

with atopic dermatitis in southern Brazil

L. LUDWIG*, T. TSUKUI†, M. KAGEYAMA† andM. FARIAS**Postgraduate Program in Animal Science, School ofLife Sciences, Pontif�ıcia Universidade Cat�olica doParan�a, Curitiba, PR, Brazil; †Central ResearchLaboratory, ZENOAQ, Koriyama, Fukushima, Japan

Allergens of the mite Dermatophagoides farinae are fre-quently associated with the sensitization of dogs withatopic dermatitis (AD). The aim of this study was toevaluate the sensitization of dogs with AD to the crudeD. farinae extract (cDf), Der f 2 and Zen 1. One hun-dred dogs of different breeds, ages and sex wereselected. All dogs had clinical diagnosis of AD and ful-filled at least five of eight pre-established criteria andexclusion of other pruritic skin diseases. Serum samplesof each dog were submitted to ELISA test (ZENOAQ,Koriyama; Fukushima, Japan) for measurement ofallergen-specific immunoglobulin (Ig)E, with a cut-offat 0.200 optical density. The results were reported indescriptive data and, for the correlation of each aller-gen, Spearman’s rank correlation coefficient was carriedout (P < 0.05). Of the 100 dogs, 92% were positive tocDf, 77% to Zen 1 and 56% to Der f 2. There was cor-relation between the sensitization to cDf and Zen 1(Spearman’s r = 0.88, P < 0.001), and not between cDfand Der f 2 (r = 0.30, P < 0.002), and between Zen 1and Der f 2 (r = 0.22, P<0.02). It is concluded that thesensitization to cDf, Der f 2 and Zen 1 is important indogs with AD in southern Brazil.Source of founding: The Coordination for the Improve-

ment of Higher Education Personnel – Brazil (CAPES)

and ZENOAQ.

Conflict of interest: TT and MK are employees of

ZENOAQ.

FC-22

Prevalence of IgE against CCD and impact of

CCD blocking in serum from cats

A. MOHAMMADDAVOODI*, C. LORENTE*, L.PANAKOVA†, E M €ULLER*, M CHRISTIAN* andR. WAGNER**Laboklin, Labor f€ur Klinische Diagnostik GMBH &CO. KG, Bad Kissingen, Germany; †Clinical Unit ofInternal Medicine Small Animals, Department forCompanion Animals and Horses, Vetmeduni, Vienna,Austria

Cross-reactive carbohydrate determinants (CCD) leadto highly multi-positive results in seasonal in vitroallergy tests. False-positive/clinically irrelevant resultsarise from the detection of immunoglobulin (Ig)Eagainst CCD (anti-CCD IgE). However, there are nostudies on this phenomenon in cats. This retrospectivestudy aimed to evaluate the prevalence of polysensitiza-tion in feline sera and the impact of the use of a CCDinhibitor (CHO blocker). A total of 472 cat sera (fromveterinary practices in the EU, Switzerland and China,

the majority from Germany) were subjected to a sea-sonal in vitro allergy test using Elisa Fc-Ԑ receptor tech-nology (Heska; Fribourg, Switzerland). Polysensitizedsamples were re-tested after adding a modified glyco-protein plant extracts (CCD inhibitor/CHO blocker;Heska). Polysensitization was present in 137 (29%)samples without a difference by season. After blocking,48% of the polysensitized sera were negative and 40%showed lower antibody levels. Forty-eight randomlyselected samples were analysed before and after block-ing to determine the impact of the CHO blocker oneach allergen. The total number of negative reactionsafter the addition of the CHO blocker were higher innettle (65%), willow (67%), mugwort-ragweed (65%)and English plantain (54%) than in a six-grass mix(31%), rye (23%) and sheep’s sorrel (25%). The 29%prevalence of anti-CCD IgE in cats is similar to thatreported in dogs and humans. The use of a CHOblocker for pollen allergy testing in cats improves theidentification of the relevant offending allergens to for-mulate allergen-specific immunotherapy.Source of funding: Self-funded.

Conflict of interest: AM, CL, EM, MC and RW work

for Laboklin.

FC-23

Prevalence of high levels of specific IgE

against mites in dogs and cats from non-

Mediterranean Europe

C. LORENTE-M�ENDEZ, E. M €ULLER and R.WAGNERLaboklin, Labor f€ur Klinische Diagnostik GMBH & CO.KG, Bad Kissingen, Germany

Presence of high levels of specific-immunoglobulin (Ig)E in serum determines the sensitization of the patientto the allergen. The objective of this study was to eval-uate the prevalence of high levels of IgE againstAlternaria/Aspergillus (A/A) Penicillium/Cladosporium(P/C), Dermatophagoides farinae (Df), Der-matophagoides pteronyssinus (Dpt), Tyrophagus putres-centiae (Tp) and Acarus siro (As) in sensitized dogs andcats. Veterinarians from non-Mediterranean Europeancountries sent serum samples from cats and dogs toLaboklin (2017 to 2019) to investigate the allergensinvolved in the allergic reaction of the patient. Serafrom cats (2,148) and sera from dogs (22,114) wereanalysed using an ELISA test based on the FceR1areceptor (Heska; Fribourg, Switzerland); 21,261(95.70%) of the dog samples and 1,814 (84.45%) of thecat samples yielded positive results to at least one aller-gen. The percentage of high levels of specific IgE in thepositive samples of dogs and cats (respectively) were10.66% and 8.88% for A/A, 4.86% and 4.91% for P/C, 91.10% and 75.91% for Df, 51.98% and 59.81% forDpt, 90.36% and 87.38% for Tp, and 82.23% and64.06% for As. Sensitization to moulds was associatedwith mite sensitization in >99.5% of positive mouldsamples. Based on these results, moulds seem not to bean important cause of sensitization in dogs and cats.Prevalence of sensitivity to all mites was high, most fre-quently to Df and Tp and least frequently to Dp.

34 Abstracts



Storage mites seem to play an important role in theallergic reactions of dogs and cats.Source of funding: Self-funded.

Conflict of interest: All authors work for Laboklin.

FC-24

Prevalence of high levels of specific IgE

against pollens in dogs and cats from non-

Mediterranean Europe

C. LORENTE-M�ENDEZ, E. M €ULLER and R.WAGNERLaboklin, Labor f€ur Klinische Diagnostik GMBH & CO.KG, Bad Kissingen, Germany

Serological allergen tests using the blocking of cross-re-active carbohydrate determinants (CCDs) have demon-strated the same reliability as the intradermal test toidentify the allergens involved in pollen-allergic dogsand cats. This retrospective study aimed to evaluate theprevalence of high levels of specific immunoglobulin(Ig)E against pollens in hypersensitised dogs and catsfrom Europe (non-Mediterranean areas). A total of10,345 sera from dogs and 1011 from cats were anal-ysed with an ELISA test based on the FceR1a receptor(Heska; Fribourg, Switzerland) together with CCDinhibition (CHO blocker; Heska) in polysensitized sam-ples. Positive results to at least one allergen were foundin 8,177 (79.04%) of the dog samples and 750 (74.28%)of the cat samples. The percentage of high levels ofspecific IgE in the positive samples of dogs and catswere (respectively) 84.50% and 83.36% for sorrel,80.80% and 75.10% for rye, 74.40% and 74.53% forsix-grass mix, 56.80% and 51.53% for English plantain,36% and 45.54% for willow, 35.10% and 43.54% formugwort/ragweed, 27.50% and 36.22% for Birch, and19.90% and 42.08% for nettle. It can be concluded thatin non-Mediterranean Europe, the principal pollenallergens for dogs and cats are sorrel, rye and grasswith similar values of hypersensitivity prevalence(>75%) for both species. Plantain is the next most fre-quent pollen. Willow, mugwort, birch and nettle seemto induce fewer hypersensitivities in dogs and cats,especially in dogs.Source of funding: Self-funded.


FC-25

Filaggrins and tight junction proteins

expression in keratinocyte cell cultures from

normal and atopic dogs

R. MARSELLA, K. AHRENS, R. WILKES, N. S.MUNGUIA and M. DORRDepartment of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA

Keratinocytes of normal and atopic dogs behave differ-ently in cell culture when creating a monolayer and dif-ferentiating. Normal keratinocytes achieve confluencefirst and then differentiate while atopics start differenti-ating, creating little “domes” even if they are not yetconfluent. In this study we grew four atopic and five

normal primary keratinocyte cell lines in chamber slidesto study differentiation and tight junction (TJ) expres-sion. Cells were fixed the first confluency day (D)0 andon D6. Immunofluorescent staining was done forzonula occludens, claudin, occludin, filaggrin and filag-grin-2. Five pictures per chamber for each antibodywere taken. Pictures were randomized and scored byfour blinded observers. Cell size and variability andcontinuity of TJ proteins were scored. ANOVA showedincreased cell size from D0 to D6 in both groups (effectof time, P = 0.0076) and no differences between groups.Size was more subject-dependent than disease-related(P = 0.0022). No difference for continuity was foundfor any of TJ proteins. Both filaggrins were nuclearearly on and later cytoplasmic. The increase of cyto-plasmic filaggrin-2 on D6 was significant in atopic dogs(P = 0.0085) as was the effect of subject (P = 0.002),indicating that it was more subject-dependent than dis-ease-related. The decrease of staining of filaggrin-2 inthe nucleus at D6 approached significance in the atopicdogs (P = 0.055). It is concluded that the cell culturebehaviour may be linked to factors other than TJ pro-teins. This is the first report of the nuclear transloca-tion of filaggrin-2 in dogs.Source of funding: Self-funded.


FC-26

Dermatophagoides farinae allergenic profiles

in atopic cats and horses: differences between

species

L. RAMI�O-LLUCH*, R. MOYA†, A. GONZ�ALEZ-RUIZ†, F. ALV�AREZ†, J. CARN�ES† and A.PUIGDEMONT‡

*Animal Health B.U., Laboratorios LETI, S.L.U.,Barcelona, Spain; †R&D Immunology and Allergy Unit,Laboratorios LETI, S.L.U., Madrid, Spain;‡Deptartment of Pharmacology, Therapeutics andToxicology, Universitat Aut�onoma de Barcelona,Barcelona, Spain

House dust mite (HDM) allergens already have beenreported as an important cause of allergenic reactionsin cats and horses, yet there are no studies about themain proteins involved in the sensitization. The objec-tive of this study was to investigate the allergenic sensi-tization profile of cats and horses with atopicdermatitis against Dermatophagoides farinae. Nineteencats and 15 horses sensitized to D. farinae [clinical diag-nosis confirmed with positive specific immunoglobulin(Ig)E levels by ELISA test (Greer; Lenoir, USA)], fourcontrol healthy cats and two control healthy horseswere included in the study. Allergenic profiles wereinvestigated by immunoblot. The D. farinae extract(Laboratorios LETI; Barcelona, Spain) was elec-trophoretically separated by SDS-PAGE, transferredand incubated with the serum samples. Membraneswere incubated with the corresponding secondary anti-body (anti-IgE) and developed by chemiluminescence.Results showed that most cats recognized high molecu-lar weight (MW) allergens. However, most also showedstrong recognition of proteins of approximately30 kDa, and only three cats showed recognition oflower MW allergens. Feline controls did not show anyband recognition. Different allergenic profiles were

Abstracts 35



found among positive horses and all sera recognizedallergens of a broad MW range (from 14 to >100 kDa).Nonetheless, proteins between 25 and 37 kDa were rec-ognized for the majority. Horse controls did not showany band recognition except for one control with a

37 kDa band. In conclusion, cats recognized mainlymedium and high MW proteins while horses did notpresent a specific HDM sensitization allergenic pattern.Source of funding: Self-funded.


36 Abstracts



ALLERGIC DISEASES: TREATMENT

FC-27

Effectiveness of a nutraceutical with

blackcurrant oil and tyndalised Lactobacillusreuteri in controlling intestinal dysbiosis and

pruritus in atopic dogs

L. CORNEGLIANI*, P. PERSICO† and A.GRAMENZI§

*Clinica Veterinaria Citt�a Di Torino, Torino, Italy;†Private practice, Milan, Italy; §Faculty of VeterinaryMedicine, University of Teramo, Teramo, Italy

Canine atopic dermatitis (cAD) is a chronic allergic dis-ease which requires a multimodal management. As wellas the impaired skin barrier, this disorder causes dys-functions of the skin and gut microbiome. The presentstudy evaluates the intestinal microbiome, faecal scoreand Visual Analog Scale (VAS) in dogs affected bycAD before, during and after the administration of anutraceutical based on blackcurrant oil, tyndallizedLactobacillus reuteri, nucleotides and zinc oxide. Forty-five dogs with cAD in maintenance therapy for at leastone year were selected and fed a diet with hydrolysedchicken and rice throughout the 120 days of the study.Throughout the study, current therapies remainedunchanged. Each dog was its own control. From day(D)0 to D60 all dogs were administered the nutraceuti-cal (Ribes Pet Symbio, NBF Lanes; Milan, Italy) oncedaily, then the administration ceased for the next 60days. Stool samples were collected on D0, D60 andD120 for microbiome analyses carried out by sequenc-ing the16S rRNA gene with calculation of the dysbiosisindex and faecal score. VAS was assessed on D0, D60and D120. Statistical analysis used the mixed-effects lin-ear model. VAS and faecal score decreased significantlybetween D0 and D60 (P<0.01), while there were no sig-nificant variations between D60 and D120. The dysbio-sis index decreased (improved) between D0 and D60(P < 0.01) and between D60 and D120 it increased(worsened) significantly (P < 0.01). The study high-lights the usefulness of this nutraceutical in the controlof intestinal dysbiosis and VAS in atopic dogs.Source of funding: NBF Lanes, Milan, Italy.

Conflict of interest: NBF Lanes paid the authors to per-

form this study. NBF Lanes was not involved in the study

design or data analysis.

FC-28

Correlation between plasmatic levels of

oclacitinib and clinical response in dogs with

canine atopic dermatitis

R. GARC�IA-FONTICOBA*,†, L. ORDEIX*,†, M.CABR�E*,† and L. FERRER*,†

*Fundaci�o Hospital Cl�ınic Veterinari, UniversitatAut�onoma de Barcelona, Barcelona, Spain; †Departmentof Animal Medicine and Surgery, Universitat Aut�onomade Barcelona, Barcelona, Spain

Oclacitinib is a selective inhibitor of Janus kinase-1approved for the treatment of allergic dermatitis indogs. Although oclacitinib has been demonstrated ashighly effective in controlling pruritus and

inflammatory lesions in atopic dogs, a minority do notshow a satisfactory response. To date, the reason forthis lack of efficacy in some dogs remains unknown.The objective of this study was to determine if the lowefficacy of the drug observed in these cases could beconsequence of pharmacokinetic factors. Twenty-fourdogs with a diagnosis of atopic dermatitis were treatedwith oclacitinib, 0.4–0.6 mg/kg orally twice daily for 14days, then once daily for 14 days. Owner-assessed pru-ritus Visual Analog Scale (pVAS) and clinician-assessedCanine Atopic Dermatitis Extent and Severity Index,4th iteration (CADESI-04) were evaluated on day (D)0and D28. The plasma concentration of oclacitinib wasdetermined by high-performance liquid chromatogra-phy at D28, 1 h after oral administration of oclacitinib.Levels of oclacitinib on D28 ranged from 1.6 to 476.6(average 226.9 � 131.5) ng/mL. Correlation was notobserved between plasma concentration of oclacitiniband reduction in pVAS score (rxy = 0.067, P > 0.05).Negative correlation was obtained when comparingdecrease in CADESI-04 score and levels of oclacitinib(rxy = –0.562, P < 0.05). Plasma concentrations ofoclacitinib were highly variable and in most dogs, evenin nonresponders, they were in the range consideredtherapeutic. These results indicate that the lack of effi-cacy of oclacitinib in some dogs does not seem to beconsequence of pharmacokinetic factors.Source of funding: Apoquel tablets were supplied by Zoe-

tis (Parsippany, NJ, USA); the manufacturer was not

involved in study design, data collection and interpreta-

tion or publication.

Conflict of interest: LF has received unrelated honoraria

for lecturing from Zoetis. No conflicts of interest have

been declared by the other authors.

FC-29

Performance of a combined application of

ophytrium-containing shampoo and mousse in

dogs with sensitive skin: a European field trial

M. GATELLET*, S. �AHMAN†, V. BRUET‡, P. M.CADOT§, R. S. MUELLER¶, C. NOLI††, T.NUTTALL‡‡, E. OLLIVIER*, T. BLONDEL* and N.SAVELLI§§

*Ceva Sant�e Animale, Libourne, France; †VetaDermVeterin€arklinik, Lomma, Sweden; ‡SkinVetDermatologie V�et�erinaire, Nantes, France; §Cliniquev�et�erinaire SeineVet, Rouen & Boos, France; ¶LudwigMaximilian University, Munich, Germany; ††ServiziDermatologici Veterinari, Peveragno, Italy; ‡‡The Royal(Dick) School of Veterinary Studies and The RoslinInstitute, Roslin, Scotland; §§Nad�ege Savelli EIRL,Villeneuve Loubet, France

Topical products can provide direct relief of the dryand irritated skin in atopic patients. The objective ofthis study was to evaluate the performance of a proto-col combining a shampoo and a mousse each contain-ing ophytrium, a purified plant extract with skinbarrier strengthening, soothing and antibiofilm proper-ties in dogs with sensitive skin (DOUXO S3 CALMShampoo and Mousse, Ceva Sant�e Animale; Libourne,France). Twenty dogs with a history of atopic

Abstracts 37



dermatitis presenting a skin flare were enrolled in aprospective, multicentric, uncontrolled study. Dogswere bathed on day (D)0 and then given eight mousseapplications every 48–72 h. Canine Atopic DermatitisExtent and Severity Index, 4th iteration (CADESI-04)scores were assessed by a dermatologist and pruritus bythe owners using the pruritus Visual Analog Scale(pVAS) score on D0, D7 and D21. Owner and veteri-narian evaluations of performance and satisfaction wererecorded on D21. Mean CADESI-04 scores improvedthroughout the study, from 18 at D0 to 6.4 at D21(58.3% mean improvement; P < 0.05 at all timepoints).On D21, 63.2% of the dogs achieved ≥70% CADESI-04 improvement; 78.9% had a score below the targetthreshold of 10. Mean pVAS scores decreased signifi-cantly after 21 days (P < 0.05 at all time-points) withmean 30.8% improvement; 42.1% of the dogs had≥50% pVAS score decrease. Global veterinary assess-ment of the protocol was satisfactory, good or excellentin 73.7% of cases. Most owners (78.9%) were satisfiedor very satisfied with the protocol. One shampoo andeight mousse applications of Ophytrium significantlyimproved skin health and pruritus in these atopic dogs,resulting in high satisfaction levels for veterinarians andowners.Source of funding: Ceva Sant�e Animale, Libourne,

France.


fees, funding or salary from Ceva Sant�e Animale.

FC-30

Evaluation of a new hydrolysed fish and rice

starch elimination diet in the diagnosis of

adverse food reactions in cats

C. NOLI* and G. BELTRANDO†

*Servizi Dermatologici Veterinari, Peveragno (CN),Italy; †Universit�a degli SAtudi di Torino, Grugliasco(TO), Italy

Diagnosis of food allergy (FA) is based on a two-month elimination diet (ED) and confirmed by positivereactions to challenge with the old food. Hydrolyseddiets are commonly used for this purpose; unfortu-nately, they may contain partially hydrolysed allergenicingredients which may bias the results. The selection ofthe right ED is mandatory for a correct diagnosis. Theaim of this study was to evaluate the suitability ofhydrolysed fish and rice starch diet (Vet-Life FelineUltra-Hypo, Farmina Pet Food; Nola, Italy) in thediagnosis of FA in cats. Cats with suspected FA wereenrolled in the study. For each cat, pruritus wasassessed by a Visual Analog Scale and lesions by Scor-ing Feline Allergic Dermatitis (SCORFAD) on day (D)0 and D56 of ED. Cats showing ≥50% pruritus and/orSCORFAD improvement were considered FA andchallenged with the old food, fish and rice. Cats notresponding to the ED were administered a second diettrial (Anallergenic, Royal Canin; Aimargues, France).Twenty-six cats were included, one was excluded fornoncompliance and two were lost to follow-up. Of theremaining 23, three interrupted the study for gastroin-testinal signs, and 14 (61%) improved and underwentthe dietary challenge. Of these, 5/14 (36%) reacted totheir old food and were diagnosed with FA; two ofthese reacted to fish and four to rice. No

nonresponding cat responded to the second ED. Farm-ina UltraHypo seems to be a useful ED when screeningcats for FA. Furthermore, it did not cause reactions incats with rice or fish hypersensitivity.Source of funding: Farmina Pet Food, Nola, Italy.

Conflicts of interest: C.N. has received consultation fee

and speakers fee from Farmina.

FC-31

Anti-inflammatory effects of oclacitinib on

skin lesion scores in a house dust mite-sensitive

colony of beagles

J. T. ROSS*, K. RUDOLPH†, J. MLECZKO†, T.BARRETT† and A. J. GONZALES**Global Therapeutics Research, Zoetis, Kalamazoo, MI,USA; †Lovelace Biomedical, Albuquerque, NM, USA

Oclacitinib is approved for the control/treatment ofatopic and allergic dermatitis in ≥1-year-old dogs andhas shown significant improvement in skin condition incontrolled field studies. To objectively compare efficacyof oclacitinib to prednisolone, a study was performedin a laboratory model of allergic dermatitis. House dustmite (HDM)-sensitive beagles were exposed to topicalHDM allergen (15 mg/mL), and erythema, papules andexcoriations were scored with a 0 to 5 scale (0, normal;5, severe). Based on total lesion scores (ery-thema+papules+excoriations), 21 animals were random-ized into treatment groups (n = 7 per treatment):placebo (0.0 mg/kg per os twice daily), oclacitinib(Apoquel, Zoetis; Kalamazoo, MI, USA; 0.4–0.6 mg/kgp.o. twice daily) or prednisolone (0.5 mg/kg p.o. oncedaily for five days, then tapered every other day). Sub-sequently, animals were exposed to HDM, then treatedfor 12 days. Oclacitinib significantly (P < 0.05)decreased all measures compared to placebo-treatedcontrols. Area under the curve (mean � SEM) for ery-thema, papules, excoriations and total lesion scores inplacebo-treated animals were 278.5 (� 22.8), 141.9(� 30.5), 111.6 (� 23.9), and 532.0 (� 76.6), respec-tively, compared to 207.5 (� 21.2), 47.2 (� 5.9), 46.8(� 13.7) and 301.6 (� 35.8) respectively, in oclacitinib-treated animals. Prednisolone did not have significanteffects on lesion scores. Pruritic behaviour also wasquantified, and both oclacitinib and prednisoloneshowed significant (P < 0.05) reductions in pruriticevents compared to placebo. These data support previ-ous findings with oclacitinib and suggest that it hasstrong anti-inflammatory properties along with antipru-ritic effects.Source of Funding: Zoetis, Kalamazoo, MI, USA.

Conflict of Interest: AJG and JTR are employees of

Zoetis.

38 Abstracts



FC-32

Is a digital pVAS equivalent to a paper PVAS

to measure itch in dogs suffering from Atopic

Dermatitis?

A. WRIGHT*, S. SPIEGEL†, K. KAMI �NSKI†, M.KARWALA† and P. HILL‡

* Zoetis, Inc, Parsipanny, NJ, USA; † Tivix, Wroclaw,Poland; ‡ Veterinary Dermatology and Immunology,Adelaide, South Australia, Australia

An ePRO (electronic patient reported patient outcome)measure that has been adapted from a paper-basedmeasure should produce data that are equivalent orsuperior to the data produced from the original paperversion. This prospective study was performed to showmeasurement equivalence of a new digital pruritusVisual Analog Scale (pVAS) in an app (Itch Tracker)to the widely accepted paper pVAS following recom-mendations of the ISPOR ePRO Good Research Prac-tices Task Force. Equivalence was determined to be ifscores from dog owners completing the PVAS on paper

were within 10 mm of the digital pVAS on a scale of1–100 mm. Comparing the mean scores obtained onthe two modes of administration from the same personcan be used to assess measurement equivalence. In2017, dog owners from 31 different veterinary clinics inAustralia with 77 unique dogs diagnosed with atopicdermatitis completed the paper pVAS at the veterinaryoffice. Then a code to the digital pVAS app (ItchTracker) was provided with an initial injection of loki-vetmab (Cytopoint, Zoetis Inc.; Parsipanny, NJ, USA)and pVAS completed upon download on a mobilephone. Mean average from the app pVAS was –1.5 mm below paper pVAS mean, much smaller thanthe allowable 10 mm. Therefore, the app-based pVASwas shown to be equivalent to the paper-based pVASand can be used by owners to track pruritus at home.This may assist veterinarians and dog owners trackresponse to pruritus therapy in between visits to theveterinary clinic.Source of funding: Zoetis, Parsipanny, NJ, USA.

Conflict of interest: AW is an employee of Zoetis.

Abstracts 39



INFECTIOUS SKIN DISEASES

FC-33

Antimicrobial resistance gene and multidrug

efflux pumps against antiseptics for

staphylococci isolated from canine superficial

pyoderma in Japan

N. MURAYAMA*, Y. OSHIMA-TERADA*, K.MORI*, H. NAKAMINAMI†, T. SAITO† and N.NOGUCHI†

*Dermatology Services for Dogs and Cats, Tokyo,Japan; †Tokyo University of Pharmacy and LifeSciences, Tokyo, Japan

Meticillin-resistant staphylococci in canine pyoderma isa serious problem. The aim of this study was to accessmecA, antimicrobial resistance genes and multidrugefflux pump genes of staphylococci from canine superfi-cial pyoderma in Japan. One hundred and thirty-sixdogs were evaluated from May 2018 to April 2019. Sta-phylococci mecA gene, antimicrobial resistance genes(including ermA, ermB, ermC, msr, inuA, mphC, aacA/aphD, tetM, tetK and fosB) and multidrug efflux pumpgenes including qacA/B and smr were identified byPCR. Of these, 72% were Staphylococcus pseudinter-medius, 25% S. schleiferi ssp. coagulans and 3%unidentified staphylococci, respectively, and 47% of allstaphylococci had mecA positive. All staphylococciwere positive for ermA, ermB, ermC, msr, inuA, mphC,aacA/aphD, tetM, tetK and fosB in 0%, 46.3%, 0.7%,0%, 0%, 0%, 50.7%, 46.3%, 1.5% and 70.0%, respec-tively. MecA-negative staphylococci were positive forermA, ermB, ermC, msr, lnuA, mphC, aacA/aphD, tetM,tetK and fosB in 0%, 22.2%, 0%, 0%, 0%, 0%,27.8%, 33.3, 2.8% and 62.5%, respectively. MecA-posi-tive staphylococci was positive for ermA, ermB, ermC,msr, lnuA, mphC, aacA/aphD, tetM, tetK and fosB in0%, 74.6%, 1.6%, 0%, 0%, 0%, 76.6%, 60.9%, 0%and 78.1%, respectively. Multidrug efflux pump genesincluding qacA/B and smr were 0.7% and 2.2% (respec-tively) in all isolates. MecA-positive staphylococci werepositive for qacA/B and smr in 1.6% and 1.6%, respec-tively. MecA-negative staphylococci were positive forsmr in 2.8%. This study indicated that mecA-positivestaphylococci had many antimicrobial resistance genes,and a few staphylococci had multidrug efflux pumpgenes.Source of funding: Self-funded.


FC-34

Survey of dermatophytes in stray dogs and

cats in Puerto Rico confirmed with MALDI-

TOF

A. HERNANDEZ-BURES*, W. A. BIDOT†, M.O’DELL‡, C. W. MADDOX‡§, W. E. SANDER* andJ.B. PIEPER**Department of Veterinary Clinical Medicine, College ofVeterinary Medicine, University of Illinois, Urbana, IL,USA; †Animal Care Services, University of Florida,Gainesville, FL, USA; ‡Veterinary DiagnosticLaboratory, College of Veterinary Medicine, Universityof Illinois, Urbana, IL, USA; §Department ofPathobiology, College of Veterinary Medicine,University of Illinois, Urbana, IL, USA

Dermatophytosis is a contagious fungal skin conditionof high zoonotic importance and this raises publichealth concern in working environments with largeinfluxes of stray animals. After hurricanes impactedPuerto Rico in 2017, there has been an increased num-ber of stray animals due to owner abandonment, whichhas led to veterinary groups visiting the island to per-form trap–spay/neuter events. The purpose of thisstudy was to establish the prevalence of stray animalswith dermatophytes and the prevalence of asymp-tomatic carriers. Hair samples were collected from 55stray dogs and 44 stray cats (total 99) during a trap–neuter–release event using the Mackenzie brush tech-nique and cultured on Derm-duet plates. Direct micro-scopy and examination of the isolates were performed.In addition, isolates were evaluated by matrix-assistedlaser desorption/ionization-time-of-flight (MALDI-TOF). This project was approved by the InstitutionalAnimal Care and Use Committee. A total of 19/99 ani-mals (19%) were positive for Microsporum canis (18/19)and Trichophyton spp. (1/19). Prevalence rates of29.5% in feline patients and 10.9% in canine patientswere observed, approaching statistical significance(OR = 2.71 feline, P = 0.096). Positive cultures wereobserved in 9.8% of adult patients and 34.2% of juve-nile patients, which was statistically significant(OR = 3.48 juvenile, P = 0.02). Asymptomatic patientswere positive in 36% of the population while animalswith clinical lesions were positive in 13.5%, which wasapproaching significance (OR = 0.38 clinical lesions,P = 0.094). This study demonstrates the importance ofestablishing safety protocols among animals admittedto trap–spay/neuter events for stray animals, regardlessof lesions.Source of Funding: Self-funded.


40 Abstracts



FC-35

Determination of resistance to antimicrobial

agents and resistance to meticillin in

Staphylococcus pseudintermedius isolates fromcutaneous infections in dogs

M. SILVA*, O. VIEIRA-DA-MOTTA†, A.FIGUEIREDO‡, F. FERREIRA‡ and M. SILVA-CARVALHO‡

*Est�acio de S�a University (UNESA), Rio de Janeiro,Brazil; †State University of Norte Fluminense, Rio deJaneiro, Brazil; ‡Federal University of Rio de Janeiro,Rio de Janeiro, Brazil

Resistance to antimicrobial agents in Staphylococcusspp. was investigated in 124 study dogs. Over 40%resistance rates were observed against erythromycin(48.3%), penicillin (45.9%), clindamycin (42.7%) andtetracycline (37.9%). Between 20 and 40% resistancewere observed for trimethoprim-sulfamethoxazole andtetracycline. The lowest resistance rates (<15%) wereobserved for oxacillin, gentamicin, ciprofloxacin, chlo-ramphenicol, rifampicin and teicoplanin. Likewise, itwas found among samples of S. pseudintermedius thatresistance to penicillin (52.3%) and erythromycin(51.1%) were greatest, followed by clindamycin resis-tance (40.6%). The lowest resistance (10%) also wasobserved for gentamicin, chloramphenicol, rifampicinand teicoplanin. Regarding the detection of meticillin-resistant (MR) Staphylococcus strains, 17 of 124 sam-ples were identified as resistant to meticillin by pheno-typic methods: six among dogs with pyoderma, fouramong animals with otitis, six with cutaneous tumourand one with surgical site infection. In addition, PCRwas used on all samples to detect the mecA gene andconfirm meticillin resistance. Of the 17 isolated MRsamples, the majority (n = 13) belonged to the genomicislands and were identified by PCR as S. pseudinter-medius. The remaining samples were identified by PCRas S. aureus (n = 4). Defining multiresistance as resis-tance to three or more classes of antimicrobials, amongthe isolated MR S. aureus (MRSA) samples, 75% (3/4)showed multidrug resistance. With regard to MRS. pseudintermedius (MRSP) samples, a high rate ofmultidrug resistance also was observed (92%, 12/13).Source of Funding: None declared.


FC-36

The comparative efficacy of different

itraconazole treatment protocols for the

treatment of feline sporotrichosis in Rio de

Janeiro, Brazil

F. CARVALHO*,†, M. MACHADO‡, S. KOCH§,R. RAMADINHA‡ and J. FERNANDES‡

*College of Veterinary Medicine, Centro Universit�ario deValenc�a, Valenc�a, Rio de Janeiro, Brazil; †College ofVeterinary Medicine, Universidade Iguac�u, Nova Iguac�u,Rio de Janeiro, Brazil; ‡Department of VeterinaryScience, Universidade Federal Rural do Rio de Janeiro,Serop�edica, Rio de Janeiro, Brazil; §College of VeterinaryMedicine, University of Minnesota, MN, USA

Sporotrichosis caused by an agent of the Sporothrixschenckii complex, is an important endemic zoonosis in

the state of Rio de Janeiro, Brazil, affecting mainlycats. Various antifungals are available to treat this dis-ease with itraconazole currently being the drug ofchoice. However, the number of cases refractory totreatment has increased, possibly because of drug resis-tance or the quality of the compounded itraconazoleoften used to treat the disease. The study objective wasto compare different protocols using itraconazole totreat feline sporotrichosis. A total of 107 cats from thecity of Vassouras diagnosed with sporotrichosisbetween 2016 and 2018 were included based on positivecytological findings and fungal culture. Disease extentand severity varied from a single skin lesion to multiplelesions affecting the skin and mucosa and/or respira-tory tract. The cats were randomly assigned to receivethe following treatment protocols: (i) generic itracona-zole (Prati Donaduzzi; Paran�a, Brazil) at 100 mg/cat/day per os; (ii) compounded itraconazole at 100 mg/cat/day p.o.; and (iii) generic or compounded itracona-zole at 100 mg/cat/day p.o. combined with 5 mg/kg/day p.o. of compounded potassium iodide. Response totreatment was evaluated after 150 days of therapy.There were no statistical differences (chi-square test,P < 0.05) in the number of cats that achieved cure, diedspontaneously or were euthanized, between the treat-ment protocols. Based on the study results, brand nameor compounded itraconazole is an effective treatmentfor feline sporotrichosis if administered for a long per-iod of time.Source of funding: Coordenac�~ao de Aperfeic�oamento de

Pessoal de N�ıvel Superior (CAPES), Brazil.Conflict of interest: None declared.

FC-37

Feline sporotrichosis outbreak in Curitiba,

Brazil

D. KUNG*, F. MONTI†, L. BALLARDIN*, S.RIBEIRO*, L. CONDAS‡, M. CASTRO§, G.FERREIRA*, S. AMADEI§ and M. FARIAS**School of Life Sciences, Pontif�ıcia UniversidadeCat�olica do Paran�a, Curitiba, PR, Brazil; †UniversidadeTuiuti, Paran�a, Brazil; ‡Self-employed veterinarian,Paran�a, Brazil; §Biotechnology, School of Life Sciences,Pontif�ıcia Universidade Cat�olica do Paran�a, Paran�a,Brazil

The present study evaluates the outbreak of 295 casesof feline sporotrichosis caused by Sporothrix brasiliensisin Curitiba, southern Brazil, from May 2016 to June2019. Cytopathological or histopathological and myco-logical tests were performed for the final diagnosis.Molecular identification of isolates was performedusing the calmodulin gene species-specific region. Thedemographic variables analysed included breed, age,sex, reproductive status, outdoors access, soil/decayvegetation contact, cohabitation and contact with out-doors animals. Clinical data included disease durationbefore diagnosis, lesion classification and topography.Zoonotic data included type of transmission, topogra-phy and classification of lesions. Demographic datashowed that 275 (93%) cats lived in urban area, 283(96%) were mixed breed of adult age (average 44-month-old), 203 (69%) were males, 160 (58%) wereintact, 223 (77%) had outdoor access and 231 (84%)had contact with soil/decay vegetation. The disease

Abstracts 41



duration before diagnosis for 275 cats was over 14weeks. The head and forelimbs were affected more fre-quently. Sixty-nine (25.3%) of 272 owners developedsporotrichosis from contact with exudate and scratchesor bites from affected cats. Lesions developed mainlyon their upper limbs and typically in a localized form.In conclusion, intact male and adult cats represent thepopulation most exposed to S. brasiliensis in Curitiba.These animals develop disseminated cutaneous lesionsand are the most important host and propagator ofsporotrichosis in urban environments in this state.Source of funding: Coordenac�~ao de Aperfeic�oamento de

Pessoal de N�ıvel Superior (CAPES).Conflict of interest: None declared.

FC-38

Antifungal susceptibility of Sporothrixbrasiliensis isolated from feline sporotrichosis

D. C. KUNG*, F. MONTI*, L. BALLARDIN*, L.CONDAS†, L. BORBA-SANTOS‡, E. MENEZES‡, S.ROZENTAL‡ and M. FARIAS**Postgraduate Program in Animal Science, School ofLife Sciences, Pontif�ıcia Universidade Cat�olica doParan�a, Paran�a, Brazil; †Freelance veterinarian, Paran�a,Brazil; ‡Laboratory of Fungal Cell Biology, Institute ofBiophysics Carlos Chagas Filho, Federal University ofRio de Janeiro, Rio de Janeiro, Brazil

This study aimed to evaluate the susceptibility of Spor-othrix brasiliensis, isolated from cats with sporotri-chosis, to itraconazole (ITZ) and relate it to diseaseseverity and outcome. The diagnosis was confirmedbased on cytopathological or histopathological andmycological test results. Molecular identification of iso-lates was performed using the calmodulin gene species-specific region. Susceptibility tests were preceded by fil-amentous to yeast conversion according to Clinical andLaboratory Standards Institute (CLSI) protocols. Fungiare here designated as wild-type (WT) for those withoutintrinsic or acquired resistance and nonWT (NWT) forthose harbouring intrinsic or acquired resistance mecha-nisms. Of the 67 cats included in the study, all wereinfected with S. brasiliensis. Itraconazole susceptibilitywas performed in 37(54.4%) isolates that convertedfrom filamentous to yeast. Complete clinical data wereavailable for only 17(46%) of these isolates. Twelve(70.5%) of these were WT and five (29.5%) NWT. Ten(83.3%) cats with WT isolates were cured; however,eight (80%) were cured with ITZ combined to potas-sium iodide (KI). Two (16.7%) cats were euthanized.Three cats with sporotrichosis caused by the NWT iso-late were cured with ITZ and KI combined, and twodied from the disease. Thirteen (76.5%) of the 17 caseshad the disseminated cutaneous form associated withrespiratory clinical signs. In conclusion, most catsinfected with ITZ-susceptible S. brasiliensis strainsrequired combination therapy (i.e. ITZ and KI) toachieve disease cure. Unfavourable prognosis was mostoften associated with ITZ-resistant S. brasiliensisstrains.Source of funding: Coordenac�~ao de Aperfeic�oamento de

Pessoal de N�ıvel Superior – (CAPES).


FC-39

Evaluation of the response to therapeutic

protocols in feline sporotrichosis

F. MONTI, L. BALLARDIN, S. RIBEIRO, D.KUNG, V. GMYTERCO, C. PIMP~AO and M.FARIASDepartment of Veterinary Medicine, School of LifeSciences, Pontifical Catholic University of Paran�a,Paran�a, Brazil

Sporotrichosis is an epidemic mycotic infection in Bra-zil that affects primarily cats living in urban areas. Theaim of this longitudinal study was to evaluate the effi-cacy of different therapeutic protocols in cats withsporotrichosis caused by Sporothrix brasiliensis. Fifty-nine cats diagnosed with sporotrichosis were enrolled inthe study. Thirty-four were treated with generic itra-conazole (ITZ) monotherapy at the dose of 10 20 mg/kg/day per os in patients ≤3.0 kg or 100 mg/cat/dayp.o. in patients >3.0 kg. Twenty-five cats were treatedwith ITZ using the same treatment protocol combinedwith compounded potassium iodide (KI) at the dose of5 mg/kg/day p.o. Among the 34 cats treated solely withITZ, 11 (32%) were cured, one (3%) was euthanizedand the other 22 (65%) required the addition of KI totheir therapeutic protocol. Of the 47 cats treated withthe combination of ITZ and KI, 24 (51%) were cured,seven (15%) died and 16 (34%) required the additionof intralesional amphotericin B deoxycholate (AMB).When AMB was added to the treatment regimen, nine(56%) of the 16 cats recovered from the disease. Theoverall cure rate was 74.5% (44/59 cats). In conclusion,the combination of ITZ and KI therapy was necessaryin most cats to achieve disease cure. In some cases,however, the addition of a third drug (intralesionalAMB) to the treatment regimen was needed to improvesuccess rate.Source of funding: Coordination for the improvement of

higher education personnel (CAPES), Brazil.


FC-40

Performance of a pad containing Ophytrium

and chlorhexidine digluconate 3% in dogs with

local bacterial and/or Malassezia overgrowth

M. GATELLET*, R. KESTEMAN*, B. BAULEZ†, F.PRADIES†, P. M. CADOT‡, H. DROPSY§, P.FIORA¶, J. L. MATHET††, E. OLLIVIER*, C.BILLY* and X. DE-JAEGER**Ceva Sant�e Animale, Libourne, France; †Cliniquev�et�erinaire VPlus, Saint Aubin de Blaye, France;‡Clinique v�et�erinaire SeineVet, Rouen & Boos, France; §

Ecole nationale v�et�erinaire d’Alfort, Maisons-Alfort,France; ¶Clinique v�et�erinaire Saint-Jacques, Agen,France; ††Clinique v�et�erinaire des Glycines, Orl�eans,France

Local disturbances of the microbiota are common indogs with underlying skin conditions. Antiseptic topicalproducts are indicated to control such superficial disor-ders. The objective of this study was to evaluate theperformance of a daily application of pads containingOphytrium and chlorhexidine digluconate 3%(DOUXO S3 PYO Pads, Ceva Sant�e Animale;

42 Abstracts



Libourne, France) in dogs with focal bacterial and/orMalassezia overgrowth. Eighteen dogs with focal skindysbiosis were enrolled in a prospective, multicentric,open study. Dogs received daily pad applications for 14days. Bacterial and/or Malassezia counts per micro-scopic field and a global score of the most affected area(0–17 scale based on extension, severity, bacterial andMalassezia cytological scores) were assessed by a veteri-narian and pruritus by the owner (pruritus Visual Ana-log Scale, pVAS) on day (D)0, D7, D14. Owner andveterinarian evaluations of performance and satisfac-tion were recorded. Eleven dogs had primarily cocciovergrowth and seven mostly Malassezia. Mean bacte-rial and Malassezia counts decreased after 14 days (6.9to 1.1; 7.6 to 1.5 respectively); 88.9% of dogs achieveda ≥ 70% microbial decrease. Mean global score of themost affected area and pVAS significantly improved atD14, respectively, from 8.6 to 2.6 and 4.5 to 1.2(P < 0.05 each; mean improvements of 70.4% and71.4%, respectively). Global veterinary assessment ofthe protocol was satisfactory, good or excellent in88.9% of cases. Most owners (94.4%) considered theprotocol efficacious. Using a pad containing Ophytriumand chlorhexidine digluconate 3% daily for 14 daysimproved the skin condition and pruritus of dogs withlocal dysbiosis, resulting in high satisfaction levels forboth veterinarians and dog owners.Source of funding: Ceva Sant�e Animale, Libourne,

France.

