CSIROREPORT OF RESEARCH July 2000 – June 2003

LIVESTOCK INDUSTRIES

Printing and publication details

Copyright CSIRO Livestock Industries

For further information contact:CSIRO Livestock IndustriesQueensland Bioscience Precinct306 Carmody Road, St Lucia Qld, 4067Ph: (07) 3214 2200Fax: (07) 3214 2900

The information contained in this document is general information provided as part of CSIRO’s statutory role in the dissemination ofinformation relating to scientific and technical matters. It is not professional, medical, technical or expert advice and is subject to theusual uncertainties of advanced scientific and technical research. It may not be accurate, current or complete and is subject to changewithout notice. It should never be relied upon as the basis for doing/failing to do something without first contacting CSIRO LivestockIndustries in writing to seek up-to-date advice.

ProductionThe preparation of this Research Review involves a wide range of staff in CSIRO Livestock Industries.

CompilationWendy Pyper - Communication Officer, CSIRO Livestock IndustriesAnne Tuppack – Executive Officer, CSIRO Livestock IndustriesPhillipa Loveridge – Assistant Executive Officer, CSIRO Livestock Industries

PhotographyCSIRO Livestock Industries staffOthers as acknowledged

Design and layoutjoltSuite 12, Plumridge House36 Agnes Street, Fortitude Valley, Qld, 4006Ph: (07) 3216 0656Fax: (07) 3216 0657www.joltstudio.com.au

Managing editorWendy Pyper – CSIRO Livestock Industries

Foreword 2Who we are 3Divisional vision statement 4Glossary 5Livestock capabilities 6

Livestock applications of biotechnology 8Livestock applications of biotechnology 9Genes and proteins 10Molecular genetics 12Genes for production species 14Pests and diseases 16Cell biology 19Ruminant microbiology 22Aquaculture viral disease 24

Diagnostic sciences 26Diagnostic sciences 27Diagnosis and epidemiology 28Fish disease research and diagnosis 30Molecular diagnostics 32Electron microscopy and emerging pathogens 35Newcastle disease 37Virology 40Tissue culture and media production 41

Tropical livestock systems 42Tropical livestock systems 43Genetics 44Livestock systems/livestock environment interactions 47Stress physiology 49Ruminant nutrition 50Parasitology 54

Infectious diseases and food safety 56Infectious diseases and food safety 57Avian virology 58Johne’s disease 60Protein biochemistry and proteomics 62Plant-associated toxins 65Molecular detection 68

Mediterranean livestock systems 70Mediterranean livestock systems 71Reducing environmental impact 73Greater productivity 76New/improved products 78

Vaccines and therapeutics 84Vaccines and therapeutics 85Development of novel therapeutics 86Gene technologies 88Viral delivery 90

Temperate livestock systems 92Temperate livestock systems 93Disease resistance and animal welfare 94Productivity and enterprise diversification 98Information systems 101

CRC involvement 104CRC for Australian biosecurity 105CRC for Australian sheep industry 105CRC for cattle and beef quality 105CRC for innovative dairy products 106CRC for plant-based management of dryland salinity 106CRC for poultry 106CRC for vaccine technology 106

Publication list 2000-2003 107

Staff list 141

of contentstable

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forewordfrom the Chief

The work of CSIRO Livestock Industries pursues four broad aims: providingimprovements in production capacities for livestock producers; developing environmentally sustainable production systems; protecting our livestock industriesfrom losses from diseases and pests; and maintaining market access.

A number of strategies are being used to improve production outputs whilereducing input costs. At one end of the scale we have made a major commitmentto biotechnology, in particular, to harnessing the potential of new genomic technologies.We have the opportunity to make huge leaps in our understandingof genetic traits, such as marbling and tenderness in beef, and disease resistance,as genome sequences for the major production species are documented. At theother end of the scale we are looking at animal nutrition, production efficienciesand environmental sustainability, and designing decision support tools for livestockproducers.

The Division plays a major role in maintaining and improving animal health inAustralia.We provide disease surveillance and diagnostic services, as well as exotic disease outbreak emergency response training. In collaboration with theDepartment of Agriculture, Fisheries and Forestry, we are the key agency for disease diagnosis, research and policy advice in animal health.

Our scientists are also focussing on protecting livestock from pests and diseases,and reducing the use of antibiotics and other chemicals through breeding forresistance traits, as well as developing innovative new products and delivery techniques.

To deliver results to industry, we work in collaboration with a range of industryparticipants, including rural investors, pastoral and animal health companies, thefarming community, state and federal development agencies, and international aidagencies. CSIRO Livestock Industries is also a member of six CooperativeResearch Centres, and works with new business partners to commercialise innovative technology.

Thank you for taking the time to read about our achievements in working for theAustralian people and Australia's livestock industries.

Shaun CoffeyChief

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Our scientific achievements andcontributions to the world of science, and the many staff whohave progressed our researchfrom strength to strength arecelebrated in this report.

whowe are

CSIRO Livestock Industries wasformed on 1 July 2000 throughthe amalgamation of the formerCSIRO Divisions of AnimalHealth and Animal Production,and parts of Tropical Agriculture.The Division works in the majortemperate, Mediterranean andtropical production zones of Australia.

The Division has specialised field andlaboratory facilities at Brisbane andRockhampton in Queensland, Armidalein New South Wales, Floreat Park inWestern Australia and the AustralianAnimal Health Laboratory in Geelong,Victoria.

At the Queensland Bioscience Precinct,opened in May 2003 in Brisbane,molecular and biochemical techniquesare being applied to tropical livestockproduction. Projects include genomemapping, understanding the geneticand biochemical basis of various facetsof growth and production, and developing new technologies for thecontrol of parasites and diseases.Strengths in bioinformatics underpinmuch of this research.

Scientists at the JM Rendel Laboratoryand the nearby Belmont ResearchStation, in Rockhampton, conductresearch into beef cattle nutrition andmetabolism to improve livestock productivity and production efficiency.

Other research includes on-farmand/or genetic techniques to increasepest and disease resistance, improvemeat quality, reduce methane emissions, and develop economically,environmentally and socially sustainableproduction systems.

At Floreat Park in Perth, theMediterranean Livestock Systems program is investigating livestock production on saline and waterloggedland, methods to reduce greenhousegas emissions from ruminants, and precision agriculture techniques toimprove pasture and livestock productivity.There is also a focus onnew or improved products from livestock, such as nutritionally enhancedmeat and milk, and better woollen garments.

At Armidale, the newly upgraded F.D.McMaster Laboratory is the site ofresearch into control strategies for arange of livestock parasites includingnematodes and sheep blowfly.The siteis also a centre for research into animalhealth and welfare, wool productionand quality, enterprise diversification,and information technology.With anew Centre for AdvancedReproductive Technologies, the site will continue building on its strengths in animal breeding and genetics.

The Australian Animal HealthLaboratory (AAHL), in Geelong, is oneof the most sophisticated laboratoriesin the world for the safe handling andcontainment of animal disease agents,and critical in Australia's efforts toremain free of major exotic diseases.Research at the site aims to enhanceAustralia's disease control capacity byimproving our understanding of viral,bacterial and plant poisoning diseaseprocesses, and mechanisms of diseasecontrol in livestock.This is achievedthrough ongoing research programs todevelop sensitive, accurate and timelydiagnostic tests that are critical to thesuccess of any eradication campaign inthe event of a disease outbreak. AAHLalso undertakes research to developnew diagnostic tests, vaccines, and therapeutics, for endemic animal diseases of national importance.

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CSIRO Livestock Industries conducts research and provideshigh quality services with Australia’s livestock and allied industries, to facilitate their sustainable competitive advantage.