Conflict of interest: All the authors received reimburse-

ments, fees, funding or salary from Ceva Sant�e Animale.

FC-41

Significance of isolation of Staphylococcusschleiferi in canine superficial bacterial

folliculitis

T. OSUMI*,†, Y. SHINOMIYA*, I. IMANISHI*, Y.SHIMAZAKI†, K. IDE*,†, K. IYORI‡ and K.NISHIFUJI*,†

*Laboratory of Veterinary Internal Medicine; †AnimalMedical Centre, Tokyo University of Agriculture andTechnology, Tokyo, Japan ‡Vet Dem Tokyo, Kanagawa,Japan

Canine superficial bacterial folliculitis (SBF) is a bacte-rial skin disease characterized by follicular papules andpustules. Although Staphylococcus pseudintermedius isthe most frequently isolated bacterial species,S. schleiferi is occasionally isolated from skin lesions ofcanine SBF. The pathogenic involvement of S. schleiferiin canine SBF has not been sufficiently investigated. Inthis study, we investigated whether S. schleiferi can besolely isolated from pustules in canine SBF. Initially,seven staphylococcal isolates were obtained from sevenpustules in a dog diagnosed with SBF and subjected tomultiplex PCR to identify the staphylococcal species.One isolate was S. schleiferi, whereas the remaining sixwere S. pseudintermedius. A total of 36 staphylococcalisolates were obtained from four pustules in the samedog three months after the first study and subjected tothe same experiment. Four of 12 isolates obtained froma single pustule were S. schleiferi, while the remaining32 obtained from all four pustules were S. pseudinter-medius. These results imply that identification ofS. schleiferi obtained from a single pustule of canine

SBF does not always represent the primary pathogencausing the disease. Bacterial culture and antibiotic sus-ceptibility testing from multiple lesions are recom-mended when S. schleiferi is isolated from a pustule incanine SBF.Source of funding: Self-funded.


FC-42

Evaluation of the effects of chlorhexidine

digluconate with and without canine b-defensin103 or canine cathelicidin against multidrug-

resistant clinical isolates of Staphylococcuspseudintermedius

L. KHER*, D. SANTORO*, V. CHALA† andC. NAVARRO†

*Department of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA; †Virbac SA Laboratories, Carros,France

Host defence peptides (HDPs) such as b-defensin (BD)and cathelicidin (Cath) play a fundamental role in theepithelial natural defence against micro-organisms.Because of their mechanism of action, HDPs haveshown very low bacterial, fungal and viral resistance.Our objective was to determine if their presence mightfacilitate the antimicrobial activity of chlorhexidineagainst multidrug-resistant Staphylococcus pseudinter-medius (MRSP) in in vitro conditions. Ten MRSP clini-cal isolates were used for this study. Two-fold serialdilutions of 2% chlorhexidine solution (1:32,000,1:64,000, 1:128,000 and 1:256,000) alone or in combina-tion with synthetic canine HDPs (cBD103 and cCath)were prepared. A concentration of 1 µg/mL of syn-thetic HDP was selected based on previous studies.Friedman’s test and a Dunn’s multiple comparison testwere used for statistical analysis. Minimal inhibitoryconcentration 90 (MIC90) after 24 h for chlorhexidine[1:64,000 dilution (0.3125 µg/mL)] alone or in combina-tion with HDPs were obtained. Minimal bactericidalconcentration was achieved only for chlorhexidine com-bined with cCath at 1:32,000 (0.625 µg/mL) and1:64,000 (0.3125 µg/mL) dilution after 30 and 60 minexposure, respectively. A significant reduction(P < 0.05) of colony forming units was seen for thesame combination compared to chlorhexidine alone formost concentrations and time points tested (5, 15, 30and 60 min). A 2-log10 reduction (synergistic effect)was achieved for the highest concentrations (1:32,000and 1:64,000) after 30 and 60 min, respectively. Thesepreliminary data suggest the potentiation of chlorhexi-dine on MRSP when associated to cCath. Potentialmechanics could involve cell membrane disruptiveproperties of HDPs.Source of funding: Virbac Corporation, Carros, France.

Conflict of interest: DS received reimbursements, fees,

funding or salary from Virbac.

Abstracts 43



FC-43

Efficacy of proprietary formulations containing

essential oils and plant extracts compared to

chlorhexidine plus miconazole shampoo in

canine superficial pyoderma

C. ROMERO*, G. SHEINBERG†, A. M.CORDERO§ and R. HEREDIA**DERMAVET, Iztapalapa, Mexico; †Centro VeterinarioMexico, Mexico City, Mexico; §VETDERM,Guadalajara, Mexico

Superficial pyoderma is a common condition in dogs.Antibiotic and potentially antiseptic resistance is agrowing problem and natural topical formulations mayprovide an alternative option. Three natural formula-tions with essential oils and plant extracts were com-pared to chlorhexidine + miconazole and microsilvershampoo. Thirty dogs with clinical and cytological evi-dence of superficial pyoderma were randomized intothree groups: 1, PYOclean shampoo (LDCA; Castres,France) twice weekly and daily rinse-free PYOcleanMousse (LDCA); 2, PYOclean shampoo twice weeklyand daily use of PYOclean Spray; and 3, Biohex sham-poo (Vetbiotek; Largo, FL, USA) twice weekly. Cyto-logical and lesion scores, and pruritus Visual AnalogScore (pVAS) were performed on day (D)1, D7 andD14. Bacterial numbers decreased by 100% on D7 andD14 in groups 1 and 2, and 88.89% on D7 and 100%on D14 in Group 3. Yeast numbers decreased on D7and D14 by 45.46% and 81.82% (respectively) inGroup 1, 76% and 92% in Group 2, and 50% and90% in Group 3. Lesion scores decreased on D7 andD14 by 70.27% and 90.61% in Group 1, 52.49% and87.95% in Group 2, and 33.55% and 62.94% in Group3. The pVAS decreased on D7 and D14 by 74.65% and95.78% in Group 1, 56.63% and 86.75% in group 2,and 71.09% and 69.88% in Group 3. We conclude thatthese plant-based products can be an effective alterna-tive to chlorhexidine for topical treatment of superficialpyoderma in dogs.Source of funding: Dermoscent Laboratoire (Laboratoire

de Dermo-cosmetique Animal), Castres, France.


FC-44

Re-evaluation of oxacillin disk diffusion

breakpoints for detection of mecA-mediated

oxacillin resistance in Staphylococcus schleiferiisolated from dogs

Y. SHIMAZAKI*, K. SHIMOIKE†, K. NISHIFUJI*,‡

and K. IYORI†

*Animal Medical Centre, Tokyo University ofAgriculture and Technology, Tokyo, Japan; †Vet DermTokyo, Kanagawa, Japan; ‡Laboratory of VeterinaryInternal Medicine, Tokyo University of Agriculture andTechnology, Tokyo, Japan

The recent guidelines for antimicrobial susceptibilitytesting for oxacillin reveal that the breakpoints of diskdiffusion inhibitory zone (≤17 mm) against Staphylo-coccus pseudintermedius also could be applied to meti-cillin-resistant S. schleiferi (MRSS). However, theprevious study used an insufficient number ofS. schleiferi isolates that showed an oxacillin disk

diffusion inhibitory zone of 15–20 mm. This studyaimed to determine the optimum breakpoint of oxacil-lin disk diffusion inhibitory zone specific to S. schleiferisubsp. coagulans (SSC) and to determine whether thevalue represents a clinical breakpoint for cefalexinagainst SSC in canine pyoderma. Receiver operatingcharacteristic curve analysis revealed that the bestbreakpoint of disk diffusion inhibitory testing againstSSC was achieved when it was approximately 22 mm(Youden’s index = 0.89, n = 93). Next, 25 dogs withpyoderma, in which oxacillin-susceptible (≥23 mm,n = 9), intermediate (18–22 mm, n = 6) and resistant(≤17 mm, n = 10) SSC strains were isolated from skinlesions, were treated with oral cefalexin for 14 days.There was a significant difference in the improvementrate of clinical scores between oxacillin-susceptible and-intermediate groups (Friedman test: P < 0.01), whereasthere were no significant differences between intermedi-ate and resistance groups (P > 0.05). These findingsindicate that the oxacillin disk diffusion breakpoint of≤22 mm is the optimum clinical breakpoint to predictthe efficacy of cefalexin in dogs with pyoderma causedby SSC.Source of funding: Self-funded.


FC-45

A case of multiresistant staphylococcal

infection of the ear successfully treated with

diode laser and photodynamic treatment with

infracyanine green

A. VERCELLI, E. GUIDI, A. CORONA and L.CORNEGLIANIClinica Veterinaria Citt�a Di Torino, Torino, Italy

Photodynamic treatment with infracyanine green anddiode laser is used in human and veterinary dentistryfor the treatment of bacterial infections. A 10-year-oldmale West Highland white terrier with chronic unilat-eral otitis and a history of unsuccessful systemic andtopical antibiotic treatments was referred. On physicalexamination, several skin fistulas were present adjacentto the ear base; the ear canal was completely stenoticand otoscopy could not be performed. Computertomography revealed calcification of the ear canal withruptured tympanic membrane, abscessation, osteomyeli-tis of the bulla and temporal condylus. Based on bacte-rial culture and susceptibility Staphylococcuspseudintermedius sensitive to pradofloxacin was iso-lated. Systemic treatment with oral pradofloxacin 3 mg/kg once daily improved the infection and the dog hadtotal ear canal ablation and bulla osteotomy per-formed. After one month, despite antibiotic treatmentnew fistulas developed at the same area. Bacterial cul-ture revealed a multiresistant S. pseudintermedius sus-ceptible to rifampicin. Under general anaesthesia, thearea was cleaned with 0.5% saline solution and 1 mLof infracyanine green (Emundo, A.R.C. GmbH; Nurn-berg, Germany) was injected into the fistulas. A treat-ment with diode laser (A.R.C. Laser 810 nm) of fourcycles 500 MW for 30 s per centimetre of affected areawas performed. No antibiotic was administered andone week later, 50% of the fistulas were closed withsignificant improvement of the patient’s clinical condi-tion. Cytological results and bacterial culture were

44 Abstracts



negative 72 h post-treatment. Total recovery occurredafter two weeks. Photodynamic treatment is a promis-ing antibacterial technique in case of localized refrac-tory bacterial skin infections.Source of funding: Self-funded.


FC-46

Adhesive tape impression cytology in

dermatophytosis and pemphigus foliaceus

L. J. VOGELNEST*,†, C. B. LUDWIG* and P. A.RAVENS**Small Animal Specialist Hospital, North Ryde, NSWAustralia; †School of Veterinary Science, University ofSydney, NSW Australia

Adhesive tape impression (ATI) cytological evaluationhas recognised value in veterinary dermatology fordiagnosis and monitoring response to therapy of bacte-rial and yeast overgrowth, and some superficial para-sitic infestations. However, suitability of ATI fordiagnosis and monitoring response to therapy of der-matophytosis or pemphigus foliaceus (PF) is poorlyevaluated. This retrospective study evaluated ATI find-ings in dogs and cats with confirmed dermatophytosisor PF. Patient records were reviewed for 10 dogs and10 cats with dermatophytosis confirmed by culture,histopathological evaluation and/or complete responseto antifungal therapy. Where culture was performed,Microsporum canis or M. gypseum were isolated. Fun-gal elements were reported on ATI in 10/10 cats and 8/10 dogs. Arthrospores were recorded as moderate tonumerous in 10 cats, located within and clusteredaround hair shaft fragments and keratinocytes. Fungalhyphae were recorded as sparse within hair shafts ofsome cats. Fungal hyphae were reported in 8/10 dogs,and typically recorded as focal or sparse and locatedon keratinocytes associated with clusters of neutrophils(most commonly) or within hair shafts. Arthrosporeswere not reported in any dog. Patient records also wereevaluated for 10 dogs and 10 cats with PF confirmedby histopathological evaluation. Acantholytic ker-atinocytes were reported associated with neutrophilclusters in all 20 cases, typically recorded as few-to-moderate in cats and moderate-to-numerous in dogs.Adhesive tape impressions appear valuable for the diag-nosis and potential monitoring of dermatophytosis andPF in dogs and cats.Source of funding: Self-funded.


FC-47

Staphylococcal species isolated and

antimicrobial resistance from canine and feline

bacterial cultures in Hong Kong

R. WESTERMEYERAnimal Allergy Specialists, Kaneohe, Hawaii, USA; andAsia Veterinary Diagnostics, Hong Kong, Hong Kong

This bacterial surveillance study was performed to char-acterize all staphylococcal isolates cultured fromSeptember 2016 to March 2017 in Hong Kong. The

objectives of this study were to find the most commonstaphylococci isolated from cats and dogs as well as theresistance patterns for these bacteria. All staphylococcalisolates submitted during the study period wereincluded. Cultured bacteria were analysed by matrix-assisted laser desorption-ionization-time-of-flight massspectrometry (MALDI-TOF MS) and identified by BIO-

TYPER (v2.0) software (Biomerieux; Marcy-l’�Etoile,France). Sensitivity was determined by disk diffusionfollowing Clinical and Laboratory Standards Institute(CLSI) guidelines M100-S26, M100-S27 and VET01S3recommendations for standard antibiotics. In total, 17species of staphylococci were identified. Staphylococcusfelis was most frequently isolated from cats at 54% fol-lowed by S. pseudintermedius at 16%. Staphylococcuspseudintermedius accounted for 64% and S. schleiferi30% of the isolates from dogs. For all species ofstaphylococci, resistance rates were as follows: penicillin61%, azithromycin 41%, doxycycline 37%, clindamycin28%, enrofloxacin 18%, marbofloxacin 18% and meti-cillin 5%. No isolates tested were resistant to baci-tracin. Amikacin resistance was 0.5% and rifampicinresistance was 1%. For S. felis isolated from cats, resis-tance rates were as follows: penicillin 50%, azithromy-cin 29%, clindamycin 16%, doxycycline 7%, meticillin0%, enrofloxacin 0% and marbofloxacin 0%. ForS. pseudintermedius isolated from dogs, resistance rateswere as follows: penicillin 77%, doxycycline 66%, azi-thromycin 61%, clindamycin 49%, enrofloxacin 28%,marbofloxacin 27% and meticillin 4%. Overall,S. pseudintermedius was the most common staphylococ-cus isolated with low meticillin resistance, and highdoxycycline and azithromycin resistance.Source of funding: Self-funded.


FC-48

Expression of antimicrobial peptides and

cytokines following bacterial stimulation in an

ex vivo skin explant model

J. E. HYUN*, Y. B. KIM†,‡ and C. Y. HWANG§

*Department of Veterinary Internal Medicine, KonkukVeterinary Medical Teaching Hospital, KonkukUniversity, Seoul, Republic of Korea; †BON AnimalMedical Center, Gyeonggi-do, Republic of Korea;‡Department of Veterinary Surgery, College ofVeterinary Medicine, Konkuk University, Seoul,Republic of Korea; §Laboratory of VeterinaryDermatology, Research Institute for Veterinary Science,College of Veterinary Medicine, Seoul NationalUniversity, Seoul, Republic of Korea

The skin protects the body by mediating variousimmune responses against exogenous substances includ-ing bacteria, fungi and viruses. In addition, it has a pre-dominant role as a physical barrier. Despite thesignificant protection offered via various mechanisms,bacterial infection of the skin is one of the most com-mon skin diseases in veterinary medicine. Here, weexamined the structural and immunological changes inthe skin during infections with Pseudomonas aeruginosaand Staphylococcus pseudintermedius using skin explantsfrom four healthy beagles. Skin structure was generallywell preserved in uninfected controls, yet defects in theskin structure, including injury of keratinocytes and

Abstracts 45



dermo-epidermal junction disruption, were identifiedwhen skin explants were exposed to P. aeruginosa andS. pseudintermedius. On exposure to P. aeruginosa,marked linear cleft formation and acantholysis alongthe basal layer was noted after 24 h of culture. In addi-tion to the skin structure defects, mRNA expressionlevels of the antimicrobial peptides (AMP) cBD103 andS100A8 were decreased, which was confirmed byimmunohistochemical staining. Conversely, mRNAexpression levels of IL-6, IL-8 and TNF-a wereincreased in S. pseudintermedius-exposed skin explants.Taken together, these results suggest that the ex vivocanine skin model used in this study is a suitableresearch tool for investigating bacterial skin infectionsin dogs.Source of funding: Self-funded.


FC-49

Change in incidence and antibiotic

susceptibility of meticillin-resistant

Staphylococcus from canine pyoderma in

referral practice in Melbourne, Australia from

2014 to 2019

D. ROBSONAnimal Skin and Ear Specialists, Melbourne VeterinarySpecialist Centre, Melbourne, Victoria, Australia

Staphylococcus pseudintermedius is the most commoncause of canine pyoderma. Meticillin-resistant Staphylo-coccus (MRS) contains the mecA gene and causes

clinical resistance to all B-lactam antibiotics. The inci-dence of MRS in cases of canine pyoderma requiringsystemic antibiotic therapy in a referral dermatologypractice in Melbourne, Australia, was examinedprospectively from September to December in 2014,2016 and 2019. Cases were classified as responders toempirical therapy (RTET), lost to follow-up (LTFU),culture-confirmed meticillin-susceptible Staphylococcussp. (MSS) or culture-confirmed MRS. Both confirmedincidence (i.e. based on culture and susceptibility) aswell as presumed incidence (i.e. included all LTFUcases on the assumption they were MRS and the RTETcases assuming they may have been successfully treatedMRS cases) were calculated. Trends in antibiotic sus-ceptibilities, as well as the incidence of non-S. pseudin-termedius MRS also were examined. Significance wasassessed using Fisher’s exact test. Ninety-four, 104 and123 dogs entered the study in 2014, 2016 and 2019,respectively. The incidence of confirmed (14.4–24.3%)and presumed (38.3–42.7%) MRS pyoderma cases wasnot significantly different in any of the three years. Anincrease in non-S. pseudintermedius MRS from 2014(0.0%) to 2019 (16.7%) also was not significant(P = 0.1435). However, the number of extensively resis-tant MRS isolates was significantly greater in 2019(26.7%) than 2014 (0.0%; P = 0.038), largely due to areduction in susceptibility to trimethoprim-sulfonamide(P = 0.0106; 2014–2019), minocycline (P = 0.0187;2016–2019) and chloramphenicol (P = 0.0283; 2016–2019). Canine pyoderma complicated by MRS contin-ues to be a challenge in clinical practice in Melbourne.Source of funding: Self-funded.


46 Abstracts



EQUINE SKIN DISEASES

FC-50

Prevalence of horses suspect of atopic

dermatitis could be higher than that of insect

bite hypersensitivity

C. LORENTE-MENDEZ and R. WAGNERLaboklin GmbH & Co. KG, Bad Kissingen, Germany

Insect bite hypersensitivity (IBH) is considered the mostcommon allergic skin disease in horses, whereas theprevalence of atopic dermatitis (AD) is unknown. Thisretrospective study aimed to evaluate the prevalence ofpositive serum immunoglobulin (Ig)E antibodies againstfour groups of allergens: insects, pollens, mites andmoulds using ELISA technology. Serum samples from2,412 horses in Europe with a possible allergic diseasewere tested. Two thousand and sixty-nine (85.78%)samples yielded positive results for at least one of thefour groups of allergens and 674 (32.58%) of thesesamples were positive to allergens representing all of

the groups. From the 2,069 positive samples, 49.73%(1,029) were positive to insects, 84.73% (1,753) tomites, 65.80% (1,360) to moulds and 75.80% (1,570) topollens. Only 39 (1.88%) samples were exclusively posi-tive to insects. The percentage of samples positive toinsects was lowest from January to April (average28.21%), and highest from May to October (average50.48%). Positive samples for mites (76.41–98.46%)and pollens (61.76–86.79%) were >60%, and formoulds (50–80%) >50% throughout the year. Theprevalence of positive reactions was higher on amonthly basis for any of the three groups of aeroaller-gens than for insects. Sole reactions to insects wererare. The results suggest that the prevalence of horsessuspected of having AD is higher than that of IBH,and serum IgE reactive to both allergen groups is com-mon.Source of funding: Self-funded.


Abstracts 47



SKIN DISEASES OF EXOTIC SPECIES

FC-51

Efficacy of sarolaner on African hedgehogs

(Atelerix albiventris) infested with Capariniatripilis

A. RANGEL*, C. ROMERO†, G. SHEINBERG‡, R.HEREDIA† and A. CORDERO§

*Hospital Veterinario Grandes Compa~neros de Torre�on,Coahuila, Mexico; †Dermavet, Edo Mex, Mexico;‡Centro Veterinario Mexico, CDMX, Mexico;§Vetderm, Guadalajara, Mexico

The most common mite in the African pygmy hedgehog(Atelerix albiventris) is Caparinia tripilis, causing lesionsand itching of the head, pinnae, dorsum and the exter-nal auditory canal. The objective of this study was toevaluate the efficacy of sarolaner as a treatment forC. tripilis infestation in African pygmy hedgehogs. TwoAfrican hedgehogs were recruited, a male and a female,nine months old, weighing 350 g, presenting with gener-alized erythema, scaling, pruritus and loss of spikes. A

Visual Analog Scale was used to score the lesions andpruritus, and microscopic observations made for thepresence of the C. tripilis mite. Both animals were trea-ted with 2 mg/kg of sarolaner (Simparica, Zoetis; Kala-mazoo, MI, USA) orally, mixed with wet food. Patientswere evaluated on pruritus level, skin lesions and pres-ence of parasites on day (D)1, D7, D14, D30 and D60.The data were analysed with the Wilcoxon signed ranktest. The severity of lesions in both hedgehogs showedsignificant decrease (P = 0.03) from D1 to D30, with ascore of zero from D30 to D60. The itching decreasedsignificantly from D1 to D15 (P = 0.02) reaching andremaining zero at that time. The presence of mitesdecreased significantly (P = 0.03) in both hedgehogsfrom D1 to D15 post-treatment, reaching zero andremaining thus until D60. This study demonstrates mitecontrol for 60 days with only one administration of sar-olaner in hedgehogs infested with C. tripilis.Source of funding: Self-funded.


48 Abstracts



NEW TRENDS IN DIAGNOSIS AND TREATMENT

FC-52

Role of extracellular vesicles in skin diseases:

potentiality of new diagnostic tests based on

the analysis of circulating extracellular vesicles

L. BONGIOVANNI*,†, A. ANDRIESSEN*, T. A. P.DRIEDONKS*, E. N. M. NOLTE-THOEN* and A.DE BRUIN*,†

*Department of Biomolecular Health Science, Faculty ofVeterinary Medicine, Utrecht University, Utrecht, theNetherlands; †Department of Pediatrics, UniversityMedical Center Groningen, University of Groningen,Groningen, the Netherlands.

Analagous to messages sent by cells to communicatewith each other, extracellular vesicles (EVs) haveemerged as important mediators of intercellular com-munication in various physiological and pathologicalprocesses. In human skin diseases such as cancer, espe-cially cutaneous melanoma, as well as inflammatorydiseases (such as acne, Malassezia, Leishmania, atopicdermatitis), EVs have been outlined as important play-ers in disease pathogenesis and progression. EVs couldpotentially be clinically applicable as drug-delivery vec-tors, cell-free vaccines and immunomodulators. In diag-nostics, circulating EVs provide a novel approach tothe development of biomarkers, carrying informationof the tissue/cell of origin and being enriched with dis-ease-relevant molecules. In particular, mRNA mole-cules are packaged into EVs in a specific manner. Thisis why we think that circulating EVs and their mRNAcargo can be exploited as a potential diagnostic test incanine patients with different type of diseases, such ascancer. We applied a protocol to isolate EVs fromcanine blood samples based on the use of size exclusionchromatography (SEC) and analyse their RNA content.First, we collected blood from healthy dogs and caninecancer patients with different types of skin cancer. EVswere isolated by using qEVoriginal/ 70 nm (Izon) SECcolumns and confirmed by Western blotting; RNA wasextracted from SEC fraction pools and analysed byquantitative reverse transcription (qRT)- PCR. Protocoldetails on how blood samples were collected and pro-cessed, EVs isolated, and RNA extracted and analysedwill be shown and explained, discussing current limita-tions and potential applications in veterinary medicine.Source of Funding: European Research Council under the

European Union’s Seventh Framework Programme (FP/

2007-2013)/ERC grant number 337581.


FC-53

Susceptibility of Australian canine otitis

externa bacterial and fungal isolates to

florfenicol and terbinafine

S. DE BURGH*, E. HICKEY† and D. TROTT†

*Bayer Australia Ltd, Pymble, Australia; †University ofAdelaide, Adelaide, Australia

Most conventional otitis externa (OE) treatments fordogs require single/multiple daily applications andcleaning by dog owners, until the condition is resolved.Florfenicol/terbinafine/mometasone (Neptra, Bayer;Leverkusen, Germany) applied as single treatment elim-inates pet-owner noncompliance and reduces the possi-bility of antimicrobial resistance arising fromincomplete treatment protocols. An Australian surveyidentified the most prevalent canine OE bacterialpathogens as Pseudomonas aeruginosa, Staphylococcuspseudintermedius, Proteus sp., beta-haemolytic strepto-cocci (e.g. Streptococcus canis) and Escherichia coli.Malassezia pachydermatis is the most common fungalpathogen. This study was undertaken to determine min-imum inhibitory concentrations (MICs) of florfenicoland terbinafine against bacterial and fungal isolatessourced from Australian clinical cases of canine OE.Isolates of P. aeruginosa were excluded. All bacterialisolates showed consistent MIC values when testedagainst florfenicol, resulting in very narrow MICranges. S. canis and S. pseudintermedius isolates showedlow MIC values of ≤4 lg/mL. Gram-negative isolatesexhibited slightly higher MIC values against florfenicoland MICs were still very low overall. Proteus sp. andE. coli isolate MIC values ranged from 4 to 16 lg/mL,whereas M. pachydermatis isolates showed a muchwider range of MIC values against terbinafine (0.125–32 lg/mL). However, most of these isolates were sus-ceptible to the lower concentrations in vitro, demon-strated by the low MIC50 and MIC90 values of 0.25and 1 lg/mL, respectively. Recent isolates from casesof canine OE in Australia are susceptible to florfenicoland terbinafine at concentrations achieved during topi-cal delivery of Neptra, at the recommended dose rate.Source of funding: This study was fully funded by Bayer

Australia Ltd.

Conflict of interest: SdeB is an employee of Bayer Aus-

tralia Ltd.

Abstracts 49



FC-54

Single myringotomy in the treatment of middle

ear otitis in the cat: a retrospective study

S. DELEPORTE* and P. PRELAUD†

*Dermatology Department, LORRAINEVET, Ludres,France; †Dermatology Department, ADVETIA CentreHospitalier V�et�erinaire, V�elizy, France

Otitis media occurs frequently cats yet data on medicaltreatment are lacking except for polyp-associated otitismedia. We studied retrospectively 24 cases of cats withotitis media confirmed by MRI or CT without polyp orneoplasia. Most cats (58%) were presented with bilat-eral involvement. Both parts of the bulla were involvedin 97% of cases. Clinical signs included head tilt 54%(11/24), Horner syndrome 29% (7/24), ataxia or otalgia9% (2/24), otitis externa 21% (5/24), nystagmus andfacial paralysis 4% (1/24). Cytological findings of mid-dle ear effusion were predominantly neutrophilic. Cocciwere identified on cytological examination in 20/38samples and rods in 2/38. Bacterial culture was positivein 50% of samples, with isolation of Pseudomonas sp.,Pasteurella multocida, Staphylococcus felis, S. schleiferi,S. pseudintermedius and Serratia marcesens. All catswere treated using single myringotomy with soft bullaflushing (0.5–2 mL saline) associated with systemic cor-ticosteroid (prednisolone 1 mg/kg/day, seven–10 days)and systemic antibiotics according to susceptibility test-ing (one month). Seventeen cats (71%) were clinicallycured 60 to 240 days after treatment. One cat washealthy with a mild head tilt. One cat was euthanisedbecause of failure to control Pseudomonas infection.Another one died because of complications of anaesthe-sia. The four remaining cats were cured after ventralbulla osteotomy. This simple technique with only onemyringotomy and mild ear flushing offers a very practi-cal, safe and efficient way to treat suppurative otitismedia in cats.Source of funding: Self-funded.


FC-55

Efficacy of narrowband ultraviolet B therapy

for eight dogs with cutaneous T-cell lymphoma

T. HOSHINO*, M. HADA*, A. SATO*, P. F.MOORE† and M. NAGATA**Dermatology, Veterinary Specialists Emergency Centre,Saitama, Japan; †Department of PathologyMicrobiology and Immunology, University of California,Davis, CA, USA

Cutaneous T-cell lymphoma (CTCL) is a malignantskin neoplasm. In humans, ultraviolet (UV) light ther-apy is an established treatment for CTCL and chosenas one of the first-tier treatments in early stage CTCL.The aim of this study was to evaluate the efficacy andsafety of narrow band (NB)-UVB therapy for canineCTCL. Eight dogs were recruited at Veterinary Special-ists Emergency Centre. CTCL was diagnosed based ontypical clinical features of CTCL and histopathologicalinfiltrations of atypical T cells in the epidermis or der-mis. The affected area was irradiated with NB-UVBusing a portable device (TARNAB, Inforward, Inc.;Tokyo, Japan), which supplies 312�11 nm centre

wavelength through the 160 x 60 mm rectangular shapeprobe. The NB-UVB dose was started from a minimum250 mJ once or twice weekly and then graduallyincreased to maximum if previous irradiation wasunsuccessful. The skin lesions were nasal erythema and/or depigmentation, nasal erosion and/or ulcer, perianalerythema and/or depigmentation, perianal erosion and/or ulcer, and erythroderma, with remission rates of75.0%, 50.0%, 33.3%, 20.0%, 20.0% and 0%, respec-tively. Effective doses were 500–850 mJ and most casesrequired once weekly proactive irradiations. Post-trea-ted skin biopsy collected from one dog demonstratedfewer infiltrations of tumour cells. No severe adversereactions appeared except for mild erythema (28.5%)or pruritis (28.5%). This study suggested that NB-UVBtherapy is safe and useful for canine CTCL.Source of funding: Self-funded.


FC-56

Detection and inhibition of IgE for cross-

reactive carbohydrate determinants evident in

an ELISA for detection of allergen-specific

IgE in the serum of dogs and cats

K. LEE*, B. MCKINNEY*, K. BLANKENSHIP*, K.ENCK* and D. MORRIS†

*Stallergenes Greer, Lenoir, NC, USA; †School ofVeterinary Medicine, University of Pennsylvania,Philadelphia, PA, USA

It has recently been demonstrated that immunoglobulin(Ig)E specific for cross-reactive carbohydrate determi-nants (CCD) is present in the serum of allergen-sensi-tized dogs and cats, and that these CCD-specificantibodies might confound serological testing. Theobjective of this study was to document the prevalenceof CCD detectable in a monoclonal cocktail-basedenzyme-linked immunosorbent assay (ELISA) designedfor the detection of allergen-specific IgE in the sera ofdogs and cats, and to define a means for successfulinhibition of these CCD. The incidence of reactivity tobromelain and a commercially available inhibitor ofcarbohydrate-specific antibodies (RIDA-CCD) wasevaluated in 100 dog sera samples before and after inhi-bition with RIDA-CCD, and a proprietary inhibitorwhich contains carbohydrates derived from bromelain(BROM-CCD). Subsequently, sera from 600 dogs and600 cats were evaluated using a serum diluent with andwithout BROM-CCD. Although both the RIDA-CCDand BROM-CCD inhibitors demonstrated successfulreduction of CCD reactivity, a more efficient profile ofinhibition was evident with BROM-CCD. Mite reactiv-ity in dog and cat sera was largely unaffected; however,substantial inhibition for pollen allergens (trees, grassesand weeds) was shown. After BROM-CCD inhibition,1% of canine samples and 13% of feline samples wererendered completely negative for allergen reactivity.The results demonstrate that BROM-CCD is effectivein reducing reactions with irrelevant carbohydrates, andthat inhibition of CCD reactivity might substantiallyalter the outcome of the in vitro reactivity profile usedfor selection of allergens to be included in animmunotherapeutic regime.

50 Abstracts



Source of funding: Funding for this study was provided

by Stallergenes Greer.

Conflict of Interest: Four authors are employees at Stal-

lergenes Greer.

FC-57

Appropriate chemotherapeutic treatment of

equine skin tumours in the field

A. M. RASHMIR*, S. I. HONKISZ† and R. S.MCCONNICOǂ

*Veterinary Surgical Services, Washington, DC, USA;†Michigan State University, East Lansing, MI, USA;ǂLouisiana Tech University, Ruston, LA, USA

Tumours of the skin are the most common form ofneoplasia in the horse. They are frequently treated infield situations where the potential for public exposureto the chemotherapeutic agents is high. The NationalInstitute for Occupational Safety and Health (NIOSH)has recently updated the requirements for handlingchemotherapeutic agents. Knowledge of these updatedrequirements combined with current recommendationsfor the use of these agents in the treatment of cuta-neous neoplasms will advance the treatment of thehorse and provide protection from inadvertent humanexposure. Therefore, our objective is to provide accu-rate information regarding the treatment of the mostcommon forms of equine tumours in the field and theappropriate management of the chemotherapeuticagents used. For example, intratumoural cisplatin hasproduced favourable long-term control of several typesof equine skin tumours and can be performed in thefield setting without general anaesthesia. Both the vet-erinarian administering the cisplatin and the handlerare required to wear NeoPro gloves, tyvek suits, faceshields, gloves and a N-95 mask. The drug should beinjected with PhaSeal leur lock connectors to avoidspray under pressure. Clean-up should be achieved inthe same attire with 10% bleach solution followed by1% sodium thiosulfate solution. Being careful not toself-contaminate, the PPE should then be disposed ofin a designated chemotherapy waste bin. Likewise,treatment with other chemotherapeutic agents that aretopical, intralesional or in impregnated beads should allbe considered a risk to the veterinary team and thepublic, and protective equipment worn.Source of funding: Self-funded.


FC-58

Owner assessment of long-term treatment of

PPID

H. SCHOTT, J. STRACHOTA and J. MARTENIUKDepartment of Large Animal Clinical Sciences, Collegeof Veterinary Medicine, Michigan State University, EastLansing, MI, USA

Owners of 29 equids enrolled in a long-term safetystudy of Prascend (1 mg pergolide mesylate tablets) fortreatment of pituitary pars intermedia dysfunction(PPID) were surveyed about their experience with thistreatment 10 years after the start of treatment. Allequids enrolled in the study were initially treated with a

dose of 2 µg/kg per os, once daily and 26/29 equidshad a dosage increase to 4 µg/kg p.o., once daily from0.5 to 6 years after onset of treatment. Over the courseof the study, 25/29 equids died (n = 4) or were sub-jected to humane euthanasia (n = 21) between 1.5 and10 years after onset of treatment. Death or euthanasiawas attributed to complications of PPID, specificallylaminitis, in five equids while death in the remaining 20equids was attributed to disorders associated withadvancing age. Eighty-six percent (25/29) of ownerscompleted the follow-up survey, and 71% and 70%strongly agreed and 25% and 30% agreed that treat-ment with Prascend improved their equid’s quality oflife and prolonged lifespan, respectively. The improve-ment in clinical signs was greatest for energy level(77%), hair coat (71%) and muscle mass (61%). Over-all satisfaction with treatment on a scale of 1–10 (10,best) was 5 (9%), 7 (9%), 8 (13%), 9 (17%) and 10(52%); 87% of owners either agreed or strongly agreedthat they would provide life-long treatment if they hadanother equid with PPID. In conclusion, owner satis-faction with Prascend as a treatment for PPID is high.Source of funding: Boehringer-Ingelheim Animal Health

USA, Inc., Duluth, GA, USA provided Prascend for par-

ticipants in this study.

Conflict of interest: HS is a consultant and has received

reimbursements, honoraria, and research funding from

Boehringer-Ingelheim Animal Health, USA.

FC-59

Comparison of oral absorption of parenteral

triamcinolone acetonide in dogs to absorption

of oral triamcinolone acetonide tablets, a pilot

study

C. ROMERO*, C. GRIFFIN†, M. G. PAPICH‡, G.SHEINBERG§, C. MARTIN¶ and R. HEREDIA**Dermavet, Edo Mexico; †Animal Dermatology ClinicSan Diego, California, USA; ‡North Carolina StateUniversity, Raleigh, North Carolina, USA; §CentroVeterinario Mexico, CDMX, Mexico; ¶Vetderm,Guadalajara, Mexico

The objective of this study was to determine if inject-able triamcinolone acetonide (TA) administered orallyis absorbed to the same level as TA oral tablets. Acomparative study was conducted with seven dogs thathad not received any medications in 90 days before thestudy. All dogs received an oral dose of oral formula-tion TA (0.11 mg/kg Cortalone, Vedco, BoehringerIngelheim; Ridgefield, CT, USA). Three weeks later,the same dogs were administered with the same oraldose of parenteral formulation of TA (Atriben, Holli-day Scott; Buenos Aires, Argentina). On both occa-sions, samples were taken at 0, 15, 30, 60, 120, 240 and480 min, and 25 and 48 h post-administration of thedrug, to evaluate plasma concentrations of TA by high-performance liquid chromatography (HPLC). CBC andblood chemistry were performed at 120 min, 25 h and36 h. For the analysis of data ANOVA was performed,the comparison of means was performed using theLSMEANS procedure of SAS (alpha = 0.05). There wasno statistical difference in plasma concentrations of TAbetween formulations (P = 0.05) in the measurementsmade at 15, 30, 60, 120, 240, 480 min, and 25 and 48 hafter oral administration of oral and parenteral TA.

Abstracts 51



Comparison of means of analysed blood biometry andblood chemistry values showed no significant differ-ences (P = 0.05. These results indicate that parenteralTA can be administered by the oral route for the treat-ment of dog dermatopathies for veterinarians who donot have access to oral TA.Funding source: Self-funded.

Conflict of interest: none declared.

FC-60

Effects of sarolaner on earthworms (Eiseniafoetida)

C. ROMERO*, R. HEREDIA*, L. MIRANDA*, G.SHEINBERG† and A. MARTIN‡

*Dermavet Hospital Veterinario, Ciudad de M�exico,Mexico; †Centro Veterinario M�exico, Ciudad deM�exico, Mexico; ‡Veterinaria Dermatolog�ıa VeterinariaEspecializada, Guadalajara, Mexico

The present study examined the effect of sarolaner onearthworms. Experiment 1: three groups of rabbits weretreated with 2.5 mg/kg sarolaner (Simparica, Zoetis;Parsippany, NJ, USA) and a fourth group was nottreated. Stool was collected for each rabbit group andthen were applied in compost sections with earthworms(three sections received sarolaner-treated rabbit faecesand one section received the control rabbit faeces).Experiment 2: organic matter with 120 mg sarolanerwas added to three groups of 100 worms each and afourth group did not receive sarolaner. The morphol-ogy and number of earthworms for both experimentswere evaluated on day (D)1, D15 and D30 post-treat-ment by separating the worms from the organic matter.Data were analysed using a Tukey’s standardized rangetest with P = 0.05 and Fisher’s exact test was used forstatistical comparisons. For the earthworms exposed torabbit faeces, there were no significant differences inthe number of worms between sarolaner-treated andcontrol groups at D15 and D30; in fact, there was anincrease in the number of earthworms in all groups.For the second experiment there were no significant dif-ferences in earthworm numbers on the same evaluationdays. Comparison between both experiments showedno differences at D15 and D30 post-treatment. Thesedata revealed that sarolaner treatment did not affectthe reproduction and growth of earthworms up to 30days post-administration through either indirect expo-sure (via the faeces of rabbits medicated with sarolaner)or direct administration of sarolaner to the vermicom-post.

Source of funding: Self-funded.


FC-61

Usefulness of a novel isolation and drug

susceptibility test media for diagnosis and

treatment of superficial bacterial folliculitis in

dogs

H. SHIMOURA, M. ESUMI, S. KANDA and K.IYORIVet Derm Tokyo, Kanagawa, Japan

Recently, an isolation and drug susceptibility testingmedium, Hifu-Quik (NIPPON ZENYAKU KOGYO,Co., Ltd.; Tokyo, Japan) was made available in Japanfor dogs with staphylococcal infection. This medium issalt agar-based with four fractions containing specificconcentrations of cefazolin, ofloxacin or minocycline,plus control. In this study, we aimed to evaluate theusefulness of the test media for treatment of superficialbacterial folliculitis (SBF) in dogs. Twenty-nine dogswith SBF were enrolled in this study. Samples were col-lected from skin lesions and inoculated onto the fourfractions of the test media and incubated at 35 for 24–48 h. Samples also were subjected to routine bacterialculture and antimicrobial susceptibility disk diffusiontesting according to Clinical and Laboratory StandardsInstitute (CLSI) guidelines. All cases showed colonyformation on the control fraction of the test mediumwithin two days. In the bacterial culture test, Staphylo-coccus pseudintermedius (n = 22) and S. schleiferi subsp.coagulans (n = 7) were isolated. Concordance ratesbetween antimicrobial susceptibility test results and col-ony formation on the test medium were 100% for cefa-zolin (24/24) and ofloxacin (26/26), and 72.2% (13/18)for minocycline. All dogs were randomly treated withsystemic cephalosporins (n = 13), fluoroquinolones(n = 9) or tetracyclines (n = 7). Overall, the predictionrate of clinical response to antibiotics, based on testmedium results, was 86.2% (25/29). This findingdemonstrates that this test medium could be a rapidand beneficial tool for determining antimicrobial sus-ceptibility and clinical response to cephalosporins, fluo-roquinolones and tetracyclines in dogs withstaphylococcal SBF.Source of funding: Self-funded.

Conflict of interests: None declared.

52 Abstracts



ACVD RESIDENTS

FC-62

Real-time PCR detection of undeclared animal

DNA in three commercial elimination diets for

canine adverse food reactions

T. M. S. A. BOEHM*, C. J. KLINGER† and R. S.MUELLER**Centre for Clinical Veterinary Medicine, LudwigMaximilian University, Munich, Germany; †TierklinikStuttgart Plieningen, Veterinary Specialist Hospital,Stuttgart, Germany

An elimination diet is currently the gold standard todiagnose canine adverse food reactions. However, mul-tiple studies have shown the contamination of novelprotein diets that have been claimed to be suitable forfood trials. It is unclear if several samples from differ-ent batches are all uniformly contaminated or if suchcontamination only occurs sporadically and how largethe percentage of undeclared protein is in contaminatedsamples. The purpose of this study was to test severalsamples of different batches of three selected proteindiets for dogs and, if present, quantify contaminatingproteins in those samples. Twelve samples of threecanned selected protein diets of one brand were chosen.Diets contained camel and parsnip, kangaroo andpumpkin, and horse with amaranth, peach and beetroot(Terra Canis; Munich, Germany). Specimens werescanned for undeclared DNA of animal origin usingquantitative real-time PCR testing for horse, turkey,chicken, beef, mutton and pork antigens (Institute forFood Safety, Austrian Agency for Health and Nutri-tional Safety; Vienna, Austria). Eleven of 12 sampleswere not contaminated with any of the tested proteinsnot listed on the label. One sample with camel andparsnip contained <1% turkey. When testing differentbatches of commercial selected protein diets, contami-nation with small amounts of proteins not listed on thelabel can occur intermittently. Whether those smallamounts are clinically relevant is currently unknown.Based on this study, commercial selected protein dietsmay be suitable as maintenance diets for dogs withdiagnosed food allergy, yet their use in diagnostic elimi-nation diets is still questionable.Source of funding: The study was funded by Nestl�eDeutschland AG.

Conflict of interest: Nestl�e Deutschland AG was not

involved in the selection of food samples nor in the choice

of commercial laboratory.

FC-63

Clinically relevant plasma concentrations of

rifampicin induce resistance in multidrug-

resistant canine Staphylococcus intermediusgroup isolates

K. HICKS*, T. HATHCOCK†, D. BOOTHE‡, R.KENNIS* and A. WHITE**Department of Veterinary Clinical Sciences, College ofVeterinary Medicine, Auburn University, Auburn, AL,USA; †Department of Pathobiology, College ofVeterinary Medicine, Auburn University, Auburn, AL,USA; ‡Department of Anatomy, Physiology and

Pharmacology, College of Veterinary Medicine, AuburnUniversity, Auburn, AL, USA

Antimicrobial resistance is a growing concern and com-plicates treatment of Staphylococcus pseudintermediusdermatitis in dogs. Treatment with rifampicin (RFP) isconsidered only in S. pseudintermedius isolates that areboth meticillin-resistant (MR) and multidrug-resistant(MDR). Recent data demonstrated that canine plasmaRFP concentrations after 6 mg/kg per os are 600–1,000-fold greater than the minimum inhibitory concen-tration [MIC (90)] of S. pseudintermedius (MIC0.008 µg/mL). The objective of this study was to deter-mine the killing properties of RFP against MR andMDR S. pseudintermedius at concentrations similar tothose achieved in plasma after oral dosing. One controlisolate (S. aureus ATCC 25923, American Type CultureCollection; Manassas, VA, USA; MIC 0.008 µg/mL)and three canine MR-MDR S. pseudintermedius isolates(MICs 0.004, 0.008 and 0.016 µg/mL) were exposed toRFP (MP Biomedicals; Santa Ana, CA, USA) at con-centrations from 32- to 1,024-fold greater than the iso-late MIC. The number of viable colony forming unitswas determined using the BacTiter-Glo kit (Promega;Madison, WI, USA) at different time points (0, 2, 4,12, 24 and 48 h) to create a time–kill curve. RFP activ-ity against all four isolates was consistent with a time-dependent and bacteriostatic response. At either 24 or48 h, each isolate demonstrated resistance to RFP atone (n = 2) or two (n = 2) of the six concentrationstested ranging from 32- to 256-fold greater than theMIC. These resistant isolates exhibited an increase inthe MIC via Etest (BioM�erieux; Durham, NC, USA)with post-exposure MIC ranging from 1 to >32 µg/mL.These findings strongly suggest that S. pseudintermediusdevelops RFP resistance at high concentrations in vitro,calling into question RFP use in dogs.Source of funding: Auburn University Department of

Clinical Sciences Research and Graduate Studies Intra-

mural Grant.


FC-64

A shortened immunotherapy dose-escalation

saves time, but is it safe? A case-control study

comparing the rates of adverse reactions

between conventional and fast-escalation

subcutaneous immunotherapy protocols

I. HERRMANN*, K. E. LOFT† and T. OLIVRY**Department of Clinical Sciences, College of VeterinaryMedicine, NC State, Raleigh, NC, USA; †MSPCAAngell Memorial Hospital, New York City, NY, USA

Allergen-specific immunotherapy (ASIT) is the onlyintervention believed to change the course of atopic dis-ease. As dogs appear to have fewer severe adverseevents (AEs) compared to people receiving ASIT, aprolonged dose-escalation might not be needed. Ourobjectives were to retrospectively compare the incidenceand characteristics of AEs induced by a fast-escalationsubcutaneous immunotherapy (f-SCIT) protocol tothose of two conventional (c-SCIT) regimens. The AEsduring the induction phase were compared between

Abstracts 53



three groups of 50 dogs treated with either f-SCIT pro-vided by ALK-Abello ([ALK]; Round Rock, TX, USA)or with c-SCIT provided by ALK or by StallergenesGreer ([Greer]; Boston, MA, USA). We graded thereported AEs using a modified World Allergy Organi-zation system (local reaction to grade I–IV systemicreactions) and assessed factors such as self-limitation.Dogs treated with the ALK f-SCIT (11/50; 22%) andGreer c-SCIT (8/50; 16%) had a significantly higherrate of total AEs compared to those receiving the ALKc-SCIT (1/50; 2%) (Fisher’s exact test; P = 0.004 andP = 0.03, respectively). Most of the AEs (10/11; 91%)in the ALK f-SCIT group were self-limiting grade I sys-temic reactions. In each of the three groups of dogs, asingle grade II systemic reaction was observed, andthere were no grade III-to-IV systemic reactions. Thisf-SCIT regimen is simpler, its maintenance phase isreached faster, and most of its associated AEs are mildand self-limiting. Whether this f-SCIT protocol leads toa faster time to efficacy needs to be determined infuture studies.Sources of funding: Self-funded.


FC-65

Oral dexamethasone sodium phosphate

solution significantly reduces pruritus and

clinical lesions in feline hypersensitivity

dermatitis: an open-label study

D. MCCLINTOCK, M. AUSTEL and F. BANOVICDepartment of Small Animal Medicine and Surgery,College of Veterinary Medicine, University of Georgia,Athens, GA, USA

Feline hypersensitivity dermatitis (HD) is a multifacto-rial, pruritic condition for which glucocorticoids are

considered a mainstay of therapy, despite the difficultyof administering oral tablets to cats and the potentiallysevere adverse effects of parenteral glucocorticoid injec-tions. We evaluated the efficacy and tolerability of dex-amethasone sodium phosphate solution 4 mg/mL(DSP, DexaJect SP, Henry Schein Animal Health; Oak-brook Terrace, IL, USA) given orally for treatment offeline HD in an open-label study. Nineteen cats withclinical signs and dermatological lesions compatiblewith feline HD were administered DSP solution at0.2 mg/kg per os once daily for 21 to 28 days. Com-plete blood cell counts, serum chemistry profiles andurinalyses were analysed for 6/19 cats pre- and post-DSP administration. After oral DSP administration,dermatological lesions evaluated through Scoring FelineAllergic Dermatitis (SCORFAD) were significantlyreduced from 7.4 (mean score) to 3.4 (54.3% reduction,Wilcoxon’s ranked sum tests, P < 0.01) and the ownerpruritus Visual Analog Score (pVAS) was reduced froman initial 7.9 to 2.1 (73.9% reduction, P < 0.01). Six of19 cats experienced mild and transient adverse effects(sneezing, lethargy, polyphagia, inappropriate elimina-tion, and polydipsia), and a single cat developed hyper-glycaemia and glucosuria, which normalized four weeksafter drug cessation. There was no reported difficultyadministrating oral DSP solution. In conclusion, oralDSP solution induced a significant and rapid improve-ment of clinical signs and pruritus in feline HD afterthree to four weeks of treatment.Source of funding: American College of Veterinary Der-

matology Resident Grant.


54 Abstracts



POSTERS

SKIN BIOLOGY AND GENETICS

P-1

Effect of topical medication on the

nasomaxillary skin fold microbiome of French

bulldogs

A. REXO*, B. HANSEN*, M. CLARSUND†, V.FADOK‡ and J. BERNSTEIN§

*Dermatology and Allergy Services for Animals,Springfield, VA, USA; †Division of Biotechnology, LundUniversity, Sweden; ‡Zoetis-US Pet Care, Bellaire, TX,USA; §Long Green Animal Dermatology Center,Baldwin, MD, USA

Host–microbe interactions play an important role inthe pathogenesis of dermatitis in the nasomaxillaryfolds of French bulldogs, as it is often complicated bysecondary bacterial and fungal infections. Next-genera-tion sequencing of DNA was used to characterize thecomposition of the microbiome inhabiting nasomaxil-lary skin folds in healthy French bulldogs. Then theeffect of two topical products on the fold microbiomewas assessed. There were seven dogs treated with anenzyme-containing product effective against biofilm for-mation without biocidal activity; six dogs were treatedwith a 2% Chlorhexidine solution with biocidal activityagainst bacteria and fungi, and six dogs were leftuntreated as a control for a total of 19 study partici-pants. Dogs were assigned randomly to each group andthe investigator was blinded. The main skin bacterialphyla inhabiting the skin fold at inclusion were Firmi-cutes, Actinobacteria and Proteobacteria. The main skinfungal phyla were Ascomycota and Basidiomycota.Topical treatment increased the diversity of bacterialand fungal compositions over time (increase in micro-bial diversity score; enzyme 42%, Chlorhexidine 11%,control <5%); these differences were statistically signifi-cant (enzyme; P = 0.028, Chlorhexidine; P = 0.048). Aclear correlation (r2 = 0.8) between abundance of clini-cally relevant pathogens and microbial diversity wasseen. We conclude from the results of this study thatthe nasomaxillary skin fold microbiome of apparentlyhealthy French bulldogs contains a high abundance ofclinically relevant pathogens (mean 36.4%) and thattopical therapy using either Chlorhexidine or enzymeproduct increases microbial diversity in the skin foldsand reduces the relative abundance of pathogens.Source of funding: Self-funded. MiDOG LLC provided

the sample collection devices, the microbiome sequencing

and preliminary microbiome analysis.

Conflict of interest: MC was involved in the development

of the enzyme formulation however no reimbursements,

fees, funding or salary were received.

P-2

The canine skin and ear microbiome: a

comprehensive survey of pathogens implicated

in canine skin and ear infections using a novel

next-generation sequencing-based assay

S. TANG*, A. PREM†, J. TJOKROSURJO†, M.SARY†, M. A. VAN BEL†, A. RODRIGUES-HOFFMANN‡, M. KAVANAGH§, G. WU†, M.VAN EDEN* and J. KRUMBECK**MiDOG LLC, Irvine, CA, USA; †Zymo ResearchCorp., Irvine, CA, USA; ‡Department of VeterinaryPathobiology, College of Veterinary Medicine &Biomedical Sciences, Texas A&M University, CollegeStation, TX, USA; §Saddleback Animal Hospital,Tustin, CA, USA

This study analysed the complex bacterial and fungalmicrobiota of healthy and clinically affected canine earand skin samples. A total of 589 canine samples wereincluded: 257 ear swab samples (128 healthy and 129clinically affected) and 332 skin swab samples (172healthy and 160 clinically affected) were analysed usingnext-generation sequencing (NGS) to determine bothrelative and absolute abundances of bacteria and fungipresent in the samples. This study showed that thecanine microbiota of clinically affected cases was char-acterized by an overall loss of microbial diversity, highmicrobial biomass and overgrowth of certain membersof the microbiota. The combination of relative andabsolute abundances best explained the observed phe-notype of these samples. Compared to healthy samples,78.3% of the clinically affected ear samples had micro-bial overgrowth. Of these, 69.8% had bacterial over-growth, 16.3% fungal overgrowth, and 7.0% bothbacterial and fungal overgrowth. The most importantmicrobial drivers of ear infections were Malasseziapachydermatis, Staphylococcus pseudintermedius,S. schleiferi, and several anaerobic bacteria such asFinegoldia magna, Peptostreptococcus canis and Porphy-romonas cangingivalis. The anaerobic microbes identi-fied previously were not commonly recognized aspotential pathogens in canine ear infections, perhapsdue to the inclusion of these organisms within biofilms.Similar observations were found for skin samples, withyeasts and anaerobes playing a less important role com-pared to clinically affected cases.Source of funding: MiDOG LLC.

Conflict of interest: MK and AR-H are on the advisory

board of MiDOG LLC.

P-3

Anti-inflammatory effect of shea butter

extracts in canine keratinocytes in vitro

D. LIM and T. OHCollege of Veterinary Medicine, Kyungpook NationalUniversity, Daegu, Republic of Korea

Shea butter (Vitellaria paradoxa) is a fat extracted fromshea tree nuts and contains relatively high concentra-tions of nonglycerides. Shea butter extract (SBE) has

Abstracts 55



been used to treat various skin problems in human suchas burns, eczema and rashes, yet little is known aboutits effect on canine skin. This study evaluated the cyto-toxicity and anti-inflammatory effect of SBE in caninekeratinocytes. The cytotoxic effects of lipopolysaccha-ride (LPS, 5–50 ng/mL) and SBE (50–200 lg/mL) wereevaluated using the Cell Counting Kit (CCK 8) assay.Noncytotoxic concentrations of LPS and SBE wereadministered to canine cell cultures. To evaluate theanti-inflammatory effect of SBE, the levels of inter-leukin (IL)-1b, IL-8, IL-12 and tumour necrosis factor(TNF) a were measured using ELISA kits. The concen-tration of each cytokine was quantified in the control,LPS-treated and LPS + SBE-treated groups. Increasedlevels of IL-1b, IL-8 and IL-12 were found in the LPS-treated groups relative to the control groups (P < 0.05).The LPS + SBE-treated groups showed lower levels ofIL-1b, IL-8 and IL-12 than the LPS-treated groups(P < 0.05). These results suggest that SBE may have anapplication as a topical agent for canine inflammatoryskin diseases. However, in vivo study is needed to evalu-ate the safety and efficacy of SBE in dogs.Source of funding: Self-funded.


P-4

Anti-inflammatory and wound-healing effects

of Calendula officinalis in canine keratinocytes

in vitro

J. H. JEON, D. H. LIM, Y. S. JIN, S. G. BAE and T.H. OHDepartment of Veterinary Dermatology, College ofVeterinary Medicine, Kyungpook National University,Daegu, Korea

Calendula officinalis (C. officinalis) is a traditionalmedicinal plant that calms skin inflammation and pro-motes wound healing. In humans, the dermatologicaleffects of C. officinalis have been demonstrated andmany products contain C. officinalis for wound repair.This study aimed to evaluate the anti-inflammatory andwound healing effects of C. officinalis in canine ker-atinocytes. Petals of C. officinalis were extracted with70% ethanol at 85°C using a Soxhlet apparatus. Todetermine the appropriate concentration of extracts, anEscherichia coli 026:B6 lipopolysaccharide (LPS) cell-viability assay was performed using a Cell CountingKit (CCK 8). The final extract and LPS concentrationswere 10 µg/mL and 10 ng/mL, respectively. To assessthe anti-inflammatory effect of C. officinalis, interleukin(IL)-1b, IL-8, IL-12, and tumour necrosis factor (TNF)a were measured using an enzyme-linked immunosor-bent assay. Statistical analysis was performed usingone-way ANOVA (Fisher least significant differencemethod) with SIGMAPLOT for Windows v12.0 (Systat;San Jose, CA, USA). All experiments were run in tripli-cate. The 10 µg/mL extracts significantly reduced theconcentrations of IL-1b, IL-8 and IL-12 compared tothe groups that were treated only with LPS (P < 0.05).The concentration of TNFa was decreased although

not to a significant degree. The wound-healing effectwas evaluated using the scratch assay. The scratchedkeratinocytes were treated with 5, 10 and 100 µg/mLextracts. The induced lesions reduced by 13.57%,74.02% and 0.42%, respectively. These results suggestthat C. officinalis has anti-inflammatory and wound-healing effects in canine keratinocytes and can be usedfor wound repair in dogs.Source of funding: Self-funded.


P-5

Anti-inflammatory effects of the conditioned

media from canine adipose tissue-derived

mesenchymal stem cells on canine

keratinocytes

Y. JIN and T. OHCollege of Veterinary Medicine, Kyungpook NationalUniversity, Daegu, Korea

Mesenchymal stem cells (MSCs) are known to haveanti-inflammatory effects and studies have shown thatMSC paracrine factors are the main contributors tothese effects. These paracrine factors are present inMSC-conditioned media (MSC-CM). In this study, weinvestigated the anti-inflammatory effects of canineMSC-CM on canine keratinocytes. MSC-CM was madewith canine adipose tissue-derived mesenchymal stemcells. The positive marketers of these cells identified bythe flow cytometer are CD44 and CD90, and the nega-tive marketers are CD29, CD34, CD45 and MHC2.Keratinocyte-conditioned media (KCM) was made withcanine keratinocytes (CNT-09, CellnTec; Bern, Switzer-land) as a control. Canine keratinocytes were stimu-lated with lipopolysaccharide (LPS; Escherichia coli026:B6) to evaluate the anti-inflammatory efficacy ofMSC-CM. Three groups of keratinocytes were cultureduntil 100% confluence was reached. The media forGroup 1 was changed to KCM for comparison withthe LPS-added groups. The media of groups 2 and 3were changed to KCM and MSC-CM with LPS added,respectively. After 4 h, each group of keratinocytes waswashed and CM was re-applied. After 20 h, the concen-tration of pro-inflammatory cytokines, such as tumournecrosis factor (TNF) a, interleukin (IL)-1ß, IL-8 andIL-12 were measured using a commercial enzyme-linkedimmunosorbent assay kit. Group 2 showed higher con-centrations of IL-1ß (P < 0.001), IL-8 (P < 0.05) andIL-12 (P < 0.001) compared with Group 1. Theseresults demonstrate that LPS can induce keratinocytesto elevate the concentrations of some pro-inflammatorycytokines. Group 3 showed lower concentrations ofTNFa (P < 0.001), IL-1ß (P < 0.001), IL-8 (P < 0.05)and IL-12 (P < 0.001) compared with Group 2. Theseresults highlight the potential efficacy of MSC-CM inreducing skin inflammation.Source of funding: Self-funded.


56 Abstracts



FOLLICULAR/CORNIFICATION FUNCTION AND PATHOLOGY

P-6

Zinc responsive dermatosis-type II syndrome

in an adult dog

M. CARDOSO*, P. VENTER†, C. HENRICH†, C.GOMES† and G. FREDO‡

*Dermalove Veterinary Dermatology Clinic, Caxias doSul, RS, Brazil; †C~ao Noia Veterinary Hospital, NovoHamburgo, RS, Brazil; ‡Diagnostic Pathology andCytology Laboratory, Caxias do Sul, RS, Brazil

Zinc is an important cofactor and modulator of manybiological functions. It plays an important role in dogdermatoses, yet its deficiency is rare and caused by diet-ary insufficiency or genetic anomalies. The goal of thisreport is to show the potential applicability of a dietaryapproach to treat type-II zinc responsive dermatosis. A2-year-old, female, mongrel dog presented with apathy,symmetrical periocular hyperkeratosis, ulceration,crusts and fissures on the nasal planum, paw pads andvulva. Skin cytological evaluation revealed coccoid bac-teria. Amoxicillin and clavulanate at a dosage of20 mg/kg per os, twice daily, was prescribed. After twoweeks, the dermatological signs worsened and a biopsywas obtained revealing epidermal orthokeratotic hyper-keratosis with areas of parakeratosis, and moderatehyperplasia with focal areas of heavy papillomatousprojections. Cellular debris and clusters of coccoid bac-teria were observed in the stratum corneum. The dogowner reported a rancid smell in the food offered tothe patient. This finding and the biopsy results sug-gested a probable dietary deficiency in zinc. A premiumdiet for adult dogs was prescribed (Golden FormulaSmall Breed Dogs, PremieRpet; S~ao Paulo, Brazil).After six weeks of the new diet, the skin lesionsresolved completely. It is likely that the rancid diet pre-viously eaten by the dog led to important nutrient defi-ciencies resulting in the development of the describedclinical signs. This case indicates that a balanced dietcan be used solely to treat canine type-II zinc respon-sive dermatosis.Source of funding: Self-funded.


P-7

Mutations in noncoding sequences of the EDAgene may be a cause of X-linked hypohidrotic

ectodermal dysplasia in dogs

E. MOURA*, S. DALTRO†, D. S�AS‡, J. ENGRACIAFILHO§, M. FARIAS§ and C. PIMP~AO§

*Service of Medical Genetics, School of Life Sciences,Pontif�ıcia Universidade Cat�olica do Paran�a, Curitiba,PR, Brazil; †Instituto Gonc�alo Moniz, Fundac�~aoOswaldo Cruz, Salvador, BA, Brazil; ‡Genotyping –Diagn�osticos Gen�eticos, Botucatu, SP, Brazil;§Postgraduate Program in Animal Science, School ofLife Sciences, Pontif�ıcia Universidade Cat�olica doParan�a, Curitiba, PR, Brazil

X-linked hypohidrotic ectodermal dysplasia (XLHED)is a rare genodermatosis that affects humans, dogs, cat-tle, mice and probably other mammals. Affected indi-viduals are usually males with characteristic alopeciaand dental abnormalities. In dogs, three mutations inthe EDA gene have shown to cause the dysplasia. Theaim of this study was to verify whether a male poodledog with clinical and histopathological diagnosis ofXLHED had one of these three mutations. All proce-dures were approved by the Institutional Committeefor Ethical Use of Animals in rResearch. The DNA ofthe affected dog was submitted for next-generationsequencing (NGS) and the result was compared to thereference sequence (CanFam3.1 NC_006621.3) and theEDA gene sequence of a normal dog. A panel with apool of multiplex primers was designed covering all theexons and their flanking regions in addition to theentire intronic region from exons 4 to 8, a large part ofthe 5’ untranslated region (UTR) region and the entire3’ UTR. The NGS was performed using the Ion S5System platform. No point mutation or indels werefound in the exons and splice sites of the EDA gene,and a transition (approx. 779–1,188C>T) and atransversion (*1,018T>A) were found in the intron 6region and 3’UTR, respectively. Genetic diseases inhumans have been reported to be caused by mutationsin introns and UTRs. Therefore, the cause of the dys-plasia of the dog in this study is probably a deep intro-nic mutation, a 3’UTR mutation or both.Source of funding: Self-funded.


Abstracts 57



ALLERGIC DISEASES: PATHOBIOLOGY AND DIAGNOSIS

P-8

Results of intradermal skin test in 58 dogs

with atopic dermatitis in Santiago, Chile

B. RIVERA and V. BALAZSOftaderm, Cl�ınica especializada en Oftalmolog�ıa yDermatolog�ıa Veterinaria, Santiago, Chile

The purpose of this study was to identify the mostprevalent allergens in atopic dogs in Santiago, Chile.Fifty-eight dogs (26 females, 32 males) with a clinicaldiagnosis of atopic dermatitis (AD) were tested usingintradermal skin test (IDST). The most prevalent aller-gens were mixture of grasses (64%) and storage mites(60%). In 25 dogs with mostly indoor lifestyle, the mostcommon positive allergen reaction was to storage mites(69.2%). In 33 dogs living outdoors, positive reactionswere most common to mixture of grasses (71.4%).Twenty-two dogs with seasonal signs of AD showed ahigher positive reaction to maple (66.7%). Thirty-sixdogs with permanent allergy had more positive reac-tions to mixture of grasses (67.6%). When comparingdogs living in urban areas with dogs living in ruralareas, results showed that in urban dogs the mostprevalent allergens were mixture of grasses (63.3%),storage mites (61.2%) and Penicillium (59.2%). In ruralareas the most frequent positive reactions were to mix-ture of grasses (66%), Cladosporium (66%) and maple(55.8%). These results show that environment and liv-ing conditions can influence the results of IDST.Source of funding: Self-funded.


P-9

Exploring the role of cytokines and

chemokines in canine allergic skin disease

S. BOWEN, M. ALEO, S. CORUM, X. ZENG, G.RINCON and A. GONZALESZoetis, Kalamazoo, MI, USA

Atopic dermatitis (AD) in dogs is clinically character-ized by chronic pruritis and skin lesions. It has beenwell-established that AD is associated with a T helpercell (TH)1/TH2 cytokine imbalance, resulting inincreased production of TH2 cytokines including inter-leukin (IL)-4 and IL-31, with the latter playing a keyrole in pruritus in certain animal species. The objectiveof this study was to characterize the role of othercytokines and chemokines in canine AD. We employeda cell model utilizing peripheral blood mononuclearcells (PBMCs) collected from beagles sensitized to acommon allergen, the house dust mite (HDM; Der-matophagoides farinae) and polarized toward a TH2phenotype. To assess gene expression within individualcells, TH2 polarized PBMCs were treated with bothHDM and staphylococcal enterotoxin B (SEB) aller-gens for 3 to 72 h followed by 10x single cell sequenc-ing. Eighteen different cell populations were identifiedwith a significant increase observed for the activatedmemory T-cell population after 72 h. Cell type-specificgene expression of several cytokines were evaluated andcompared between allergen-treated and untreated cellsat each timepoint. Several cytokine transcripts were

induced in HDM/SEB-treated cells compared tountreated cells, including IL-4, IL-13, IL-6, IL-33,tumour necrosis factor (TNF)a and IL-17A. In addi-tion, IL-31, GM-CSF, interferon-c, IL-6, IL-8, ker-atinocyte chemotactic (KC)-like, monocytechemoatractant protein-1 (MCP-1), TNFa and IL-10protein levels were induced over 72 h. Taken together,these results highlight that numerous cytokines andchemokines are induced after allergen exposure andlikely play a key role in triggering clinical signs associ-ated with canine AD.Source of funding: Zoetis, Kalamazoo, MI, USA.

Conflict of interest: All authors are employees of Zoetis.

P-10

Allergic sensitization profile from prick test in

dogs with atopic dermatitis in Colombia

R. FERREIRA*, J. DUE ~NAS†, J. GUZMAN† and J.S�ANCHEZ§

*Dermatovet Cursos, Porto Alegre, Brasil;†Dermalatam, Cali, Colombia; §Dermasur, Bogot�a,Colombia

Skin prick test (SPT) is the first choice for the detectionof type I hypersensitivity in human atopic dermatitis(AD). Canine AD resembles several clinical signs of thedisease in the human counterpart. In canine AD, intra-dermal testing is the test of choice, and there are fewreports on the use of SPT in dogs. The purpose of thisstudy was to evaluate SPT reactions in atopic dogs anda healthy control group to six environmental allergens.The antigen panel consisted of common allergens inColombia and more specifically house dust mites andgrass pollens. Six allergen extracts were applied on thelateral thorax of forty atopic dogs (22 females, 18males) and 10 healthy dogs (five females, five males).The skin was pricked with a feather lancet (Duotip-Test, Lincoln Diagnostics, Inc.; Decatur, IL, USA) andevaluated for the positive percutaneous reaction at20 min post-application of the allergens. None of thedogs in the control group showed a positive reaction tothe allergens included in the test. In the group of aller-gic animals a total of 26 dogs (65%) were polysensi-tized and 13 (32.5%) were monosensitized. Overall, themost frequently sensitizations in decreasing order wereDermatophagoides farinae (57.5%), D. pteronyssinus(40%), Blomia tropicalis (37.5%), Cynodon dactylon(22.5%) Paspalum notatum (20%) and Lolium multiflo-rum (17.5%). This was the first work published onprick testing on dogs with AD in Colombia.Source of funding: Self-funded.


58 Abstracts



P-11

Pruritic area and lead time after food

challenge test

K. KAWANO* and H. SHIMAKURA†

*Tokyo Animal Allergy Center, Adachi-Ku, Tokyo,Japan; †Department of Veterinary Microbiology, Schoolof Veterinary Medicine, Azabu University, Sagamihara-shi, Kanagawa, Japan

Elimination diet trial and food challenge testing remainthe gold standards for diagnosis of cutaneous adversefood reactions in dogs. These tests are difficult to per-form due to the risks involved with food challenge test-ing. We reported the characteristics of food challengetesting, aiming to promote it in general veterinary prac-tice. Forty-six dogs with cutaneous adverse food reac-tions were enrolled in this study. Elimination diet trialwas performed for four weeks using food composed ofnovel proteins. Subsequently, the dogs were fed thefood that had previously been eaten during food chal-lenge testing. The pruritus Visual Analog Scale scorewas recorded before and after the elimination diet trial.The time and the site of onset after food challenge test-ing were recorded. Significant symptomatic improve-ment was observed after the elimination diet trial(P < 0.0001). All dogs exhibited pruritus and noneexhibited anaphylaxis after the food challenge testing.Among these dogs, 60.9% developed pruritus within12 h; notably, 23.9% of dogs developed pruritus in 3–6 h. The limbs exhibited the highest frequency of pruri-tus (56.5%) followed by the face (26.1%). The presentstudy is the first to investigate the detailed status afterfood challenge testing in dogs. These results will assistthe veterinarians and the owners in the food challengedecision-making process.Source of funding: Self-funded.


P-12

Microarray analysis of differential gene

expression in atopic beagles undergoing acute

and chronic allergen challenges

R. MARSELLA*, A. RIVA†, K. AHRENS*, R.WILKES*, N. S. MUNGUIA* and M. DORR**Department of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA; †Interdisciplinary Center forBiotechnology Research and Bioinformatics Center,University of Florida, Gainesville, FL, USA

This study aimed to investigate differential gene expres-sion of chronic reactions as prior studies have onlyexamined acute reactions. Five atopic beagles allergicto Dermatophagoides farinae were epicutaneously chal-lenged daily for three days and twice weekly for threemore weeks. Biopsies were taken before [Day (D)0] and24 h after allergen exposure (D1, D2, D8, D15 andD29). Dermatitis was scored those days using theCanine Atopic Dermatitis Extent and Severity Index,3rd iteration (CADESI-03). Total mRNA wasextracted, complementary DNA was made and hybri-dized to a Canine Genome 2.0 Array (GenechipHybridization Kit, Affymetrix; Santa Clara, CA, USA).The chip did not include interleukin (IL)-31 so real-

time PCR was done for IL-31 gene expression. Onedog was removed on D8 due to severity of flare.Another dog was removed from analysis as an outlier.L-amino-acid oxidase-like gene expression increased (5-fold changes on D1 and D8, 4.5-fold on D15 and 2-fold on D28). Gap-junction protein had >3-fold changeincreases on D1, D8 and D28. Calumenin had 2.5-foldincrease on D1 and D8. Keratin-5 had >3-fold increaseon D8. Trichohyaline and keratin-associated-proteinsgene expression had 3.3- to 4-fold change decrease atD15 and D28. Compared to D0, IL-31 gene expressionwas significantly increased on D8 (P = 0.0098), D15(P = 0.0068) and D28 (P = 0.0187). Correlation of IL-31 mRNA and clinical scores was not significant(R = 0.33; P = 0.078). It is concluded that allergenchallenge decreases gene expression of markers of kera-tinization and increases keratins associated with prolif-eration. Gene expression of IL-31 is increased inchronic allergen exposure.Source of funding: Self-funded.


P-13

Filaggrin expression in the skin of normal and

atopic dogs challenged with house dust mites

R. MARSELLA, K. AHRENS, R. WILKES, N. S.MUNGUIA and M. DORRDepartment of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA

Filaggrin and filaggrin-like protein (filaggrin 2) havebeen described in dogs. This study aimed to investigatethe expression of these proteins in normal and atopicdogs, and to investigate the effect of allergen exposurein both populations. Ten atopic beagles and five nor-mal research beagles were challenged daily with allergen(Dermatophagoides farinae) for three days. Biopsieswere taken before allergen exposure and after the thirdexposure [Day (D)3] in both groups. Biopsies also weretaken from atopic dogs on D10, a week after last aller-gen exposure. Immunohistochemical evaluation wasperformed for both filaggrins. Five pictures of each sec-tion were imaged, then scored by four observers (una-ware of group and time) for intensity of staining andepidermal thickness. A mixed-effects model indicatedthat, upon allergen exposure, the intensity of filaggrinstaining was significantly increased in the atopic dogscompared to the normal dogs (P = 0.01). Atopic sam-ples also showed a significant increase in epidermalthickness after allergen exposure (D3 versus D0,P = 0.005) while the normal dogs did not show change.There were no significant differences between normaland atopic dogs in the staining appearance for filag-grin-like protein in the stratum granulosum and cor-neum. In summary, atopic dogs appear to be morereactive to allergen exposure than normal dogs basedon increased epidermal thickness and filaggrin expres-sion. Further studies should address the potential dif-ferences in function between these two filaggrins asthey pertain to the skin barrier function.Source of funding: Self-funded.


Abstracts 59



P-14

Flea or not flea? Portuguese water dog atopic

dermatitis phenotype

T. CASEMIRO, H. L. PEREIRA, T. NUNES, E.DELGADO and A. M. LOURENC�OCentre for Interdisciplinary Research in Animal Health,Faculty of Veterinary Medicine, University of Lisbon,Lisbon, Portugal

Dogs with flea allergy dermatitis (FAD) typically havelesions distributed on the dorsolumbar area. Many Por-tuguese water dogs (PWDs) with atopic dermatitis(AD) appear to have lesions in this area as well. Theobjective of this study was to establish if the dorsolum-bar area could be included in the PWD AD phenotype.Ninety-nine client or breeder-owned PWDs wereenrolled and AD diagnosis was made in 26. Clinicaldiagnosis of AD was made according to accepted crite-ria and after ruling out other causes of pruritus. Serol-ogy for flea saliva-specific IgE antibodies wasperformed (LETIPharma; Barcelona, Spain) and bodyareas showing lesions were recorded for each dog.Results were compared with a generic population ofatopic dogs using the Z-test for proportions. Resultsshowed that PWDs with AD have a special predisposi-tion to lesions on the dorsolumbar area not related toFAD (38.5%; P < 0.05). Additionally, significant differ-ences were found in the percentage of atopic PWDsshowing lesions in certain areas compared with the gen-eric population of affected dogs: front feet (96.2%;P < 0.05), pinnae (92.3%; P < 0.001), eyelids (69.2%;P < 0.001), development of otitis (92.3%; P < 0.001),dry skin (69.2%; P < 0.001) and conjunctivitis (57.7%;P < 0.001). Dry skin may have resulted from a primarydefect. In conclusion, the PWD is predisposed towardsdevelopment of lesions on the dorsolumbar area. Thiscan be misleading as this region is classically associatedwith FAD.Source of funding: Self-funded; Flea-specific IgE serology

testing was funded by LETIPharma, Barcelona, Spain.


P-15

First report of allergic disease distribution

dogs in a dermatology specialty hospital in

Mexico City

A. ROMERO*, G. SHEINBERG*, C. ROMERO† andA. M. CORDERO§

*Centro Veterinario Mexico, Mexico City, Mexico;†Dermavet, Estado de Mexico, Mexico; §Vetderm,Guadalajara, Mexico

Canine dermatological allergic manifestations are com-mon causes of veterinary consultation. Canine atopicdermatitis (cAD) and food allergy dermatitis (FA) areconsidered clinically indistinguishable, requiring foodtrials for diagnosis. Several studies have attempted todescribe the affected dog populations. The presentstudy evaluates the presence of cAD, FA or both dis-eases in a population of dogs treated in Centro Veteri-nario M�exico. Retrospective data of 368 diagnosedallergic dogs were obtained from August 2018 to Octo-ber 2019. Of patients, 58.4% presented cAD, 27.7%FA and only 13.8% were diagnosed with both.

Yorkshire terriers were most likely to present cAD(OR: 3.246, 95% CI: 1.301, 8.104; P = 0.008), Frenchbulldogs FA (OR: 2.622, 95% CI: 1.444, 4.760;P = 0.001) and pugs both CAD with FA (OR: 2.922,95% CI: 1.035, 8.282; P = 0.035). Age was shown to bea risk factor, with more animals diagnosed with CADat 3–6 years of age (OR: 1.813; 95% CI: 1.261, 2.605;P = 0.001), and at six months to 3-year-old for animalswith FA (OR: 3.198; 95% CI: 2.051, 4.988;P = 0.0001). The group with both diseases showed noage predisposition. Animals with FA also exhibited anincreased risk factor for developing otitis (OR: 1.891;95% CI: 1.175, 3.043; P = 0.008), compared to animalswith cAD. The presence of cAD and food allergy indogs in this study is similar to what has been reportedpreviously, with a higher risk factor in French bulldogsand Yorkshire terriers.Source of funding: Self-funded.


P-16

Durability of the atopic dermatitis phenotype

in a laboratory colony of beagles

K. RUDOLPH, T. R. YELLOWHAIR, J. J.DEARING and E. G. BARRETTLovelace Biomedical Research Institute, Albuquerque,NM, USA

Lovelace Biomedical Research Institute (Albuquerque,NM, USA) has been maintaining a colony of allergi-cally sensitized beagles for the past 20+ years. Hereinwe characterized the durability of the phenotype of thisunique colony of dual-sensitized [ragweed (RW) andhouse dust mite (HDM)] beagles and their response tosteroid treatment. The colony was established by breed-ing only allergically sensitized females and then chal-lenging their offspring to a series of subcutaneous,epicutaneous and inhalation challenges with crudeHDM and RW extracts (Greer Laboratories; Lenoir,NC, USA) beginning within the first week after birthand continuing until 16 weeks of age. Subsequently,dogs were challenged by epicutaneous HDM or inhaledRW to induce either a dermatitis or asthmatic-likeresponse while receiving five daily pre-treatments oforal steroid at 1 mg/kg (Prednisolone, Lloyd, Inc.;Shenandoah, IA, USA). The dermatitis phenotype ischaracterized by an increase in the clinical modifiedCanine Atopic Dermatitis Extent and Severity Index(CADESI) scoring of papules, excoriation and ery-thema, and an increase in pruritus, as measured byvideo recording of pruritic behaviour. The dermatitisphenotype is attenuated by prophylactic treatment withoral steroids. By contrast, the asthmatic phenotype isonly partially responsive to oral steroid treatment witha reduction in lung eosinophils and lack of effect onlung neutrophilia and bronchoconstriction. This uniqueallergically sensitized beagle colony exhibits both arobust complex dermatitis and asthmatic phenotypethat shares many features similar to both clinical dis-ease in companion pets and humans with differentialresponsiveness to steroids.Source of Funding: Lovelace Biomedical Research Insti-

tute, Albuquerque, NM, USA.

Conflict of interest: Authors are employees of Lovelace

Biomedical.

60 Abstracts



P-17

Expression of canine b-defensin 103 mRNA in

skin biopsies from atopic and healthy dogs

after exposure to different bacteria

C. SIMOU*, K. KOKKINAKI†, M. CHATZIS†, L.ARCHER‡, V. NTAFIS§, E. VINGOPOULOU¶, V.SIARKOU¶, P. XENOULIS†, M.SARIDOMICHELAKIS† and D. SANTORO‡

*Small Animal Clinic, Athens, Greece; †Clinic ofMedicine, Faculty of Veterinary Science, University ofThessaly, Karditsa, Greece; ‡Department of SmallAnimal Clinical Sciences, College of VeterinaryMedicine, University of Florida, Gainesville, FL, USA;§Faculty of Animal Science and Aquaculture,Agricultural University of Athens, Athens, Greece;¶Laboratory of Microbiology and Infectious Diseases,Faculty of Veterinary Medicine, Aristotle University ofThessaloniki, Thessaloniki, Greece

Defective production, activity, or both, of host defencepeptides are postulated to be a cause of staphylococcalskin infections in dogs with atopic dermatitis (AD).The aim of this study was to compare canine b-defensin103 (cBD103) mRNA expression between dogs withAD and healthy controls, after incubation of skin biop-sies with a meticillin-susceptible or a meticillin-resistantStaphylococcus pseudintermedius clinical isolate (MSSPand MRSP, respectively) and a normal member of theskin microbiota (Kocuria rhizophila; formerly Micrococ-cus luteus). One 8 mm punch biopsy was obtained fromthe noninfected skin of the abdominal area from 10dogs with AD and 10 healthy controls. The sample wassectioned into four equal-sized pieces and each incu-bated at 37°C with either phosphate buffer saline (PBS)or one bacterial (MSSP, MRSP or K. rhizophila) sus-pension (2 x 108 colony forming units/mL) for 24 h.Biopsy samples were then washed in PBS; epidermiswas trimmed off the dermis and stored at –80°C untilmeasurement of cBD103 mRNA with quantitativereverse transcription PCR. cBD103 mRNA was unde-tectable in most samples. There was no differencebetween dogs with AD and healthy controls or betweenPBS, MRSP, MSSP and K. rhizophila regardless ofwhether all samples or only the samples with at leastone positive result were analysed. In conclusion, underthese experimental conditions, the incubation of atopicor healthy canine skin with different bacteria failed toupregulate the expression of cBD103 mRNA.

Source of funding: European Society of Veterinary Der-

matology Practitioner Grant 2012.