DIVISION LOCATIONS

Queensland Biosciences Precinct306 Carmody RoadSt Lucia, Qld 4067Ph: (617) 3214 2200Fax: (617) 3214 2900

ArmidaleFD McMaster laboratoryChiswickNew England HighwayLocked Bag 1Armidale, NSW 2350Ph: (612) 6776 1300Fax: (612) 6776 1333

RockhamptonJM Rendel LaboratoryBruce Highway (Ibis Avenue)PO Box 5545Rockhampton Mail Centre, Qld 4702Ph: (617) 4923 8100Fax: (617) 4923 8222

PerthCentre for Environment and LifeSciencesUnderwood AvenueFloreat Park,WA 6014Ph: (618) 9333 6000Fax: (618) 9387 8991

Australian Animal Health Laboratory(AAHL)5 Portarlington RoadEast Geelong,Vic 3220Ph: (613) 5227 5000Fax: (613) 5227 5555

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divisionalvision statement

Rockhampton

Brisbane

Armidale

Geelong

Perth

AAHL Australian Animal Health LaboratoryACIAR Australian Centre for International Agricultural ResearchAFLD AAHL Fish Diseases LaboratoryARGT annual rye-grass toxicityAPP Actinobacillus pleuropneumoniaeAWI Australian Wool Innovation LimitedBAC bacterial artificial chromosomecDNA complimentary deoxyribonucleic acidChIFN-γ chicken interferon gammaCSIRO Commonwealth Scientific and Industrial Research OrganisationCRC Cooperative Research CentreDRDC Dairy Research and Development CorporationDNA deoxyribonucleic acidEGF epidermal growth factorELISA Enzyme-linked Immunosorbent AssayEST expressed sequence tagFMD foot and mouth disease FOO food on offerGAV gill associated virusGRDC Grains Research and Development CorporationHSP heat shock proteinIBDV infectious bursal disease virusIBV infectious bronchitis virusIVF in vitro fertilisationkg/day kilograms per dayMLA Meat and Livestock AustraliaMoV mourilyan virusNDV Newcastle disease virusNIRS near infra-red reflectance spectroscopyOIE Office International des Epizooties% percentPAs pyrrolizidine alkaloidsPATs plant-associated toxinsPCR polymerase chain reactionPGR pasture growth ratepST porcine somatotrophinRAPD random amplified polymorphic DNARIRDC Rural Industries Research and Development CorporationRNA ribonucleic acidRNAi ribonucleic acid interferencedsRNA double stranded ribonucleic acidmRNA messenger ribonucleic acidSARS CoV severe acute respiratory syndrome coronavirusSNP single nucleotide polymorphismTSE transmissible spongiform encephalopathyWSSV white spot syndrome virusYHV yellow head virus

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glossary

Advanced reproductive technologiesModern reproductive technologiesinclude artificial insemination, semenfreezing, semen sexing, nuclear transfer,embryo transfer, embryo micromanipulation and cloning. Inrecent years, these techniques havebeen applied to produce animals withnew genes, which are designed to bebeneficial to agriculture and to animaland human health. CSIRO LivestockIndustries is conducting research toincrease the efficiency of advancedreproductive technologies as well asresearch to develop new methods for transgenic animal production.

Animal welfareCSIRO Livestock Industries is at theforefront of research aiming to developobjective methods for assessing animalwelfare. Across-site research into theresponses of cattle, sheep and pigs todifferent stresses will inform welfaremanagement decisions in live export,feedlot and intensive farming situations.

BioinformaticsThe close alignment of bioinformaticswith biomolecular science researchactivities will play a key role in theDivision’s success. Our bioinformaticsgroup is developing a suite of web-accessible databases and tools forthe storage, management, and analysisof information generated by the molecular and genomics projects in the division.

Decision support systemsCSIRO Livestock Industries has developed a range of decision supporttools to help industry improve production and profit.These toolsinclude AUSPIG, for improved pig production, SelectGene and SelectBreeding Services, which togetherenable fine wool breeders to optimisetheir breeding programs, and Nemesis,which assists breeders in incorporatingnematode resistance into their breeding program.Web-based tools forpasture production estimates have alsobeen successful in assisting producerswith pasture management decisions inWestern Australia.

Disease diagnosis and testingCSIRO Livestock Industries is at theforefront of disease diagnosis and thedevelopment of diagnostic tests forviral and bacterial diseases that affectanimals and humans. At the AustralianAnimal Health Laboratory (AAHL) inGeelong – a unique, national facility forinfectious and exotic disease researchand diagnosis - diagnostic tests havebeen developed for a range of nationally and internationally importantdisease organisms.These includeHendra and Nipah viruses, Australianbat lyssavirus, Newcastle disease,infectious bursal disease viruses ofpoultry, and diseases of aquaculture.The facility also acts as a trainingground for field and laboratory basedanimal health professionals in diseaserecognition and the methods and technologies used to diagnose disease.

Our work helps protect Australia’s livestock and aquaculture industriesfrom major economic losses in theevent of emergency disease outbreaks,and supports national and internationaltrade by providing scientific supportand information on animal diseaseissues.The work is closely supportedby our capabilities in vaccine and therapeutic development, and drawson skills in virology, pathology, serology,electron microscopy, bio-molecularengineering and monoclonal andrecombinant antibody technology.

Electron microscopyAdvanced transmission and scanningelectron microscopy techniques areused for disease diagnosis, identificationand characterisation of viruses, and forunderstanding viral structure, functionand replication.

Environmental managementThe Division is undertaking a variety ofprojects that address environmentalissues relating to livestock and livestockmanagement.These include reducingmethane emissions from ruminantsusing antimethanogenic compoundsand vaccine technology, and improvingthe economic viability of salt-affectedpasture for livestock production, whilemanaging the effects of salinity andwaterlogging. Our livestock research ingeneral, strives to assist industries to beeconomically viable, ecologically sustainable and capable of contributingto the social and natural resourceobjectives of the nation.

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livestockcapabilities

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Microarray technologyMicroarrays consist of a collection ofcDNA (complimentary DNA) from tissues of interest (fat, muscle, blood orskin for example), attached in a gridpattern to a glass microscope slide.Changes in gene expression can thenbe detected by hybridising mRNA orcDNA from the cells or tissues ofinterest, to the array.We have developed a range of microarrays tomeasure changes in gene expressionfor different aspects of sheep, cattleand pig production.

Molecular genetic technologiesThese technologies include genomemapping, gene sequencing, understandinggene function, and genetic modificationof organisms. Our genomic capabilitieshave resulted in, for example, thedevelopment of a bovine genome map,the identification of genes for improvedproduction traits and cane toad development, and the genomic characterisation and diagnosis ofemerging and exotic viruses. NewRNA interferance technology is beingused to develop genetic constructs targeting genes for viruses that causedisease in chickens, pigs, cattle and sheep.

Protein biochemistry and proteomicsThe identification, purification and characterisation of viral and bacterialproteins and antigens, aid the development and improvement ofdiagnostics and vaccine developmentmethodologies in the areas of animaland human health.Targets includenewly emergent diseases such asHendra, Nipah and Menangle viruses,avian and mammalian cytokines, andantigenic proteins of Mycobacteriumavium subspecies paratuberculosis(Johne’s disease) and Moraxella bovis(Pinkeye).

ToxicologyBy combining our expertise in organicchemistry, immunochemistry,biochemistry, toxicology and animalhusbandry, CSIRO Livestock Industriesstudies all aspects of plant-associatedtoxins. Our research aims to lessen theimpact of plant-associated toxins onfarmers, animals, livestock feed industries and human food-relatedindustries; help safeguard domestic and international markets for Australianagricultural produce; and control contamination of human food.

Vaccines and therapeuticsCSIRO Livestock Industries uses cuttingedge science to produce vaccinesagainst diseases of economic significance to livestock industriesaround the world. Our portfolio ofvaccines ranges from simple inactivatedvaccines (vaccines that contain organisms that have been killed orinactivated with heat or chemicals), tobacteria that have been attenuatedthrough manipulation of virulence factors on the chromosome, and vaccines made from viral surface proteins. In partnership with industry,universities and veterinary pharmaceutical companies, the Division has a range of products in development, from those that are inthe early concept phase through to the later stages of commercial development.

The need to find alternatives to chemicals and antibiotics used in livestock industries has also stimulatedresearch into the use of cytokines –natural modulators of the immune system – to control diseases and promote the welfare and growth ofproduction animals.The potential toexploit bacteriocins – small bioactivepeptides produced by bacteria – astherapeutics, is also under investigation.

We have made a major

commitment to biotechnology,

in particular, to harnessing

the potential of recent

developments in genomic

technologies

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Genes and proteins 10Molecular genetics 12Genes for production species 14Pests and diseases 16Cell biology 19Ruminant microbiology 22Aquaculture viral disease 24

livestockapplications of biotechnology

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Program Leader Dr Peter Willadsen

Ph (07) 3214 2467Fax (07) 3214 2480Email peter.willadsen@csiro.au

The Livestock Applications ofBiotechnology Program, based atthe Queensland BiosciencesPrecinct, St Lucia, uses the latestmolecular and biochemical techniques to improve productsderived from sheep, cattle, pigsand prawns, to understand disease resistance in these animals, and to develop moresustainable production systems.The program also features anumber of projects with applications for human medicine.