P-18

Evaluation of skin prick test to identify

sensitization to house dust mite allergens in

dogs

V. CUNHA*,†, C. SOUZA‡, E. CONTRERAS§, T.NUNES†, R. ROCHA† and F. BERNARDES‡

A/V Clinica Veterinaria, Rio de Janeiro, RJ, Brazil;†FDA Allergenic, Rio de Janeiro, RJ, Brazil;‡Department of Veterinary Clinical Medicine, Universityof Illinois, Urbana, IL, USA; §Department of ClinicalSciences, Colorado State University, Fort Collins, CO,USA

Prick tests have been used in human medicine for dec-ades for the evaluation of skin sensitization towardsenvironmental allergens. The objective of this studywas to evaluate the prick test to identify sensitizationto house dust mites (HDM) in dogs with clinically diag-nosed atopic dermatitis (AD). Ten healthy dogs and 45dogs with AD were enrolled. Histamine and saline solu-tions were used as positive and negative controls,respectively. Three solutions of Dermatophagoides fari-nae (DF), D. pteronyssinus (DP) and Blomia tropicalis(BT) (FDA Allergenic; Rio de Janeiro, RJ, Brazil) weretested. Prick tests were performed in duplicate accord-ing to Duotip-II manufacturer recommendations (Lin-coln Diagnostics Inc.; Decatur, IL, USA). A positiveresponse to an antigen was defined as wheals having amean diameter of ≥3 mm larger than the mean diame-ter of negative control wheals. One of the 10 healthydogs had a positive response on the prick test whereas30 of the 45 dogs with AD had a positive response.The odds of having a positive antigen response on theprick test were greater among dogs with AD (OR 18.0;95% CI, 2.1 to 155.6; P = 0.002). Of the 30 positivedogs with AD, three had positive responses to oneallergen (1 DF, 2 DP). Eighteen to two different mites’allergens (10 DF and DP, six DF and BT, two DP andBT). Nine of the 30 dogs had positive responses to allthree mites’ allergens. Duotip-II prick test is a poten-tially safe tool to evaluate HDM sensitization in dogswith AD.Source of funding: Self-funded.


Abstracts 61



ALLERGIC DISEASES: TREATMENT

P-19

Topical therapy increases the efficacy of

lokivetmab in canine atopic dermatitis and

delays disease flares: a single-blinded,

randomized, controlled study

E. BENSIGNOR* and E. VIDEMONT†

*Veterinary Dermatology Referral Service, Paris,France; †Veterinary Hospital, Annecy, France

Lokivetmab is an effective treatment for canine atopicdermatitis (cAD). The aim of this prospective studywas to evaluate the effect of combining the administra-tion of lokivetmab with a topical therapy to betterimprove cAD skin lesions and extend the duration ofthe efficacy of the injectable treatment. Thirty atopicdogs were included and randomly allocated to one oftwo treatment groups: A, a single injection of lokivet-mab (Cytopoint, Zoetis; Louvain la Neuve, Belgium);or B, a single injection of lokivetmab combined with aweekly topical therapy of a spot-on (ATOP 7, Der-moscent, LDCA; Castres, France) and a shampoo(ATOP 7; Dermoscent, LDCA), specifically designed toimprove atopic skin barrier defect. Clinical parametersevaluated included pruritus (Visual Analog Scale), skinlesions using Canine Atopic Dermatitis Lesion Index(CADLI), cosmetic evaluation and owner and investi-gator global assessment of efficacy (OGATE). Dogswere followed-up at Day (D)10, D17 and D31 and untila clinical relapse. An improvement was noted for alldogs as soon as day 10 without difference between thetwo groups. At D17 and D31, CADLI, cosmetic andOGATE scores were significantly more improved inGroup B than in Group A (P < 0.05, Wilcoxon–Mann–Whitney U-test). A longer lasting remission alsowas noted in Group B compared to Group A. Dogsbeing treated by the combination treatment showed alonger extended time to flare compared to dogs treatedwith lokivetmab alone (P < 0.05). This study suggeststhat the combination of lokivetmab with topical thera-pies is of great value for treating cAD.Source of funding: Laboratoire de Dermo-Cosm�etiqueAnimale (LDCA).

Conflict of interest: Both authors are consultants for

Zoetis Animal Health and LDCA.

P-20

Long-term adherence to topical treatments in

canine atopic dermatitis: a randomized

intervention study

E. BENSIGNOR*, M. LARHER* and J. PATTYN†

*Veterinary Dermatology Referral Service, Paris,France; †Veterinary Clinic, Cesson Sevigne, France

Estimates from the World Health Organization indicatethat patients in developed countries only take about50% of prescribed medicine for chronic diseases suchas hypertension and diabetes. It may be the same forowners of atopic dogs. The aim of this randomizedintervention study was to evaluate the value of a closefollow-up monitoring programme to improve the com-pliance of pet owners for treating their dog. Thirty

atopic dogs were included and randomized in a 1:1ratio to a standard of care or a weekly e-mail contactgroup. Control visits were performed after three andsix months for evaluation of skin lesions (Canine Ato-pic Dermatitis Lesion Index) and pruritus scoring(Visual Analog Scale) and for compliance estimation.Six dogs were lost to follow-up and 24 dogs completedthe study. Greater compliance was observed in theintervention group (62% versus 34% after threemonths, and 54% versus 31% after six months;P < 0.05). CADLI and pruritus decreased more in theintervention group at all time points (P < 0.05). Thisstudy underlines the challenge of long term complianceto treatments in canine atopic dermatitis. Compliance islow in the short term and decreased in the long term,yet can be increased by a close follow-up. This is a con-siderable challenge for both general practitioners andveterinary dermatologists.Source of funding: Self-funded.


P-21

Use of a probiotic blend in 42 dogs diagnosed

with canine atopic dermatitis associated with

recurrent superficial bacterial infection

M. CARDOSO* and F. OLIVEIRA†

*Dermalove Veterinary Dermatological Office, Caxiasdo Sul, RS, Brazil; †Formula Bichos VeterinaryManipulation, Novo Hamburgo, RS, Brazil

Probiotics are live bacteria that, when ingested, mayhave beneficial effects in humans or animals. The maingoal of this study was to test the efficacy of a probioticfor control of recurrent bacterial skin infections, a com-mon condition in dogs with atopic dermatitis (cAD).For this study, a probiotic blend was used that con-tained the following: Lactococcus lactis, Lactobacilluscasei, Lactobacillus salivarius, Lactobacillus acidophilusand Bifidobacterium bifidum (F�ormula Bichos; NovoHamburgo, RS, Brazil). Forty-two patients with cADand recurrent bacterial infection, of both sexes, differ-ent breeds and weights, that used the probiotic blend ata dose of 8.04 x 108 colony forming units/kg for ≥90days in a row were monitored for two years. Adminis-tering the probiotic blend while fasting in the eveningimproves the adhesion of strains to the intestine.Patients were monitored for nine months after startingsupplementation through clinical exams and skin cyto-logical evaluation once monthly. Of the 42 patients, 14(33.3%) did not respond to treatment and a new sys-temic antibiotic cycle was administered and 28 (66.6%)showed a positive response and the infection did notreturn. Twenty of the 42 (47.6%) needed topical treat-ment with associated antibacterial shampoo, six(14.3%) needed topical and systemic antipruritic treat-ment, and two (4.8%) systemic antipruritic treatment.It was observed in two (4.8%) that when probioticblend was suspended, the bacterial infection returnedwithin 14 days. Using these probiotics appears to aid inthe control of recurrent bacterial infection in cases ofcAD as systemic antibiotics are reduced.Source of funding: Self-funded.


62 Abstracts



P-22

Investigation of the efficacy of a topical k-

opioid agonist in a model of canine atopic

dermatitis

R. MARSELLA*, K. AHRENS*, R. WILKES*, A.TRUJILLO*, M. SOEBERDT† and C. ABELS†

*Department of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA; †Sudbrackstrasse, Bielefeld,Germany

The aim of this prospective, four week, placebo-con-trolled, cross-over study was to investigate efficacy of1% topical k-opioid agonist gel in a model of canineatopic dermatitis (AD). Fourteen beagles were chal-lenged by epicutaneous application of house dust mitesevery three to four days for a total of nine challenges.Severity of dermatitis was assessed (Canine Atopic Der-matitis Extent and Severity Index, 3rd iteration) beforethe first challenge (baseline) and 6 h post-allergen chal-lenge. Pruritus was monitored using HERO cameras(GoPro; Santa Barbara, CA, USA) that recorded epi-sodes of pruritus over a 75 min period. Pruritus scoringwas evaluated at 10 time periods [baseline, 4 h post-allergen challenge and the last day of the study on Day(D)28]. Scoring was done blindly by personnel usingBORIS software (DBios; Torino, Italy) for 60 min afterthe dogs had settled for 15 min. A global subjectivescore was given using Visual Analog Scale. A four weekwashout period occurred, dogs were crossed-over andthe study was repeated. Topical k-opioid agonist gel(Wolff GmbH; Bielefeld, Germany) was applied oncedaily on the inguinal area (0.6 mL/dog). The resultswere analysed using combined data. ANOVA showedsignificant effect of time (P < 0.0001) and group(P = 0.0001) on dermatitis scores. Overall, no statisti-cally significant effect on pruritus was found owing toa crossing of scores on D17. Over time, the placeboscores increased while the active ingredient showeddecrease after the first three weeks. It is concluded thatthis approach is promising in dogs with AD and longerstudies with more frequent application may be benefi-cial.Source of funding: Wolff GmbH & Co. KG Arzneimittel.

Conflict of interest: RM has received grant funding from

Wolff for the completion of this project and MS and CA

are employees of Wolff GmbH & Co. KG Arzneimittel.

P-23

Evaluation of safety and efficacy of a new

ciclosporin A in the therapy of naturally

occurring canine atopic dermatitis

R. LUCAS*, S. Z. MACHADO‡, J. LOPES†, M. C.A. TEIXEIRA‡, M. MASCARENHAS†, M. D.LUSTOZA‡, P. BRIANTAIS§, D. BEVIANI*, T. M.ALBUQUERQUE*, C. PELEGRINI* and V. M. C.CUNHA**Dermatoclinica, S~ao Paulo, SP, Brazil; †Promove, Riode Janeiro, RJ, Brazil; ‡Virbac, S~ao Paulo, SP, Brazil;§Virbac, Carros, France

For several years ciclosporin A (CsA) has been recog-nized as a major component of canine atopic dermatitis(cAD) multimodal therapy. A new oral liquid

formulation of CsA (Cyclavance, Virbac; Carros,France) is now available for dogs in many countriesand it facilitates precise dosing and owner compliance.The purpose of this study was to evaluate the safetyand efficacy of this CsA for the control of cAD.Thirty-nine client-owned dogs with nonseasonal cADreceived an oral solution of Cyclavance (5 mg/kg), oncedaily for 60 days. Before treatment and at Day (D)30and D60 after therapy, skin lesions were graded by clin-icians using the Canine Atopic Dermatitis Extent andSeverity Index, 4th iteration (CADESI-04). Prurituswas assessed by the owners using a Visual Analog Scale(pVAS). The percentage of dogs showing reduction ofpruritus from baseline on D30 and D60 was 44.27%and 60.85%, respectively, and the CADESI-04 scoredecreased by 50% and 69.84%, respectively. The clini-cal improvement reported was 87.2% of the cases. Astatistically significant improvement was seen in thedog’s quality of life (P < 0.001). Haematological andserum chemical mean values remained within the nor-mal reference ranges. Mild gastrointestinal signs wereseen in 15% of the dogs. Our study provides evidencethat Cyclavance is safe and reduces the severity of clini-cal signs in dogs with nonseasonal cAD. Thus, Cycla-vance should be considered a valuable alternativetherapy in dogs with cAD.Source of funding:Dermatology Research grant from Vir-

bac.

Conflict of interest: RL received reimbursements, fees

and funding from Virbac for this study.

P-24

Evaluation of a new ciclosporin A compared to

prednisolone for treatment of canine atopic

dermatitis in a randomized controlled trial

R. LUCAS*, M. C. A. TEIXEIRA‡, J. LOPES†, S. Z.MACHADO‡, M. D. LUSTOZA‡, P. BRIANTAIS§,D. BEVIANI*, T. M. ALBUQUERQUE*, C.PELEGRINI* and V. M. C. CUNHA**Dermatoclinica, S~ao Paulo, SP, Brazil;†Dermatoclinica, S~ao Paulo, SP, Brazil; ‡Virbac, S~aoPaulo, SP, Brazil; §Virbac, Carros, France

Ciclosporin A, a calcineurin inhibitor, has been usedfor the long-term treatment of canine atopic dermatitis(cAD). A new oral liquid formulation of ciclosporin A(Cyclavance, Virbac, Carros, France) is now availablefor dogs and it facilitates precise dosing and ownercompliance. The purpose of this study was to comparethe efficacy of Cyclavance with prednisolone in thetreatment of cAD and evaluate the possible adverseevents from each therapy. Ninety-four dogs with non-seasonal cAD were randomly allocated to receive anoral solution of either Cyclavance (5 mg/kg daily) orprednisolone (Prederm, Ourofino Sa�ude Animal; Cravi-nos, SP, Brazil) at 1 mg/kg once daily for 15 days, thenonce every two days for 15 days, and then once everythree days hours for 30 days. Dogs treated with pred-nisolone manifested more adverse events when com-pared to ciclosporin. Pruritus Visual Analog Scale scoreand Canine Atopic Dermatitis Extent and SeverityIndex, 4th iteration decreased by 58.9% and 54.1%,respectively, in the prednisolone group and by 49.2%and 65.7%, respectively, in the Cyclavance group. Theveterinarians who oversaw the trial reported a clinical

Abstracts 63



improvement in 89.1% (prednisolone) and 86.4%(Cyclavance). According to the owners, quality-of-lifeimprovement after 60 days of treatment ranged from36.6% to 93.5% in the dogs treated with prednisolone.By contrast, animals treated with Cyclavance showed a70.5–95.5% improvement by the end of treatment. Thepresent findings demonstrate the effectiveness of Cycla-vance for the treatment of cAD, resulting in improvedQoL and fewer adverse events than prednisolone.Source of funding: Dermatology Research grant from

Virbac.

Conflict of interest: DL received reimbursements, fees

and funding from Virbac.

P-25

Control of pruritus in atopic dogs with

lokivetmab in Rio de Janeiro, Brazil

R. RAMADINHA and J. ALMEIDAAnimalia Clinica Veterinaria, Rio de Janeiro, Brazil

Itching is an uncomfortable sensation that induces self-mutilation in atopic dogs. There are several medicationsto control itching and some have undesirable adverseeffects. Lokivetmab (Cytopoint, Zoetis; Kalamazoo,USA) was chosen because it can control pruritus in ato-pic dogs with minimal to no adverse effects. BetweenJune 2019 and February 2020, 46 dogs diagnosed withcanine atopic dermatitis (cAD), of different breeds andages, were treated at Animalia Clinica Veterinaria. Mostdogs were <7-year-old (63.0%) and shih tzus were morefrequent. All animals with Malassezia spp. or Staphylo-coccus spp. infections were treated previously with 3%chlorhexidine shampoo, antifungal (itraconazole) orantibiotic (cefalexin) as needed. Pruritus was treated withprednisolone 0.5 mg/kg orally for 10–15 days, withregressive doses. Once the infections resolved and pruri-tus was controlled and they were off therapy for five to15days, the dogs were administered lokivetmab subcuta-neously at a dose of 2 mg/kg. One dog presented sterilepustular dermatitis three days post-administration, withspontaneous resolution. The pruritus was controlled in80.4% (37/46) of the animals and 32/37 of these main-tained control of their cAD with repeat dosing of lokivet-mab. Five owners considered the results to be excellent,yet lacked financial ability to maintain control. Threeowners preferred to return to previous therapy withoclacitinib. In conclusion, lokivetmab is considered aneffective therapy for cAD.Source of funding: Self-funded.


P-26

Sublingual immunotherapy (SLIT) is beneficial

in long-term atopic Shiba dogs in Japan: two

case reports

M. IMAMOTO*, S. IMAMOTO*, Y. FUKUI† and K.YAMADA‡

*Shinjo Animal Hospital, Nara, Japan; †KomachiAnimal Hospital, Ibaraki, Japan; ‡Yamada AnimalHospital, Tokyo, Japan

Sublingual immunotherapy (SLIT) is a therapeuticoption in canine atopic dermatitis (cAD). The Shibabreed in Japan is predisposed to cAD, yet the efficacyof SLIT remains unknown in atopic Shiba dogs. Thisstudy describes the treatment of two Shiba dogs inJapan with SLIT. The first case was a 10-year-oldShiba male, who had four years of therapy with pred-nisolone combined with antihistamines, prednisolonecombined with ciclosporin, oclacitinib, and a combina-tion of oclacitinib and ciclosporin without significantimprovement. SLIT (Diamond Animal Health Inc.; DesMoines, IA, USA) containing dust mites based onserum immunoglobulin (Ig)E testing was started. Forthe first two years of SLIT, prednisone and ciclosporinsupport was provided. Six months after the initiation ofthe SLIT treatment, the patient recovered from the skinlesions and the alopecia. The second case was a 7-year-old female showing seasonal allergic pruritus aroundthe eyes and muzzle from summer to autumn. The firstclinical signs were treated with prednisolone and anti-histamines. Because of the marginal effect, the patientwas switched to prednisolone and oclacitinib, and clini-cal signs worsened. The serum IgE test revealed grass(perennial rye and timothy) and weed (sweet vernal)reactivity. SLIT was started while the patient continuedprednisolone and oclacitinib treatment. After a fewmonths, the pruritus decreased, and after three years ofSLIT treatment, it was possible to decrease the dose ofsystemic treatments. No adverse effects were observedin either case from the sublingual treatment. It is con-cluded that the SLIT treatment was beneficial in thesetwo Shiba dogs.Source of funding: Self-funded.


P-27

Hypoallergenicity of an insect-based diet in the

treatment of canine cutaneous adverse food

reaction

K. LEE, T. YUN, Y. KOO, M. YANG and B. KANGLaboratory of Veterinary Internal Medicine, VeterinaryTeaching Hospital, Chungbuk National University,Cheongju, Republic of Korea

The objective of this study was to evaluate the benefi-cial effect of short-term use of an insect-based diet indogs with cutaneous adverse food reaction (CAFR).Nineteen dogs with CAFR were enrolled and classifiedinto three groups. The insect-based diet group (n = 7)and the salmon-based diet group (n = 6) were fed withmealworm- and salmon-based diets, respectively, andthe control group (n = 6) was given commercial diet for12 weeks. In all groups, the skin lesions were evaluatedbased on the Canine Atopic Dermatitis Extent and

64 Abstracts



Severity Index, 4th iteration (CADESI-04). Transepi-dermal water loss (TEWL) was measured to assess skinbarrier function. The pruritus level and alteration ofmedication usage also were evaluated. All indices wereassessed every four weeks after the initial administra-tion of diets. In the pruritus level and medicationscores, there were no significant between- and within-group differences at all time-points. However, in theinsect-based group, a significant decrease in theCADESI-04 score was observed at eight weeks com-pared to that of the baseline score (P = 0.031). TheCADESI-04 score of the insect-based group was

significantly lower when compared with the controlgroup at eight weeks (P = 0.008) and 12 weeks(P = 0.012). The TEWL of control group was signifi-cantly higher than that of insect-based diet group at 12weeks (P = 0.022) and that of salmon-based diet groupat every time-point. These results demonstrate a poten-tial hypoallergenicity of the insect-based diet.Source of funding: Basic Science Research Program

through the National Research Foundation of Korea

(NRF) funded by the Ministry of Education

(2017R1D1A3B03028863).


Abstracts 65



IMMUNE-MEDIATED DISEASES

P-28

Probable pemphigus foliaceus in a 4-month-old

kitten due to a vaccine

N. A. MCEWANUniversity of Liverpool, School of Veterinary Science,Liverpool, Cheshire, UK

The purpose of this report is to detail pemphigus foli-aceus (PF) in a 4-month-old kitten associated with vac-cinations and to emphasise that some cats will respondbetter to dexamethasone than prednisolone. Four daysafter routine vaccination, the patient showed crustingskin lesions on the face, feet and a caseous paronychia.There also was hair loss and devitalisation of the skinof the tail tip. Grossly the skin lesions were consistentwith PF. The lesions on the tail tip were suggestive ofvasculitis. Cytological results and skin biopsies wereconsistent with PF. The patient previously had beentreated with prednisolone per os at 5.3 mg/kg/day withno improvement, so this was changed to dexametha-sone p.o. at 0.5 mg/kg/day. Marked improvement wasseen after 14 days and then the dose was tapered andstopped at the end of three weeks. A six month follow-up did not show any recurrence. Most cases of PF areidiopathic and typically are a life-long condition. In thiscase the condition resolved and did not return. Usingthe Narajo scoring system the score for this patient wasprobable adverse drug reaction. McEwan published areport on a Siamese cat that developed PF associatedwith the administration of cimetidine. In that case thehistopathological findings showed changes consistentwith PF and also vasculitis. In conclusion this wouldappear to be a case of vaccine-induced PF and possiblevasculitis.Source of funding: University of Liverpool, Liverpool,

UK.


P-29

Skin scrotal nodules in three dogs:

immunophenotyping of the infiltrating cells

F. ALBANESE*, F. ABRAMO†, S. L. BENALI*, L.LUCIANI‡, G. CALZAVARA§, A. PIRONE† and V.MIRAGLIOTTA†

*Veterinary Diagnostic Laboratory MyLav-La Vallonea,Passirana di Rho (MI), Italy; †Department ofVeterinary Sciences, University of Pisa, Pisa, Italy;‡Free practitioner, Cattolica (RN), Italy; §AmbulatorioVeterinario San Giorgio, Padova, Italy

Solitary and multiple skin fibro-histiocytic nodules areuncommon findings in young dogs. Lesions are foundon face, scrotum and legs. Histologically they arenonencapsulated confluent nodules composed of solidsheets of plumped and spindle-shaped cells morphologi-cally consistent with histiocytes or fibroblasts. Theimmunophenotype of the cells composing the lesion hasnever been investigated. Because differential diagnosisincludes a range of infectious and noninfectiousimmunologically driven diseases, establishing theimmunophenotype of the infiltrating cells might be ofvalue for therapeutic purposes. We describe three cases

of canine scrotal nodules where the infiltrate was char-acterized with specific histiocytic markers. Skin biopsiesfrom scrotal multiple nodules were obtained forhistopathological evaluation. Periodic acid Schiff (PAS)stain and immunofluorescence were performed by com-bined use of three markers: Iba-1 for histiocytes (ab-cam, ab5076), Macrophage Marker (CD11b, CD68,CD163, CD14, CD16) antibody Panel (abcam,ab254013) and vimentin (SanBio, MONX10662).Merged images were used to localize and co-localize thepositive signals. All cases were negative for PAS stainand showed mixed immunoreactivity to the three anti-bodies. Most of the infiltrating cells were Iba-1 positive;the macrophage antibody panel immunostained many,not all, of the Iba-1 positive infiltrate thus documentingthe presence of a double population. Vimentin waslocalized around the nodules in spindle-shaped cellsand also co-localized with the other infiltrating cells.Immunomodulatory therapy was ineffective in two dogswith lesions spontaneously regressing; one dog withouttherapy had stable lesions. This study documents thatat least a double population of histiocytic cells was pre-sent within the scrotal nodules.Source of funding: Local institutional funding, University

of Pisa.


P-30

Toxic epidermal necrolysis in a dog after a

triamcinolone injection

V. BALAZSOftaderm, Cl�ınica especializada en Oftalmolog�ıa yDermatolog�ıa Veterinaria, Santiago, Chile

Corticosteroids are used for treating dogs with Stevens–Johnson syndrome/ toxic epidermal necrolysis (SJS/TEN). In human medicine, there have been reports inwhich the administration of high-dose corticosteroidshas induced SJS/TEN. In veterinary medicine suchevents have not been reported. We present a case of a4-year-old, 4.9 kg female pug that was injected with1 mg/kg triamcinolone for allergic responses; no otherdrug was administered. The day after the injection thedog presented with depression, lethargy and anorexia.The skin showed macular and patchy erythema thatprogressed to confluent erythema on the trunk, ven-trum, groin, face and inner pinna. On the third day,there were vesicles on the inner pinna and ventrum.Epidermal detachment started on the trunk and beganto generalize showing pseudo-Nikolsky sign. The dogwas sent to an intensive care unit. Complete bloodcount and differential white blood count tests werewithin normal limits. Clinical chemistry panel revealedthe following concentrations: creatinine 2.5 mg/dL(normal 0.4–1.8); alanine aminotransferase (ALT)113 U/L (normal 22.0–35.0); total bilirubin 12 mg/dL(normal 0.15–1.45); albumin 1.6 mg/dL (normal 2.8–4.0). Blood gas test indicated 4.5 mmol/L (normal <2.5)lactate. Systemic arterial pressure was 60 mmHg (nor-mal 120–130 mmHg). Five days after the triamcinoloneinjection the dog died with disseminated intravascularcoagulation due to systemic failure. A skin biopsy was

66 Abstracts



not performed. The diagnosis was toxic epidermalnecrolysis triggered by an overdose of triamcinolone.Veterinarians should be aware of the possibility of life-threatening adverse reactions by overdoses of systemiclong-lasting corticosteroids.Source of funding: Self-funded.


P-31

Canine perianal fistulas in a German shepherd

cross dog

A. L. BOOPet First Veterinary Centre, Kuala Lumpur, Malaysia

This is a case report of canine perianal fistulas in a 3-year-8-month-old male German shepherd cross dog. Hewas presented for second opinion of nonhealing peri-anal wound. The dog was first treated by the primaryveterinarian with metronidazole and papase for a week,with no improvement seen in the perianal lesions. Thereported clinical signs included perianal wound, fre-quent scooting, perianal licking, perianal pain,increased frequency of defaecation and aggressive beha-viour when lifting up the tail. Physical examinationsrevealed swollen perianal area with fistulas and mucop-urulent discharge at the fistulas. Impression smear ofthe perianal lesions showed the presence of intracellularcocci and degenerated neutrophils indicating infection.A tentative diagnosis of canine perianal fistulas wasmade based on the history, clinical signs and physicalexamination findings. Immunomodulatory therapy wasinitiated with ciclosporin, 4.5 mg/kg per os once dailyand cefalexin, 22.7 mg/kg p.o. twice daily, to treat thebacterial infection. The perianal lesions showed markedimprovement after initiation of treatment and contin-ued to improve when the ciclosporin regimen was mod-ified and combined with ketoconazole, 5 mg/kg/dayp.o. during the subsequent follow-up. One month afterinitiation of ciclosporin therapy 0.1% tacrolimus oint-ment was introduced as a topical immunomodulatoryagent. The patient showed continuous improvementand was lost to follow-up two months after the initia-tion of treatment.Source of funding: Self-funded.


P-32

Cutaneous neutrophilic panniculitis and

vasculitis associated with shar-pei

autoinflammatory disease – a case report

M. CABR�E*, L. ORDEIX*,†, R. GARC�IA-FONTICOBA* and L. FERRER*,†

*Fundaci�o Hospital Cl�ınic Veterinari, UniversitatAut�onoma de Barcelona, Bellaterra, Spain;†Departament de Medicina i Cirurgia Animals,Universitat Aut�onoma de Barcelona, Bellaterra, Spain

Skin lesions described in shar-pei autoinflammatory dis-ease (SPAID) are clinically unspecific and histopatho-logically poorly characterized. The objective of this casereport was to describe a shar-pei dog with SPAID andcutaneous neutrophilic panniculitis and vasculitis. An

8-year-old spayed female shar-pei was presented withtwo symmetrical firm plaques in the caudal part ofboth thighs, a smaller plaque and two circular ulcerswith well-defined borders and reticulated hyperpigmen-tation in the right flank. The patient was under treat-ment with 0.5 mg/kg of prednisone every 48 h owing torecurrent bouts of fever. General physical examinationwas normal. Complete blood cell count, serum bio-chemistry, urinalysis and serological testing for Leish-mania, Anaplasma, Ehrlichia and filaria were normal ornegative. A deep skin biopsy for histopathologicalexamination and bacterial culture was performed. Bac-terial culture was negative and the dermatopathologicalexamination revealed a neutrophilic panniculitis andvasculitis. This patient harboured one mutant allele ofthe mis-sense mutation of the MTBP gene. Completeclinical response was observed after three weeks admin-istration of 1 mg/kg prednisone once daily, with subse-quent reduction of the dose during the followingmonth. Relapses were not seen during the following sixmonths. To the best of the authors’ knowledge, this isthe first case of SPAID reported with cutaneous neu-trophilic panniculitis and vasculitis.Source of funding: Self-funded.


P-33

A case of sterile eosinophilic pustulosis and

subcorneal pustular dermatosis in a canine

patient

M. CARDOSO*, G. FREDO† and G.CASAGRANDE‡

*Dermalove Veterinary Dermatological Office, Caxiasdo Sul, RS, Brazil; †Diagnose Pathology and CitologyLaboratory, Caxias do Sul, RS, Brazil; ‡ConfortoAnimal Veterinary Clinic, Caxias do Sul, RS, Brazil

The aim of this report was to demonstrate the effi-ciency of ciclosporin in treating sterile eosinophilic pus-tulosis, which is a rare idiopathic dermatosis in dogs. A3-year-old, female neutered Chihuahua arrived at theveterinary dermatological service for severe itching clas-sified at level 5 intensity of Hill’s Scale. The examina-tion revealed erythematous follicular and nonfollicularpapules and pustules, annular erosions and epidermalcollarettes in the ventral, dorsal and face regions.Antibiotic therapy was cefalexin 30 mg/kg twice dailyper os and prednisolone 1 mg/kg once daily for 14days, with improvement and then worsening whentreatment was suspended. Allergic exclusion tests wereperformed using topical ectoparasiticide and dietarymanagement with a hypoallergenic ration, and thepatient did not tolerate suspending the corticosteroidbeyond 48 h. Blood tests were performed with nochanges; skin biopsy revealed multifocal epidermalhyperplasia, formation of subcorneal cracks with pre-dominance of eosinophils. In the superficial dermisinfiltrates included a moderate presence of eosinophils,and occasional mastocytes, neutrophils and histiocytes.The initial treatment was cetirizine 2 mg/kg twice daily,without response. Treatment was changed to pred-nisolone 1 mg/kg once daily for two weeks. After that,prednisolone 1 mg/kg every 48 h for one week, in com-bination with ciclosporin at 7 mg/kg once daily. After60 days of treatment, the lesions did not return, and

Abstracts 67



the pruritus was classified as intensity level 2. Ciclos-porin appeared to assist in the control of sterile eosino-philic pustulosis, producing a corticosteroid-sparingeffect and likely increasing the patient’s life expectancy.Source of funding: Self-funded.


P-34

Erythema multiforme as a paraneoplastic

syndrome in a dog with chronic lymphocytic

leukaemia: case report

V. GMYTERCO, B. PACHECO, L.KLUTHCOVSKY, F. NOGUEIRA and M. FARIASPostgraduate Program in Animal Science, School of LifeSciences, Pontif�ıcia Universidade Cat�olica do Paran�a,Curitiba, PR, Brazil

Erythema multiforme is an uncommon dermatopathyin which the keratinocyte is the target of an immuneresponse and induction of apoptosis. Herpes virusescan trigger the disease in humans, whereas in dogs ithas been associated with drugs, infections, idiopathicrashes and rarely with lymphoreticular neoplasia. Thisstudy aims to report a case of erythema multiformetriggered by chronic lymphocytic leukaemia. One 6-year-old, female shih-tzu was evaluated owing tochronic papular-crusted and intensely pruritic lesionswith multifocal distribution not responding to previoustreatment with antibiotics and topical antiseptic ther-apy. There were arciform macules, papular and erythe-matous lesions covered by crusts on the abdomen,trunk, limbs, interdigital and periorbital areas.Histopathological examination revealed keratinocyteapoptosis in all layers of the epidermis that formednecrosis. Complete blood count (CBC) revealed mildanaemia and lymphocytosis (12,496 lymphocytes/lL).Serum biochemical results showed hyperproteinaemiadue to hyperglobulinaemia. Myelogram showed ery-throid hypoplasia and an increased number of maturelymphocytes compatible with chronic lymphocytic leu-kaemia (CLL). Quantitative real-time PCR of bloodsamples indicated the prevalence of clonal populationof T lymphocytes. Based on clinical and histopathologi-cal signs, the diagnosis established was paraneoplasticerythema multiforme. Treatment included chlorambucil(0.2 mg/kg per os, once daily for seven days and thencontinuously at 0.1 mg/kg every 48 h) combined withprednisolone (1 mg/kg p.o. once daily). After sixmonths of treatment, CBC normalized and skin lesionsimproved, although recurrences were common. Thiscase report suggests that erythema multiforme, whenchronic and difficult to control, may have a paraneo-plastic origin.Source of funding: supported by the coordination for the

improvement of higher education personnel – Brazil

(CAPES).


P-35

Extra-auricular lesions of proliferative and

necrotizing externa otitis in two kittens

P. PANZUTI, O. JONGH, M. MOSCA, S. VIAL andD. PINVetAgro Sup, Marcy l’ �Etoile, France

Proliferative and necrotizing otitis externa (PNOE) is arare disease affecting the external ear canals and con-cave pinnae of cats. We report two cases of PNOE withextra-auricular lesions. Case 1, an 8-month-old domes-tic short hair (DSH) kitten was presented with bilateralchronic otitis, suggestive of PNOE, and had unusualhyperkeratotic skin lesions along the eyelids. Case 2, a6-month-old DSH kitten, was presented with a fourmonth history of bilateral otitis, suggestive of PNOE,associated with numerous arciform to nummular hyper-keratotic skin lesions overlying erosions disseminatedover the entire body. In both cases, histopathologicalexamination of skin biopsies from ears and extra-auri-cular lesions showed a marked orthokeratotic andparakeratotic hyperkeratosis and papillomatous hyper-plasia of the epidermis and of the outer root sheath ofhair follicles. Numerous individually shrunken andhypereosinophilic keratinocytes with pyknosis wereobserved at all levels of the epidermis and outer rootsheath of hair follicles. A marked lymphocytic exocyto-sis was present in all levels of the epidermis along withsatellitosis. Case 1 resolved spontaneously after twomonths. In Case 2, topical tacrolimus (Protopic 0.1%ointment, LEO Pharma A/S; Ballerup, Danemark) wasapplied into ears. Topical application to the dissemi-nated lesions was impractical so oral ciclosporin (Cycla-vance, Virbac; Carros, France; 7 mg/kg/day) wasadded. Clinical remission was observed after twomonths for ear lesions and other skin lesions are stillongoing at the time of writing. To the best of theauthors’ knowledge, this is the first report of extra-auri-cular lesions associated with PNOE in cats.Source of funding: Self-funded.


P-36

Efficacy of dietetic food on generalized

idiopathic ischaemic dermatopathy in three

miniature dachshunds

K. SHIBATA* and Y. KAGAWA†

*DVMs Animal Medical Center Yokohama, Kanagawa,Japan; †North Lab, Hokkaido, Japan

Generalized idiopathic ischaemic dermatopathy (GIID)is a type of ischaemic dermatopathy that is adult-onsetand has no correlation to vaccines. Treatment ofischaemic dermatopathy includes immunosuppressive orimmunomodulatory agents and vitamin E. We hypothe-sized that Derm Defense (DD, Hill’s-Colgate (Japan)Ltd; Nibancho Chiyoda-Ku, Japan) dog food enrichedwith vitamin E may have potential as treatment for thiscondition, similar to other types of vitamin E adminis-tration. The onset with GIID on average is in 7- to 15-year-old dogs. The dogs in this study had oedematousalopecia with scale or crust on the pinnae. Other skinlesions included necrosis of the pinna, alopecia on theface, on the tail tip and over bony prominence areas.

68 Abstracts



They also had weight loss and mild myoatrophy.Histopathological findings of two dogs biopsied showedfollicular atrophy with mild fibrosis around the hairfollicles. Two cases were fed DD without any drugs. Inone case, oedema on the pinnae decreased over timeand no other treatment was needed. In another case,although crusts on the pinnae disappeared after twomonths of consuming DD, the dog needed concurrenttreatment to control scratching behaviour and oedemaon the pinnae. The last case was treated with DD and

oclacitinib. Oedema and crusts on the pinnae improvedwithin two months of treatment, at which time oclac-itinib was stopped. Four months later, the skin lesionson the pinnae were resolved. The dogs regained lostweight. These findings support DD as having efficacyas an adjunctive treatment in GIID cases.Source: Derm Defense was gifted by Hill’s-Colgate

(Japan).

Conflict of interest: KS has lectured for Hill’s-Colgate

(Japan).

Abstracts 69



INFECTIOUS SKIN DISEASES

P-37

Canine cutaneous pigmented viral plaques: a

clinicopathological and molecular study

M. MAZZEI*, M. ORLANDI†, M. FORZAN*, M.MEI*, F. ALBANESE†, F. LEONE†, M.VASCELLARI‡ and F. ABRAMO**Department of Veterinary Sciences, University of Pisa,Pisa, Italy; †Veterinary Diagnostic Laboratory LaVallonea, Passirana di Rho (MI), Italy;‡Histopathology Department, Istituto ZooprofilatticoSperimentale delle Venezie, Legnaro (PD), Italy

Currently, 23 types of canine papillomavirus (CPV)belonging to Chi, Lambda and Tau genera have beenidentified as responsible for oral papillomatosis, exo-phytic cutaneous papilloma, inverted papilloma andpigmented viral plaques. Twelve types of PV, allbelonging to the Chi genus, are reported as aetiologicalagents of pigmented viral plaques. Our objectives wereto: (i) investigate the clinical and histopathologicalcharacteristics of canine pigmented viral plaques; (ii)evaluate the location of viral antigen in epithelial cellsby immunohistochemical (IHC) analysis; and (iii) iden-tify the CPV type using PCR. Cases were retrieved ret-rospectively from histopathology laboratories andclinical history and signalment data were collected.IHC analysis was performed using the human PVmouse monoclonal antibody (ab-2417, AbCam; Cam-bridge, UK). Extracted DNA was amplified by PCRwith generic or specific sets of primers. Sixteen caseswere collected. Lesions were circular with irregularmargins, flat or raised, hyperpigmented, single or multi-focal and often grouped. Histologically all samplesshowed a scalloped surface configuration with hyper-plasia, hyperkeratosis, hyperpigmentation and presenceof few koilocytes. IHC identified the viral capsid pro-tein L1 in 14/16 cases. PCR with generic and CPV-spe-cific primers, and sequencing allowed identifying theviral genotype CPV-4, CPV-8 and CPV-9 in 2/14, 2/14and 10/14 cases, respectively. In the two PCR-negativesamples, IHC scored positive. The combined diagnosticmethods used in this study confirmed the role of differ-ent PVs in all selected cases. Differently from other PVtypes, CPV8 was found in young female dogs on theface and flank.Source of funding: University of Pisa.


P-38

Successful resolution of a case of canine oral

papillomatosis with cryotherapy and a 70%

solution of trichloroacetic acid

V. BALAZS and B. RIVERAOftaderm, Cl�ınica especializada en Oftalmolog�ıa yDermatolog�ıa Veterinaria, Santiago, Chile

Canine papillomaviruses can affect the skin andmucous membranes of dogs. Most lesions regress spon-taneously; however, severe cases may be refractory totreatment. There is scarce published information on theuse of cryotherapy or trichloroacetic acid for treatingoral papillomatosis in dogs. A 2-year-old, 9.4 kg,

female neutered beagle presented with oral papillomasthat began at six months of age after recovering fromdistemper. The lesions were multifocal verrucous nod-ules localized to the lips, gingiva and buccal mucosa.Complete blood cell count and differential white bloodcell count were within normal limits. The chemistryprofile only revealed a two-fold elevation of alkalinephosphatase and a slight elevation of alanine amino-transferase. Other values were within normal limits.Distemper PCR was negative. A biopsy was not per-formed. There was no response to therapy with azithro-mycin nor autologous papillomavirus vaccine. The dogwas anaesthetized and the lesions were treated usingcryotherapy with six freeze–thaw cycles per lesion(freeze times 6 s). One month later, the size and num-ber of the papillomas had reduced significantly. A smallamount of 70% trichloroacetic acid was applied onceto each lesion and allowed to dry. One month later thelesions had resolved completely. Treatment success inthis case suggests that cryotherapy complemented witha solution of trichloroacetic acid may be an effectivetherapy for persistent oral canine papillomatosis. In thiscase, owing to the long-lasting period of this disease, itis unlikely that the oral papillomas could have resolvedwithout treatment.Source of funding: Self-funded.


P-39

Ineffectiveness of marbofloxacin to treat

canine leproid granuloma: a case report

R. SILVA*, B. MANIER*, T. COSTA*, C.BOTELHO* and J. FERNANDES†

*Postgraduation program, Universidade Federal Rural doRio de Janeiro, Rio de Janeiro, Brazil; †Department ofVeterinary Science, Universidade Federal Rural do Riode Janeiro, Rio de Janeiro, Brazil

Canine leproid granuloma is caused by alcohol-acidresistant bacillary bacteria, which does not stain withroutine stains such as Diff-Quick. It mainly affects ani-mals with short coats and body regions such as thepinna with superficial vascularization and low hair den-sity. The diagnosis should be made via histopathologi-cal evaluation using Ziehl–Neelsen stain.Immunohistochemical analysis and PCR can be used inorder to identify the bacterial species involved. Thetreatment involves the use of antibiotics and most com-monly fluoroquinolones, doxycycline and clar-ithromycin, as monotherapy or combination. A dogwith a nodular, alopecic, nonpruritic lesion located tothe convex aspect of the pinna was presented at theVeterinary Hospital of the Universidade Federal Ruraldo Rio de Janeiro. Cytological examination showed apyogranulomatous inflammation and the presence ofunstained filamentous structures (negative image) sug-gestive of Mycobacterium organisms. Histopathologicalexamination identified the presence of bacillary struc-tures within macrophages, confirming the diagnosis ofcanine leproid granuloma. Oral marbofloxacin was ini-tiated at a dose of 2.75 mg/kg every 24 h for 30 days.After 35 days, the dog owner reported the appearance

70 Abstracts



of new lesions; therefore, it was decided to change themedication to enrofloxacin combined with doxycycline,which led to complete resolution of the lesions in 30days. Despite canine leproid granuloma being consid-ered a self-limiting disease, the lesions improved onlyafter changing the treatment. Oral administration ofmarbofloxacin at 2.75 mg/kg every 24 h was not effec-tive for the treatment of canine leproid granuloma inthis case.Source of funding: Partially funded by the Coordenac�~aode Aperfeic�oamento de Pessoal de N�ıvel Superior-Brasil(CAPES).