On the products front, genetic techniques have been used to improvemeat tenderness and marbling in beefcattle. Breeders can now select animalswith favourable marbling and tendernessgenes using GeneSTAR® marbling andtenderness tests. Other projects lookat the biochemistry of marbling, andmolecular techniques to modifyexpression of the trait. Genes and/orproteins involved in better wool qualityand larger prawns are also being sought.

As disease is a problem in all intensivelyfarmed animals, the program aims toidentify genes for disease resistance.These include genes for resistance tomastitis in dairy cows, tick fever in cattleand pneumonia in pigs.Work onendemic and exotic aquaculture viraldiseases has resulted in commercialdiagnostic tests for three viruses.

Genetic maps are an important part ofthe gene discovery process and suchmaps are being developed for the blacktiger prawn Penaeus monodon and thecow Bos taurus. Our increasingbioinformatics capacity will help copewith the influx of information fromthese mapping projects and willimprove management and analysis ofresearch data.

Bioinformatics has also aided the production of a bovine genome database - which will assist the searchfor new bovine genes – and identifica-tion of peptides that block the interac-tion of proteins involved in bacterial replication and repair. Such peptidescould provide leads for the developmentof broad-spectrum antimicrobials forhuman use.

On the production front, we are working on methods to improve woolproduction, protect sheep against internaland external parasites, deliver drugsand other molecules to livestock, andmanage aquaculture ponds.We arealso looking at ways to improve digestion in ruminants, using moleculartechniques to enhance ruminantmicrobe digestion of fodder and todetoxify certain forage plants.

Human applications of our work includereducing enteropathogenic bacteria on beef carcases, developingelastomeric proteins for medical andindustrial use, and producing bovine cartilage for use in cancer treatment trials.

livestockapplications of biotechnology

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GENES AND PROTEINS

Through the CRC for Innovative Dairy Products and the CRCfor Cattle and Beef Quality, scientists in this sub-program areidentifying genes associated with mastitis resistance in dairycows, and marbling in beef cattle.The sub-program also supports the bioinformatics needs of the Division.

Sub-program LeaderDr Ross TellamPh (07) 3214 2476Email ross.tellam@csiro.au

MASTITIS RESISTANCE

Mastitis is an infection of the mammary(udder) tissue in cattle, sheep andgoats, caused by a range of bacteriaand sometimes yeast or fungi.Infections reduce milk output and altermilk quality, which manifests as ‘offflavours’, poor shelf life, and otherundesirable characteristics. As antibiotictreatment is ineffective against someorganisms, and leads to residues in themilk, research efforts aim to identifygenes associated with resistance tomastitis.These could one day be usedto select dairy cattle with inherentresistance to mastitis, or which couldbe modulated in a way that enhancestheir protective activity in cattle.

To identify candidate genes, a bovineimmune gene microarray – derivedfrom mRNA in mammary epithelialcells and circulating white blood cells –is being developed, which will allowassessment of the gene expressionprofile in infected and mastitis-resistantcattle.The function of the genes identified by these immune-screening

procedures will then be establishedthrough sequencing, and cell and biochemical techniques

MEAT MARBLING

The eating quality of beef is improvedby ‘marbling’ - the accumulation of fatin skeletal muscle. As a result, marblingis a highly desired trait in Japan. InAustralia, Europe and America, however,consumers prefer leaner cuts. By identifying and regulating the genes and proteins involved in marbling, we maybe able to control the amount and distribution of marbling, and improvereturns to industry through animalsadapted to specific markets.

Research is focussed on a collection ofgenes involved in adipogenesis (fat cellformation) and in particular, the Dlk-1gene. Dlk-1 is a primary regulator ofadipogenesis.When it is present in pre-adipocytes (fat progenitor cells)there is suppression of adipogenesis,and when it is absent in pre-adipocytesthere is promotion of the cells towardsadipogenesis.

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By controlling Dlk-1 expressionthrough ‘immuno-modulation’, we aimto increase or decrease the productionof adipose tissue and therefore marbling.We have developed a recombinantDlk-1 protein and raised antibodies toit. It is thought that when cattle arevaccinated with the recombinant protein or infused with the antibodiesto Dlk-1, the inherent biochemicalactivity of Dlk-1 will be altered, and thepropensity for adipogenesis modified.

BIOINFORMATICS

An international consortium thatincludes CSIRO Livestock Industries,aims to sequence the bovine genomeby the end of 2005.The sequence willhelp scientists identify genes for desirableproduction traits such as meat marblingand tenderness, growth efficiency, anddisease resistance. It will also enableproduction of animals adapted to specific markets.

Members of the Genes and Proteinssub-program have committed to theend sequencing of 40 000 bacterialartificial chromosome (BAC) clones(containing fragments of the bovinegenome) in phase one of the project.End sequencing of the clones will assistconstruction of a physical map of thebovine genome, that is, the ordering ofBAC clones from one end of thegenome to the other. Appropriate BACclones will then be fully sequenced.

With the recent purchase of a Beowulfcluster – a 64 dual processor system –the team is manoeuvring into a positionto manage and analyse the huge volumeof internal and external data that willarise from the bovine genomesequencing project and associatedresearch efforts.This increasing bioinformatics capacity will also betransferred to the management andanalysis of the ovine and other eukaryoticgenomes in the future.

Data management within CSIRO isexpected to improve with the uptakeof LIMS (Laboratory InformationManagement System).This system willtrack and standardise information generated in laboratory experiments,and improve the statistical power ofscientific analyses, particularly microarray experiments.

Bioinformatics has also been used toidentify peptides that block the interaction of proteins involved inbacterial replication and repair. Suchpeptides could lead to the developmentof broad-spectrum antimicrobials.

By identifying and regulating the genes and proteins involved in marbling,scientists aim to improve returns to industry through animals adapted to specific markets. (Photo Greg Bortolussi, CSIRO Livestock Industries)

As antibiotic treatment is ineffective against some organisms,and leads to residues in the milk, research efforts aim toidentify genes associated with resistance to mastitis“ “

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MOLECULAR GENETICS

The Molecular Genetics sub-program uses molecular techniques to identify genes for commercially important traits in cattle, prawns and sheep.

Sub-program LeaderDr Bill BarendsePh (07) 3214 2444Email bill.barendse@csiro.au

BEEF QUALITY

Beef tenderness and ‘marbling’ - theaccumulation of fat in skeletal muscle – enhance the eating experience of Australian beef. Bothqualities are heritable and, through theCRC for Beef Quality, genes associatedwith each have been identified.To dothis, the human genomic map wascompared with the early stages of thebovine genome map, and the mostlikely location of the marbling and tenderness genes identified.Associations between these genesand the tenderness and marbling phenotypes were then identified usingpedigree information, blood samplesand carcasses from thousands of animals across seven breeds.

The GeneSTAR® Marbling test,commercialised by Genetic Solutions, isnow available to breeders.The DNAtest is based on the thyroglobulin gene,which encodes a protein that indirectlyaffects metabolism.The more recentGeneSTAR® Tenderness test is based

on two different forms of the calpastatin gene, which encodes anenzyme that inhibits tenderising post-mortem. One form of the gene isassociated with increased tendernessand the other with increased toughness.

Cattle breeders can now select cattlebased on their GeneSTAR® rating:2-STAR, 1-STAR or 0-STAR. Animalswith a 2-STAR rating for tenderness,for example, have two copies of thefavourable form of the gene and a 10%difference in tenderness compared to0-STAR (no copies of the favourableform) cattle. Further research isunderway to identify more genes fortenderness, marbling, parasite resistanceand other traits, in order to improvethe power and economy of thesegenetic tests.

Work has recently begun looking atthe genetic basis of tick resistance indairy cattle and retail beef yield – theamount of saleable meat on an animal.

Research into beef quality has resulted in commercialtests for meat tenderness and marbling.Photo: Genetic Solutions

livestockapplications of biotechnology

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BOVINE GENOME DATABASE

In collaboration with the bioinformaticsgroup a database of all publicly available bovine sequences has beendeveloped.The Interactive Bovine InSilico SNP database (IBISS), will assistthe search for new bovine genes.Thedatabase identifies single nucleotidepolymorphisms (SNPs), which havebeen filtered to distinguish possiblesequencing errors from putative SNPs.The database also provides gene structure information by identifyingintron/exon boundaries.