P-40

Isolation of strains of Sporothrix schenkiicomplex subjected to freezing

F. CARVALHO*,†, P. VITOR‡, M. MACHADO§ andJ. FERNANDES§

*College of Veterinary Medicine, Centro Universit�ariode Valenc�a, RJ, Brazil; †College of Veterinary Medicine,Universidade Iguac�u, RJ, Brazil; ‡Private VeterinaryClinic, Veterin�aria Montese, Resende, RJ, Brazil;§Department of Veterinary Science, Universidade FederalRural do Rio de Janeiro, RJ, Brazil

Sporotrichosis is a deep mycotic infection caused byfungi belonging to the Sporothrix schenckii complex.The zoonotic nature of sporotrichosis results in greatpublic health concerns. Domestic cats are an importantsource of fungal dissemination through their nails andoral cavity, and can infect humans and other animalsmainly by the traumatic implant of the agent throughthe skin. From the moment that an animal dies, properhandling of the carcass represents a critical point toavoid the dissemination of the agent. This work aimedto investigate the presence of viable and potentiallyinfectious forms of the fungus in carcasses of cats diag-nosed with sporotrichosis kept under –18°C tempera-tures. Skin and nail samples of 20 frozen carcasses werecollected and plated onto Mycosel (HiMedia; Mumbai,India) agar. Half of the samples were cultured in anoven at 37°C and half at room temperature (25°C). Allsamples were incubated for seven days and then anal-ysed in the laboratory. Fungi belonging to theS. schenckii complex were isolated from five of 20 sam-ples. The mycelial form of the fungus was identified infour carcass samples and the yeast form in one. Thestudy findings demonstrate that Sporothrix organismscan still be viable in carcasses even under freezing con-ditions. Therefore, carcasses should be considered animportant source of zoonotic infection and environmen-tal contamination.Source of funding: Self-funded.


P-41

Efficacy of mupirocin or chlorhexidine in the

topical treatment of canine superficial

pyoderma caused by resistant and

multiresistant bacteria

R. GAERTNER, V. GMYTERCO and M. FARIASPostgraduate Program in Animal Science, School of LifeSciences, Pontifical Catholic University of Paran�a(PUCPR), Paran�a, Brazil

Superficial pyoderma (SP) is a frequent cutaneousinfection of dogs, usually caused by Staphylococcuspseudintermedius. Its recurrence is commonly associatedwith allergic, seborrhoeic or endocrine skin diseases,and the regular use of systemic antibiotics for its con-trol has been associated with meticillin-resistant (MR)and multidrug-resistant (MDR) infections. This studyevaluated the efficacy of topical 0.2% mupirocin and4% chlorhexidine mousse in the treatment of SP indogs with MR and MDR strains of Staphylococcusintermedius group (SIG). We selected eight dogs withrecurrent multifocal SP characterized by impetigo andfolliculitis, secondary to atopic dermatitis. Culture andantibiogram were performed in samples collected fromprimary lesions. Meticillin-resistant SIG was isolated infive dogs and MDR SIG in three. Five dogs (two MRSIG and three MDR SIG) were treated topically with0.2% mupirocin spray and three (MR SIG) were trea-ted with 4% chlorhexidine mousse, twice daily for 21days. Lesion severity was assessed by a numerical lesionscore ranging from zero to six and the degree of pruri-tus by a validated pruritus Visual Analog Scale(pVAS), weekly for 21 days. Response to treatment wasevaluated by ANOVA and P < 0.05 considered signifi-cant. Reduction of lesion scores and degree of prurituswas significant (P < 0.05) in both treatments, suggest-ing that both treatments were effective in controllingrecurrent MR SP. Moreover, treatment with 0.2%mupirocin was effective in controlling MDR strains.Source of funding: Drogavet Veterinary Handling Phar-

macy provided the aqueous spray of 0.2% mupirocin and

the 4% chlorhexidine mousse.


P-42

Cutaneous protothecosis caused by Protothecawicherhamii infection in a dog

V. GMYTERCO*, L. LUDWIG*, T. JAGIELSKI†,M. RIBEIRO‡ and M. FARIAS**Postgraduate Program in Animal Science, School ofLife Sciences, Pontifical Catholic University of Paran�a(PUCPR), Paran�a, Brazil; †Department of AppliedMicrobiology, Institute of Microbiology, Faculty ofBiology, University of Warsaw, Poland; ‡Department ofVeterinary Hygiene and Public Health, School ofVeterinary Medicine and Animal Sciences, UniversidadeEstadual Paulista, Botucatu, SP, Brazil

Protothecosis is an infectious disease caused by unicel-lular, achlorophyllous microalgae ubiquitous in nature.Opportunistic infections occur in humans and animals.In dogs, Prototheca zopffi is the most common patho-genic species, causing enteric, ocular and/or neurologi-cal signs. Cutaneous lesions are less frequent, often

Abstracts 71



misdiagnosed or unnoticed, and caused by P. wicker-hamii. We report an uncommon case of protothecosisin a 2-year-old, mixed-breed dog with a history of cuta-neous lesions of four months duration and contact withwastewater. Physical examination showed exudativenasolabial plaques and ulcerated, erythematous andpainful lesions on all foot pads associated with enlarge-ment of regional lymph nodes. Haematological and bio-chemical examinations were unremarkable. Histologicalexamination revealed a severe cutaneous inflammatoryreaction, with numerous periodic acid Schiff-stainedround or oval encapsulated structures, compatible withPrototheca species. Microbiological culture of cutaneousexudate on Sabouraud agar kept in aerobic conditions,at 37°C, revealed white colonies after 48 h. Phenotypic-based identification using a newly described PCR-restriction enzyme analysis (PCR-REA) based on thecytB gene, confirmed the organism identity as Pro-totecha wicherhamii. Oral itraconazole was initiated at10 mg/kg once daily and partial improvement wasobserved after eight weeks of treatment, with total reso-lution after six months. However, recurrence occurredafter two months of therapy discontinuation and thetreatment was changed to oral terbinafine at 20 mg/kgtwice daily, unsuccessfully. Itraconazole was re-startedwith partial improvement of lesions at this time. Wehighlight the difficulty in treating P. wicherhamii infec-tion and the public health concern given that cutaneousand systemic P. wicherhamii infection has been reportedin humans.Source of funding: Self-funded.


P-43

Antifungal susceptibility of ravuconazole

against dermatophyte isolates from animals

R. KANO, H. FURUTA and H. KAMATACollege of Bioresource Sciences, Nihon University,Fujisawa, Japan

Dermatophytosis is a common cutaneous mycosis insmall animals and frequently causes human infection.Ravuconazole (RVCZ) is a newly available (approvedfor human use in Japan in 2018) broad-spectrum anti-fungal azole that exhibits excellent activity against der-matophytes. To the best of the authors’ knowledge, thisis the first study to investigate the in vitro susceptibilityof RVCZ against dermatophyte isolates from animalsand a terbinafine (TERB)-resistant strain of Microspo-rum canis. The Clinical and Laboratory StandardsInstitute (CLSI) M38-A2 test was used in the study.The minimal inhibitory concentrations (MICs) ofRVCZ for 17 clinical isolates of M. canis ranged from0.03125 to 0.5 mg/L; for three clinical isolates ofM. gypseum from <0.03125 to 0.03125 mg/L; for oneclinical isolate of Trichophyton benhamiae <0.03125 mg/L; and for nine clinical isolates of T. mentagrophytes<0.03125 mg/L. The MICs for the TERB-resistantM. canis strain (47C) were 0.03125 mg/L for RVCZ,0.025 mg/L for itraconazole (ITCZ) and >32 mg/L forTERB. The results of the present study indicate thatRVCZ might be more effective than ITCZ and TERBagainst clinical isolates of dermatophytes.Source of funding: Self-funded.


P-44

In vitro evaluation of the bactericidal activity

and residual effect of biocidal sprays on canine

hair against meticillin-resistant Staphylococcuspseudintermedius

A. ALVA*, D. MEZA*, C. CHUQUIZUTA*, E.ESCOBEDO† and G. MONTOYA†

*Department of Microbiology, Universidad Cient�ıfica delSur, School of Veterinary Medicine, Lima, Peru;†Department of Dermatology, Universidad Cient�ıfica delSur, School of Veterinary Medicine, Lima, Peru

There is an urgent need to investigate alternative andeffective topical treatments against skin infectious dis-eases. The study aim was to assess the efficacy andresidual effect of the following biocidal sprays:chlorhexidine 2% (Durvet, Durvet), sodium hypochlo-rite 0.01% (Clorox, Clorox Per�u SA), hypochlorousacid 0.65% (Ecaderm, Eca Control SAC) and N-acetyl-cysteine 0.04% (Fluimucil, Zambon SpA). Sodiumchloride 0.9% was used as negative control. Each spraywas applied once on the flank of 30 healthy dogs cover-ing an area of 25 cm2 and 2 cm distant from eachother. Dog hairs (0.02 g) were collected daily for thefirst four days post-treatment and placed equidistantfrom each other onto a Mueller–Hinton agar platestreaked with meticillin-resistant Staphylococcus pseud-intermedius (MRSP) isolated from a dog with pyo-derma. The size of the inhibition halo was measuredevery 24 h. Chlorhexidine 2% had a residual effect ofthree days, forming an inhibition halo of 2.4 � 0.5 cmon day (D)1, 1.7 � 0.5 cm on D2 and 1.0 � 0.6 cm onD3. The other sprays showed an inhibition halo; how-ever, focal areas of bacterial growth surrounding thehairs were observed after D1. On D1, the halo for thesodium hypochlorite spray was 1.6 � 0.5 cm, forhypochlorous acid 1.6 � 0.5 cm and for N-acetylcys-teine 1.5 � 0.4 cm. In conclusion, chlorhexidine 2%spray was more effective in inhibiting MRSP growthin vitro than the other products and had a longer resid-ual effect.Source of funding: Self-funded.


P-45

Association of prior antibiotic exposure to

isolation of meticillin-resistant Staphylococcuspseudintermedius from dogs with skin and ear

infections

K. K. MWACALIMBA, V. FADOK, S. CAMPBELLand D. AMODIEZoetis LLC, Parsippany, NJ, USA

This study examined the association between antibioticexposure and clinical isolation of meticillin-resistantStaphylococcus pseudintermedius (MRSP) from dogsusing the electronic database of a hybrid practicelocated in the United States, from August 2011 toMarch 2018. Antibiotics included fluoroquinolones,cefalexin, cefovecin, cefpodoxime, clindamycin or‘other’, used within 180 days before culture. Compar-isons were based on proportion analysis (SAS 9.4;Cary, NC, USA) using a two-sided Student’s t-test(P ≤ 0.05). Five hundred and seventy-eight

72 Abstracts



S. pseudintermedius clinical isolates (20, abscess; two,cyst; 69, ear; 487, skin) from 503 dogs were reviewed;50% of the isolates (289) were MRSP. Clinical isolatesfrom skin had the highest rate of MRSP (51.5%, 251/487). Dogs received between none to six antibioticcourses in the 180 days before culture. Of the 578 iso-lates, 33% were from dogs with no history of antibioticexposure, while 35% had received antibiotics once.Fifty dogs had more than one culture: 21 were previ-ously MRSP but not MRSP from next isolate; 14 werepreviously non-MRSP and MRSP on a subsequent cul-ture. Dogs with at least one exposure to antibiotics had44–58% MRSP isolates. Forty-seven percent (140/300)of isolates from dogs that received one or two coursesof antibiotics were MRSP (P = 0.1025) and 60% (50/83) of isolates from dogs that received three or fourcourses of antibiotics were MRSP (P = 0.0083). Theseresults support prior literature that previous antimicro-bial therapy is an important risk factor for the isolationof MRSP.Source of funding: Zoetis.

Conflict of interest: KM, VF, SC and DA are employees

of Zoetis, Inc.

P-46

Effect of repeated use of suboptimal

concentration of enilconazole on sensitivity of

Malassezia pachydermatis to ketoconazole and

posaconazole

P. BOURDEAU, C. THIBAULT, C. DUMONT andC. ROUXLaboratory LDPM LabOniris, Department of ClinicalSciences, Unit DPM, Veterinary School (Oniris),University of Nantes, France

The frequent use of antifungal medications in dogs canresult in changes in the sensitivity of Malassezia organ-isms to these drugs. This study was conducted to assessthe effects of suboptimal use of enilconazole on the sen-sitivity of Malassezia organisms to ketoconazole andposaconazole. Twenty healthy beagles with bilateralaural populations of M. pachydermatis were included.On day (D)0 and D5 one ear of each dog received0.5 mL of a sub-therapeutic concentration of enilcona-zole (0.05%). Evolution of sensitivity of the yeastorganisms to two azoles (ketoconazole and posacona-zole) was evaluated before treatment on D0 and D10using ETEST (BioM�erieux; Marcy-l’Etoile, France)strips. Minimum inhibitory concentration (MIC; µg/mL) ranges in untreated ears included: ketoconazoleD0, 0.125–0.25 µg/mL (median 0.19) and D10, 0.125–0.38 µg/mL (0.19); posaconazole D0, 0.0125–0.25 µg/mL (0.125) and D10, 0.047–0.38 µg/mL (0.125). Sensi-tivity of Malassezia to ketoconazole in treated ears was0.094–0.25 µg/mL (0.19) before treatment and 0.125–0.25 µg/mL (0.19) on D10. Likewise the sensitivity toposaconazole was 0.0125–0.25 µg/mL (0.125) beforetreatment and 0.047–0.25 µg/mL (0.125) on D10. TheMIC remained unchanged post-treatment in 16/20 earsfor ketoconazole and 6/20 for posaconazole; it washigher in 2/20 ears for ketoconazole and 7/20 forposaconazole and equally lower for each antifungal. Inthis study, the repeated application of suboptimal dosesof enilconazole did not induce any changes in the

sensitivity of M. pachydermatis to ketoconazole andposaconazole.Source of funding: Self-funded.


P-47

Control of dermatophytosis in guinea pigs:

comparison of procedures in 83 cases

P. BOURDEAU, A. LE FRIEC, C. DUMONT, M.MUKOKO, F. ALEXANDRE and C. ROUXLaboratory LDPM LabOniris, Department of ClinicalSciences, Unit DPM, Veterinary School (Oniris),University of Nantes, France

Although guinea pigs have been widely used in experi-mental models for studying antifungals, there is virtu-ally no comparative information on methods forcontrol of dermatophytosis in practice. The informationin this study was obtained from the authors’ labora-tory. The protocols used in practice and their efficacy(post-treatment negative cultures) were compared usingchi-squared tests. From a series of 83 cases, enilcona-zole 0.2% was used in 83% of animals; every four daysduring six weeks in 66% and as the only therapy in39%. Itraconazole (5 mg/kg/day) was used in 90% ofthe 83 cases; in an alternate week therapy in 89%, threetimes in 91% and with 60% (6/10) efficacy when usedalone (a prolonged daily treatment cured 3/3 guineapigs). Griseofulvin was used in 10% of the 83 cases at20–40 mg/kg (usually 25) daily over four weeks in60%. The tolerance was always excellent (occasionaldifficulties in administering enilconazole or griseoful-vin). Combined systemic and topical treatment (n = 36)was the most effective [86% versus 81% (only topical)and 69% (only systemic)] (P = 0.494). All long-hairedanimals (4/4) were cured when clipped (versus 5/8unclipped). Both disinfection of environment (69% cureversus 31% without disinfection; P = 0.277) and treat-ment of conspecifics (86% cure versus zero cure if nottreated) helped. From these results we propose thecombination of enilconazole and itraconazole for sixweeks with clipping of long-haired animals, treatmentof conspecifics and disinfection of the environment asthe most effective management of dermatophytosis inguinea pigs.Source of funding: Self-funded.


P-48

Application of the ETEST technique to

evaluate the sensitivity of Malasseziapachydermatis to ketoconazole, posaconazole

and itraconazole in normal dogs

P. BOURDEAU, C. THIBAULT, F. HUBERT and F.ALEXANDRELaboratory LDPM LabOniris, Department of ClinicalSciences, Unit DPM, Veterinary School (Oniris),University of Nantes, France

Little is known about the sensitivity to antifungal medi-cations of resident populations of Malassezia pachyder-matis in dogs. The objective of this study was to

Abstracts 73



evaluate the activity of ketoconazole, posaconazole anditraconazole in ear canal populations of this yeast.Malassezia pachydermatis was obtained from swabbingthe ear canals of 20 healthy adult beagles. Isolatedcolonies were used to produce suspensions (2.5 McFar-land) that were swabbed onto Sabouraud’s-Tween 20agar. ETEST (BioM�erieux; Marcy-l’Etoile, France)strips of ketoconazole, posaconazole or itraconazolethen were applied to the agar. After incubation (32°Cfor 48 h) the minimal inhibitory concentration (MIC;µg/mL) were read directly from the strips. The MICranges included ketoconazole 0.094–0.25 µg/mL (me-dian 0.19), posaconazole 0.0125–0.25 µg/mL (0.125)and itraconazole 0.008–0.047 µg/mL (0.032). Left andright aural yeast populations expressed identical MICsfor ketoconazole and posaconazole in 9/20 dogs anditraconazole in 2/7 dogs. Bilateral sensitivity profileswere identical for posaconazole and ketoconazole in 5/20 dogs and for all three antifungals tested in 1/7 dogs.There was no predictability of MIC values betweenketoconazole and posaconazole (i.e. susceptibility toone medication neither correlated with nor implied sus-ceptibility to the other). ETEST was shown to be rapidand easy to evaluate sensitivities of M. pachydermatisto antifungal medications in dogs. Malassezia popula-tions in ear canals express a wide range of in vitro sen-sitivity profiles including differences between affectedears of the same animal in a high proportion of dogs.Source of funding: Self-funded.


P-49

Diagnosis of dermatophytosis in guinea pigs:

comparative results of fungal culture and

trichoscopy from 400 cases

P. BOURDEAU, A. LE FRIEC, F. HUBERT, M.MUKOKO, F. ALEXANDRE and C. ROUXLaboratory LDPM LabOniris, Department of ClinicalSciences, Unit DPM, Veterinary School (Oniris),University of Nantes, France

The objective of this study was to describe informationobtained from fungal culture or trichoscopy in cases ofdermatophytosis in guinea pigs. Samples (carpetmethod) sent to the authors’ laboratory were culturedunder laboratory conditions for fungal identificationand growth quantification (colony forming units). Theresults were compared to trichoscopy using the chi-squared test. In the period 2016–2017, 400/740 (54%)samples were positive for dermatophytes. When com-bined with 261 positive samples from 2014–2015, thefrequencies of 661 dermatophyte species included: Tri-chophyton porcellae 82.8%, T. mentagrophytes 14.5%,Microsporum canis 1.4% and two concurrent species1.4%. The respective times for identification (days �SD) were 6.2 � 1.61, 5.7 � 1.61, 7.1 � 1.97 and6.9 � 1.45 (P = 0.108–0.49). The cfu total was signifi-cantly higher for T. porcellae and T. mentagrophytes(56% and 61% cfu > 50) than M. canis (67% cfu = 1)(P = 0.0078). Post-treatment evaluation indicated that21.2% T. porcellae and 9.5% T. mentagrophytes casesremained infected, mainly when initial cfu > 50 (27%)(P = 0.599). Trichoscopy (n = 584) had a limited sensi-tivity (17%) and negative predictive value (43%) yetwas highly specific (99%) with a high positive

predictive value (98%), independent of the fungal spe-cies (P = 0.843). When positive, trichoscopy was indica-tive of severe infection (92% cfu > 50) (P = 2.001e-7).These results indicate that fungal culture is rapid, speci-fic and sensitive for the diagnosis of dermatophytosisand essential to evaluate mycological cure.Source of funding: Self-funded.


P-50

Cowpox virus dermatitis in a cat: a case report

I. OTROCKA-DOMAGAŁA*, K. PA�ZDZIOR-CZAPULA*, M. MIKIEWICZ*, M. GESEK*, W.KRASI �NSKA-PRZEWORSKA† and B.TWORKOWSKI†

*Department of Pathological Anatomy, Faculty ofVeterinary Medicine, University of Warmia and Mazuryin Olsztyn, Poland; †Veterinary Clinic in Olsztyn,Poland

Cowpox virus (distributed throughout Europe andAsia) sporadically infects outdoor hunting cats becausewild rodents are the reservoir of this pathogen. Theinfection is manifested by skin lesions located predomi-nantly on the head, neck and forelimbs, and some indi-viduals develop fatal necrotizing pneumonia. We reporta case of cowpox virus infection in a cat with atypicaldistribution of the cutaneous lesions. A 7-year-old, neu-tered male European short hair, outdoor cat presentedwith fever (40°C), severe erythema with crusting andulceration of the skin in the abdominal area. The catreceived antibiotics and glucocorticoids with subsequentprogression of the skin lesions. Therefore, multiple full-thickness skin biopsies were sampled for histopathologi-cal evaluation, which revealed acanthosis, severe epider-mal spongiosis and ulceration. Large, irregular,polygonal eosinophilic inclusion bodies were seenwithin the cytoplasm of keratinocytes in both the epi-dermis and hair follicles. Massive, multifocal to coalesc-ing infiltrations of macrophages, eosinophils,neutrophils and plasma cells were observed in thesubepidermal and perifollicular areas, extending to theinterstitial dermis. These microscopic changes werecompatible with cowpox dermatitis. Cowpox virus(Orthopoxvirus) infection was confirmed by PCR test-ing of the epidermal crusts sampled after initialhistopathological diagnosis. Within a few weeks, skinlesions started to heal with nearly complete recoveryduring the following three months. Owing to the zoo-notic potential of cowpox virus, this infection shouldalways be regarded as a differential diagnosis when ahunting cat presents with ulcerative skin lesions locatedanywhere on the body.Source of funding: Self-funded.


74 Abstracts



P-51

Tongue lesions as a clinical manifestation of

canine leishmaniosis in an endemic area

R. LIMA* and C. SOUZA†

*Clinica Veterinaria Canis & Catus, RN, Brazil;†Department of Veterinary Clinical Medicine, Universityof Illinois, IL, USA

Canine leishmaniosis is a vector-borne protozoan dis-ease endemic to >70 countries. The disease can havevarious clinical presentations. This report describes fourcases of canine leishmaniosis presenting with tonguelesions as the main clinical sign. The dogs were exam-ined at a veterinary clinic in an area endemic for Leish-mania infantum in North-eastern Brazil. The age of thedogs at presentation ranged from four to nine yearsand they were of four breeds: boxer, Rottweiler, minia-ture pinscher and pit bull. All dogs presented with ton-gue lesions characterized by ulcers (two), coalescingpapules and plaques (two), loss of papillae appearanceon the dorsal and lateral aspects of the tongue (two),erythema and papules on the ventral tongue surface(one), and irregular tongue margins (one). One dogshowed bilateral periocular ulcers and scaling with lym-phadenomegaly, in addition to tongue lesions. Anotherdog was previously diagnosed with leishmaniosis andhad cutaneous lesions that resolved with initial therapy.Maintenance therapy with allopurinol and domperi-done was administered for two years before the occur-rence of tongue lesions. Cytological findings fromtongue lesions of all four dogs showed amastigotesinside macrophages, cytoplasmic vacuoles and extracel-lular amastigotes. Two dogs had a negative serologicalrapid test (immunochromatography) result before thecytological confirmation of leishmaniosis. Two dogswere euthanized after the diagnosis of leishmaniosis.The remaining two dogs were treated with a combina-tion of miltefosine, allopurinol, and domperidone. Ton-gue lesions resolved after 30 days of therapy. Tonguelesions should be considered as a clinical manifestationof canine leishmaniosis.Source of funding: Self-funded.


P-52

Topical application of a proprietary blend of

essential oils and plant extracts is associated

with fewer relapses of pyoderma

V. A. FADOK*, F. SECKERDIECK†, E.BENSIGNOR‡, C. NOLI§, A. OLIVEIRA¶ and R. S.MUELLER†

*Zoetis, Parsippany, NJ, USA; †Ludwig MaximillianUniversity, Munich, Germany; ‡Clinique Veterinaire,Paris, France; §Servizi Dermatologici Veterinari,Peveragno, Italy; ¶University Lusofona de Humanidadese Tecnologias, Lisboa, Portugal

Dogs with cutaneous disorders can develop recurrentstaphylococcal pyoderma. Addressing the underlyingcause is necessary yet not always sufficient to controlrecurrences. The purpose of this study was to determinethe effects of weekly use of topical PYOspot (Der-moscent Laboratorie; Castres, France) on pyodermarelapse. Dogs admitted to the study had at least four

pyoderma outbreaks in the previous year. Allergy medi-cations were permitted if the dogs were treated for atleast three months previously; allergen immunotherapywas permitted if started one year previously. Antibioticsand topical therapy were permitted as needed. Dogsadmitted into the study received either topical PYOspotor a topical placebo applied once weekly. Owners anddermatologists were blinded, and the dogs were ran-domly assigned to either arm. Each dog was evaluatedby a dermatologist at least four times over the follow-ing year: after first visit (V1) at the start, V2 at approx-imately three to four weeks to ensure pyodermaresolution, V3 at six months or relapse of pyodermaand V4 at 12 months. Dogs were assessed by physicalexamination and cytological evaluation. Twenty-eightdogs from multiple centres were enrolled; two withdrewbefore V4. Fourteen dogs were treated with PYOspotand 12 with placebo. Both groups had fewer episodesof pyoderma after one year (PYOspot 5.0 � 1.69 to1.07 � 1.3; placebo 5.3 � 1.49 to 2.67 � 1.43); how-ever, the difference between the groups at year end wassignificant (P = 0.008). We conclude that the regularuse of PYOspot, in addition to treatment for the under-lying condition, can reduce pyoderma relapses.Source of funding: PYOspot and placebo were provided

by Dermoscent Laboratorie.


P-53

Necrotizing soft tissue infection: A case report

A. VILLARREAL*, D. CAMPOS†, D. BORGES† andJ. FERNANDES†

*Pet Skin Care Services Panama, Panama, Panama;†Department of Veterinary Science at UniversidadeFederal Rural de Rio de Janeiro, RJ, Brazil

Necrotizing soft tissue infections (NSTI) are severeinfections of the skin, subcutaneous tissue, superficialfascia, and occasionally, deep fascia, muscle and adi-pose tissue. They can lead to systemic toxicity, septi-caemia and high mortality rates, if not treated in time.In this report, we describe a case of NSTI caused by abite, in a 4-year-old female beagle. The patient pre-sented with small puncture wounds on the sacral regionand hind legs. These areas were associated withoedema, erythema and loss of sensitivity. An extensivearea of crepitation at the base of the tail also wasnoted. One of the most relevant clinical signs wasintense pain during examination, not correlated withthe small wounds observed. After surgical debridement,a large zone of deep necrotic tissue was observed in thelumbosacral region, as well as purulent exudate anddevitalized tissue. Therapy with ceftriaxone, metronida-zole and tramadol chlorhydrate was initiated. In addi-tion, thorough cleaning using warm pressurized salinesolution and 0.01% iodopovidone solution two times aday, as well as compression bandaging were established.A silver sulfadiazine paste was applied to the woundsuntil granulation tissue was observed, approximately 20days after the initial debridement. The patient had agood recovery, with optimal healing and cicatrizationof the involved areas. Recognizing signs of a possibleNSTI can lead to an early diagnosis and treatment,which are fundamental factors for a good prognosis.Source of funding: Self-funded.


Abstracts 75



P-54

The efficacy of wipes containing 0.1%

chlorhexidine gluconate and EDTA for canine

superficial pyoderma: a pilot study

Y. TAKAHASHI*, K. MORI†, Y. OSHIMA-TERADA† and N. MURAYAMA†

*Minamino Animal Hospital, Tokyo, Japan;†Dermatology Services for Dogs and Cats, Tokyo, Japan

Therapy with chlorhexidine-based shampoos can beefficacious in canine superficial pyoderma yet the sham-pooing process is often laborious especially in largedogs. The purpose of this study was to assess the effi-cacy of wipes containing 0.1% chlorhexidine gluconateand EDTA in dogs with superficial pyoderma. Thestudy included seven dogs from three months to 11years of age. These dogs were diagnosed with superfi-cial pyoderma based on typical clinical signs and skincytological results. The owners gently wiped the lesionson the ventral abdomen once daily for three weeks.The following subjective evaluation was performed attreatment end: excellent (complete resolution), good(some improvement), fair (no improvement) and poor(exacerbation). Erythema, papules/pustules, scales/crusts and pruritus were scored 0–3 by the investigatorsbefore and at treatment end (0, none; 1, occasional ormild; 2, frequent or moderate; 3, severe). CutaneouspH and transepidermal water loss (TEWL) were mea-sured before and after the trial. These data were non-categorical, thus the Wilcoxon signed rank test wasused to analyse differences. The subjective evaluationshowed five excellent, one good and one fair response.Three, five, six and five dogs showed a reduction inscores for erythema, papules/pustules, scales/crusts andpruritus, respectively. Erythema and pruritus improvedsignificantly (P < 0.01). Skin pH (7.81 before, 7.46after: P < 0.01) and TEWL (3.13 before, 3.92 after:P < 0.01) changed significantly. No adverse effects werenoted in any of the dogs. This study suggest that oncedaily 0.1% chlorhexidine gluconate and EDTA wipescan improve canine superficial pyoderma.Source of funding: Self-founded.


P-55

Antimicrobial resistance gene and multidrug

efflux pumps against antiseptics for

staphylococci isolated from canine superficial

pyoderma in Japan

N. MURAYAMA*, Y. OSHIMA-TERADA*, K.MORI*, H. NAKAMINAMI†, T. SAITO† and N.NOGUCHI†

*Dermatology Services for Dogs and Cats, Tokyo,Japan; †Tokyo University of Pharmacy and LifeSciences, Tokyo, Japan

Meticillin-resistant staphylococci in canine pyoderma isa serious problem. The aim of this study was to assessmecA gene, antimicrobial resistance genes and mul-tidrug efflux pump genes of staphylococci from caninesuperficial pyoderma in Japan. Bacterial isolates from136 dogs were evaluated from May 2018 to April 2019.Staphylococci, mecA gene, antimicrobial resistancegenes including ermA, ermB, ermC, msr, inuA, mphC,aacA/aphD, tetM, tetK and fosB, and multidrug effluxpump genes including qacA/B and smr were identifiedby PCR. Of the staphylococci, 72% were Staphylococ-cus pseudintermedius, 25% S. schleiferi subsp. coagulansand 3% unidentified species, and 47% of all staphylo-cocci were mecA-positive. The staphylococci positivefor ermA, ermB, ermC, msr, inuA, mphC, aacA/aphD,tetM, tetK and fosB were: all staphylococci – 0%,46.3%, 0.7%, 0%, 0%, 0%, 50.7%, 46.3%, 1.5% and70.0%, respectively; MecA-negative staphylococci –0%,22.2%, 0%, 0%, 0%, 0%, 27.8%, 33.3, 2.8% and62.5%, respectively; and MecA-positive staphylococci –0%, 74.6%, 1.6%, 0%, 0%, 0%, 76.6%, 60.9%, 0%and 78.1%, respectively. Multidrug efflux pump genesincluding qacA/B and smr were in 0.7% and 2.2% (re-spectively) of all isolates. MecA-positive staphylococciwere positive for qacA/B and smr in 1.6% and 1.6%,respectively. MecA-negative staphylococci was positivefor smr in 2.8%. This study indicated that mecA-posi-tive staphylococci had many antimicrobial resistancegenes, and a few staphylococci had multidrug effluxpump genes.Source of funding: Self-funded.


76 Abstracts



OTITIS

P-56

Relationship between cytological,

bacterioscopic and clinical features in 64 cases

of chronic bacterial otitis externa in dogs


The aim of this study was to characterize the relation-ships between bacterioscopic, clinical and cytologicalfeatures of chronic bacterial otitis in dogs. Sixty-fourdogs (104 ears) with chronic bacterial otitis externa(positive bacterioscopy confirmed by bacteriologicalevaluation) were included. Information on chronicity,primary factors, previous topical antibacterial treat-ments and pain was collected. Clinical scoring (OTIS-3)and type (Er, erythematous; Cer, ceruminous; ErCer,erythemato-ceruminous; Sup, suppurative) wasrecorded. Cytological pattern [bacterial overgrowth(BOG), suppurative, pyogranulomatous], presence ofMalassezia and bacterioscopic results (C, cocci; R,rods) also were collected. Chi-squared or Fisher’s exacttests were used (P < 0.05). Bacterial population (mor-phological) results were: majority C, 40/104 ears(38.4%); majority R, 22/104 (21.2%); or equivalent C:R ratio (B), 42/104 (40.4%). No correlation wasobserved between the bacterioscopic results and pri-mary factors, pain, OTIS-3 or presence of Malassezia.A trend was observed with chronicity: more than onemonth in 35% C, 40% B and 57% R. Statistical corre-lations were observed between the bacterial morphologyand the number of previous antibacterial treatments [0(55% C, 27% R) versus 1, 2 or 3 (45% C, 63%R)(P = 0.03)], the type of otitis (Er, Cer, ErCer 15% R,Sup 13% C) (P = 9.10-5) and the type of reaction(BOG, 53% C, pyogranulomatous 17% C)(P = 8.9 9 10-3). Morphology of bacteria is the firstelement for identification and this study brings newinformation on possible links with clinical and cytologi-cal significance.Source of funding: Virbac Corporate, France.


P-57

Bacterial chronic otitis: correlation between

cytological inflammatory patterns, clinical

aspect and bacteriology in 64 dogs


One hundred and four ears (of 64 dogs) with chronic(minimum two weeks) bacterial otitis externa (bacteriaobserved then identified by bacteriological evaluation)were included in this study. Primary factors, chronicity,previous topical antibacterial treatments, pain, clinicalscoring (OTIS-3) and classification (Er, erythematous;Cer, ceruminous; ErCer, erythemato-ceruminous; Sup,suppurative) were collected. Otitis were classified byinflammatory pattern at cytological evaluation:

bacterial overgrowth (no inflammatory cell) (BOG; 43ears, 41.4%), suppurative (>90% neutrophils) (Pyo; 55ears, 52.9%), pyogranulomatous (>50% neutrophilsand macrophages) (PyoG; six ears, 5.8%). Chi-squaredor Fisher’s exact tests were used (P < 0.05). No correla-tion was observed between the cytological pattern andchronicity (more than two months: 76% BOG, 85%Pyo, 100% PyoG), pain, OTIS-3 or the presence ofMalassezia. There was a statistical correlation betweencytological pattern and primary factors (BOG observedin 89.3% of allergic otitis) (P = 0.02); number of pre-vious antibacterial treatment: 0 (58% BOG, 31% Pyo,17% PyoG) versus 1, 2 or 3 (P = 4.6 9 10-2); clinicaltype of otitis [BOG with ErCer (61.9%), Pyo and PyoGwith Sup (59.2% and 66.7%, respectively; P = 5 9 10-3];. There also was a statistical correlation with bacteriaisolated: Staphyloccocus and Corynebacterium withBOG (53.8% and 50%, respectively), Pseudomonas withPyo/PyoG (80%) (P = 0.04); abundance of Malassezia[moderate, high level with BOG (84.6%);P = 2.4 9 10-3]. These results highlight the importanceof cytological inflammatory patterns to describe otitisand possible relationships with bacterial causal agents.Source of funding: Self-funded.


P-58

Bacterial chronic otitis: clinical and cytological

data in 64 dogs


The aim of this study was to improve knowledge onbacterial chronic otitis. For one year, dogs with chronic(more than two weeks) bacterial externa (CBO) wereprospectively analysed. Sixty-four dogs were included(104 ears). The mean age was 6.5 years with a distribu-tion of age older than the overall canine population ofONIRIS hospital. There was no sex predisposition.Seven breeds represented 55% of cases: French bulldog,cocker spaniel, Labrador retriever, bichon frise, Ger-man shepherd dog, beagle and West Highland whiteterrier. Except for Labrador retriever and beagle, allbreeds were statistically over-represented compared tothe hospital population. The duration was over twomonths in >70% and the number of topical bacterialtreatments before consultation 1.6. The suspected pre-disposing factors were hypertrichosis (9%), physiologi-cal glandular hyperplasia (cocker spaniel) 9%,swimming (3%) and abnormal ear conformation(1.5%). Primary factors were identified in 61% of cases:hypersensitivity (48%), foreign body (4.8%), nodule(4.8%) and endocrinopathy (2.9%). Perpetuating fac-tors were stenosis (40%) and induration (35%). InCBO, the otitis was erythematous in 7%, ceruminousin 7%, cerumino-erythematous in 40% and suppurated46%. Cytologically, bacterial overgrowth was observedin 41%, suppurative in 53% and pyogranulomatous in6%. Bacteria were mainly cocci in 38%, mainly rods in21% and equivalent C:R ratio in 41%. Malassezia

Abstracts 77



often were present (>50%) as both cocci and rods (nostatistical difference) in low concentration (>75% ofcases). This study brings new data about clinical andcytological features in bacterial chronic otitis in dogs.Source of funding: Virbac Corporate, France.


P-59

Lagochilascaris as a cause of unilateral

suppurative otitis in two cats: a case report

M. CARDOSO*, F. CAGOL†, K. MATTE† and L.ANTONIO†

*Dermalove Veterinary Dermatology Office, Caxias doSul, RS, Brazil; †S~ao Francisco Veterinary Center,Bento Gonc�alves, RS, BrazilOtitis is characterized by an acute or chronic multifac-torial inflammatory disease of the auditory system.Lagochilascariasis is a zoonosis limited to the Americancontinent caused by a nematode helminth of the Ascari-didae family of the genus Lagochilascaris and is anuncommonly reported parasite in mammals. The infec-tion occurs through the ingestion of raw meat fromhosts containing the larvae. The parasite infects subcu-taneous regions, usually in the cervical region and sur-rounding tissues of the face. The aim of this report isto provide information on the occurrence of lagochilas-cariasis as cause of otitis media in felines and on therelevance of otoendoscopy as an efficient diagnosticmethod. In the city of Bento Gonc�alves (RS, Brazil),one intact 8-year-old female and an intact 1-year-oldmale cat, both of undefined breed, were presented withsuppurative right unilateral otitis resulting in a putridsmell and pain. In the female, an otoendoscopic examrevealed a polypoid, multilobulated, elongated tissuethat extended from the external ear towards the middleear. After traction removal of the polyp, cylindricalfusiform worms of a milky-white colour were observed.Forty nematode parasites of the genus Lagochilascariswere found in tissue from the female patient and fourwere isolated from the male patient. Otoendoscopy, inthese cases, was fundamental in diagnosis, treatmentand prognosis of both patients.Source of funding: Self-funded.


P-60

In vitro antimicrobial activity of Epiotic SIS

and Easotic against pathogens associated with

canine otitis externa in Australia

W. Y. CHAN*,†, A. G. KNOWLES‡ and D. J.TROTT**Australian Centre for Antimicrobial ResistanceEcology, School of Animal and Veterinary Sciences, TheUniversity of Adelaide, South Australia, Australia;†Faculty of Veterinary Medicine, Universiti PutraMalaysia, Serdang, Selangor, Malaysia; ‡Virbac(Australia) Pty Ltd, Milperra, New South Wales,Australia

Otitis externa (OE) in dogs is caused by a variety ofaetiological agents. OE is commonly treated with

antibiotics to control the underlying infection, in con-junction with ear cleansers to remove waxy debris andexudate that may encourage further microbial growth.We conducted studies to determine the antimicrobialproperties of the ear cleanser, Epiotic SIS (Virbac; Car-ros, France) against clinical isolates from Australiandogs, and whether Epiotic SIS could enhance theantimicrobial effect of Easotic (miconazole nitrate andgentamicin sulfate, Virbac), an antibiotic treatment forOE. Minimum inhibitory concentration (MIC), mini-mum bactericidal concentration (MBC) and time–killkinetic assays were performed with Epiotic SIS andEasotic against multiple isolates of meticillin-resistantStaphylococcus pseudintermedius (MRSP), meticillin-sus-ceptible S. pseudintermedius (MSSP), Pseudomonasaeruginosa, Proteus mirabilis, Escherichia coli andMalassezia pachydermatis. Epiotic SIS inhibited micro-bial growth in all isolates (MIC 0.0156–0.125); thiseffect was greatest against Malassezia, MSSP andMRSP. Epiotic SIS exhibited microbicidal activityagainst all isolates (MBC 0.0312–1.0). Epiotic SIS killedPseudomonas within 4 h of exposure, and all other iso-lates except for Proteus within 24 h. When Epiotic SISand Easotic were tested in combination, an additive tosynergistic interaction was observed against MSSP andMRSP isolates, while an additive interaction was seenagainst Malassezia. Epiotic SIS exhibited antimicrobialproperties against clinical isolates of bacteria and yeastrepresenting the most common causes of canine OE,including MRSP. Epiotic SIS was shown to enhancethe effect of Easotic against common aetiologicalagents of OE isolated from Australian dogs.Source of funding: Virbac, Australia.

Conflicts of interest: WYC received salary from

ACARE. DJT has received research funding from Luoda

Pharma, Neoculi, Zoetis, Bayer, Boehringer Ingelheim,

Virbac, and Elanco.

P-61

Resolution of antibiotic resistant bacteria seen

in canine chronic otitis externa cases

M. FUJIMURAFujimura Animal Allergy Hospital, Osaka, Japan

Otitis is a frequent complication of repeat-occurrenceallergic disorders in canines and steroids and/or antibi-otics are commonly used treatment options. Because ofsuch remedies, microbes are becoming more drug-resis-tant, and the frequency with which stronger drugs arebeing used is increasing. A study was performed look-ing at the effects of an alternative, ZYMOX Otic, anenzyme-based ear solution containing three naturalenzymes (lactoperoxidase, lactoferrin and lysozyme) onantimicrobial activity and effectiveness in biofilm eradi-cation. Presented are two of five clinical cases of dogswith chronic dermatitis and subsequent otitis externa(OE). Each case presented with allergic skin disordersand concurrent ear infections that were difficult totreat. These two case studies evaluated ear conditionspre-application and post-application of ZYMOX Oticsolution. Using a numerical scale of 0–3 (3, severe),redness, pruritus, cerumen and purulence were graded.Swab specimens were taken for cytological examinationand bacteria were counted and recorded on a scale of 0(rare) to 3+ (numerous). Results showed that chronic

78 Abstracts



ear infections were being controlled at approximatelytwo to three weeks into these clinical trials. Further-more, ZYMOX Otic solution was effective in improv-ing clinical signs associated with infectious OE. Thegrade of redness, pruritus, cerumen and purulencedecreased within the first week, along with decrease ofOE infection. Therefore, ZYMOX Otic solution wasshown to be a viable treatment option for reducing bac-terial and yeast infections and providing clinicalimprovement of OE in dogs.Source of funding: Self-funded.