PRAWNS

The Kuruma prawn (Penaeus japonicus)is a high value export product forsouth-east Queensland, fetching $80-$120/kg on the Japanese market.As large, live prawns fetch top prices,our research, in collaboration withCSIRO Marine Research, Food ScienceAustralia, Rocky Point Prawn Farm, andTomei Australia, is focussed on geneticimprovement.

A genetic map identifying two quantitative trait loci regions associatedwith body weight, body length (head-tail) and carapace (head) lengthhas been produced. Using genomewalking techniques, these regions willbe characterized and searched forgrowth-related genes. Once genes are

identified, the DNA markers associatedwith these genes will be validated incommercial populations. Ultimately,these markers could become part of acommercial test for industry selectionof prawns with enhanced growth.

Markers associated with sex have alsobeen identified.These could one dayallow aquaculturalists to stock theirponds with female prawns. Fightingamongst male prawns, and otherreproductive stresses associated withmixed sex ponds are thought to be amajor factor in poor pond performance.Also, female prawns generate greaterprofits for farmers than males, due totheir larger size and better feed-conversion rates. As little is known about the mechanisms of sex determination in prawn species, theidentification of markers associatedwith gender could lead to elucidationof the developmental aspects of sexdetermination in prawns.

SHEEP

Through the Australian Sheep IndustryCRC’s Strategic Research Program, asheep tissue bank is being establishedto store a range of tissue samples,including blood, skin and muscle.Thebank will also include access to databases of associated phenotype,genotype and pedigree data.Thisresource will be essential for thedetection, confirmation and

comparison of genes for commerciallyimportant traits. Such genes will beidentified through the Sheep GeneMarkers project examining the sheepgenome for genes influencing traitssuch as wool and meat quality,reproduction, and disease resistance.Once identified, and associated withmarkers and screening tests, thesegenes can be rapidly selected for oragainst. A coarse map showing the estimated position of genes associatedwith a range of important wool qualitytraits is expected by 2004.

Recessive pigmentation genes causedark fibres in the Australian Merino,which can contaminate the wool clipand reduce its value. Contaminationmay be due to pigmented fibres on anotherwise white animal, or from thephysical transfer of dark fibres fromcoloured sheep to white sheep in theflock. Currently, the only way to reducethis problem is to remove colouredanimals and their white coloured carrier parents from the flock.Thesheep pigmentation project, fundedthrough the Sheep CRC, aims to identify recessive gene(s) responsiblefor pigmented wool and developDNA-based tests to detect white carrier sheep. A coarse map of theestimated position of the gene(s) forpigmented recessive self-colour blackwill be available in 2003. A feasibilitystudy of mapping other recessivecoloured phenotypes has also begun.

The genetic map of the Kuruma Prawn (Penaeus japonicus) will help scientistsidentify genetic markers for production traits and sex. Photo: Genetic Solutions

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GENES FOR PRODUCTION SPECIES

The genes for production species sub-program aims to characterise the biochemical and genetic interactions thatcontribute to beef quality and tick resistance in cattle, diseaseresistance in livestock, and the growth of wool and hair.

Sub-program LeaderDr Sigrid LehnertPh (07) 3214 2445Email: sigrid.lehnert@csiro.au

BEEF QUALITY

Research aims to identify genes andbiochemical pathways that are important in beef quality by looking atchanges in gene expression under different nutritional conditions and indifferent breeds. In research funded bythe CRC for Cattle and Beef Quality,and Meat and Livestock Australia, abovine microarray, based on fat andmuscle tissue cDNA libraries, is beingused to investigate gene expressionchanges in muscle and fat tissue samples from cattle on different diets,or from breeds with different fatmetabolism or meat marbling traits.Changes in gene expression, in culture,as pre-adipocytes (fat progenitor cells)differentiate into adipocytes, are alsobeing studied. Interesting genes identified in this comparative analysiswill be investigated using bioinformaticsand functional studies. Applying geneexpression surveys to these systemshas the potential to provide novelinsights into the biochemical processes

under investigation, to pinpoint novelgene candidates for the traits underinvestigation, and ultimately, to contribute to novel biotechnology for use by the industry.

TICK RESISTANCE

The cattle tick, (Boophilus microplus),is a significant parasite of beef cattle.Using a microarray of skin cDNA,gene expression in tick resistant and susceptible cattle will be compared.Improved knowledge of the basis oftick resistance may lead to the development of genetic or other management strategies to be incorporated into an integrated system of parasite control.

DISEASE RESISTANCE

Pneumonia caused by Actinobacilluspleuropneumoniae (APP), is a commonrespiratory disease amongst intensivelyfarmed pigs, causing significant economiclosses for the industry. As some pigsperform better in an infectious and

Changes in gene expression in muscle and fat tissue indifferent breeds of cattle, or in cattle on different diets,can be detected using a bovine microarray.

livestockapplications of biotechnology

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WOOL QUALITY

This project aims to identify the genesand biochemical pathways involved inwool formation and quality.This willprovide a clearer understanding of thebasic cell biology of the wool follicleand enable manipulation of woolgrowth rates and fibre characteristics.The first step in understanding thesecontrols has involved the generation ofthree EST skin libraries, from RNAexpressed during follicle developmentin the foetus, or during wool growth inthe adult Merino.The ESTs weresequenced, analysed, and their expression patterns determined inwool follicles by in situ hybridisation.This has provided functional informationabout the candidate genes and identifieda subset of follicle specific genes forgene profiling studies.

With funding from the AustralianSheep Industry CRC, a combinedovine/bovine microarray based on

challenging environment than others,we are looking for genes that conferdisease resistance in those animals.With funding from Australian PorkLimited, a microarray containing cDNAfragments from subtracted libraries ofpig immune cells and tissue was developed.This microarray was probedusing RNA from white blood cells (circulating leukocytes) of pigs thatperformed well after immune challengewith APP and those that performedpoorly. About three hundred candidategenes were identified and sequenced.Other molecular techniques will beused to validate the identity and functionof the most promising candidates infurther investigations.

Similar techniques will be used to identifygenes responsible for innate immunityin sheep and cattle, and variation inresponse to vaccination.

sheep and beef skin cDNA librarieshas been developed.The microarray is being used to compare gene expression between sheep breeds with different hair and wool traits,and differences in gene expressionbetween the wool, haired and inguinal(fibre deficient) regions of sheep during wool follicle development.Expression levels of genes up- ordown- regulated will be quantified by quantitative real time PCR.

Information from these experimentswill allow molecular strategies to bedevised to reduce fibre diameter, orchange fleece characteristics such asfibre growth rate, fibre strength, crimpor pigmentation. Genes controlling thehair cycle can be targeted to removewool or hair in regions where it isunwanted or causes health problems,such as the breech.The genes are alsopotential markers that can be used inselective breeding programs.

By identifying genes and biochemical pathways involved in wool formationand quality, strategies to improve fibre characteristics, such as staplestrength, may be devised.

This project aims to identify the genes and biochemical pathways involved in wool formation and quality“ “

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PESTS AND DISEASES

The search for new and improved controls over human diseases and animal pests forms the basis of this sub-program.

Sub-program LeaderDr Kritaya KongsuwanPh (07) 3214 2512Email kritaya.kongsuwan@csiro.au

NEW PHARMACEUTICALS

Bacterial replication involves many proteins, organised into a complexmultifunctional machine known as the‘replisome’. In most bacteria the DNAPolymerase III enzyme forms the coreof this machine, with its ß-subunitinteracting with a range of other proteins to ensure efficient DNA replication and repair. For this reason,the ß-subunit acts as an ideal target forthe development of new antimicrobialcompounds.These compounds couldinhibit function of the protein byblocking the critical protein-proteininteractions of the subunits.

Bioinformatics was used to identify afive-amino-acid motif (QL[SD]LF)essential for protein-protein interactionon the ß-subunit.The importance ofthis motif in binding was confirmedusing conventional biochemical techniques.

As a result of this work a range ofpeptides that block protein-proteininteraction in vitro have been developed.The next step is to demonstrate inhibition in vivo. Peptidesthat achieve in vivo inhibition will actas lead compounds for drugs targetingHelicobactor pylori (responsible forstomach ulcers),Vancomycin resistant

Enterococcus sp., and Methycilin resistant Staphylococcus aureus, amongothers.This work will continuethrough a spin-off company,BetaBiotics, a collaborative venturebetween CSIRO and the Institute of Molecular Biosciences at theUniversity of Queensland.