P-62

Video-otoscopy-guided tympanostomy tube

placement for treatment of primary secretory

otitis media in cats

R. LUCAS*, I. QUIRICO*, M. MASCARENHAS†

and D. BEVIANI**Dermatoclinica, S~ao Paulo, SP, Brazil; †PromoveCl�ınica Veterin�aria, Rio de Janeiro, Brazil

Primary secretory otitis media is an uncommon diseasein cats. Current treatment recommendations includemanual removal of the mucoid effusion from the tym-panic cavity through a myringotomy incision and topi-cal or systemic corticosteroids. Although some cats stillpresent a recurrent mucoid effusion from the middleear. The aim of this study was to describe video-oto-scopy-guided tympanostomy tube placement in fourcats with middle ear effusion and assess the clinical out-come. Diagnosis of primary secretory otitis media wasbased on history, physical examination, video-otoscopyand CT or MRI findings. Three cats presented bilateraland one unilateral otitis media. One showed concomi-tant respiratory signs and all cats presented with neuro-logical signs. The decision to add tympanostomy tubeplacement to the treatment was the recurrence of theotitis media 45 to 50 days after a prior myringotomy.A tympanostomy tube with 1.2 mm internal diameter(Shepard ventilation tube, Medicone; Cachoeirinha,Brazil) was placed in the myringotomy site undervideo-otoscopic guidance using malleable grasping for-ceps (Karl Storz Veterinary Endoscope; Goleta, CA,USA). Clinical signs resolved rapidly in all cases fol-lowing the procedure and all cases were asymptomaticat follow-ups at 30, 60, 90, 120 and 180 days. Noadverse effects were noted. Results of this study indi-cate that tympanostomy tubes provide continual tym-panic cavity ventilation and drainage, and may be aneffective alternative to repeated myringotomy for thetreatment of primary secretory otitis media in cats.Source of funding: Self-funded.


P-63

An atypical presentation of ototoxicity caused

by an ear medication containing florfenicol,

terbinafine and betamethasone acetate

M. MASCARENHASPromove Cl�ınica Veterin�aria, Rio de Janeiro, Brazil

Florfenicol, terbinafine and betamethasone acetate inan adaptable gel (Osurnia, Elanco Animal Health;Greenfield, MA, USA) may be ototoxic and shouldonly be applied with an intact tympanic membrane.Adverse effects associated withear medications mayinclude otitis media, Horner’s syndrome, facial nerveparalysis and vestibular clinical signs (ataxia, nystag-mus, head tilt). The objective of this report was todescribe a case of an atypical presentation of ototoxic-ity caused by Osurnia not previously reported in the lit-erature. A 3-year-old male Maltese dog presented to anemergency service with severe exudate, swelling, painand oedema of the left eye. The signs appeared approx-imately 24 h after the dog received an application ofOsurnia in both ears for treatment of acute otitisexterna. The dog was diagnosed with neurogenic kera-toconjunctivitis sicca (KCS) and an ipsilateral dry nose.There were no signs of other neurological deficits.Video-otoscopy was performed and the tympanic mem-brane of the left ear was ruptured. For treatment ofthis patient it was essential to clean and remove all ofthe medication instilled within the external ear canal.When neurogenic KCS is associated with an ipsilateraldry nose, it is likely that damage has occurred to thepreganglionic parasympathetic fibres proximal to thepterygopalatine ganglion. This is the first report of anototoxic medication that led to KCS and an ipsilateraldry nose. Considering that most cases of ear disease indogs are treated topically, veterinarians should beaware of all possible signs of ototoxicity.Source of funding: Self-funded.


P-64

In vitro activity of Copaiba oil against

Malassezia pachydermatis strains from canine

otitis

A. PACHECO, S. SOUZA, L. MEDEIROS, E.SOUZA and T. SILVACenter for Biological and Natural Sciences, FederalUniversity of Acre, Rio Branco, AC, Brazil

Otological infections caused by Malassezia spp. are fre-quent and the use of topical antimicrobials is usedwidely to control these infections. However, the devel-opment of resistance to antimicrobials has beenreported widely and research for new therapy options isnecessary. In this context, medicinal plants such ascopaiba oil offer a promising alternative to controlinfections caused by several micro-organisms includingMalassezia strains. Thus, we hypothesized that copaibaoil could inhibit yeast strains isolated from canineinfections. For this study, a total of 23 strains ofM. pachydermatis obtained from the culture of canineotitis were used. Initially, inhibition of growth was con-firmed by the well diffusion technique using 100%copaiba oil. Subsequently, different concentrations of

Abstracts 79



copaiba oil were assessed using the microdilutionmethod to obtain the minimum inhibitory and fungici-dal concentrations (MIC and MFC). Fluconazole wasused as a positive control. The MIC of copaiba oil inthe evaluated samples was 0.61%, confirmed by theabsence of colonies through visual analysis. The MFCwas 1.85% because there was no yeast growth afterinoculation. Thus, we can confirm that copaiba oil hasthe capacity to inhibit strains of M. pachydermatisin vitro; however, clinical studies should be carried outin order to confirm in vivo effectiveness.Source of funding: Self-funded.


P-65

Reacidification of the ear canal with an ear

cleanser in a model of alkalinization of the

external auditory canal in dogs

P. PANZUTI*, O. FANTINI†, M. MOSCA*, G.NOEL* and D. PIN**VetAgro Sup, Marcy l’Etoile, France; †V�etoquinol,Lure, France

Cerumen plays a protective role in the external auditorycanal (EAC), especially through bactericidal and fungi-cidal functions. In humans, changes towards alkalinityof the EAC is one of the local factors in the progres-sion of acute to chronic otitis. Higher EAC pH isfound in dogs with otitis externa. We developed amodel of alkalization of the EAC in dogs in order toevaluate the acidic effect of the ear cleaner Sonotix(Vetoquinol; Lure, France). The study was approvedunder Ethic Committee no. 1812. Five mL of a pH10.1 phosphate-buffered saline (PBS) solution wereinstilled into both ears of 10 healthy beagles in order toreach pH 9. Then 2 mL of Sonotix was instilled intothe right ear with the left ear kept untreated. Using askin pH probe applied on the cavum conchae, EAC pHmeasurements were performed before the pH 10.1 PBSsolution application, immediately after Sonotix applica-tion and at 1 h, 2 h, 3 h, 4 h, 9 h, 24 h and 48 h post-application. Overall, pH values were significantly lowerfor Sonotic-treated ears versus untreated ears (general-ized linear mixed model, P < 2 9 1016). Immediatelyafter Sonotix application, mean pH values droppedabruptly to 6.38 + 0.27 in treated ears and then slowlydecreases over the next 48 h until reaching pre-alkalini-zation of 5.87 + 0.66. Sonotix elicited a quick reacidi-fication of the EAC. This procedure appears suited forevaluating the acidic effects of ear cleansers.Source of funding: Vetoquinol.

Conflict of interest: OF is an employee of Vetoquinol.

P-66

Variation in biofilm production by

Pseudomonas aeruginosa isolates from canine

otitis using different media and incubation

times

M. L. PICCIONE and D. J. DEBOERSchool of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, USA

Pseudomonas aeruginosa is commonly isolated fromdogs with otitis and frequently produces biofilms. Thereare vast differences in reported prevalence of biofilm-forming ability of P. aeruginosa isolated from canineotitis. Differences could be explained by strain, incuba-tion time, temperature, media or regional variation. Wesought to compare biofilm production of P. aeruginosaisolated from canine otitis using different media (mini-mal versus enriched) at different incubation times. TheP. aeruginosa isolates were assessed for biofilm produc-tion using a crystal violet microtiter plate assay. Biofilmproduction in Lennox Broth (LB) and in M63 and M9minimal media was assessed after 24 h, 48 h and 72 hof incubation at 30°C. All isolates tested (n = 29) wereclassified as biofilm formers; however, the amount ofbiofilm produced varied greatly between strains. At24 h, all strains produced biofilm in LB. One strain didnot produce biofilm in minimal media (M9, M63).Incubation times >24 h in LB impeded accurate quan-tification of biofilm production. Our study demon-strates that the amount of biofilm produced varies withtime and medium used for incubation. We determinedthat optimal production of P. aeruginosa biofilm wasachieved with LB at 24 h. Although minimal media canbe used, strains may be inaccurately categorized as non-biofilm formers. Further studies are needed to assessfactors increasing formation of biofilm and investigatebiofilm-disrupting compounds for successful treatmentof canine Pseudomonas otitis.Source of funding: Self-funded.


P-67

Clinical efficacy of Neo-Burow’s solution in

six dogs with suppurative otitis media

A. ZAIKE*, K. MORI†, Y. OSHIMA-TERADA† andN. MURAYAMA†

*Ishii Animal Hospital, Chiba, Japan; †DermatologyServices for Dogs and Cats, Tokyo, Japan

Suppurative otitis media is common in dogs, yet diffi-cult to control owing to antimicrobial-resistant bacteria.Burow’s solution exhibits antibacterial activity for sup-purative otitis externa and otitis media in human. How-ever, it takes several days to make and quality-controlis challenging. Recently, Neo-Burow’s solution has beenreported that has an aluminium acetate base; it is easierto prepare and has equivalent efficacy as Burow’s solu-tion in humans. This case report assesses the efficacyfor treating suppurative otitis media in dogs. The studyincluded six dogs aged from three months to 14 years.They were diagnosed with suppurative otitis mediabased on typical clinical features with video otoscopeand ear-discharge cytology. We irrigated ear canals andmiddle ears with Tritz-EDTA (Tris-EDTA Otic PLUS,

80 Abstracts



Kirikan Ltd.; Tokyo, Japan) and 0.05% chlorhexidinegluconate solution (0.05% MASKIN WATER, Mar-uishi Pharmaceutical; Osaka, Japan) using a tube untildischarge disappears, and then rinsed them with Neo-Burow’s solution on the first day, and instilled fivedrops of five-fold diluted Neo-Burow’s solution intothe ear canals once daily for 14 days. Four ears of fourdogs completely resolved and there were no bacteriawith ceruminous cytological results after the treatments.

However, four ears of two dogs did not improve andthe ceruminous cytological results showed bacteria. Noadverse effects were observed during the trial. Ourresults indicated Neo-Burow’s solution could be effec-tive and safe in the treatment of chronic suppurativeotitis media in dogs.Source of funding: Self-funded.


Abstracts 81



PARASITES AND PARASITOLOGY

P-68

Larvicidal efficacy of nitenpyram against

infestation by the human bot fly in a dog: a

case report

P. ANDRIOTTI*, P. OLIVEIRA*, R.TORTORIELLO*, C. BOTELHO*, R. MELO† and J.FERNANDES†

*Post Graduation Program, Universidade Federal Ruraldo Rio de Janeiro, RJ, Brazil; †Department ofVeterinary Science, Universidade Federal Rural de Riode Janeiro, RJ, Brazil

Injuries caused by human bot fly Dermatobia hominis(Diptera: Cuterebridae) are extremely uncomfortableand painful for dogs. This species of fly causes nodularand furunculous primary cutaneous myiasis, which mayresult in opportunistic bacterial infections and predis-pose toward secondary myiasis. In spite of the wideoptions of chemical treatment for large animals, caninedermatobiosis therapy remains a challenge for veteri-narians. The aim of this case report was evaluation oflarvicidal efficacy of nitenpyram against infestation byhuman bot fly in a dog. A canine from Brazilian ruralarea was examined with a total of 16 oedematous, ery-thematous and ulcerated nodules. For confirmation ofparasitism, all nodules were examined through digital-digital compression for visualization of D. hominis lar-vae. The treatment instituted was nitenpyram, at a doseof 1 mg/kg per os as a single dose. A 6 h post-adminis-tration assessment confirmed that all larvae were dead.However, they were not expelled from the furuncularlesions, likely due to their shape, large size relative tothe opening of the tracts and the presence of rows ofspines on the larvae. Therefore, the dead larvae weremechanically removed with minimal discomfort shownby the dogs at the time of removal. It is concluded thatnitenpyram is effective in killing D. hominis and is use-ful as an adjunct in treating dogs with myiasis causedby these larvae.Source of funding: This study was financed in part by

Coordenac�~ao de Aperfeic�oamento de Pessoal de N�ıvelSuperior – Brazil.


P-69

Current status of canine straelensiosis

(Straelensia cynotis) in France, its place of

origin

P. BOURDEAU*, F. DEGORCE†, C. DOUINE* andF. CARREZ**Ecole Nationale Veterinaire de Nantes - ONIRIS,University of Nantes, France; †Laboratoire d’AnatomiePathologique du Sud-Ouest LAPVSO, Toulouse, France

Since the first observations in the mid-1990s in Franceand presentation of the disease at WCVD 2000, strae-lensiosis has been noted in several European countries.This study evaluated the current situation in Franceusing two mechanisms: (i) a questionnaire sent fromLaboratory DPM to veterinary practices and (ii) a ret-rospective analysis of diagnostics performed inLAPVSO, both covering the period 2011–2017. Of 616

collected questionnaires, only 96 clinics (15.6%) hadknowledge of Straelensia and 36 (6.7%) had diagnosedcases. Of 27 clinics reporting case numbers, 11 onlysaw one case while two had >20 cases. Of 26 clinicscommenting, disease incidence was considered stable in69%, decreasing in 12% and increasing in 19%. Thefirst diagnosis of straelensiosis was made before 2011 innine clinics and after 2011 in 15. The distribution ofcases covered 21/95 departments (autochthonous in≥18). The mentioned techniques for diagnosis were deep“excisional” scrapings in 63% and histopathological in76%. During 2011–2017, 46 canine cases were identifiedin LAPVSO (versus 49 during the 11 previous years).In this study, 13 new locations (departments) weredetected (seven from questionnaire, five from the labo-ratory and one from both). Straelensia remains poorlyknown by veterinarians and thus is likely underdiag-nosed. It continues to be present in its initially reportedarea and is spreading peripherally with detection in fivedistant areas: since 2017, two new areas have beendetected resulting in its presence in at least 39/95departments of France.Source of funding: Self-funded.


P-70

Fleas in dogs: yearly evolution through

prospective/retrospective evaluation over a 24-

year period (1995–2018)

P. BOURDEAU*, P. BLUMSTEIN†, A. ROUSSEL‡,F. TRAVERS§, L. JEANNEAU* and V. BRUET**Ecole Nationale Veterinaire de Nantes - ONIRIS,University of Nantes, France; †Veterinary ClinicVetania, Vigneux de Bretagne, France; ‡VeterinaryClinic Armonia, Villefontaine, France; §Charles Lechat,Nantes, France

The aim of this study was an analysis of the prevalenceof flea infestation in dogs in western France. Data werecollected as recorded from the clinic of the Oniris DPMUnit. Every dog was assessed systematically for fleainfestation (gross examination, combing methods) forthe presence of fleas and/or flea faeces, possibly associ-ated with hypersensitivity. Cases of dogs suggestive ofhaving flea bite hypersensitivity (no flea detected =FBH) also were collected in a prospectively validatedGood Clinical Practice system. We present hereinyearly variations over a retrospective 24-year period(1995–2018). Of 32,238 dogs, 2,804 were found infested(8.69%) and 3,858 (11.96%) were diagnosed with FBHresulting in a global prevalence of 20.6%. This wasassociated with a global flea:no flea ratio (f:nf) of 0.72.The annual infestation ranged from 4.8% (2005) to17.3% (1997) and, for FBH, from 17% (2000) to34.8% (2012). The global impact of flea-related der-matoses decreased from 1997 to 2006 (minimum 8.2%),then increased (maximum 41.2% in 2012). A decreasein flea detection was observed during the last period2014–2018 (8.2 to 3.2%; possibly an isoxazolineseffect?) and not for FBH frequency (23.8 to 22.3%).Until 2006, f:nf was >1 and remained �0.35 after 2013.Although fleas have become less often detected in dogs,

82 Abstracts



their impact remains high with approximately 30% ofdogs having fleas and/or signs of FBH. These resultsquestion the quality of use in the field for moderninsecticides and the real clinical benefits obtained.Source of funding: Self-funded.


P-71

Fleas in cats: yearly evolution through

prospective/retrospective evaluation over a 24-

year period (1995–2018)

P. BOURDEAU*, P. BLUMSTEIN†, A. ROUSSEL‡,F. TRAVERS§, L. JEANNEAU* and V. BRUET**Department of Clinical Sciences, Unit DPM,Veterinary School (Oniris), University of Nantes,France; †Veterinary Clinic Vetania, Vigneux deBretagne, France; ‡Veterinary Clinic Armonia,Villefontaine, France; §Charles Lechat, Nantes, France

The objective of this study was to determine the preva-lence of flea infestation in cats in western France. Datawere collected as recorded from the clinic of the OnirisDPM Unit. Every cat was systematically assessed forflea infestation (gross examination, combing methods)for the presence of fleas and/or flea faeces possiblyassociated with hypersensitivity. Cases of cats sugges-tive of having flea bite hypersensitivity (no flea detected= FBH) also were collected in a prospectively validatedGood Clinical Practice system. We present here yearlyvariations over a retrospective 24-year period (1995–2018). Of 5,219 cats, 1,245 were infested (23.8%) and2,109 (33.5%) were diagnosed with FBH resulting in aglobal prevalence of 64.6%. The range of annual infes-tation varied from 10.2% (2018) to 44% (2008) and,for FBH, from 2.6% (2001) to 59.6% (2012). The glo-bal impact of flea-related dermatoses progressivelydecreased from 1995 to 2005 (22.7%), then increased to72.7% in 2014 and then decreased to 59.7% in 2018again (with a decrease of flea-positive cats). Interest-ingly, from 1995 to 2008 (apart from two years) amajority of cats were positive for fleas, yet during thelatter 11 years fleas were not detected on most cats(maximum 83.2% in 2018). These results show thatfleas persist as a dominant component of feline derma-tology (almost two thirds of cats examined). They alsosuggest that the flea–cat relationship may have beenmodified, resulting in a higher proportion of sensitizedcats presented to dermatologists.Source of funding: Self-funded.


P-72

Efficacy of sarolaner on the treatment of

myiasis caused by Cochliomyia hominivorax in

dogs

F. CARVALHO*,¶, M. SILVEIRA†, M. SOUZA§, M.MACHADO‡, C. ROCHA§ and J. FERNANDES‡

*College of Veterinary Medicine, Centro Universit�ariode Valenc�a, Valenc�a, RJ, Brazil; ¶College of VeterinaryMedicine, Universidade Iguac�u, Nova Iguac�u, RJ, Brazil;†Hospital Veterin�ario S~ao L�azaro, Volta Redonda, RJ,Brazil; §Policl�ınica Veterin�aria SOS Animal, Vassouras,RJ, Brazil; ‡Department of Veterinary Science,Universidade Federal Rural do Rio de Janeiro,Serop�edica, RJ, Brazil

The aim of this study was to evaluate efficacy of saro-laner on the treatment of myiasis caused by New Worldscrewworm Cochliomyia hominivorax in naturallyinfested dogs. Selected for study were 12 dogs, of sev-eral breeds, aged 2- to 10-years old, with diagnosedwith myiasis by observation of maggots in wound(s).The dogs received the isoxazoline sarolaner (Simparica,Zoetis; Parsippany, NJ, USA) as a single dose per os,following the manufacturer’s recommended dose of2 mg/kg (range: 2–4 mg/kg) for control of the cat fleaand the brown dog tick (Rhipicephalus sanguineus) indogs. For 72 h post-treatment, the dogs were kept inindividual kennels to evaluate the number of dead lar-vae. The regions most affected were: dorsum (43%),nasal cavity (15%), ear (14%) and mammary tissuewith tumour (7%). A single dose of sarolaner had anefficacy of 100% after 16 h post-treatment. No adverseeffects were observed in treated animals. Followingtreatment the animals returned to the environment inwhich they lived, and no new infestations wereobserved for a period of ≥30 days. Sarolaner is aneffective treatment option in myiasis caused by C. ho-minivorax in dogs.Source of funding: Self-funded.


P-73

Efficacy of imidacloprid plus moxidectin spot-

on and selamectin spot-on formulations against

the ear mite Otodectes cynotis in dogs

T. FUKASE* and Y. NAKAMURA†

*Department of Veterinary Medicine, Faculty ofVeterinary Medicine, Okayama University of Science,Imabari, Japan; †Katsuragi Institute of Life Sciences,Tokyo, Japan

The ear mite Otodectes cynotis inhabits the external earcanal of many species of mammals including dogs, catsand ferrets. We evaluated and compared the efficacy oftwo ectoparasiticides formulated as spot-on productsagainst O. cynotis in dogs. Eighteen dogs naturallyinfested by O. cynotis were randomly assigned to threetest groups of six animals each; an unmedicated con-trol, one group medicated with imidacloprid + mox-idectin spot-on and one group medicated withselamectin spot-on. The drugs were administered to thedogs six times at monthly intervals, with doses of10 mg/kg for imidacloprid, 2.5 mg/kg for moxidectin,and 6 mg/kg for selamectin. The existence or absence

Abstracts 83



of the mites in earwax was examined microscopically atweekly intervals. The dogs were all treated under theGuidelines for Proper Conduct of Animal Experimentsand Related Activities in Katsuragi Institute of LifeSciences. Mites were detected from all of the dogs ofunmedicated control throughout the six month researchperiod and had completely disappeared from 6/6 dogsof the imidacloprid + moxidectin-medicated group byfour months after the first treatment, and from 4/6dogs of the selamectin-medicated group during theresearch period. No adverse effects were observed inany of dogs. The imidacloprid + moxidectin productwas considered to be much more effective againstO. cynotis in dogs than the selamectin products.Source of funding: Self-funded.


P-74

Leishmania mexicana in a central Texas cat:

clinical presentation, molecular identification,

sand fly vector collection and novel

management

K. HOPKE*, A. MEYERS†, L. AUCKLAND†, S.HAMER†, D. FLORIN‡, A. DIESEL* and A.PATTERSON**Department of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, Texas A&M University, CollegeStation, TX, USA; †Department of VeterinaryIntegrative Biosciences, College of Veterinary Medicine,Texas A&M University, College Station, TX, USA;‡Independent Medical Entomologist, New York, NY,USA

Feline leishmaniasis is a rare parasitic disease that car-ries both therapeutic challenges and zoonotic concerns.This case report documents the clinical appearance,diagnosis and novel treatment of a cat from centralTexas with cutaneous leishmaniasis. The cat presentedwith a linear erosion on the right pinnal margin, anulcerated exophytic nodule on the right hock and aswelling in the right nostril. Cytological and histopatho-logical findings were consistent with leishmaniasis. PCRconfirmed the presence of Leishmania mexicana, a spe-cies endemic to Texas. An epidemiological investigationwas conducted by trapping sand flies from the cat’senvironment. Sand flies collected were identified as Lut-zomyia species, known vectors of Leishmania. Surgicalexcision of lesions is typically considered ideal, yet thelesion location in this case and the owner’s reluctanceto pursue invasive treatments deemed surgical removalimpractical. Given the lack of validated medical thera-pies for L. mexicana in cats, treatments typically pre-scribed for canine leishmaniasis were administered.Although allopurinol achieved clinical success, it wasdiscontinued owing to suspected drug-related neutrope-nia. Topical imiquimod also improved lesional skin yetwas unsustainable owing to application difficulty.Alternative therapies were explored including an anti-malarial Chinese herb known as artemisinin. Oraladministration of artemisinin resulted in nearly com-plete clinical resolution of cutaneous lesions withoutreported adverse events. The cat remained systemicallyhealthy with stable lesions nearly eight months after ini-tiation of artemisinin therapy. This case report demon-strates feline leishmaniasis in central Texas and

provides the clinician with therapeutic options for med-ical management.Source of funding: Self-funded.


P-75

Search for cat fleas with low sensitivity

against fipronil in Japan

Y. NAKAMURA* and T. FUKASE†

*Katsuragi Institute of Life Sciences, Tokyo, Japan;†Department of Veterinary Medicine, Faculty ofVeterinary Medicine, Okayama University of Science,Imabari, Japan

The phenylpyrazole-class compound fipronil has beenused widely as an insecticide. However, occurrence offipronil-resistant cat fleas is suspected. The presentstudy was done to find fipronil-resistant cat fleas inJapan. Cat fleas were collected from 10 dogs and 10cats every other year during 2010–2018. The fleas ofeach isolate were examined for their sensitivity againstfipronil through contact with the compound impreg-nated on round filter paper, diameter 9 cm, in of 1, 5,10, 50, 100 or 500 lg amounts. Knock-down of thefleas was observed over time for 24 h. Knock-down offleas from many isolates occurred dose-dependentlywith 1–500 lg fipronil. However, a small number of thefleas of some isolates showed relatively low-sensitivitywith no knock-down with 1–10 lg (one in 2010, two in2012, one in 2014, two in 2016 and three in 2018).However, even the fleas of low-sensitivity isolates werethought to be eliminated from body surfaces of dogsand cats after administration of a spot-on formulationof fipronil, based on the results of another study whichexamined the amounts of fipronil distributed on theskin of the medicated animals. Accordingly, a fipronilspot-on product should be considered still effectiveagainst cat fleas on dogs and cats in Japan.Source of funding: Self-funded.


P-76

Efficacy of some spot-on-formulated

ectoparasiticides against the ear mite

Otodectes cynotis in cats



Otodectes cynotis inhabits the external ear canal ofmany species including dogs, cats and ferrets. We eval-uated and compared the efficacy of some ectoparasiti-cides formulated as spot-on products against O. cynotisin cats. Thirty cats naturally infested by O. cynotis wererandomly assigned to five test groups of six animalseach; (i) an unmedicated control and four groups medi-cated with spot-on formulations of (ii) imidacloprid(10 mg/kg) + moxidectin (1 mg/kg), (iii) selamectin(6 mg/kg), (iv) selamectin (6 mg/kg) + sarolaner (1 mg/kg), and (v) fluralaner (40 mg/kg). Medications were

84 Abstracts



administered to cats six times at monthly intervals,except for the fluralaner product which was adminis-tered twice at three month intervals. The existence orabsence of mites in earwax was examined microscopi-cally at weekly intervals. The cats all were treatedunder the Guidelines for Proper Conduct of AnimalExperiments and Related Activities in Katsuragi Insti-tute of Life Sciences. Mites existed in all of the unmedi-cated control cats through the test period and haddisappeared completely from all cats of the imidaclo-prid + moxidectin- and the fluralaner-medicated groupsby three and two months after the first medication,respectively. In both the selamectin- and theselamectin + sarolaner-medicated groups, mites wereonly eliminated from three cats during the research per-iod. No adverse effects were observed in any of cats.The imidacloprid + moxidectin and fluralaner productswere considered to be much more effective againstO. cynotis in cats than the selamectin and theselamectin + sarolaner products.Source of funding: Self-funded.


P-77

Cat fleas with low sensitivity against

isoxazoline compounds in Japan



Some isoxazoline-class compounds such as afoxolanerhave been used recently as active ingredients in ectopar-asiticides for dogs and cats. We found a cat flea popu-lation showing low sensitivity against isoxazolines andexamined the drug sensitivity of this flea population.Two dogs, kept in the same house and infested by catfleas, were treated with an afoxolaner tablet at the rec-ommended dose and their flea burdens did notdecrease. A sarolaner tablet, subsequently administeredto the dogs at the recommended dose, also was ineffec-tive. In order to clarify the reason why these isoxazoli-nes were ineffective, we carried out two experimentsunder the Guidelines for Proper Conduct of AnimalExperiments and Related Activities in Katsuragi Insti-tute of Life Sciences. In the first experiment, anothertwo dogs that had previously been administered afipronil spot-on product were artificially infested by thefleas collected from the above-mentioned case dogs. Inthe second experiment, the two case dogs were cleanedby shampooing to remove all fleas and then artificiallyinfested by cat fleas collected from other dogs. In bothexperiments, the artificially infested fleas were con-firmed to be completely eliminated. These results meanthat the fleas on the case dogs were sensitive to fiproniland also that the blood of the case dogs containedafoxolaner and/or sarolaner. Accordingly, it was con-cluded that the cat fleas on the two case dogs showedlow sensitivity against isoxazoline compounds.Source of funding: Self-funded.


P-78

Pathological characterization of buffalo fly

lesions in Queensland cattle

M. N. NASEEM*, A. RAZA*, C.CONSTANTINOIU‡, M. MCGOWAN†, R.ALLAVENA†, A. TABOR* and P. JAMES**Queensland Alliance for Agriculture & Food Innovation(QAAFI), The University of Queensland, Australia;†School of Veterinary Science, The University ofQueensland, Australia; ‡College of Public Health,Medical & Vet Sciences, James Cook University,Queensland, Australia

Haematobia irritans exigua, commonly known as buf-falo fly (BF), is the major ectoparasite of cattle in trop-ical and subtropical areas of Australia. BF also cantransmit a filarial nematode, Stephanofilaria spp., whichhas been speculatively implicated in the aetiology of BFlesions. Lesions associated with BF infestation rangefrom raised, dry, alopecic, hyperkeratotic or scab-encrusted to variably haemorrhagic areas of ulcerationwhich also represent a major animal welfare concern.Lesions appear different in northern and southernQueensland cattle. This could be a consequence of dif-ferences in cattle breeds, differences in the prevalenceor epidemiology of the nematode, or differences in theecology of their vector. Skin samples were collectedfrom cattle slaughtered at abattoirs in northern andsouthern Queensland. Lesions were scored (0–4) macro-scopically for the presence of haemorrhage, ulceration,exudation, alopecia and skin thickness. Skin strips rep-resenting the centre, edge and peripheral normal tissue,were dissected from each lesion, preserved in 10% neu-tral buffered formalin and processed by paraffin sec-tioning and staining with haematoxylin & eosin.Microscopically, lesions were scored (0–4) for the epi-dermal and dermal damage, and inflammatory charac-teristics. This poster describes differences in the grossand microscopic pathology of BF lesions from northernand southern Queensland cattle, with and withoutStephanofilaria spp. present. The results will help toclarify the pathology and pathogenesis of BF lesionsand provide a basis for investigating genetic differencesbetween cattle in susceptibility to lesion development.Source of funding: Meat and Livestock Australia.


P-79

Pemphigus foliaceus-like drug reaction to

canine leishmaniosis treatment: a case report

H. L. PEREIRA, C. PELETEIRO and A. M.LOURENC�OCentre for Interdisciplinary Research in Animal Health,Faculty of Veterinary Medicine, University of Lisbon,Lisbon, Portugal

This case report describes a pemphigus foliaceus (PF)-like drug reaction to allopurinol and meglumine anti-moniate therapy in a dog treated for canine leishmanio-sis (CL). A 2-year-old, male neutered mongrel dogpresented with scaling alopecia, mild pruritus, weightloss, pale mucosae membranes and lymphadenomegaly.Laboratory findings included severe nonregenerative,normocytic and normochromic anaemia and leucocyto-sis with neutrophilia, hyperglobulinaemia without

Abstracts 85



hypoalbuminaemia, elevated urea and proteinuria.Leishmania ELISA test was positive for leishmaniosis(titre >1/320). Both allopurinol (10 mg/kg twice daily)and meglumine antimoniate (100 mg/kg once daily)were initiated. Five days later the dog started develop-ing dermatological signs compatible with a cutaneousadverse drug reaction. The patient presented febrilewith multiple pustules and vesicles in the abdominalregion, thighs and thorax, and also crusts and pustulesin both ears and on head. Cytological smear revealedneutrophils with acantholytic cells. Skin biopsy wasperformed and drugs were discontinued. Thehistopathological report described skin hyperplasia withspongiosis and subcorneal pustule formation containingacantholytic cells among hypersegmented nondegener-ated neutrophils. A similar pustule was identified in afollicular sheath. The histological changes were compat-ible with PF possibly triggered by an adverse drug reac-tion. Treatment for PF was started with a low dose ofprednisolone (1 mg/kg) due to leishmaniosis. Nucleo-tides and active hexose correlated compounds also wereinitiated for CL treatment. With minimal response afterseven days and a poor prognosis, owners choose toeuthanize the dog. To the best of our knowledge, this isthe first case report of a PF-like drug reaction to allop-urinol and meglumine antimoniate therapy in a dog.Source of funding: Self-funded.


P-80

The effectiveness of neem (Azadirachta indica)shampoo against dermatitis caused by brown

dog tick (Rhipicephalus sanguineus) in dog

N. I. SITEPUAnimal Teaching Hospital, Udayana University,Denpasar, Bali, Indonesia

Ectoparasites are a common cause of skin problems indogs; the brown dog tick (Rhipicephalus sanguineus) isthe most common cause in Bali, Indonesia. Treatmentwith synthetic chemical drugs may have harmfuladverse effects. The use of traditional medicine hasbecome an option in some parts of the world includingIndonesia. Neem leaves are used as a traditional medi-cine and allegedly have antiparasitic activity as theycontain sulfur, flavonoids and tannins. This study useddogs that presented to the Animal Teaching Hospital atUdayana University, Denpasar, Bali, Indonesia. Eachtreatment was replicated two times a week, over arange of two weeks to two months. The results showedthat in the first week and second week, neem shampoodid not have a significant result. This is because thepharmacokinetics of neem shampoo require a longertime to achieve its maximum effect. A significant effectof neem shampoo started to be seen in the third weekof treatment and in the fourth week, showed more sig-nificant results. Assessment criteria included observa-tion of skin problems and number of ticks found bycombing the dogs.Source of Funding: Self-funded.


86 Abstracts



EQUINE SKIN DISEASES

P-81

IL-31: a mediator of pruritus in horses

N. CRAIG, R. WILKES, N. S. MUNGUIA, M.DORR and R. MARSELLADepartment of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA

Interleukin (IL)-31 is a mediator of pruritus and mayhave a role in equine allergic reactions although nostudy has demonstrated that the injection of IL-31directly triggers pruritus in horses. Recombinant equineIL-31 was produced and its effect evaluated in fournormal horses. A dose-escalation study was done inject-ing intradermally either saline (0.2 mL) or increasingdoses of recombinant horse IL-31 (hIL-31) on separatedays (0.2 mL containing 3 µg or 5 µg, or 0.4 mL con-taining 10 µg hIL-31). Seconds spent itching at the siteof injection were recorded by live observers for 1 h pre-injection and 4 h post-injection. After saline injection,one horse itched for 2 s while the other three did not.After the lowest hIL-31 (3 ug, n = 1) dose, 6 s of itch-ing were observed at 180 min post-injection. After 5 ug(n = 1), 317 s of itching were recorded (peaks at 40 and180 min). After 10 ug (n = 2), one horse itched at theinjection site for 15 s and spent a longer time appearingirritated without rubbing the injection site, and theother horse itched for 79 s. Two peaks were detected(at 60 and 260 min for one horse, and 125 and 165 minfor the other horse). In conclusion, IL-31 can triggerpruritus in horses and its effect is not immediate. Theobserved delayed effect possibly is linked to the releaseof other mediators. Further studies are needed to inves-tigate the cascade of events triggered by IL-31 inhorses.Source of funding: Intramural grant competition, Univer-

sity of Florida.


P-82

Investigation on IL-31 levels and correlations

with pruritus in allergic horses

N. CRAIG, R. WILKES, N. S. MUNGUIA, M.DORR and R. MARSELLADepartment of Small Animal Clinical Sciences, Collegeof Veterinary Medicine, University of Florida,Gainesville, FL, USA

Interleukin (IL)-31 is a mediator of pruritus in humans,mice and dogs. Injection of recombinant equine IL-31has elicited pruritus yet it is unknown whether theseverity of pruritus in allergic horses is correlated tocirculating levels of IL-31 and whether leucocytes ofallergic horses intrinsically (in the absence of allergenicstimulation) release more IL-31 than normal horses. Inthis study, blood from 20 healthy and actively pruritic(n = 20) allergic horses was collected and peripheralblood mononuclear cells were isolated. Global pruritusVisual Analog Scale score (pVAS, 0–10, 10 most severe)was obtained for allergic horses. IL-31 mRNA fromlysated leucocytes was measured by quantitative PCR.IL-31 transcription was normalized to previously

established reference genes for equine leucocytes: glyc-eraldehyde-3P-dehydrogenase, hypoxanthine ribosyl-transferase and succinate dehydrogenase complexsubunit A. Differences in IL-31 transcription betweenhealthy and allergic horses were tested using a one-tailed Wilcoxon–Mann–Whitney U-test. The Spearmanrank correlation test was used to evaluate the correla-tion between pVAS and IL-31 transcription. No signifi-cant difference in IL-31 transcription was identifiedbetween groups (P = 0.48006) and no significant corre-lation was found between pVAS and IL-31 transcrip-tion in the cultured leucocytes (P = 0.87851). Inconclusion, although IL-31 plays a role in pruritus,transcription is no different in the absence of allergicstimulation between allergic and normal horses, andthus currently cannot be used as a disease severity mar-ker. Further studies are needed to evaluate cytokineresponse upon allergen stimulation in horses.Source of funding: Intramural grant competition, Univer-

sity of Florida.


P-83

Histological and molecular classification of

equine sarcoid: a pattern-based approach

G. TURA*, L. GALLINA*, F. SAVINI*, G.AVALLONE*, A. E. DURHAM†,‡, R. M. LARAGIONE† and B. BACCI**Department of Veterinary Medical Sciences, Universityof Bologna, Italy; †School of Veterinary Science,University of Surrey, UK; ‡Liphook Equine Hospital,Liphook, UK

Although the histological features of equine sarcoidsare well-characterized, there is no documented associa-tion between the clinical types and their histologicalfeatures. The study goal was to characterize the mor-phological spectrum of equine sarcoids and propose ahistological classification based on the growth patternin 112 sarcoids diagnosed in 86 horses from the UK.Clinical subtypes were as follows: nodular (n = 35), ver-rucous (16), fibroblastic (12) and occult (five). Caseswere classified histologically as verrucous (epidermalfinger-like projections), nodular (dermal nodule sepa-rated from the overlying epidermis) and mixed (combi-nation of both). Immunohistochemical (IHC)investigation with S100, a-SMA, p53 and PCR forbovine papillomavirus (BPV) 1, 2 and 13 were per-formed in all cases. Chromogenic in situ hybridization(CISH) was performed in 44 cases. Tumours were clas-sified histologically as verrucous (n = 25), nodular (32)and mixed (55), and further categorized based onfibroblasts architecture into interstitial (75), and fascic-ular (37). The following histological features were iden-tified: “picket fence” formation (58), hyperkeratosis(60), epidermal hyperplasia (53) and rete pegs (27).Myofibroblastic differentiation was confirmed by IHCfor a-SMA in 44 cases and positivity was subepithelial(13), intratumoural (23) or peritumoral (8). All caseswere negative for S-100 and p53. The presence of BPV-DNA was confirmed by PCR in 107 cases. Forty-twocases were positive for CISH in fibroblast nuclei and

Abstracts 87



cytoplasm, revealing strong signal intensity and diffusepattern. The proposed histological classification isintended to improve the diagnostic accuracy of equine

sarcoids. Future studies will aim at correlating this clas-sification with tumour behaviour.Source of funding: Self-funded.


88 Abstracts



SKIN DISEASES OF EXOTIC SPECIES

P-84

Efficacy of oral fluralaner for the treatment of

the Caparinia tripilis mite in African pygmy

hedgehogs

J. GUZMAN*, D. DACAK†, R. ORDO ~NEZ‡, M.LUCERO§ and R. FERREIRA¶

*Animal House Dermatolog�ıa y Spa, Cartagena,Colombia; †Cl�ınica Veterinaria Diego Dacak, Asunci�on,Paraguay; ‡Cl�ınica Hospital Animalopolis, Guayaquil,Ecuador; §Cl�ınica Veterinaria Tacuary, Asunci�on,Paraguay; ¶Dermatovet Cursos, Porto Alegre, RS,Brazil

Fluralaner is an orally administered isoxazoline thatprovides protection against fleas, ticks and mites indogs and cats; however, studies in African pygmyhedgehogs (Atelerix albiventris) are very limited. Thiscase series evaluates the efficacy and safety of fluralanerin hedgehogs with Caparinia tripilis. Ten registeredhedgehogs (four males, six females) were chosen, withages between five months and 1-year-old, and weightrange 225–440 g. The main clinical signs were intensepruritus (100%), seborrhoea (100%), erythema (90%),scabs (60%) and loss of spikes (60%). Caparinia tripilisinfestation was confirmed in all patients by taking skinsamples with scotch tape and direct observation undera microscope. All hedgehogs were treated with a single-dose oral fluralaner at 25–56 mg/kg (Bravecto, MSDAnimal Health; Beaucouz�e, France). The response totreatment was monitored by dermatological examina-tion and sampling on day (D)7, D14, D21, D30, D60and D90. All animals showed decreased pruritus andimproved lesions from D1 onward. On D7 and D14dead mites were observed microscopically; at D21,D30, D60 and D90 no mites were observed. No adverseeffects of the medication were observed. The presentstudy confirms the efficacy of fluralaner in hedgehogsinfected with C. tripilis.Source of funding: Self-funded.


P-85

Use of afoxolaner on two Burmese pythons

(Python molurus bivittatus) with a snake mite

(Ophionyssus natricis) infestation

C. ROMERO*, B. A. FUANTOS†, G. SHEINBERG§,A. CORDERO‡ and R. HEREDIA**Dermavet, Iztapalapa, Mexico; †Fauna VeterinaryClinic, Nuevo Leon, Mexico; §Centro VeterinarioMexico, Mexico City, Mexico; ‡Vetderm,Guadalajarara, Mexico

Ophionyssus natricis is considered an important parasitein reptiles that has zoonotic potential. The use of isoxa-zolines in reptiles has not been reported. This studyobjective was to evaluate the efficacy of afoxolaner(NexGard, Boehringer Ingelheim; Duluth, GA, USA)in two Burmese pythons (Python molurus bivittatus)

with O. natricis infestation. We reviewed two specimensfrom a private collection: an albino male and female,both 2-year-old, weighing 1,139 and 1,694 g, respec-tively. Both animals presented poor quality scales,mobile black spots, depression, moulting difficulty andloss of appetite. A direct collection of mites was per-formed with a cotton swab and acetate tape. Morpho-logical characteristics observed were identifiedas O. natricis. Both patients were medicated by orogas-tric tube with afoxolaner 2 mg/kg. A 28.3 mg tabletwas macerated in a ceramic mortar with 20 mL oforganic coconut oil, and the correct liquid dose wasadministered. Observations were made on day (D)1,D3, D10 and D30 post-medication. On D3 ectopara-sites were no longer found on the snakes, and deadectoparasites were observed in the enclosures. No miteswere observed on D10, dead ectoparasites were foundin one of the enclosures on D20, and no mites werepresent on the snakes or in the environments by D30.No adverse events were reported in either animal. Asingle oral dose of 2 mg/kg afoxolaner was effective forthe treatment of O. natricis in these Burmese pythons.Source of funding: Self-funded.