SHEEP PARASITES

Intestinal nematodes, sheep blowflyand sheep lice are major causes ofproduction losses in sheep. As parasiteresistance to chemical control methodsincreases, there is an urgent need toidentify new chemical and other control methods. A three-prongedapproach to the problem is underway.

Firstly, using molecular biology techniques, a small number of genesinvolved in the establishment of intestinal nematodes in sheep havebeen identified.The genes enable the nematodes to overcome the ‘oxidativeburst’ from the immune system of thesheep. By interfering with the regulationof this genetic system, we hope toswitch the parasites’ anti-oxidant capabilities off.

Antimicrobial compounds targeted towards the subunitof DNA Polymerase III, could lead to drugs for humanbacterial infections.

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Secondly, a drug-screening assay foradult nematodes has been developed.In the past, new drugs were usuallytested on the free-living larval stage ofthe parasite, which is easy to handleand culture in the laboratory.We nowhave a nutrient medium that keeps theparasitic adult nematodes alive,enabling targeted testing of new drugs.About 200 new chemicals have beentested against adult nematodes, and sixhave significant activity. More advancedlead compounds are being tested againstnematodes known to be resistant toexisting chemicals.

Finally, the use of various Bacillusthuringiensis (Bt) endotoxins asbiocontrol agents for sheep blowfly,lice and intestinal nematodes is beinginvestigated. In collaboration withCSIRO Entomology, a number of newBt strains that are toxic to theseparasites have been identified.The mainfocus now is increasing the efficacy ofthese Bt strains using molecular techniques, and developing methods to deliver them to the sites of parasiteinfestation.

BREECH-STRIKE PROTECTION

Merino sheep are particularly prone toa painful, wasting condition known asbreech-strike.The condition occurswhen the sheep blowfly (Lucilia caprina) lays its eggs on wet or irritated patches of skin around thebreech, or ano-genital region. Afterhatching, the larvae burrow into theskin and eat the flesh for two to threedays, before dropping off into the grassto pupate.The condition tends tooccur during prolonged periods ofdamp weather, when the folds of skinaround the breech trap moisture andfaeces.Techniques to control breechstrike have focused on surgicallyextending the bare patch of skinaround the anus, with good success.However, we are developing an alternative, painless procedure that killswool follicles in the treatment area.

The technique involves application ofaminolevulinic acid - an amino acidpresent in the skin of all mammals – tothe breech.This is absorbed into themitochondria in the wool follicle and,at high concentrations, causes a buildup of protoporhyrin 9 (a haem precursor). As protoporphyrin 9 isphoto-labile, application of high intensitywhite light results in the transfer ofelectrons from the porphyrin to oxygen,

and the production of cytotoxic oxygenfree radicals.This results in targeted celldeath and cessation of wool growth inthe treated area.

Field trials have shown that the treatment offers the same level of protection as the surgical method.Aspects of the treatment process arenow being optimised, to make it fasterand easier to use in the field.

We are also studying the gene for epidermal growth factor (EGF) insheep and other animals. Human EGFis currently used for ‘biological woolharvesting’, a technique that shutsdown wool follicles for 24 hours,causing a break in the wool fibre.Eight to ten days later, the fleece canbe removed without shearing.We haveisolated and characterised the sheepEGF gene in an effort to determineand compare its action. However, thegene appears to be silent, and the EGF genes of other ruminants are also silent. Characterisation of the EGF gene for whales, dolphins and hippos is underway, to see if there is aphylogenetic link between thesespecies and ruminants.

The sheep blowfly (Lucilia caprina) causes breech-strike in sheep.Photo: CSIRO Entomology

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TICK FEVER

Outbreaks of tick fever, caused by thebacterium Anaplasma marginale, arewidespread across tick-infested areas inthe northern part of WesternAustralia, the Northern Territory, andQueensland.The bacterium destroysthe red blood cells of cattle, causinganaemia, weakness, respiratory distress,reduced fertility and milk production,and loss of appetite and condition. Ifleft untreated, the animals may die.

Through the CRC for VaccineTechnology, two proteins have beenidentified from the bacterium, whichcould form the basis of a vaccine inthe next few years.When injected intocattle, these recombinant proteins(produced after cloning and expressingeach protein gene in Escherichia coli),protect the animals from subsequentchallenge with the bacterium. Eachprotein currently confers a 50% dropin parasitemia and red blood celldeath.We hope to increase this to80% - enough to reduce the clinicalsymptoms of the disease and itsimpact on productivity, and preventmortality.To do this, two promisingproteins will be combined in a singlevaccine. If this confers increased protection larger scale vaccination trials will be conducted.

NANOTECHNOLOGY

The elastomeric protein, resilin, isfound in insects, where it is used tostore energy for jumping and flight. Itdisplays outstanding resilience andfatigue properties, likely due to the 3-dimensional structure of the protein,which is stabilised by intermolecularcross-links.

In a new collaborative project involvingfour CSIRO Divisions and threeAustralian Universities, a range of techniques will be used to gain adetailed understanding of resilin’sstructural, molecular and mechanicalproperties.This understanding mayenable synthesis of synthetic polymers(biomimetics) displaying similarresilience and fatigue properties.These could be used for medical (drug delivery, vascular prostheses),micromechanical (actuators, hinges),and industrial (accelerometers) applications.

To understand the different physicalfunctions of resilin in insects, the spatialand temporal expression of the genein Drosophila will be investigated, andthe resilin gene from the common catflea (Ctenocephaledes felis) will becloned and expressed.The propertiesof these two forms of resilin can thenbe compared.

To date, we have expressed and purified a recombinant resilin protein,and tested a variety of enzymes fortheir ability to catalyse intermolecularcross-links.These enzymes may also beused to cross-link other organic molecules with a view to producingmaterials such as plastics in a moreenvironmentally friendly way.They mayalso be used to upgrade cheap, lowvalue protein by-products into highervalue food or agricultural products.

Dr Chris Elvin and his team are employing a range oftechniques to understand the properties of resilin.

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CELL BIOLOGY

The cell biology sub-program studies known genes and askshow they regulate the structural and metabolic aspects of celland tissue growth and development. This understanding isbeing applied to improve meat quality, and animal welfaremanagement, and the discovery of novel bioactive moleculesfor human and animal health.

Sub-program LeaderDr Gregory HarperPh (07) 3214 2281Email gregory.harper@csiro.au

MEAT TOUGHNESS

Meat toughness reflects the structureof proteins within muscle fibres, andthe connective tissue that links themthrough tendons and ligaments, to theskeleton. As connective tissue is amajor contributor to meat toughness,we are interested in understanding thebiochemistry of its development.To do this we are studying:• normal age development of

connective tissue;• effects of weight loss and weight gain

on connective tissue;• development of connective tissue in

extreme genetic variants of cattle and sheep (Callipyge or transgenic sheep, as well as double muscled cattle);

• effects of other environmental stressors on connective tissue.

One goal is to identify genes thatincrease muscle growth rates, but don’tincrease muscle toughness.

MEAT MARBLING

Through a collaborative project withthe CRC for Cattle and Beef Quality,and Kobe University, Japan, the geneticpredisposition and environmental factors that cause marbling in beef cattle are under study.The JapaneseBlack breed of cattle develops high levels of intramuscular fat (marbling)without extensive subcutaneous fat.Genetics plays a role in this trait, butenvironmental factors such as ß-carotene in the diet, and animal factors such as age, also contribute.Tissue samples taken from JapaneseBlack steers slaughtered at the end ofa breed comparison experiment arebeing examined for the developmentand distribution of fat cells. A bovinemuscle/fat microarray will be used tostudy gene expression.This work willhelp us understand the variation ingene expression that occurs betweenanimals and within each animal as it

Mr. George Riding uses SDS polyacrylamide gel electrophoresis to separate andvisualise proteins that could form the basis of a tick vaccine.

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develops and ages. It is hoped variationin marbling could be reduced, as consistency and predictability areessential in the beef market.

NOVEL BIOACTIVES INANIMAL CARTILAGE

Shark cartilage is a popular but unsustainable source of bioactive molecules that is used in traditionalChinese medicine to treat malignanttumours and ageing diseases such asarthritis. Western medicine is also trialing these bioactives for their abilityto inhibit the spread of cancer byblocking blood vessel growth.