P-86

Efficacy of oral sarolaner for treatment of

sarcoptic mange in three naturally infested

rabbits: a case report

R. VENTURO*, J. BAJWA†, A. ARCILA‡ and C.ROMERO¶

*Private Practice, Lima, Peru; †VetDERM Clinic –Veterinary Dermatology & Ear Referral Medical Clinic,Vancouver, Canada; ‡Private Practice, Lima, Peru;¶Veterinary hospital DERMAVET, Mexico City,Mexico

Sarcoptes scabiei infestation is uncommon in rabbits,yet this parasite has zoonotic potential and increasingnumbers of cases have been reported recently. Macro-cyclic lactones have proven effective against S. scabieiinfestation in rabbits. We report the effectiveness oforal sarolaner (Simparica Chewable Tablets, Zoetis;Parsippany, NJ, USA) treatment in three unrelated rab-bits, naturally infested with sarcoptic mange. Theaffected rabbits were all intact male rabbits thatbelonged to different owners and were diagnosed withS. scabiei upon microscopic identification by superficialskin scraping. Presenting clinical signs included intensepruritus (3/3), ear pinna scabs (3/3), nasal hyperkerato-sis (2/3), paronychia (3/3), periocular scaling (1/3),anorexia (1/3) and alopecia (3/3). Cohabiting animalsincluded dogs (2/3) and a cat (1/3). Owners also haddermatological lesions (2/3). The rabbits were treatedwith sarolaner from 4 to 4.7 mg/kg per os once, result-ing in the resolution of clinical signs within three weeks.All three rabbits relapsed on days 50, 53 and 56,respectively, after the single sarolaner dose administra-tion. The three rabbits then were treated with two doses

Abstracts 89



of sarolaner p.o. administered 30 days apart, resultingin resolution of clinical signs within two weeks withoutrecurrence for ≥11 months. The oral sarolaner formula-tion labelled for dogs was considered palatable to allthree rabbits. No patient exhibited adverse reactions tothe medication. Based on this pilot report, sarolaner is

effective for treatment of S. scabiei in rabbits, at doserange 4–4.7 mg/kg p.o. administered twice, 30 daysapart.Source of funding: Self-funded.


90 Abstracts



NEOPLASTIC SKIN DISEASES

P-87

Nonepitheliotropic cutaneous lymphoma of

immunophenotype T in a domestic feline: a

case report

M. ANDRADE*, H. SOUZA†, D. ARAUJO‡, T.COSTA‡, F. PAIVA‡, S. SILVA*, C. BOTELHO‡ andJ. FERNANDES†

*Autonomous Veterinary, RJ, Brazil; †Post GraduationProgram, Universidade Federal Rural do Rio de Janeiro,RJ, Brazil; ‡Department of Veterinary Science,Universidade Federal Rural de Rio de Janeiro, RJ,Brazil

Lymphoma is the most common malignant neoplasmin cats; however, cutaneous involvement occurs in only0.2–1.7% of affected animals. Cutaneous lymphomamay occur in the epitheliotropic or nonepitheliotropicforms. Epitheliotropic lymphoma is mostly comprises Tlymphocytes and the nonepitheliotropic form usuallycomprises B lymphocytes. This report describes a caseof nonepitheliotropic T-immunophenotype lymphomain a cat in Brazil. The animal presented with multiple,nodular, ulcerated lesions on the right flank and innerright thigh, reporting a one-month evolution, associatedwith systemic clinical signs of hyporexia, weight lossand prostration for three months. Histopathologicalfindings from biopsies from the affected areas werecompatible with cutaneous lymphoma. The treatmentwas based on chemotherapy protocols, starting withLomustine as sole agent, later changing to CHOP pro-tocol, using vincristine sulfate, doxorubicin, cyclophos-phamide and prednisolone. A partial response wasachieved, with improvement of the condition up to thebeginning of the second protocol cycle. After a periodof absence of the tumour, a severe worsening of thegeneral state of the animal and the appearance of anew lesion was observed and the animal was eutha-nized. Subsequently, immunohistochemical evaluationconfirmed nonepitheliotropic cutaneous lymphoma andidentified immunophenotype T cells. With a small num-ber of published reports, the treatment represents achallenge and has no clear recommendations. The pro-tocol generally described with the longest remissiontime is the Madison–Wisconsin protocol, which isadapted to the CHOP protocol used for this cat.Source of funding: This study was financed in part by

Coordenac�~ao de Aperfeic�oamento de Pessoal de N�ıvelSuperior-Brazil.


P-88

A case report on nonseminomatous malignant

germ cell testicular tumour induced dermatitis

M. AWALEPet Wellness Clinic, Pimple Saudagar, Pune,Maharashtra, India

A 9-year-old male Labrador retriever with a history ofprevious cryptorchidism and castration, was presentedwith inability to rise, joint pain, erythematous and mul-tifocal alopecia, crusts, scaling, mild pruritus, hyperpig-mentation, pendulous prepuce and gynaecomastia.

Complete blood count and biochemistry test resultswere normal. Skin scraping, tape test, hair pluck andWood’s lamp examination were done. Skin scrapingand tape test revealed Demodex canis and cocci;Wood’s lamp examination was negative and KOHmicroscopy was positive for Trichophyton spp. Initially,we stabilized the dog with carprofen and Cosequin HA(Bioiberica; Barcelona, Spain); primary treatmentincluded antibiotics, and ketoconazole and ivermectinper os. However, there was minimal improvement.Hence, abdominal ultrasound was suggested to searchfor any underlying cause, although a recent ultrasoundand magnetic resonance imaging performed by the pre-vious veterinarian did not identify any abnormalities.The new abdominal ultrasound revealed two pelvicmasses. Exploratory laparotomy was performed. A tes-ticular tumour was excised and the second mass wasfound to be prostatic tumour for which finasteride p.o.was recommended. Biopsy of the testes confirmed anonseminomatous malignant germ cell tumour. Post-surgery, the dog showed remarkable improvement inskin and hair coat as well as overall health. This caseillustrates a possible relationship between dermatitis,demodicosis and a testicular tumour in older dogs.Source of funding: Self-funded.


P-89

Intratumoural treatment of a high-grade

perianal mast cell tumour with the novel small

molecule tigilanol tiglate (EBC-46).

G. K. BROWN, J. E. CAMPBELL, P. D. JONES, T.R. DERIDDER and P. W. REDDELLQBiotics Group Limited, Yungaburra, Queensland,Australia

A 12-year-old Staffordshire bull terrier was cytologi-cally diagnosed with a high-grade perianal mast celltumour. This tumour is associated with a poor progno-sis and surgical excision was likely to result in anincomplete excision with a high risk of local recurrence.The main objective was to treat this patient using intra-tumoural tigilanol tiglate (1 mg/mL in buffered 40%propylene glycol). Two injections were administered 32days apart to locally treat the tumour. On treatmentday, the modified ellipsoid method was used to calcu-late tumour volume which was then used to determinetigilanol tiglate dose (50% v/v; 0.5 mg/cm3). Efficacywas assessed on the basis of presence/absence of a com-plete response (full tumour resolution) at day (D)28and (D)60 using the Response Evaluation Criteria inSolid Tumours (RECIST) developed by the VeterinaryCooperative Oncology Group (VCOG). Before treat-ment, the tumour was 2.7 cm in diameter and had atumour volume of 5.1cm3. A wound with a maximumsurface area of 3.6 cm2 had formed by D7. At D28 apartial response was recorded and a tumour volume of0.6 cm3, a volume reduction of 87%. On D32, a secondtreatment was administered and a wound had formedwith a maximum surface area of 6.3 cm2 four dayslater. By D60 after first treatment, wound healing wascomplete and a complete response was recorded. This

Abstracts 91



case report highlights the treatment of an intractabletumour on a patient with an otherwise poor prognosisusing intratumourally administered tigilanol tiglate.Name and reference number of ethics overview commit-

tee: AEC Queensland, CA 2018-08-1223.

Source of funding: QBiotics Group Limited.

Conflict of interest: GKB, JEC, PDJ, TRD and PWR

are employed by QBiotics Group Limited.

P-90

A preliminary study of intratumoural

administration of novel small molecule tigilanol

tiglate (EBC-46) for treatment of canine soft

tissue sarcomas

G. K. BROWN, J. E. CAMPBELL, P. D. JONES, T.R. DERIDDER and P. W. REDDELLQBiotics Group Limited, Yungaburra, Queensland,Australia

The primary treatment option for soft tissue sarcomasis surgical resection, yet this can be hampered by thesize and location of the tumour. The main objective ofthis study was to provide an assessment of the safetyand efficacy of intratumoural tigilanol tiglate (1 mg/mLin buffered 40% propylene glycol) for the treatment ofcanine soft tissue sarcomas. This study was conductedunder permit from the Australian pesticides and veteri-nary medicines authority. On the day of treatment,tumour volume was determined using the modifiedellipsoidal method and tigilanol tiglate dosing was cal-culated based on this tumour volume (40% v/v;0.4 mg/cm3). The required volume was drawn up into aLuer lock syringe and injected in a fractionated manneras evenly as possible throughout the tumour. Dogswere excluded if the required tigilanol tiglate doseexceeded 0.25 mg/kg bodyweight or was >5 mg.Twenty-two dogs were treated and efficacy was assessedon the basis of presence/absence of a complete response(full tumour resolution) at day (D)28 and D42 usingthe Response Evaluation Criteria in Solid Tumours(RECIST) developed by the Veterinary CooperativeOncology Group (VCOG). A second tigilanol tiglatetreatment was administered if a complete response wasnot achieved. Sixty-eight percent (15/22) of dogsachieved a complete response by D42. A second injec-tion improved overall efficacy to 82% (18/22 dogs).This preliminary study demonstrated that tigilanoltiglate was well tolerated with good efficacy for localtreatment of soft tissue sarcomas.Source of funding: QBiotics Group Ltd.

Conflict of interest: GKB, JEC, PDJ, TRD and PWR

are employed by QBiotics Group Ltd.

P-91

Leucotrichia of epitheliotropic lymphoma in

five dogs

T. NARITA*, K. SHIBATA*, K. IYORI† and N.MURAYAMA‡

*DVMs Animal Medical Center Yokohama, Kanagawa,Japan; †Vet Derm Tokyo, Kanagawa, Japan;‡Dermatology Services for Dogs and Cats, Tokyo, Japan

Canine epitheliotropic lymphoma has a poor prognosisand consists of several types of dermatological lesionsincluding erythema, plaques, scaling, nodulation, ulcer-ation, alopecia and depigmentation of skin. Leu-cotrichia, not currently a clinical feature ofepitheliotropic lymphoma, is caused by a variety ofmetabolic and inflammatory diseases and has beenreported in association with vitiligo, alopecia areata,lymphocytic folliculitis, post-inflammatory folliculitisand drug reactions. In this study, we observed leu-cotrichia in five dogs with epitheliotropic lymphoma.The dogs were patients of three referral hospitals andseen between 2016 and 2017. The breeds were twoWelsh corgis, two mixed breeds and a miniature dachs-hund. The ages at onset were 6–14 years (mean 8years). The dogs had several typical skin lesions ofepitheliotropic lymphoma including erythema, scaling,alopecia and nodules. In addition, all dogs had macularleucotrichia, which appeared especially on the muzzle,extremities and trunk. Histopathological findingsrevealed neoplastic lymphocytes infiltrating the epider-mis and adnexal structures, follicular walls (five) and anodular infiltration of the dermis (one). There were noobservations of infiltration into anagen hair bulbs asseen in alopecia areata. Immunohistochemical evalua-tion of neoplastic cells revealed CD3-positive T cells intwo dogs. Although the mechanism underlying thedevelopment of leucotrichia remains unknown, it wasexpected that the regrowth of hair could turn white atthe site of early lesions in which epitheliotropic lym-phoma cells infiltrated into the hair follicle. Therefore,it is suggested that leucotrichia could be one of the clin-ical features of epitheliotropic lymphoma.Source of funding: Self-funded.


P-92

Linear arrector pili muscle hamartoma on the

tail of a Sphynx cat

M. ORLANDI*, N. FANTON† and F. ABRAMO‡

*Laboratorio MyLav-La Vallonea, Passirana di Rho(MI), Italy; †Clinica Veterinaria Tibaldi, Milano, Italy;‡Department of Veterinary Sciences, University of Pisa,Pisa, Italy

Arrector pili muscles are specialized adnexal structuresmade of smooth muscle, rarely involved by disease;piloleyomioma and piloleyomiosarcoma are the uniqueexamples of tumours of these muscles in cats. Arrectorpili muscle hamartoma is described in humans andrarely in dogs. The aim of this presentation is todescribe clinical and microscopic features of an arrectorpili muscle hamartoma in a cat. A 2-year-old male Sph-ynx cat presented with nonpruritic, nonpainful tail nod-ules, noticed one month previously. Clinically, seven

92 Abstracts



5 mm, alopecic, dermal nodules were arranged linearlyon the dorsal surface of the tail, surrounded by hyper-trichotic skin. FNA cytological evaluation showed aninflammatory process. No therapy was instituted. Aftertwo months, two new small nodules were observed.Surgical excision of three nodules was performed undergeneral anaesthesia and submitted for histopathologicalevaluation. Histologically, unencapsulated proliferationof mature smooth muscle cells, organized in well-defined, randomly oriented and abnormally large bun-dles was seen, expanding pre-existent appendages andsurrounded by lymphocytes. There was no evidence ofcytological atypia or mitoses within the muscle compo-nents. The diagnosis was arrector pili muscle hamar-toma. The pseudo-Darier sign, characteristic of musclehamartoma, was negative. Two weeks later, biopsy siteshad healed uneventfully and the other nodules hadundergone progressive reduction, yet hypertrichosis per-sisted. We hypothesize an origin from arrector pili mus-cle because of the anatomical relationship with hairfollicles. Linear distribution of arrector pili hamartomasand associated hypertrichosis have been described inhumans. To the best of our knowledge, this is the firstcase reported in a cat.Source of funding: Self-funded.


P-93

Use of toceranib phosphate as an adjuvant

treatment for squamous cell carcinoma: a case

report

R. RAMADINHA, J. ALMEIDA and D. CAICOARAUJOClinical and Surgery Department - Rural FederalUniversity of Rio de Janeiro, Seropedica, RJ, Brazil

The authors describe the use of toceranib phosphate(Palladia, Zoetis Animal Health; Parsippany, NJ, USA)as an adjuvant therapy for squamous cell carcinoma ofa 12-year-old female pitt bull dog. The tumour startedas multiple small and scaly nodules on the abdomenand pelvic limbs. The dog had several surgical excisionsof nodules and was treated with topical imiquimod. Alarger tumour in the inguinal region grew and had anextensive surgical procedure. Subsequently, treatmentwith carboplatin (230 mg/m2) was done with noresponse followed by radiation therapy (three fractionsof 8 Gy each) that controlled the neoplasia for a shorttime. In June 2019, the authors decided to initiate anexperimental and palliative therapy with intralesionaltoceranib phosphate (2.75 mg/kg) every 48 h. Thegrowth of the neoplasia slowed and the inguinaltumour reduced in volume by approximately 50%. Thedog was still in treatment, at the time of writing withreduced doses (2.50 mg/kg) and palliative control ofthe neoplasia. Normochromic normocytic anaemia wasobserved, and also episodes with diarrhoea occurredfour and eight months after starting therapy. The dogrecovered after treatment was suspended for two weeks

each time. Because of the good results, the authors sug-gest that further studies with toceranib phosphate as anadjuvant treatment in squamous carcinomas should bedone.Source of funding: Self-funded.


P-94

Multicentric squamous cell carcinoma in situin a dog resembling Bowen’s disease

B. L. SANZ RESSEL*,†, M. J. DUARTE‡, G. C.BROGLIA¶,§, C. G. BARBEITO*,† and A. R.MASSONE***Laboratorio de Histolog�ıa y Embriolog�ıa Descriptiva,Experimental y Comparada (LHYEDEC), Facultad deCiencias Veterinarias, Universidad Nacional de La Plata,La Plata, Buenos Aires, Argentina; †Facultad deCiencias Veterinarias, Consejo Nacional deInvestigaciones Cient�ıficas y T�ecnicas (CONICET),Universidad Nacional de La Plata, La Plata, BuenosAires, Argentina; ‡Hospital Escuela de Peque~nosAnimales, �Area de Cl�ınica M�edica en Peque~nosAnimales, Facultad de Ciencias Veterinarias, Universidadde Buenos Aires, Buenos Aires, Argentina; ¶HospitalEscuela de Peque~nos Animales, Facultad de CienciasVeterinarias, Universidad Nacional de La Plata, LaPlata, Buenos Aires, Argentina; §C�atedra deEnfermedades de Caninos y Felinos Dom�esticos,Facultad de Ciencias Veterinarias, Universidad Nacionalde La Plata, La Plata, Buenos Aires, Argentina;**Laboratorio de Patolog�ıa Especial Veterinaria Dr.Bernardo Epstein, Facultad de Ciencias Veterinarias,Universidad Nacional de La Plata, La Plata, BuenosAires, Argentina

Multicentric squamous cell carcinoma in situ (MSCCIS)is very uncommon in dogs, with only one case reportedin the veterinary literature. We describe an 8-year-oldfemale mixed-breed dog that developed multifocal well-circumscribed, pigmented, crusted, proliferative or ero-sive cutaneous lesions, measuring 0.5–3 cm in diameter.Lesions were found in pigmented and nonpigmentedskin. The most frequently affected areas were dorsalthorax, abdomen and proximal limbs. Metastasis wasnot observed. The dog was otherwise healthy.Histopathologically, lesions were characterized by well-circumscribed areas of neoplastic keratinocytes withoutinvasion through the basement membrane into the der-mis. Irregular epidermal and superficial follicularhyperplasia and dysplasia were observed. Moderateparakeratotic hyperkeratosis also was detected. Lesionson pigmented areas showed marked hyperpigmentationof the epidermis. Papillomavirus-associated cytopathiceffects in the keratinocytes were not observed. The casereported here is the second reported case of MSCCISin a dog with clinical and histological features ofBowen’s disease.Source of funding: Self-funded.


Abstracts 93



MISCELLANEOUS DISEASES

P-95

Mini review and update of lethal

acrodermatitis of bull terriers

N. A. MCEWANUniversity of Liverpool, School of Veterinary Science,Liverpool, Cheshire, UK

Lethal acrodermatitis (LAD) is an autosomal recessivegenetic condition in bull terriers. The objectives of thispaper were to give an overview of LAD and to informthat a DNA test is available. The first cases of LAD inbull terriers were reported in 1986. The largest study of28 cases was reported by McEwan, who discovered lowimmunoglobulin (Ig)A levels in these cases and pro-posed that LAD was hereditary and immunosuppres-sive. LAD causes growth stunting with most cases onlyachieving half their body size. The skin lesions consistof crusting and hyperkeratosis mainly over pressurepoints such as the elbows and hocks, the paws aresplayed with hyperkeratosis of the foot pads and oldercases have deformed claws. Bacterial and yeast skininfections (including Candida spp.) and upper respira-tory disease with nasal discharge are common withLAD. The clinical signs and histopathological confir-mation of diffuse parakeratosis suggest zinc deficiency,and the disease was compared to lethal trait A46 of cat-tle and acrodermatitis enteropathica of humans. Bothof these conditions are responsive to zinc, yet attemptsto treat LAD with zinc have all failed, including byperitoneal injections. Recently a DNA test for the genesplicing defect of MKLN1 has become available forLAD (Animal Health Trust; Lanwades Park, Kentford,Newmarket, UK). In conclusion, LAD in bull terriersis caused by a MKLN1 gene splicing defect and a DNAtest is now available.Source of funding: University of Liverpool, Liverpool,

UK.


P-96

Clamminess in Yorkshire terriers caused by

perspiration

Y. OSHIMA-TERADA, K. MORI and N.MURAYAMADermatology Services for Dogs and Cats, Tokyo, Japan

Few studies have been reported on primary hyperhidro-sis in dogs. The aim of this study was to report clammi-ness on the trunk which may occur as a primarydisease in Yorkshire terriers. Three pure-bred Yorkshireterriers and one Yorkshire terrier mix were presentedwith clamminess on the trunk with pruritus. Three dogsalso showed papules and erythema on trunk caused bysuperficial pyoderma, and 2/3 had recurrent pyoderma.Pruritus and clamminess were persistent whether or notthe pyoderma existed. The age when the clinical signsfirst became apparent ranged from six months to 2years. The mean pH of the skin on the dorsal thorax ofall four dogs ranged from 9.0 to 9.7. Histopathologicalresults showed sebaceous gland hyperplasia and dilatedapocrine glands. Two dogs were treated with oralantibiotics and topical 2% chlorhexidine acetate

(Nolvasan Surgical Scrub, Fort Dodge Animal Health;Fort Dodge, IA, USA) every other day. Another dogwas treated with the same topical 2% chlorhexidineacetate twice weekly. The dog without pyoderma wasgiven a moisturizing shampoo twice weekly (Afloat VetMoisturize Foam, QIX Corporation; Tokyo, Japan).All dogs’ pruritus and clamminess were resolved bythese treatments, with no anti-inflammatory agents nec-essary. Hyperhidrotic skin has been reported to have ahigh pH and the clamminess seen in this study wasresolved without using anti-seborrhoeic shampoo. Inconclusion, the clamminess of the skin of Yorkshire ter-riers may occur as a primary disease and may be moreattributed to hyperhidrosis than seborrhoea.Source of funding: Self-funded.


P-97

Partial and full-thickness burns in three dogs

from an electric heating pad

V. BALAZS*, A. L�OPEZ† and B. RIVERA**Oftaderm, Cl�ınica especializada en Oftalmolog�ıa yDermatolog�ıa Veterinaria, Santiago, Chile; †CentroVetera-Terapis, Temuco, IX Regi�on, Chile

Prolonged exposure to electric heating pads in veteri-nary clinics frequently causes thermal injuries in smallanimals. Three cases are described. A 1-year-old maleGerman shepherd doe (Case 1) was anaesthetized forgastrointestinal surgery with a heating pad under theback. On Day 7 post-surgery, the dog showed a largearea of devitalized skin on the dorsal trunk that pro-gressed to well-demarcated serpiginous ulcerations withcentral necrosis. These partial thickness lesions compro-mised 10–20% of the body surface. Treatment was suc-cessful with Clindamycin 11 mg/kg per os every 12 hand a topical product made of a combination of honeyand essential oils (Centro-Vetera; Temuco,Chile). A 1-year-old female mixed-breed dog (Case 2) presentedeight days after ovariohysterectomy with demarcatednecrotic lesions on the right flank surrounded by ery-thematous skin. The dog had been kept on lateralrecumbency on a heating pad after surgery. Necroticskin was resected and subcutaneous tissue approxi-mated with sutures. Partial-thickness lesions compro-mised 10% of the total body surface. Healing wasachieved using DuoDERM Hydroactive Gel (Uru-farma; Montevideo, Uruguay). A 6-month-old femalePomeranian (Case 3) presented 14 days after an ovario-hysterectomy with necrotic lesions on both flanks, dor-sum, and under the neck. The dog had been kept on aheating pad for a long time after surgery. Full-thicknessinjuries affected 20–30% of the body surface. The dogdied due to systemic decompensation. The use of elec-tric thermal pads in veterinary clinics can lead to severeinjury and should be avoided.Source of funding: Self-funded.


94 Abstracts



P-98

Sebaceous adenitis: effectiveness of ciclosporin

treatment, regardless of disease stage

A. BLANCO*, M. LOIZA*, A. WOLBERG*, E.TONELLI*, M. DUARTE*, M. SCARPA* and A.DUCHENE†

*Dermatology, Small Animals Hospital, School ofVeterinary, University of Buenos Aires, Argentina;†Pathobiology Service, Small Animals Hospital, Schoolof Veterinary, University of Buenos Aires, Argentina

There are different therapeutic options for the long-term management of sebaceous adenitis (SA) in dogswith variable results. This purpose of this study was todemonstrate the effectiveness of ciclosporin regardlessof the stage of the disease. This is a retrospectivereview of dogs with biopsy-confirmed sebaceous adeni-tis treated with ciclosporin from January 2009 toDecember 2019. Eighteen dogs were included. The fol-lowing breeds were represented: akita inu (7), standardpoodle (3), chow chow (1), Lhasa apso (1), shih tzu (1),German shepherd dog (1) and four mixed-breeds (13males, five females). Ages ranged from 1 to 8 years.The predominant skin lesions were scales, follicularcasting and hypotrichosis. Three skin biopsies weretaken from each patient. Histopathological resultsshowed various stages of the disease, including absenceof the sebaceous glands in 10/18 cases (55.55%),inflammation surrounding the glands in 5/18 (27.77%)and both characteristics in 3/18 (16.66%). Treatmentwith ciclosporin was initiated at 5 mg/kg per os oncedaily for 90 days, accompanied by baths every fourdays with a sulfur and 2% salicylic acid shampoo (Der-mosedan champ�u antiseborreico, Inst. Dermatol�ogico;Buenos Aires, Argentina), and daily sprays with 50%propylene glycol. Thirteen cases (72%) showed disap-pearance of scales and follicular casting with hairregrowth, four cases (22%) showed reduction in scaleand follicular casting and partial hair regrowth, andone case (5%) had a partial response. It is concludedthat ciclosporin and topical therapy are highly effectivein the treatment of sebaceous adenitis, regardless of thedisease stage.Source of funding: Self-funded.


P-99

Cryosurgery in the treatment of cutaneous

horn associated with papillomatosis: a case

report

F. PAIVA*, M. SOUZA*, L. JUNIOR†, T. COSTA*,D. ARAUJO*, C. BOTELHO* and J. FERNANDES‡

*Post Graduation Program, Universidade Federal Ruraldo Rio de Janeiro, RJ, Brazil; †Autonomous Veterinary,RN, Brazil; ‡Department of Veterinary Science,Universidade Federal Rural de Rio de Janeiro, RJ,Brazil

Cutaneous horns are keratinized acellular formations,manifesting in association with other skin diseases. Theoccurrence is rare, with a frequency of 0.1% of skinbiopsies in dogs. This report describes a case of cuta-neous horn associated with papillomatosis treated withcryosurgery. A 5-year-old male mixed-breed dog was

presented with a keratinized horn measuring3.0 x 4.0 cm protruding from the snout, with multiplewarty papillomatosis lesions of the face and oral cavity.The animal was concomitantly diagnosed with Ehrlichiacanis and treated with doxycycline hydrochloride,5 mg/kg per os every 12 h for 30 days. After resolutionof this haemoparasitosis, under general anaesthesia theapical portion of the horn was resected and the base ofthe surgical wound was frozen, in two freezing cyclesapproximately 1 min each, with direct spray applica-tion. Cryosurgery was used on the other verrucouslesions, with two cycles of freezing. After a 14 dayrecovery, the cryosurgery was repeated on the remain-ing warty lesions and the remaining portion of the cuta-neous horn, in two freezing cycles similar to the firstapplication. After 10 days from the second session, alllesions were in complete remission. Six months afterthe procedure, the animal remained in complete remis-sion. The treatment typically indicated for cutaneoushorns is surgical excision; however, in this case, cryo-surgery was chosen because in the affected region surgi-cal access was difficult. In conclusion, cryotherapy maybe an effective tool for the management of cutaneoushorn secondary to papillomatosis if surgical removal isless desirable.Source of funding: This study was financed in part by

Coordenac�~ao de Aperfeic�oamento de Pessoal de N�ıvelSuperior-Brasil (CAPES).


P-100

Evaluation of a plant-based balm in preventing

pedal lesions on sled dogs

A. BOUVIER*, C. THORIN†, H. POULIQUEN‡ andP. BOURDEAU**Department of Clinical sciences, Veterinary School(Oniris), University of Nantes, Nantes, France;†Department of Management and statistics, VeterinarySchool (Oniris), University of Nantes, Nantes, France;‡Department of Biology, Pathology and NutritionSciences, Veterinary School (Oniris), University ofNantes, Nantes, France

Sled dogs suffer from painful pododermatitis duringthe training season. The objective of this study was toevaluate the efficacy of Dermoscent BIO BALM (Labo-ratoire Dermo Cosmetique Animale; Castres, France)on foot pads of sled dogs. Fifty-two dogs were includedin two groups: in A (22 dogs) all feet were treated; andin B (30 dogs), the treatment was applied on right footpads only (left legs as control), for a total of 148 footpads treated and 60 control. The balm was appliedbefore and after daily running (15–50 km) during 15consecutive days. Evaluation of erythema, abrasions orcracks on all foot pads was made twice daily on days(D)1–2 and D15 in Group A; on D1–2, 4–5, 8–9, 12–13and 15 in Group B. Unpaired Student’s t-test was usedfor statistical analysis. During the study, erythema/abrasions and cracks were detected in respectively 285/2,000 and 43/2,000 feet examined, representing inGroup B 7.3% of treated (versus 14.1% untreated) forerythema/abrasion and 0.5% of treated (versus 2.3%untreated) for cracks. The overall risk of erythema/abrasion was significantly lower with treatment [oddsratio (OR) = 0.2984, P = 1.35e-7] and this difference

Abstracts 95



increased with time, from D5 (OR = 0.0638,P = 0.00741) up to D15 (OR = 0.0231, P = 9.50e-7).The treatment also reduced cracks (OR = 0.1272,P = 0.00143). The balm was well-tolerated by dogs andalso reduced “snow balls”, a frequent cause for sleddogs’ foot irritations. Dermoscent BIO BALM appearseffective in limiting foot lesions on sled dogs duringintensive training.Source of funding: Self-funded. Laboratoire Dermo Cos-

metique Animale provided free product upon request by

authors and played no role in the study otherwise.


P-101

Healing effect of a plant-based balm on pedal

lesions on sled dogs

A. BOUVIER*, C. THORIN†, H. POULIQUEN‡ andP. BOURDEAU**Department of Clinical Sciences, Veterinary School(Oniris), University of Nantes, Nantes, France;†Department of Management and Statistics, VeterinarySchool (Oniris), University of Nantes, Nantes, France;‡Department of Biology, Pathology and NutritionSciences, Veterinary School (Oniris), University ofNantes, Nantes, France

Pododermatitis is a frequent cause of pain and lame-ness of sled dogs due to training conditions. Regardlessof protective boots that dogs wear to prevent lesiondeterioration, additional treatment is often necessary.The objective of this study was to evaluate the healingeffect of Dermoscent BIO BALM (Laboratoire DermoCosmetique Animale; Castres, France) on lesionsinduced during training. Thirty included dogs receivedan application of balm on right foot pads (left ones ascontrol) for a total of 60 treated and 60 untreated footpads. Dermoscent BIO BALM was applied before andafter daily running (15–50 km) during 15 consecutivedays and erythema/abrasion was evaluated on all footpads twice daily, before and after training, on days (D)1, D2, D4, D5, D8, D9, D12, D13 and D15. Whenbilateral lesions occurred on a dog on the same day(BL), the time for healing was compared between trea-ted and untreated pads (chi-squared test). Erythema/abrasion was detected on 16/60 treated pads and 19/60untreated pads among which (respectively) 93.7% (15/16) and 42.1% (8/19) were healed (P < 0.0015). Among15 BL dogs, 14/15 (93.3%) treated pads healed muchfaster than the nontreated ones. More than half of trea-ted pads (8/15, 53.3%) were healed within one day ofapplication versus 0 for untreated pads (P < 0.001).After D4, 11/15 (73.3%) of treated feet healed versus 6/15 (40%) of untreated feet (P < 0.067). DermoscentBIO BALM appears to be effective in aiding and

speeding the healing of feet during intensive training ofsled dogs.Source of funding: Self-funded. Laboratoire Dermo Cos-

metique Animale provided free product upon request by

authors and played no role in the study otherwise.


P-102

The use of interactive technology to enhance

learning in clinical veterinary dermatology

A. PUTRA*, W. GRAM*, C. STEFANOU*,† and D.SANTORO**Department of Small Animal Clinical Science,University of Florida, College of Veterinary Medicine,FL, USA; †Department of Family, Youth, andCommunity Sciences, University of Florida, College ofAgricultural and Life Sciences, FL, USA

Clinical teaching in veterinary medicine, including der-matology, is very challenging. In several disciplines,there is an increasing interest in the use of novel inter-active technology-based techniques, such as adaptivelearning, to provide students with more opportunitiesto learn. Few studies have evaluated the use of thistechnique in veterinary medicine. We hypothesized thatadaptive learning modules given during dermatologyrotation would improve students’ dermatology testscores, and students would regard these modules asbeneficial to their overall learning experience. Ten inter-active dermatology modules were developed using anadaptive learning platform. Using a quasi-experimentaldesign, incoming veterinary students to the dermatol-ogy rotation who agreed to participate were randomlyassigned to treatment (given the modules during therotation) or control group (given the modules after therotation ends). Students completed a pre-test two weeksbefore their rotation start date and a post-test aftertheir rotation end date. A questionnaire was developedto assess students’ learning experience using adaptivelearning modules. There was a statistically significantdifference (P = 0.019) in the post-test scores betweenstudents in the treatment and control groups. Studentsin both groups received a significant increase from pre-test to post-test (P < 0.001 for treatment group,P = 0.002 for control group). Students’ views regardingthe adaptive learning modules was mostly positive. Thisstudy shows that the adaptive learning platform is aneffective novel method to augment clinical teaching inveterinary dermatology. This study also indicates thatstudents perceive the use of adaptive learning modulesas being beneficial for their education.Source of funding: Self-funded.


96 Abstracts



RARE AND NOVEL CONDITIONS

P-103

First report of an epidermolytic foot pad

hyperkeratosis in a dog

R. GARC�IA-FONTICOBA*,†, L. ORDEIX*,†, M.CABR�E*,† and L. FERRER*,†

*Servei de Dermatologia, Fundaci�o Hospital Cl�ınicVeterinari, Universitat Aut�onoma de Barcelona,Barcelona, Spain; †Departament de Medicina i CirurgiaAnimals, Universitat Aut�onoma de Barcelona, Barcelona,Spain

Hereditary foot pad hyperkeratoses (HFH) represent agroup of canine genodermatoses most commonlydescribed in the dogue de Bordeaux, Irish terrier andKromfohrl€ander. They are defined histologically asnonepidermolytic hyperkeratosis. Mutations in theKRT16 (dogue de Bordeaux) and FAM83G (Irish ter-rier, Kromfhorl€ander) genes have been associated withthe disease. We report the first case of epidermolyticfoot pad hyperkeratosis in a dog. A 12-year-old neu-tered male, mixed-breed dog was presented with a his-tory of severe painful foot pad hyperkeratosis from sixmonths old. Thick papilliform projections and fissureswere observed in all foot pads. Histopathological exam-ination revealed severe papillated epidermal hyperplasiawith orthokeratotic hyperkeratosis, intense hypergranu-losis, focal dyskeratosis and epidermolysis. TheFAM83G and KRT16 gene mutations reported to beassociated with this disease were not detected in thiscase. Human epidermolytic palmoplantar hyperkerato-sis has been associated with mutation of the gene cod-ing for keratin 9 (KRT 9). We fully sequenced this geneand found six single nucleotide polymorphisms (SNPs),four of which cause amino acid change. However, noneof these SNPs seemed to explain sufficiently the pheno-type of the dog. This is the first report of epidermolyticfoot pad hyperkeratosis in a dog. Mutations in KRT16,FAM83G or KRT9 genes do not seem to be the causeof this phenotype.Source of funding: Self-funded.


P-104

Acquired perforating dermatosis in two French

bulldogs

R. GARC�IA-FONTICOBA*,†, P. J. SANCHO‡, D.FONDEVILA†, T. LIMA*,†, L. FERRER*,† and L.ORDEIX*,†

*Servei de Dermatologia, Fundaci�o Hospital Cl�ınicVeterinari, Universitat Aut�onoma de Barcelona,Barcelona, Spain; †Department of Animal Medicine andSurgery, Universitat Aut�onoma de Barcelona, Barcelona,Spain; ‡Cl�ınica Veterinaria Dres. Sancho. Servicio deDermatolog�ıa Veterinaria Dermasantboi. Sant Boi deLlobregat, Barcelona, Spain

Acquired perforating dermatosis (APD) is a group ofhuman skin disorders characterized by collagen extru-sion from the dermis. The pathogenesis is poorly under-stood and it is commonly associated with systemicdiseases. A similar clinical and histopathological condi-tion has been reported in cats. We describe APD for

the first time in the dog. Two geriatric, male Frenchbulldogs with pituitary-dependent hyperadrenocorticismand on trilostane treatment were presented with bilat-eral symmetrical alopecia and calcinosis cutis. More-over, multiple mildly pruritic, firm, reddish and tightlyadherent crusting papules were observed on the flank.The removal of this protruding material gave rise todeep ulcers. Histopathological results (haematoxylin &eosin and Masson’s trichrome stains) showed transepi-dermal extrusion of degenerated collagen fibres perpen-dicularly orientated and associated with an intenseneutrophilic inflammatory infiltrate, crusting and necro-tic debris. The histopathological findings were compati-ble with APD. The cause of these lesions remainunknown. In one case, trilostane administration wasdiscontinued and the lesions resolved spontaneouslyafter six weeks. The other dog was euthanized fourmonths after the diagnosis owing to seizures associatedwith the growth of the pituitary adenoma and rapidworsening of neurological signs. At the time of eutha-nasia the lesions were still present and had improved tosome extent. To the best of the authors’ knowledge,this is the first report of canine APD.Source of funding: Self-funded.


P-105

Use of topic growth factors and minoxidil

sulfate in a female dog with black hair

follicular dysplasia

K. GAVAZZONI*, L. MARRA†, E. FERRARINI‡

and V. KALTBACH§

*Veterinary Clinic Santa Lucia, RS, Brazil; †SmallAnimal Clinic and Surgery, Universidade Federal deGoi�as, GO, Brazil; ‡Pharmacy Farmec, Caxias do Sul,RS, Brazil; §Pharmacy Bella Salute, Flores da Cunha,RS, Brazil

Canine black hair follicular dysplasia (BHFD) is a raredisorder limited to the black coat regions affectingbicolour or tricolour dogs during the first few weeks oflife. This case report describes the use of growth factorsas a treatment modality for canine BHFD. A 5-year-old, spayed, mixed-breed dog developed alopecia ateight months of age. The dog was otherwise healthy.Histopathological evaluation of affected areas showedfollicular dysplasia and clumps of melanine in the hairfollicle canal and peri-follicular area compatible withBHFD. The dog was unsuccessfully treated with mela-tonin at 10 mg/day orally for three years. Five yearsafter the diagnosis the dog had complete dorsal alope-cia and dry skin. Trichography was performed andshowed macromelanoses obscuring the normal hairshaft architecture supportive of BHFD. A compoundedmoisturizing shampoo containing 2% colloidal oatmeal,2% hydroviton, 2% D-panthenol, 2% aloe vera, 2%Jaborandi tincture, 1% lactic acid and 0.2% pepper-mint essential oil was prescribed every seven days. Inaddition, a compounded spray formulation containing3% basic fibroblast growth factor (BFGF), 3% nanocopper peptide and 5% minoxidyl sulfate was appliedonce daily to the dorsal region. During 40 days of the

Abstracts 97



treatment, hair growth was noted covering the previ-ously alopecic areas. The haircoat was less dense andhad a different quality. Maintenance therapy with thecompounded formulations included weekly baths andthree times a week spray application. In conclusion, thecombination of BFGF, nano copper peptide andminoxidil should be considered as a treatment optionfor canine BHFD.Source of funding: Self-funded.


P-106

Nodular dermatofibrosis and cystic renal

disease in a beagle without FLCN mutation: a

case report

B. RIVERA*, V. BALAZS*, C. TORO† and S.STUBBS‡

*Oftaderm, Cl�ınica especializada en Oftalmolog�ıa yDermatolog�ıa Veterinaria, Santiago, Chile; †Departmentof Veterinary Medicine, University of Cambridge, UK;‡London School of Hygiene and Tropical Diseases,London, UK

Nodular dermatofibrosis (ND) is a rare syndrome ini-tially described in German shepherd dogs. The diseaseis characterized by the presence of multiple dermal andsubcutaneous nodules primarily affecting the legs, headand ears. It also is associated with polycystic kidneys,renal cystadenomas or cystadenocarcinomas, and lesscommonly with uterine tumours. This condition hasbeen linked to an autosomal dominant mode of inher-ence and to mutations of the gene encoding folliculin(FLCN). An 8-year-old, intact male beagle presentedwith several well-circumscribed skin nodules located onthe extremities, head and pinna that progressed over aperiod of two years. No other clinical signs were pre-sent and haematological as well as biochemical parame-ters were unremarkable. Abdominal ultrasound showedbilateral cortical renal cysts suggesting a degenerative/neo-proliferative inflammatory process. One of the cystsin the left kidney was large and it did not appear toaffect the renal parenchyma significantly. A PCR bloodtest did not reveal germline mutations in the FLCNgene. These findings correlate with other reported casesof ND. This dog has lived for two years since diagno-sis, exceeding by far the life expectancy found in casesof malignant renal cysts, suggesting the absence of neo-plasia. It is the first time that this disorder has beendescribed in beagles. The fact that FLCN mutationswere not detected suggests that alterations in differentgenes may be involved in this particular form of ND.Future studies are still required to determine the aetiol-ogy of this FLCN-independent nonmalignant ND, nowdescribed in beagles.Source of funding: Self-funded.

Conflict of interest: None declared

P-107

Idiopathic linear pustular acantholytic

dermatosis in a border collie

T. NUNES RODRIGUES and S. VANDENABEELESmall Animal Department, Faculty of VeterinaryMedicine, Ghent University, Merelbeke, Belgium

Idiopathic linear pustular acantholytic dermatosis(ILPAD) is a very rare condition with an unknownaetiology or predisposition. A 10-month-old, femalespayed border collie was presented with a six week his-tory of a pruritic linear raised solitary lesion. The lesioninvolved the fifth pad and partially the metacarpal pad,and extended midway between the carpal pad and theolecranon of the left forelimb. It consisted of coalescentcrusts. The affected paw pads presented with moderateerythema and erosion. Previous treatments with shortcourses of oral antibiotics, antifungals, anti-inflamma-tories and glucocorticoids did not improve the lesions.Cytological examination of impression smears of thelesion revealed high numbers of Malassezia yeast. Twopunch biopsies were taken for histopathological exami-nation. Topical treatment with orbifloxacin, mometa-sone furoate monohydrate and posaconazole (Posatex,Intervet; Boxmeer, the Netherlands) and oral ketocona-zole at 5 mg/kg/day was prescribed until the biopsyresults became available. Histopathological resultsshowed neutrophilic mural and luminal folliculitis withpronounced acantholytic cells, and epidermal exocytosisof neutrophils. There was a significant improvementwith topical medication after two weeks. Ketoconazolewas discontinued and oral prednisolone at 0.7 mg/kgonce a day was prescribed at the time of histopatholog-ical results. The lesions resolved after two weeks oftherapy. The prednisolone was weaned-off and topicaltreatment discontinued. The case remained controlledfor nearly three years with occasional use of topicaltherapy and short courses of prednisolone. In a similarway to the previous cases reported in the literature,ILPAD affects only one area of the body and respondsvery quickly to glucocorticoids.Source of funding: Self-funded.