In collaboration with a biotechnologycompany and Food Science Australia,we have identified the same bioactivemolecules in bovine cartilage. Methodsto purify these molecules and producethem on an industrial (10,000 litre)scale have been developed.The product will provide an alternative andsustainable source of these therapeuticmolecules, and will also add value toabattoir waste, which is currently usedfor blood and bone fertiliser.

BIOACTIVE MOLECULEDELIVERY

A ‘cell encapsulation’ technique hasbeen developed that delivers drugsand other bioactive molecules eitherunder the skin or deep within tissues.The technique promises to reduce theneed for regular injections or providemore targeted molecule delivery inboth the livestock production andmedical instrument industries.

In the pig industry, for example, farmersfinish pigs with daily injections of thegrowth hormone, porcine somatotrophin(pST).We encapsulated rat musclecells containing a DNA ‘construct’encoding the pST gene, in alginatebeads coated with poly -L-lysine.The coating ensures the spherical capsules remain intact, but is porousenough to allow molecules of up to100 000 molecular weight, to diffuseout. This molecular weight selectivityalso excludes immunoglobulins,protecting the rat cells from the host’simmune system.The capsules werethen injected under the skin of pigs’ears. After six weeks the pigs showed a30% increase in growth and about a 5 fold decrease in back fat.The capsuleswere still intact and functional after 6 weeks in the animal.These results suggest a standard pST-mediatedgrowth response.

Other cell types have been successfullyencapsulated, including fat and ovariancells, and we are now developing animmortal rabbit muscle cell line toimprove acceptability of the techniqueto the food industry (given that rabbitsare eaten by humans). Future workincludes injection of capsules expressing various molecules into themuscles of cattle to study changes inmeat toughness and marbling, and a collaborative project with a medicalresearch group interested in enzyme-supplementation therapy for human genetic disease.

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ANIMAL WELFARE

Pigs are exposed to a variety of environmental and psychosocial stresses on a daily basis, including temperature, air quality and sharingpens with unfamiliar pigs. Under optimal experimental managementconditions, pigs are 15-30% more productive than they are in standardpiggeries, so breeding pigs forincreased stress tolerance is commercially advantageous.

In a project funded by Australia PorkLimited, a number of genes have beenidentified that are switched on or off inresponse to acute stress, such as a sudden increase in temperature.We are now looking to see if thisexpression is heritable and to correlateit with production traits.

Genes that are up-or down-regulated in response tostress could provide an objective measure of chronicstress for animal welfare purposes.

Whether these same genes can provide an objective measure ofchronic stress, is also being investigated.Production parameters and variousstress responsive blood proteins arebeing measured in pigs from piggeries,and compared to pigs housed underoptimal experimental conditions (positive control) and those housedunder long-term high temperatures(stressful negative control).Theseexperiments may assist developmentof a tool that will allow industry tocertify that animals are housed underconditions that are appropriate froman animal welfare perspective.

The capsules were then injected under the skin of pigs’ears. After six weeks the pigs showed a 30% increase ingrowth and about a 5 fold decrease in back fat“ “

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RUMINANT MICROBIOLOGY

The ruminant microbiology group has three main researchareas; on-farm control of pathogens affecting beef, improvingruminant digestion, and detoxifying important forage plantsusing microbial-based strategies.

Sub-program LeaderDr Chris McSweeneyPh (07) 3214 2665Email chris.mcsweeney@csiro.au

PATHOGENS

Bacterial pathogens associated with theconsumption of meat products,particularly hamburgers and salami, area serious human health concern.Enteropathogenic Escherichia coli, suchas serotypes 0157:H7, 0111 and 026,have caused outbreaks of haemorrhagiccolitis and haemolytic uremic syndromein Australia and overseas. Although theincidence of these infections is low,they can prove fatal.

Infection usually occurs as a result ofeating undercooked beef, derived fromcarcasses contaminated with low levelsof these pathogens.While proceduresare in place to minimise contamination(from faeces), our research has shownthat on-farm feeding practices couldbe managed to further reduce thechances of contamination. Such stepsare essential if Australia is to maintainits export markets.

The focus of our current work is to study the effect of different commercial finishing diets on total andpathogenic E. coli populations in thedigestive tract of cattle. In a series offeeding trials in Rockhampton, cattlewere fed diets based on pasture, grainor molasses. Analysis of faecal samplesthrough biochemical and serologicaltests, virulence profiles and microbialcounts, has shown total E. coli numbersto be highest in cattle fed on grain andlowest in those fed molasses androughage diets. Future work will lookat implementing suitable diets withoutcompromising productivity.We will alsoinvestigate the complexity of ruminantE. coli populations and factors thatcontribute to the pathogenesis andspread of virulence genes using molecular biology techniques, to distinguish between closely related E. coli strains and characterise strain-specific virulence properties.

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The use of probiotic organisms toreduce pathogenic E. coli load is alsobeing investigated. Lactic acid bacteria,for example, have been isolated fromthe gut of cattle and screened forinhibitory action against serotypes0157:H7, 026 and 0111. Suitably robustprobiotic organisms could be added tocattle feed just prior to slaughter, tohelp reduce specific E. coli serotypesthat are associated with disease.

RUMINANT DIGESTION

The major limitation to ruminant production in many tropical regions ofAustralia, Africa and Asia, is poor nutrition. Increasing the ability of ruminant microbes to digest fodder is one way of boosting production.This requires an understanding of thetypes of microbes present in therumen, the enzymes they produce fordigesting feed, and the nutritionalconditions under which the microbes flourish.

As 90-95% of ruminant microbes cannot be cultured, new techniquesare needed to identify these organismsand their functions. One such technique is ‘metagenomics’ – genomicanalysis of a complex microbial

community.To do this we constructeda bacterial artificial chromosomelibrary from rumen digesta samplesand then cloned large DNA fragmentsinto E. coli.These clones can now bescreened for the uncultured microbialdiversity and abundance, and novelgenes and gene activity. In the long-term, this information will be used to develop strategies to improvefeed utilization through probiotics,prebiotics, specific nutrients, or enzymeadditions to diets.

With the same objective in mind, waysto increase populations of rumen fungiare being considered. Previous workhas shown fungi can be stimulated withspecific micro-nutrients to improve thedigestion of plant fibre. By supplementingruminants with different micro-nutrients,growth factors that stimulate fungalproliferation and improve the digestionof tropical forages, may be identified.

In a project soon to begin in China,white rot fungi will be used to degradelignin in poor quality straw andimprove its digestibility in cattle.

DETOXIFYING FORAGEPLANTS

Dietary supplementation with leguminous plants is another way toboost ruminant production. Howevermany such plants contain toxins andanti-nutritional factors. Acaciageorginae, for example, produces fluoroacetate, which causes ‘Gidgeepoisoning’ in sheep and cattle. As theplant is widespread in northernAustralia, farmers are forced to lock uplarge tracts of land to protect their animals. By introducing microbes thatdegrade or neutralise such toxins, intothe rumen, animals may potentiallysafely graze on these plants. In a collaborative project with theUniversity of Western Australia, we aimto introduce a Butyrivibrio sp. into therumen that that has been geneticallymodified with a fluoroacetate-degradinggene from the soil microbe, Moraxella sp.

We are also searching for naturallyoccurring rumen microorganisms thatdegrade toxic non-protein amino acidsthat are present in some forage Acaciaspp.These plants would become morevaluable as protein supplements if therumen can adapt to protect animals fromthe adverse effects of these compounds.

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AQUACULTURE VIRAL DISEASE

The Aquaculture Viral Disease sub-program focuses on diseases of aquatic animals, in particular the viral pathogensthat threaten the sustainability of the prawn aquacultureindustry in Australia and Asia.These include Australia’s endemic gill-associated virus (GAV) and Mourilyan virus(MoV), and the exotic white spot syndrome virus (WSSV)and yellow head virus (YHV).

Sub-program LeaderDr Peter WalkerPh (07) 3214 2664Email peter.walker@csiro.au

ENDEMIC DISEASES

The endemic diseases projects haveresulted in the commercialisation oftwo PCR-based diagnostic kits, one forGAV and YHV, and a second for MoV.The kits are produced and distributedby the Taiwan-based company, FarmingIntelliGene Technology Corporation,and are used by industry to testbroodstock obtained from the wild, orfrom domestication programs, toensure the animals are disease-free.In Australia, the GAV-YHV test is alsoused in association with managementguidelines to ensure stock remain disease free during grow-out in ponds,or to allow farmers to deal rapidly andsafely with disease outbreaks and subsequent cleanup.

In an effort to reduce industry relianceon wild black tiger prawn (Penaeusmonodon) broodstock, we are collaborating on a project to domesticate the species. Our role willinvolve the development of ‘high

health’ or ‘specific pathogen free’animals and maintaining biosecurity.

We will also study vertical diseasetransmission (parents to progeny), andseek treatments that will produceprogeny from infected broodstock.As GAV and MoV infect the gonads of prawns, understanding the effect ofviral load on spawning efficiency isanother priority.

A better understanding of the structure, genetics and biology of GAVand YHV, has led to their classificationin the order Nidovirales, and establish-ment of a new family (Roniviridae) andgenus (Okavirus).The viruses are thefirst members in the order known toinfect invertebrates. Previously, onlyviruses infecting mammals and birds,such as coronaviruses and toroviruses,were classified in the Nidovirales.Thegenome of these invertebrate viruses is primitive in comparison to their mammalian and avian counterparts and may provide a window into their evolution. Comparative studies of

To reduce industry reliance on wild black tiger prawn(Penaeus monodon) broodstock, CSIRO LivestockIndustries is collaborating on a project to domesticatethe species. Photo: CSIRO Marine Research

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human and shrimp nidovirus enzymesmay also lead to the development ofnew drugs to treat disease.

The endemic diseases group has also participated in a national survey to determine if WSSV or YHV were present in Australian waters, inresponse to a disease incident in aDarwin aquaculture facility. Afteranalysing more than 3000 crustaceansamples, an absence of infection in thewild was confirmed.

EXOTIC DISEASES

Our exotic diseases work has focusedon providing research and training in molecular diagnostics with collaborators in Thailand,Vietnam and India. By helping such countriesovercome their disease problems, wehope to build a greater capacity tomanage any possible exotic diseaseoutbreak in Australia.

Research projects in Thailand haveresulted in a long-term strategicalliance with the Centre of Excellencefor Shrimp Biotechnology at MahidolUniversity.Through this alliance, the firstgenetic markers to distinguish individualisolates of WSSV and YHV have beendeveloped, and used to trace sourcesof infection and the origins of diseaseoutbreaks.Work in Thailand andVietnam has also led to identification of two new genotypes of yellowhead-related viruses (type 3 and type 4) -

indicating that YHV is a family of viruses, rather than one virus as previously thought. As the currentdiagnostic test kits do not detect thesenew genotypes, the kits will be updated to identify and quantify them.

A recent project in India, aims to usethe variable markers in individual isolates of WSSV and YHV to trace themovement of these viruses betweenhosts, farms and countries.

The sub-program also seeks to informpolicies that limit the spread of animalsand pathogens around the world. Asfish health activities in many developingcountries are poorly resourced, currentguidelines developed by the OfficeInternational des Epizooties (OIE) are often impossible to implementeffectively.We are working with theOIE, FAO (Food and AgricultureOrganisation) and other experts in fishdisease and epidemiology, to developmore relevant and easily applied policies for the Asian region.

BIOTECHNOLOGY

The biotechnology applications grouphas been building links with the Asianregion through a cooperative effortthat aims to understand prawns’responses to disease.With a consortium of researchers in Thailand,genomic and proteomic techniques arebeing used to study the response ofprawns to viral disease and compare

differences in gene expression inprawns with and without WSSV infection.

In collaboration with Mahidol Universityand the National Center for GeneticEngineering and Biotechnology inThailand, and the National TaiwanUniversity, a genetic linkage map of theblack tiger prawn (Penaeus monodon)genome is being developed.The mapwill be used to identify markers associated with commercially important traits such as disease resistance and fecundity. Informationfrom the Kuruma prawn (P. japonicus)linkage map, completed by CSIROLivestock Industries and the AustralianInstitute of Marine Science in 2001, willassist the development of this new map.

Our bioinformatics capability isexpanding to cope with the influx ofinformation from the linkage map projects.This capability will allow management and analysis of data froma large prawn genome project that willbe developed in collaboration with anetwork of Asian researchers over thenext five years.

The biotechnology group is also investigating the role of RNA interference (RNAi) in the naturaldefensive response of shrimp andother invertebrates to viral infection.This is a new and exciting field ofresearch with potential for significantscientific and commercial outcomes.

Capacity building programs in Vietnam,Thailand and India will help prawn farmers in these countries overcome disease problems.Photos: Nigel Preston, CSIRO Marine Research

Because of its biocotainment

capacity and its breadth of

expertise in viral diseases, the

program delivers scientific

support to public health

authorities on diseases that

affect animals and people

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Diagnosis and epidemiology 28Fish disease research and diagnosis 30Molecular diagnostics 32Electron microscopy and emerging pathogens 35

Newcastle disease virus 37Virology 40Tissue culture and media production 41

diagnosticsciences

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Program Leader Dr Peter DanielsPh (03) 5227 5272Fax (03) 5227 5555Email peter.daniels@csiro.au

The Diagnostic Sciences programat the Australian Animal HealthLaboratory (AAHL) in Geelongdiagnoses diseases of nationalsignificance to Australia’s farmedlivestock, aquatic animals, andfauna.The program helps protectAustralia's livestock and aquaculture industries frommajor economic losses in theevent of emergency disease outbreaks, and supports nationaland international trade by providing scientific support andinformation on animal disease issues.

The program’s skill base includes virology and serology, diagnostic andexperimental pathology, epidemiology,electron microscopy, molecular virology,bio-molecular engineering, and monoclonal and recombinant antibody technology.

The program has a strong researchprogram in diagnostic test developmentand application, in the pathogenesis ofdisease, and in the spread and management of diseases in populations.Other key functions of the programinclude training of field and laboratorybased animal health professionals indisease recognition and in methodsand technologies used to diagnose

diseases; and the provision of consultancy and advisory services tofederal and state government agencies.The program keeps at the cutting edgeof diagnostic sciences and, because ofits national role, is involved in technology transfer of materials,techniques and training to national andinternational laboratories.

Because of its biocontainment capacityand its breadth of expertise in viral diseases, the program also delivers scientific support to public healthauthorities on diseases that affect animals and people, and in assessingagents that may be useful for the biological control of pest species.

A number of scientific projects originating in the Diagnostic SciencesProgram are now expanding andevolving under the umbrella of thenew Australian Biosecurity CRC.

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DIAGNOSIS AND EPIDEMOLOGY

The Diagnosis and Epidemiology project has three mainobjectives:• To develop, maintain and apply diagnostic procedures for

emergency (exotic) and emerging diseases of livestock that affect international trade;

• To provide diagnostic testing of animals and biological products for Australian quarantine purposes, and advice to government agencies on quarantine protocols and exotic disease control;

• To conduct research on emerging diseases.

Project LeaderDr Deborah MiddletonPh (03) 5227 5016Email deborah.middleton@csiro.au

DISEASE DIAGNOSIS ANDQUARANTINE TESTING

Emergency diseases include foot andmouth disease (FMD) and transmissiblespongiform encephalopathies (TSE),such as bovine spongiformencephalopathy (BSE). Emerging diseases are those caused by novel disease agents, and once included therecently identified Nipah virus, whichcaused a disease outbreak in pigs andhumans in Malaysia in 1999.

To perform our diagnostic functions,laboratories involved in the projectdevelop diagnostic reagents, such asantigens and antibodies, to emergencydiseases, and maintain supplies of these

reagents for dispatch to regional diagnostic laboratories during outbreaks.These reagents are alsoused for routine quarantine testing ofanimals or animal-derived productsfrom overseas. Diagnostic test procedures are developed and maintained according to internationalquality control standards and accreditation procedures.

Due to individual staff expertise, theproject also supports OfficeInternational des Epizooties (OIE) reference laboratories for bluetonguevirus, Newcastle disease virus and virulent avian influenza.This functionrequires development, maintenance

AAHL is an OIE reference laboratory for a number of viruses,including bluetongue, an insect borne viral disease of ruminants.

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and distribution of internationallyaccepted diagnostic reagents and testsfor these diseases.The project alsomaintains the Australian and NewZealand Rabies Reference Laboratory,the National Brucella ReferenceLaboratory, the National Serum Bankand the TSE Reference Laboratory.