P-108

P-1A case of ulcerative neutrophilic dermatitis

resembling human bromoderma in a dog

C. DEDOLA*, A. GALLUCCI†, F. IBBA‡, F.ENNAS§, I. PORCELLATO¶ and C.BRACHELENTE¶

*Centro Veterinario Ichnos, Cagliari, Italy; †VeterinaryCentre La Fenice - Neurology Section, Cagliari, Italy;‡Animal House Clinic, Capoterra, Italy; §KarelVeterinary Clinic, Cagliari, Italy; ¶Department ofVeterinary Medicine, University of Perugia, Perugia,Italy

Bromoderma is a cutaneous adverse reaction describedin humans and caused by ingestion, inhalation or directcontact with bromides. Lesions are characterized bypustular and acneiform eruptions as well as nodulesand ulcers affecting the face, scalp and limbs. A 2-year-old, female Maremma sheepdog was referred for thepresence of six months duration painful ulcers with

98 Abstracts



violaceous borders and multiple dermal and subcuta-neous haemorrhagic nodules on the bridge of the nose,the dorsal aspect of the front paws and on all four legs.Lesions had not responded to antibacterial andimmunomodulatory therapy. Nine months before, thedog had been diagnosed with suspected idiopathic epi-lepsy and treated with potassium bromide (KBr) eversince. Histopathological examination of lesions revealedan interstitial neutrophilic dermatitis extending multifo-cally to the subcutaneous tissue. All special stains werenegative for infectious agents, and owing to the lack oftropism for follicular structures as well as negative bac-terial and fungal cultures, a diagnosis of sterile neu-trophilic process similar to pyoderma gangrenosum wasmade. A cutaneous drug reaction to KBr was sus-pected, because it was the only continuous drug in thedog’s history before the first appearance of lesions.Potassium bromide was discontinued and after sixweeks the ulcerative lesions were completely healed.The present report describes a case of an ulcerativeneutrophilic dermatitis associated with KBr administra-tion in an epileptic dog, resembling human bromod-erma. Corticosteroid treatment was ineffective incontrolling skin lesions and only drug withdrawal ledto clinical resolution.Source of funding: Self-funded.


P-109

Skin fragility in a cat with an ACTH-

producing pituitary neuroendocrine tumour

(PitNET)

B. BRUNETTI*, L. V. MUSCATELLO*, F.CLEMENTE†, S. ASIOLI‡, B. BACCI§ and F.ABRAMO§

*Department of Veterinary Medical Sciences, Universityof Bologna, Italy; †Associate Veterinary Clinic “SanLuca”, Bologna, Italy; ‡Department of Biomedical andNeuromotor Sciences, University of Bologna, Italy;§Department of Veterinary Sciences, University of Pisa,Italy

Feline acquired skin fragility syndrome is a rare condi-tion with multiple underlying causes, including sponta-neous and iatrogenic hyperglucocorticism and diabetesmellitus amongst others. A 14-year-old, female Euro-pean cat was presented for severe sudden and diffuseskin lacerations, either spontaneous or induced byminor trauma. The cat had a two year history of glyco-suria and untreated polydipsia and polyuria. On clinicalexamination, it had poor physical condition, dehydra-tion, severe plantigrade stance, thin, fragile skin, andlacerations without skin hyperextensibility. The suspi-cion was of acquired skin fragility, and owing to thepoor condition of the patient and severity of skin dam-age, compassionate euthanasia was performed. Atnecropsy, the adrenal glands were both enlarged andthe pituitary gland had a tumour measuring10 x 10 x 12 mm. Histopathological examination of theskin revealed a very thin epidermis with a single layerof keratinocytes, atrophic dermis and sparse and thincollagen. Transmission electron microscopic investiga-tion showed no structural alterations of the collagen.Both the adrenal glands had cortical hyperplasia. In thepancreas, there was amyloidosis of the Langerhans

islets. The pituitary tumour showed a loss of normalreticulin stromal pattern and multifocal periodic acidSchiff-positive neoplastic cells. Mitoses were six per 10HPF. The tumour was PIT1-negative, TPIT- andACTH-positive. P53 was negative and Ki67 index was7%. According to human classifications, this was apituitary neuroendocrine tumour (PitNET) grade 1b, acorticotroph adenoma with high Ki67 labelling index(based on the WHO 2017 classification). This tumourcaused cortical hyperadrenocorticism, skin fragility anddiabetes mellitus.Source of funding: Self-funded.


P-110

Alopecia and suprasellar secreting neuro-

endocrine tumour in a dog

C. GIMENEZ-ACOSTA*, B. HUBERT*, K.MATIASEK†, L. BLOND*, C. BAUDIN*, F.DEGORCE-RUBIALES‡ and S. PIAZZA**Veterinary Hospital Medicine Languedocia,Montpellier, France; †Clinical & ComparativeNeuropathology Laboratory, Ludwig-Maximilians-University, Munich, Germany; ‡Laboratoire d’AnatomiePathologique du Sud-Ouest, Toulouse, France

Most of the hormone-active adenomas secrete prolactin(50%) and more rarely somatotropin (10%) or corti-cotropin (5%). Secretion from another tumour type,especially in case of suprasellar extension, has not beenreported previously. A 9-year-old, intact female beardedcollie was presented with a major alopecia, especiallyinvolving the trunk, neck and tail, and sparing thehead. The dog was diagnosed with hypothyroidism andtreatment with levothyroxine was unsuccessful. Anadrenocorticotropic hormone stimulation test was con-sistent with increased adrenal gland response suggestiveof hyperadrenocorticism. No clinical improvement wasvisible after oral trilostane treatment (1 mg/kg daily for30 days). Microscopic examination of skin biopsiesrevealed a severe atrophic dermatosis, lack of inflam-mation, an extremely thin dermis, marked orthokera-totic hyperkeratosis, follicular plugging, emptyfollicular ostia, atrophic sebaceous and apocrine glands,and the preponderance of telogen and kenogen follicles.An endocrinopathy was suspected. High-dose dexam-ethasone suppression testing was consistent with anadrenal secreting neoplasia or pituitary-dependenthyperadrenocorticism. Abdominal ultrasound revealedmild symmetrical increase of adrenal gland size andhepatomegaly. The clinical profile of this patient wasambiguous with no polyuria-polydipsia, no cutaneousor muscular calcinosis and normal alkaline phosphataseplasmatic activity. The dog developed neurological clin-ical signs and was euthanized. A pituitary mass wasconfirmed during necropsy as a suprasellar neuroen-docrine-type cell tumour from pars distalis and theadrenal glands were slightly hyperplastic. This casereport describes an unusual form of hyperadrenocorti-cism in a dog.Source of funding: Self-funded.


Abstracts 99



P-111

Skin necrosis in a cat likely caused by

subcutaneous injection of enrofloxacin

A. MIYA*, E. HIRATA†, N. MURAYAMA‡ and Y.OSHIMA-TERADA‡

*Okusawa Subaru Animal Clinic, Setagaya, Tokyo,Japan; †Misaki Animal Hospital, Suginami, Tokyo,Japan; ‡Dermatology Services for Dogs and Cats, Koto,Tokyo, Japan

Although there is one case report on injection sitenecrosis due to enrofloxacin, no detail was provided.The aim of this study was to report the histologicalexamination of the injection site after subcutaneousenrofloxacin caused injection site necrosis in a cat. A 2-year-old, neutered mixed-breed cat presented for ulcera-tions over the dorsal neck. The cat had been given aninjection of enrofloxacin (5 mg/kg subcutaneously oncedaily, Baytril 2.5% Injectable Solution, Bayer, Yaku-hin, Ltd.; Osaka, Japan) and prednisolone 1–2 mg/kgs.c. once daily (Prednisolone Injection Solution KS,Kyoritsu Seiyaku Corp.; Tokyo, Japan) in the dorsalneck over the course of two months as a treatment ofanaemia. Three weeks after these treatments, a skinlesion of 1 cm full-thickness ulceration was observed.However, at the time of presentation, ulceration(8 9 5 cm) had spread over the dorsal neck. Our histo-logical examination revealed full-thickness coagulationnecrosis of the epidermis, dermis and adnexa withextension into the panniculus. The lesion was almostcompletely resolved after nine months of simply rinsingthe affected area with normal saline solution andchanging the moist wound dressing every day. Basedon the medical history of and histological examinationon this cat, we have concluded that the high pH ofenrofloxacin (approximately 11) may have caused thefull-thickness coagulation necrosis in the cat.Source of funding: Self-funded.


P-112

Hyperoestrogenism, sertoli cell tumour and

pyometra in a male schnauzer with persistent

M€ullerian duct syndrome

G. MONTOYA*, E. ESCOBEDO*, S. PIZARRO†, G.SALAS† and R. RAMIREZ†

*Department of Dermatology, Scientific University ofthe South, School of Veterinary Medicine, Lima, Peru;†Department of Pathology, Scientific University of theSouth, School of Veterinary Medicine, Lima, Peru

Persistent M€ullerian duct syndrome is a rare form ofpseudohermaphroditism in male dogs, mainly describedin toy schnauzers. This report describes a case with thiscondition. A 10-year-old male schnauzer with gynaeco-mastia, comedones, linear erythema of the preputialskin, and only one descended testicle was sent for anultrasound examination, revealing the other testicle tobe abdominal and enlarged to 10 cm. Needle aspirationand cytological evaluation were suggestive of a Sertolicell tumour and so the dog was castrated. During sur-gery, it was observed that the neoplastic testicle wasnodular, whitish coloured with dark red areas and wascranially attached to a uterine horn that was slightly

enlarged and had a moderately thick wall. The otheruterine horn had a thin wall, and was expanded, filledwith purulent brown liquid. The histopathologicalresults of the neoplastic testicle revealed a parenchymaformed by many tubules, separated by a very densestroma. These had elongated and palisading cells, themajority with multiple nucleoli. The other testicle washypoplastic. In relation to the uterus, the largest hornhad a reduced mucosa and submucosa, with infiltrationof active neutrophils and macrophages, a thin muscularlayer, marked congestion and angiogenesis. The hornthat had the neoplastic testicle attached showed evi-dence of endometrial cystic hyperplasia along withinflammation. In conclusion, this is the first report inPeru of persistent M€ullerian duct syndrome with hyper-oestrogenism, sertoli cell tumour, endometrial cystichyperplasia and pyometra in a male schnauzer.Source of funding: Self-funded.


P-113

Post-clipping hair follicle arrest and

glucocorticoid-induced alopecia treated with

malt extracts made in presence of Rhizopusoryzae in dogs



In the treatment of one dog with post-clipping hair fol-licle arrest and one dog with glucocorticoid-inducedalopecia, we evaluated the efficacy of a product con-taining water extracts of the malt made in the presenceof Rhizopus oryzae (R&U 30, Ushikoshi Research Insti-tute for Physiology; Chiba, Japan). This product isknown to develop a regulatory effect on steroid hor-mones. The first case was a 13-year-old, castrated long-haired miniature dachshund, with body weight 6.7 kg,showing alopecia of the flanks and ventrum post-clip-ping. One tablet (120 mg) of the product was adminis-tered orally twice a day. Hair regrowth occurred by 40days after starting the medication. The second case wasa 13-year-old, castrated toy poodle, with body weight5.0 kg, showing alopecia in the right flank resultingfrom topical glucocorticoid use over three months. Onetablet of the tested product was administered orallyonce a day. After 28 days, the hair began to regrow,with the colour of the newly growing hair changing tobrown instead of cream. The hair regrowth in this caseis thought to have been achieved earlier than in thecase without treatment after stopping a topical gluco-corticoid. The tested product may be helpful for somekinds of canine alopecia.Source of funding: Self-funded.


100 Abstracts



P-114

Sensory neuropathy (acral mutilation

syndrome) in three miniature pinschers

L. ORDEIX*,†, T. LIMA‡ and L. FERRER*,†

*Servei de Dermatologia, Fundaci�o Hospital Cl�ınicVeterinari, Universitat Aut�onoma de Barcelona,Barcelona, Spain; †Departament de Medicina i CirurgiaAnimals, Universitat Aut�onoma de Barcelona, Barcelona,Spain; ‡Hospital Veterinari Canis, Girona, Spain

Hereditary sensory neuropathies are uncommonly diag-nosed in dogs and usually reported as “acral mutilationsyndromes” (AMS). Similar to humans, genetic causeshave been described. A point mutation in a lincRNAupstream of the GDNF gene has been associated withAMS in sporting dog breeds (pointers, English springerspaniel, French spaniel). Although AMS has beendescribed previously in one miniature pinscher, at thistime it is not known if the disease also has a geneticorigin in this breed. The present report describes threeadditional cases of AMS in miniature pinschers. Threeunrelated dogs, two females and one male miniaturepinscher were presented with intense licking and bitingof the hind limbs leading to severe self-mutilation ofseveral claws, digits and foot pads. The age of diseaseonset was between four and 15 months. Neurologicalexamination confirmed acral analgesia of the hind feetin all cases. Mutation of the lincRNA upstream of theGDNF gene was not found. One case was lost to fol-low-up after diagnosis, while the other two showed par-tial response to gabapentin and fluoxetine. Theminiature pinscher seems to be one of the breeds wherethis specific neuropathy occurs, although it is not asso-ciated with the mutation described for other breeds.Therefore, this breed may constitute an interestingmodel for the identification of new gene mutationscausing AMS in dogs and potentially for further insightinto human neuropathies as well.Source of funding: Self-funded.


P-115

Cloned alopecia X in a Pomeranian dog

S. DELEPORTE*, P. PRELAUD† and C. LAFFORT‡

*Dermatology department, Lorrainevet, Ludres, France;†Dermatology department, Advetia Centre HospitalierV�et�eriniare, V�elizy, France; ‡Dermatology department,Clinique V�et�erinaire Alliance, Bordeaux, France

Alopecia X is a frequent cause of alopecia in Pomera-nian dogs. The pathomechanism is still obscure and agenetic basis suspected. This report of a clonedPomeranian argues strongly for a genetic basis for thiscondition. A 2-year-old male Pomeranian was presentedearly 2014 with alopecia X that had been progressingover a year. It was neutered in April 2014 and hairregrowth was observed and quickly followed by arelapse of the hair cycle arrest. The patient died acci-dentally in September 2014. His owner immediatelyarranged for his dog to be cloned in South Korea(Sooam Biotech; Seoul, South Korea). A viable clonewas born in August 2015 and was returned to its ownerin France in March 2016. It also presented with thefirst clinical signs of alopecia X at one year of age andwas immediately neutered. Partial hair regrowth was

noted followed by a relapse and worsening of thelesions three months later. This observation suggeststhe likelihood of a genetic component in the develop-ment of this disease as both the dog and its clone haddeveloped the disease at the same age regardless oftheir living conditions. Moreover, neutering, despiteallowing an identical temporary improvement, was notmore effective when performed earlier than later.Source of funding: Self-funded.


P-116

Conformational abnormality as a frequent

cause of pododermatitis in canines

A. TIWARI, B. KHAJURIA, V. VERMA and K.KANTSuvidha Vet Care and Research Centre, Jammu &Kashmir, India

Canine pododermatitis is a common clinical presenta-tion that causes paw licking or lameness. This studywas conducted to analyse the causes of pododermatitisin three breeds of dogs. Labrador retrievers (labs), bea-gles and golden retrievers (n = 15 of each breed) over 1year of age with pododermatitis were recruited for thisstudy. All dogs had hair plucks, skin scraping and cyto-logical evaluation performed. Biopsies were taken fromeight labs and six beagles, bacterial culture and sensitiv-ity tests were performed in 10 labs, 12 beagles and sixgolden retrievers with deep pyoderma. The cause ofpododermatitis was found to be demodicosis in fourlabs, four beagles and two golden retrievers, Malasseziaspp. in three labs, two beagles and four golden retriev-ers, atopic dermatitis in five labs, six beagles and sixgolden retrievers, orthopaedic pain in three labs, onebeagle and three golden retrievers, and a conforma-tional defect of the foot in five labs and nine beagles.This conformational defect caused ventrally fused padsto form a horseshoe bridge, which leads to weight bear-ing on interdigital skin. Cytological results of cases withthe conformational defect revealed degenerate neu-trophils and macrophages, and biopsy showed no neo-plastic growth. Increased body weight with splayed feetpredisposes to this painful condition, causing inflamma-tion of hair follicles and results in chronic inflammationin the feet. Conformational pododermatitis was mostcommonly seen in labs and beagles, and these patientswere the most difficult to treat. Cases of conforma-tional pododermatitis are rarely cured and requirelong-term management.Source of funding: Self-funded.


Abstracts 101



P-117

A case report of sterile granuloma/

pyogranuloma syndrome in a cat

K. MORI*, Y. OSHIMA-TERADA*, H. KONDO†

and N. MURAYAMA**Dermatology Services for Dogs and Cats, Tokyo,Japan; †Nihon University College of BioresourceSciences, Kanagawa, Japan

Sterile granuloma/pyogranuloma syndrome (SGPS) is avery rare disease in cats. The pathogenesis and treat-ment have not been fully evaluated. This case reportdescribes SGPS in a cat and the efficacy of essentialfatty acids (EFA) for this disease. A 3-year-old, cas-trated male American short hair cat, kept indoors, pre-sented with alopecic nodules on the nasal bridgewithout pruritus. The physical examination revealed noabnormalities. Trichogram, skin scrapings and impres-sion smears were unremarkable. Results of completeblood cell count and serum chemistry analysis werewithin normal limits. The histopathological findingsrevealed pyogranulomatous dermatitis consisting ofneutrophils and macrophages extending from the der-mis to the subcutaneous layers. Foreign bodies, infec-tious agents and atypical cells were not detected byperiodic acid Schiff, Ziehl–Neelsen, Giemsa and tolu-idine blue staining. Based on the clinical andhistopathological findings, the cat was diagnosed withSGPS. Two capsules once daily of EFA (Antinol, Vand P; Tokyo, Japan) were administered and the lesionsimproved after one month. However, two months afterthe initial treatment, the alopecic nodules relapsed. Thecat was treated with two capsules once daily of anotherEFA (Moegicap, Veterinarian Medical Development;Saitama, Japan) containing higher concentrations ofomega-3 fatty acids. The lesions resolved one monthlater and did not relapse for one year after the initialvisit. To the best of our knowledge, this is the firstreport of SGPS in a cat treated with EFA. This reportindicates that omega-3 fatty acids, which have anti-inflammatory effects, could be an effective treatmentfor SGPS.Source of funding: Self-funded.


P-118

Junctional epidermolysis bullosa in a domestic

cat

K. SHIBATA*, K. OKADA†, Y. KAGAWA† and M.TAKANOSU‡

*DVMs Animal Medical Center Yokohama, Kanagawa,Japan; †North Lab, Hokkaido, Japan; ‡NasunogaharaAnimal Clinic, Tochigi, Japan

Junctional epidermolysis bullosa (JEB) is a disordercharacterized by subepidermal blistering at the laminalucida. In humans, mutations of LAMA3, LAMB3,LAMC2, COL17A1 and ITGB4 have been identified asresponsible for JEB. Our objective was to examine thecause of JEB in an 11-month-old, female mixed-breedcat referred for recurrent ulcerative dermatitis. Clinicalexamination revealed ulcerations and erosions associ-ated with crusts on the pinna, head and axilla. In addi-tion, erosions with scales of multiple foot pads andonychomadesis of all claws were present. The cat alsohad severe stomatitis. Histopathological examinationrevealed subepidermal bullae. Periodic acid Schiff stainand immunostaining for collagen type IV were positiveon the bottom of the bullae, indicating that the dermo-epidermal separation occurred at the lamina lucida.These clinical and histopathological findings were con-sisted with JEB. Real-time PCR of skin cDNA revealedabnormal splicing events in exons 22–26 of LAMA3.Although the whole genome sequencing of the catrevealed no mutations in open reading frames ofLAMA3, LAMB3, LANC2 and ITGB4, mutations wereidentified in eight of 191 splicing factors that may haveinduced aberrant splicing variants of laminin 332. Thesefindings suggest that a partial defect in the laminin 332a3 chain might be the cause of JEB in this cat. Furtherstudy is need to demonstrate if mutated splicing factorscaused the aberrant splicing variants of laminin 332 inthis case.Source of funding: Self-funded.


102 Abstracts



ADDITIONAL FREE COMMUNICATION ABSTRACTSADVERSE FOOD REACTIONS

FC-66

Restrictive diet and dietary challenge guided

by prick test in the diagnosis of food allergy in

dogs with chronic pruritus associated with

atopic dermatitis

C. ALCAL�A, J. POSSEBOM, V. GMYTERCO andM. R. FARIASEscola de Ciencias da Vida, Pontif�ıcia UniversidadeCat�olica do Paran�a, PUCPR, Curitiba, Brazil

The aim of this study was to evaluate the prick testwith extracts of food proteins in the diagnosis of foodallergy in dogs with atopic dermatitis. To this end, 34dogs (Group 1) with atopic dermatitis syndrome and 10healthy dogs (Group 2) were screened, which were sub-jected to a prick test with extracts of bovine protein,chicken, egg, pork, fish, milk, soy and wheat and envi-ronmental allergens, considering positive papules largerthan 3 mm from the negative control. Dogs with a pos-itive food test received a homemade diet based on theirresult, PVAS and CADESI-4 being evaluated on days0, 30, 60 and those who improved were challenged withpositive protein for 7 days. In Group 2, one dogreacted to wheat and another to milk. In Group 1, four(12%) reacted to environmental allergens, four (12%)to food and 26 (76%) to both, 30 of which were startedon diet trials and 25 of 30 remained in the study. Per-centage improvement of PVAS (P = 0.0001) andCADESI-4 (P = 0.0001) between days 0 and 60 wasobserved in 21 (84%) dogs, with no difference betweendays 30 and 60, and all increased PVAS (P = 0.0001)and CADESI-4 (P = 0.039) after the challenge. Bovine,wheat and chicken proteins were the most reactive. Aconclusion is that dogs with atopic dermatitis are gener-ally polysensitized and the prick test can serve as aguide for establishing dietary protocols and assessingfood allergy in them.Source of funding: Coordenac�~ao de Aperfeic�oamento de

Pessoal de N�ıvel Superior (CAPES).Conflict of Interest: None declared.

FC-67

Evaluation of clinical signs related to adverse

food reaction (AFR) in dogs using a new

behaviour-recognition approach

S. M. WERNIMONT*, J. L. WEEMHOFF*, J. M.MACLEAY*, J. J. BREJDA† and K. L. GROSS**Hill’s Pet Nutrition, Inc. Topeka, KS, USA; †AlphaStatistical Consulting, Lincoln, NE, USA

Quantifying intervention response in canine adversefood reactions (AFR) is a challenge. A device providingquantitative scratching, shaking, resting and sleep datacould assist management. We paired this innovativetool with the COSCAD’18 and examined behavioursover time in an IACUC-approved prospective study.Forty-seven dogs diagnosed by food elimination trial atgeneral veterinary practices and consuming a novel orhydrolysed protein food provided complete data. Dogswore a collar-mounted activity monitor and veterinaryCADLI scores and owner PVAS scores were performedon day (D) and D21. Machine learning-based beha-viour recognition algorithms quantified the total dura-tion of scratching (s/day), shaking (s/day), resting (h/day) and sleeping (h/day); sleep quality was evaluatedwith an algorithm based on the absence of night-timedisturbance (0–100: highly disturbed-undisturbed sleep).Behaviours, CADLI and PVAS scores were analyzed inSAS using linear mixed-models with diet, time, andDietxTime as fixed-effects and significance P < 0.05.Duration of behaviours (mean � SD) were: scratching:108 � 136 s/day; shaking: 39.7 � 27.6 s/day; resting:9.1 � 2.0 h/day; sleeping: 12.6 � 2.2 h/day; and sleepquality: 90.1 � 8.4. There were no statistically signifi-cant differences between the CADLI and PVAS scorescollected at study entry and day 21 (CADLI: 4.43/50vs. 3.65/50, P = 0.09; PVAS: 3.69/10 vs. 3.49/10,P = 0.50). Quantitative, objective behaviour data fromwearable activity monitors complements the COSCAD‘18 and is free of recall bias. Quantitative behaviourdata provides a new reference for clinical studies ofdogs with AFR.Source of Funding: Hill’s Pet Nutrition.

Conflict of Interest: Authors are employees of or were

compensated (JJB) by Hill’s Pet Nutrition, Inc.

Abstracts 103



FC-68

Quantification of behaviours in dogs with

atopic dermatitis with a machine learning-

based approach

J. L. WEEMHOFF*, S. M. WERNIMONT*, J. M.MACLEAY*, J. J. BREJDA† and K. L. GROSS**Hill’s Pet Nutrition, Inc. Topeka, KS, USA; †AlphaStatistical Consulting, Lincoln, NE, USA

Quantifying intervention response in canine atopic der-matitis (CAD) is a challenge. A device providing quan-titative scratching, shaking, resting and sleep datacould assist management. We paired this tool with theCOSCAD’18 and examined behaviours in an IACUC-approved prospective study. Dogs with CAD and vet-erinarian-assessed CADLI scores ≥8/50 and an owner-reported PVAS scores ≥3.5/10 were enrolled and worean activity monitor. Machine learning algorithms quan-tified the total duration of dermatitis-related behavioursper day: scratching (s/day), shaking (s/day), resting (h/day) and sleeping (h/day). Ninety-five dogs had com-plete data. Mean � SD CADLI (13.8 � 0.78) andPVAS (6.86 � 3.6) scores confirmed clinical signs con-sistent with entry criteria. Activity data reported asmean � SD (median, interquartile range (IQR):scratching: 254 � 319.1 (131.0, 210.0) s/day, shaking:44.1 � 31.7 (35.5, 32.5) s/day, resting: 9.0 � 1.8 (9.0,1.8) h/day, and sleeping: 12.9 � 2.1 (13.0,2.3) h/day.Scratching duration was greater and more variablethough shaking duration was similar to each of twocomparator groups: a group of pruritic dogs at generalpractice clinics (mean � SD scratching: 96.9 � 11.4and shaking: 49.2 � 5.5 s/day) and a large populationof dogs with various health states (median, IQRscratching: 46, 80 and shaking: 31, 31). Quantitative,objective behaviour data from wearable activity moni-tors complements the COSCAD ‘18 and is free of recallbias. Quantitative behaviour data provides a new refer-ence for clinical studies of dogs with CAD.Source of Funding: Hill’s Pet Nutrition.



FC-69

Successful management of clinical signs related

to adverse food reaction in dogs using a food

with a new approach

J. M. MACLEAY*, J. L. WEEMHOFF*, S. M.WERNIMONT*, J. J. BREJDA† and K. L. GROSS**Hill’s Pet Nutrition, Inc. Topeka, KS, USA; †AlphaStatistical Consulting, Lincoln, NE, USA

Quantifying intervention response in canine adversefood reaction (AFR) is a challenge. Devices providingquantitative scratching, shaking, resting and sleep datacould assist management. We paired this tool with theCOSCAD’18 in an IACUC-approved randomized, con-trolled, double-masked prospective study. Dogs diag-nosed with AFR, consuming a novel or hydrolysedprotein food were enrolled. CADLI and PVAS wererecorded on day (D)0, D21 and D42 and dogs woreactivity monitors throughout. Dogs consumed theirfood (0–21 days) and then were randomized to test(TF) or control (CF) food for 22–42 days. The TF con-tained egg, n3-6-9 fatty acids, and the CF hydrolysedanimal protein. Duration of scratching (s/day), shaking(s/day), resting (h/day), sleeping (h/day), and sleepquality (0–100:highly disturbed-undisturbed sleep).Analysis was linear mixed-models with diet, time, anddiet 9 time as fixed-effects and a significance thresholdof P < 0.05 in SAS. Twenty-three TF and 21 CF com-pleted study. Baseline and D42 CADLI and PVASscores were similar (P > 0.16). In 35 dogs stratified onCADLI (> or <8/50) there were no differences betweenTF and CF (P > 0.60). Dogs beginning with CADLIscore ≤8/50 remained in that category. In activity datafrom 32 dogs there were no differences between TF &CF in change from baseline for any endpoint(P > 0.30), and no linear trends (P > 0.20). Pet ownersatisfaction was similarly positive between groups.Feeding a food formulated for AFR resulted in success-ful transition and comparable control of clinical signsvs. feeding a hydrolysed protein-based food.Source of Funding: Hill’s Pet Nutrition.



104 Abstracts



FC-70

Diagnosis of feline cutaneous adverse food

reactions with extensively hydrolysed protein

diet

U. MAYER*,1, K. BERGVAL†, L. BECO‡, J.FONTAINE§, T. BORD¦, S. AHMAN**, R.ALMELA*, L. BOUTIGNY†† and I. LESPONNE††

*Kleintierspezialisten, Augsburg, Deutschland;†Department of Clinical Sciences, University ofAgriculture, Uppsala, Sweden; ‡Cabinet v�et�erinaireMonVt, Spa, Belgium; §Animal Clinic, Bruxelles,Belgium; ¶Clinique v�et�erinaire de l’Arche, Salon-de-Provence, France; **DjurAkuten AB, Stockholm,Sweden; ††Royal Canin, Aimargues, France

The diagnosis of cutaneous Adverse Food Reactions(cAFR) in pets relies on performing an elimination trialfollowed by re-challenge. The objective of this prospec-tive multicenter study was to assess the diagnostic effi-cacy of an extensively hydrolysed protein-based food incats. Adult cats with suspected cAFR were includedinto the study by dermatology specialists, if also meet-ing criteria for pruritus (Pruritus Visual Analog Scaleequal or greater to 1) and/or lesions (Scoring FelineAllergic Dermatitis (SCORFAD) equal or greater to 4).

Amongst non-inclusion criteria were other diseases, ges-tation or lactation, secondary infections, impossibilityto stop flavored medications. Drug usage was allowedonly in precise circumstances e.g. needing oral or localantibiotics during the elimination phase. After inclusionvisit V0, cats were fed the test diet (extensively hydrol-ysed protein and purified corn starch) for eight weeks,with follow-up visits at day 30 (V1) and 60 (V2). Thencats were challenged with previous diet untill V3 (flareup, 14 days otherwise), and back to test diet until studycompletion. Thirty-four cats were enrolled, ten com-pleted both phases. Eight of these had lesions improvedwith the test diet (SCORFAD decreasing from median3.5 [2;5] at visit (V)0 to 1.5 [0;6] at V2, owners mention-ing skin improvement after 12 days in average). Two ofthem, also experiencing worsening during re-challenge,were diagnosed as cAFR cases. Owner-assessed fecalscores confirmed good digestive tolerance of the food.The results show that this food may constitute a valu-able tool for feline cAFR diagnosis. Further studies arewarranted to confirm it.Source of Funding: Royal Canin SAS.

Conflict of Interest: IL is employee of Royal Canin SAS.

LB is a former employee of Royal Canin SAS.

Abstracts 105



CUTANEOUS ECOSYSTEM

FC-71

Prevalence of methicillin resistant

Staphylococcus pseudinteremedius in dogs with

skin and ear infections in South Africa

C. PRIOR*, M. KARAMA†, A. MOODLEY‡ andA. LEISEWITZ**Department of Companion Animal Clinical Studies,Faculty of Veterinary Science, University of Pretoria,Onderstepoort, South Africa; †Department ofParaclinical Sciences, Veterinary Public Health, Facultyof Veterinary Science, University of Pretoria,Onderstepoort, South Africa; ‡Department of Veterinaryand Animal Science, Faculty of Health and MedicalSciences, University of Copenhagen, Copenhagen,Denmark; ‡International Livestock Research Institute,CGIAR AMR Hub, Nairobi, Kenya

Staphylococcus pseudintermedius (SP) is an importantopportunistic pathogen, frequently associated with der-matitis and otitis in small animals. The emergence andrapid expansion of methicillin resistance is problematicdue to multi-drug resistance and reduced treatmentoptions. We aimed to 1) confirm that standard labora-tory methods used at five regional veterinary labs inSouth Africa (RSA) can accurately identify SP; 2)

determine if there is association between resistance tofirst and second tier antibiotics and presence of mecA;and 3) determine if there is an association betweenMRSP and previous antibiotic use. Sixty-eight clinicalsamples from five geographically dispersed laboratoriesin RSA were collected over a 24-month period. SP weredetected by a standard laboratory method and antimi-crobial susceptibility testing by disc diffusion. PossibleMRSP isolates were flagged when resistance to oxacillinwas observed. All isolates were confirmed as SP andgenotyped for the mecA gene by PCR. Fifty-three iso-lates were confirmed to be SP (78%) and only 44(83%) were MRSP. Thirty-two MRSP were isolatedfrom pyoderma (32/44, 73%) and 8/44 from otitis(18%). All mecA positive samples were recovered fromdogs that had been exposed to antimicrobials previ-ously (50/68, 73.5%). MRSP isolates are commonamong dogs with pyoderma and otitis in South Africa.There is an urgent need for better laboratory diagnosisof MRSP and surveillance of dogs presenting pyodermaand otitis in South Africa. Moreover, diligent antibioticstewardship will be crucial to prevent a deterioration inthis situation in South Africa.Source of funding: Zoetis Animal Health.


106 Abstracts



ATOPIC DERMATITIS

FC-72

Evaluation of owner conduct in the

management of dogs with atopic dermatitis, in

the municipality of Rio de Janeiro-RJ

M. SILVA* and M. SILVA-CARVALHO†

*Est�acio de S�a University (UNESA), Rio de Janeiro,Brazil; †Federal University of Rio de Janeiro, Rio deJanerio, Brazil

Dermatological problems are currently one of the mainreasons for going to veterinary clinics for small ani-mals. Canine Atopic Dermatitis is an inflammatoryskin disease generated by an exacerbated reaction ofthe immune system to different types of allergens. It isa disease of genetic origin, in which the epidermal bar-rier undergoes changes, being weakened as a defensesystem. Treatment involves controlling secondary infec-tions, managing itching and excluding any potentialirritating agents. The present work had as objective tomake a survey of data, in order to better understandthe profile of owners of dogs diagnosed with CAD. Aquantitative questionnaire consisting of 16 questionsinvolving the daily lives of animals and their caregiverswas applied virtually through Google Docs, in the cityof Rio de Janeiro. A total of 100 responses wereobtained. Results show many caregivers still fail tounderstand the importance of a complete therapeuticapproach. Although most prevent ectoparasites (50%),many have the habit of offering varied snacks (51%) totheir dogs, in addition to bathing in a pet shop (50%).The awareness of caregivers of atopic animals regardingoptimal care must be made to improve the collabora-tion that is essential for successful treatment of atopicdogs.Funding Source: Self-funded.


FC-73

Topical corticosteroids and urea for the

treatment of canine atopic dermatitis

T. MATSUDA*, N. NUNOKAWA*, S. HIRANO*and K. IYORI†

*Nunokawa Dogs and Cats Hospital, Kanagawa, Japan;†Vet Derm Tokyo, Kanagawa, Japan

This study investigated the efficacy of topical corticos-teroids (TCSs) of varying potencies with and withouttopical moisturizer in dogs with atopic dermatitis (AD).Seven dogs with chronic AD were enrolled. Three offour digital skin lesions of each dog were randomlyselected for treatment with one of three topical regi-mens: (1) 0.1% mometasone furoate cream, (2) 0.1%triamcinolone acetonide cream, or (3) 0.1% triamci-nolone acetonide cream mixed with a topical 20% ureacream. Triamcinolone acetonide and 20% urea creamwere mixed in equal amounts. Each topical agent(0.25 g per digit referred to finger-tip-unit in humans)was applied twice daily for 14 days. The investigatorsand dog owners were blinded to the treatment alloca-tion. Clinical scores were determined by the CanineAtopic Dermatitis Extent and Severity Index(CADESI-4) on experimental days 0, 7, and 14. Ondays 7 and 14, mean CADESI-4 score significantlyreduced in the lesions treated with mometasone (32%on day 7 and 50% on day 14), triamcinolone (34% onday 7 and 57% on day 14), and triamcinolone plusurea (29% on day 7 and 54% on day 14) comparedwith day 0 (p < 0.05). There were no significant inter-group differences in CADESI-4 scores on days 7 and14. Thus, triamcinolone acetonide cream, classified as amedium potency TCS in Japan, may be as effective forAD as the very high potency mometasone furoate. Mix-ing triamcinolone with 20% urea cream ultimatelyresults in reduction in TCS amount required to achievean appropriate clinical response.Source of Funding: Self-funded.


Abstracts 107



ADDITIONAL POSTER ABSTRACTSMISCELLANEOUS

P-119

Bromelain based enzymatic debridement of

deep burns on the paws of a cat

L. WALDMANDermatology and Allergy Clinic, Haifa, Israel

A bromelain based proteolytic enzyme preparation,NexobridTM (MediWound, Yavne, Israel) is used totreat moderate and deep burns in humans. It removeseschar while preserving healthy dermis, thereby,improving healing and reducing the need for repeatedgeneral anesthesia, surgery and autografting. A 13-year-old, male neutered, domestic medium hair cat sufferedsmoke inhalation, lower urinary tract disease, and sev-ere burns to all paws in a forest fire. After cleaning thepaws, the bromelain based proteolytic enzyme prepara-tion was applied under occlusive dressing for 4 h result-ing in complete debridement down to healthy dermis.To prevent drying, the wounds were treated, withoutdressings, with sea-buckthorn oil three times daily forsix weeks. The day after the proteolytic enzyme treat-ment, the cat began to walk cautiously to the litter tray.Pseudoeschar formed within two days. Granulation tis-sue appeared after six days and clobetasol propionate0.05% cream was then applied three times daily for sixweeks. Pain was controlled with buprenorphine0.025 mg/kg orally four times daily for the first twoweeks, then meloxicam 0.2 mg/kg orally for one dayfollowed by 0.05 mg/kg for six days. One week afterdebridement, the cat was alert, walking in the houseand eating well, despite signs of painful paws. Fiveweeks later, adhesions in the left hind paw were surgi-cally released. In conclusion, a bromelain based prote-olytic enzyme preparation successfully treated deep pawburns in a cat, mitigating the need for intensive woundcare.Source of Funding: NexoBrid

TM

was donated by Medi-

Wound.


P-120

Vetch toxicoses in Angus cows fed Vicia sativasubsp. sativa (Common Vetch) in New South

Wales – a case report

A. JORDAN*, B. WATT†, W. BERRY‡ andP. PINCZOWSKI**EMAI, NSW Department of Primary Industries; †LLS,NSW Department of Agriculture; ‡Blayney VetHospital, NSW, Australia

Vetch toxicoses is a well-recognised syndrome of granu-lomatous eosinophilic dermatitis and systemic diseaseassociated with ingestion of Hairy Vetch (Vicia villosa),which has also been reported with Narrow-leaf Vetch(Vicia sativa subsp. nigra). In December 2019, approxi-mately 10 cows from a herd of 50 developed cutaneousulcers and crusts over the head, neck, and perineum,which were erythematous and exudative when lifted.There was also some alopecia and scaling. A skinbiopsy was submitted, and histopathology showedexudative and ulcerative, perivascular and periadnexal,eosinophilic and histiocytic dermatitis with folliculitisand multinucleate giant cells, including Langerhanscells. The cattle were being fed Morava hay, variety ofCommon Vetch (Vicia sativa subsp. sativa), and in lightof histopathology findings, feeding of Vetch hay wasceased. Skin lesions largely resolved within two weeks.Varieties of Common Vetch have an increasing role incropping systems and as livestock fodder in NSW, andawareness of a possible role in toxicoses should beraised.Source of Funding: Local Land Services provided

resources to facilitate the investigation, and AJ is

employed by the Department of Primary Industries, both

within the New South Wales Government Department of

Industry, Agriculture and Environment, Australia.


108 Abstracts



ALLERGIC DERMATITIS

P-121

A masked, randomized clinical trial evaluating

the efficacy and safety of lokivetmab

compared to saline control in client owned

dogs with allergic dermatitis

L. VAN BRUSSEL*, H. MOYAERT*,M. ESCALADA*, S. P. MAHABIR† andM. R. STEGEMANN**Zoetis Belgium SA, VMRD, Zaventem, Belgium;†Zoetis Inc., VMRD Kalamazoo, MI, USA

Interleukin (IL)-31 is an important mediator in atopicdermatitis and is, as recently reported in humans, alsodysregulated in other allergic diseases. To demonstratethe efficacy and safety of lokivetmab (an anti-IL-31monoclonal antibody) for treatment of pruritus associ-ated with allergic dermatitis in dogs.For this study,dogs (n = 123) were enrolled from 14 practices in Por-tugal, Hungary, France and Germany. Animals were atleast moderately pruritic at enrolment and a presump-tive diagnosis of allergic dermatitis was made. Animalswere randomized to placebo (saline, n = 62) or lokivet-mab (1–3.3 mg/kg, n = 61). Dogs were treated once onday 0. Pruritus was assessed daily by the owner untilday 7 and weekly thereafter through a Visual AnalogueScale (VAS) by Hill. Skin condition was assessed by a

veterinarian dermatitis VAS on days 0, 7, 14 and 28.Starting from one day after treatment, the percentagereduction from baseline of Owner-assessed pruritusVAS LS means was significantly higher in the group ofanimals treated with lokivetmab versus the controlgroup, with 57.71% reduction compared to 21.78% onDay 28. For all timepoints, the percentage reductionfrom baseline of Investigator-assessed skin conditionVAS means was significantly higher in the lokivetmabgroup compared to the control group, with 57.05%reduction versus 20.49% on Day 28. Overall, the occur-rence of health events during the in-life phase was com-parable between both treatment groups. In conclusion,in this study lokivetmab was a safe and efficacioustreatment for dogs with allergic dermatitis.Source of Funding: Zoetis Inc, Parsipanny, NJ, USA.

Conflict of Interest: All authors are employees of Zoetis

Inc.

Acknowledgements: We thank the clinicians who enrolled

dogs in this study and performed the clinical investiga-

tions: Jean-Michel Arquilliere, Emmanuel Bensignor,

Laurent Bourdenx, Vincent Mah�e, Isabelle Papadopoulo,

Annette Feische, Elisabeth Koldt, Petra Wittmann,

Bal�azs Cap�ari, L�aszl�o Kir�aly, Szil�ard Kraxner, Krisztina

P�alfi, Joao Araujo, Catarina Duarte, Adriana Estrela

and Manuela Mangas.

Abstracts 109



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