Recent diagnostic advances includeinstallation of a robotics system toimprove our capacity to test samplesin the event of an FMD outbreak. Ourlaboratory information managementsystem is also being upgraded toimprove access to veterinary caseinformation on a national scale.

EXPERT ADVICE

Provision of expert advice to government agencies may relate to the development or implementation ofquarantine protocols, diagnostic or epidemiological advice in emergencysituations, and advice to national orinternational livestock producer ortrade delegations. It also includes participation in the management ofnational exotic disease monitoring andsurveillance programs such as theNorthern Australian QuarantineStrategy and the National ArbovirusMonitoring Program, and membershipof ad hoc national committees such asthose responding to emerging diseasesituations.

The project has an important role intraining programs to support nationalexotic disease preparedness. Coursesinvolving substantial practical components are provided for key government veterinarians with responsibilities in this area. Lecturesand demonstrations are provided toveterinary practitioners and students.

In 2002, project staff participated inand assisted Exercise Minotaur, a desktop simulation of an FMD outbreak in Australia, involving morethan 1100 people from governmentagencies and livestock industry bodies.

RESEARCH

Our research programs aim toimprove diagnostic tests and produceinformation of epidemiological significance.This includes description ofthe pathogenesis of emerging diseasessuch as Australian bat lyssavirus andvery virulent infectious bursal disease,and studies of vaccines for Japaneseencephalitis. Our involvement in theAustralian Biosecurity CRC will investigate transmission and pathogenesis of serious pathogens,including the Nipah, Hendra and WestNile viruses.

To develop staff experience with exotic diseases and increase the disease control capability in othercountries, the project has a technologytransfer component as well.Thisinvolves collaborative research with,and training of, scientists in South EastAsia.Through an AusAID program inVietnam, for example, veterinarians arebeing briefed on the latest diagnosisand vaccination strategies for classical swine fever.The transfer ofinformation, such as the need to vaccinate pigs at six weeks of age inorder to confer protection againstswine fever, will help reduce futureoutbreaks of the disease and protectan important source of meat for thelocal people.

A CSIRO/AusAID project will help control outbreaks of classical swine fever inVietnam and protect an important source of meat for the local people.

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FISH DISEASE RESEARCH AND DIAGNOSIS

Australia is free of many of the aquatic pathogens (viruses,bacteria and parasites) that plague overseas fisheries andaquaculture industries.

Project LeaderDr Mark CranePh (03) 5227 5118Email mark.crane@csiro.au

The AAHL Fish Diseases Laboratory(AFDL) is a key player in maintainingnational fish health through researchinto infectious diseases of finfish,crustaceans and molluscs, and the provision of diagnostic and health surveillance services for quarantine and industry.

DISEASE DIAGNOSIS,SURVEILLANCE,ANDEXPORT CERTIFICATION

The Office Internationale desEpizooties (OIE) maintains a list of finfish, crustacean and mollusc diseases,many of which are exotic to Australia.The AFDL has, or aims to acquire, thecapability to detect and identify thesediseases for import/export testing andsurveillance, using skills in histopathology,molecular biology, immunology andAAHL’s electron microscopy capability.

SalmonidsAmong the OIE listed diseases arethree viral pathogens of salmonids;infectious pancreatic necrosis virus,viral haemorrhagic septicaemia virus,and infectious haematopoietic necrosisvirus. AFDL conducts a surveillanceprogram for these diseases for theTasmanian farmed salmon industry, andexport certification of salmon eggsfrom Tasmania for use overseas. Exportcertification testing is based on internationally accepted protocols forvirus detection.

AFDL has also developed a diagnostictest for a major bacterial pathogen ofsalmon, Piscirickettsia salmonis, andestablished tests for other exoticpathogens such as infectious salmonanaemia virus, spring viraemia of carpvirus, Myxobolus cerebralis,Renibacterium salmoninarum andAeromonas salmonicida.

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CarpSpring viremia of carp, caused byRhabdovirus carpio or spring viraemia ofcarp virus, can kill up to 80% of farmedcarp in Europe, when the rise in watertemperatures between winter andsummer is slow.The virus has not beendetected in Australia, but it has beendetected in ornamental fish, such asgoldfish. As Australia imports largenumbers of goldfish, AFDL has established a diagnostic capability forthe virus.

PilchardsPilchard herpes virus has been implicated in the deaths of pilchardsduring disease outbreaks in Australia in1995 and 1998.The tuna industryimports pilchards for feed and,although not proven, concerns havebeen raised that imported pilchardsmay have transferred the virus toAustralia’s wild stocks.The AFDL is amember of the ConsultativeCommittee on Emergency AnimalDiseases joint pilchard scientific working group, which aims to improveAustralia’s diagnostic capability for thedisease and assist development ofmanagement strategies should the disease strike again.

PrawnsIn collaboration with the AquacultureViral Disease sub-program in Brisbane,AFDL is working to improve diagnostictools available for the major exotic diseases of prawns, including whitespot syndrome virus and yellowhead virus.

OystersIf the Australian oyster industry beginsto export product overseas, AFDL willneed to provide export certification. Asmollusc diseases, such as Bonamiosis(caused by the protozoan parasiteBonamia), have been poorly studied inAustralia, the laboratory aims to workwith the OIE reference laboratory formollusc diseases, to develop moleculardiagnostic tools. Relationships betweenBonamia infections in different countries will also be investigated, toenable development of diagnostic toolsthat can distinguish between endemicand exotic strains.

Red sea bream iridovirusWork recently started on a diagnostictest for red sea bream iridovirus, whichinfects more than 30 species of fish,including snapper, tuna, red sea bream,sea bass, mackerel and yellowtail kingfish.The virus has caused seriouslosses in Japan’s marine aquacultureindustry, but has not been detected inAustralia. However, current tests available in Australia do not reliablyidentify the virus. Samples of the virus

and diagnostic antibodies have beenimported from the OIE iridovirus reference laboratory in Japan. Usingthese samples AFDL will establishreagents and protocols for bothimmunodiagnostic and molecular(PCR) diagnostic tests.

TunaNo viruses of tuna have yet been identified. However, based on experience in other fish species it islikely that young fish are susceptible to viral infection and disease.To assistdetection and identification of viralagents, AFDL aims to develop tuna celllines for use in disease surveillance forthe Southern bluefin tuna industry, aswell as identification of disease-freebroodstock, should these be requiredin an industry domestication program.

OTHER WORK

The AFDL provides diagnostic reagentsand conducts diagnostic tests for arange of other exotic pathogens. It alsoprovides laboratory training in infectiousdisease diagnosis and technology transfer to other fish health laboratories.

Red sea bream iridovirus infects more than 30 species of fish, including thejack mackerel.

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MOLECULAR DIAGNOSTICS

The primary function of the Molecular Diagnostics project isto support the investigation of emergency animal diseases(both exotic and endemic), and exclusion testing (presence or absence of a specific disease agent).

Project LeaderDr David BoylePh (03) 5227 5018Email david.boyle@csiro.au

To do this, the project uses cuttingedge molecular diagnostic technologies,including the polymerase chain reaction(PCR), gene cloning, sequencing andrecombinant antibodies.The tests areunderpinned by various research activities, including the development ofrecombinant antibodies for poultrypathogens, characterisation of theMenangle virus genome and its pathogenesis in pigs, and studies oncapripox viruses that affect sheep,goats and cattle.

DIAGNOSTIC TESTS

The acquisition of a robotic system forDNA extraction and testing, and implementation of real timePCR/sequence detection tests usingTaqMan chemistry and associatedinstrumentation, will boost our diagnostic capacity and accuracy in theevent of an emergency disease outbreak.Tests that we have developedand/or implemented allow the detection of disease agents within four

hours of receipt of the samples. Morethan two of these hours involve unattended instrument time, freeing upstaff for other activities.

Tests are available for diseases such asfoot and mouth disease (FMD),Newcastle disease, Nipah virus,Hendra virus and Australian batlyssavirus.The system can also detectall infectious bursal disease virus(IBDV) pathotypes, and differentiatebetween exotic very virulent strainsand endemic non-pathogenic strains.These tests are supported by international quality assurance standards (ISO17025). Samples fortesting come from the Chief VeterinaryOfficer in each state.

The project has the diagnostic capabilityfor some 50 other pathogens and alsoacts as the Australian reference laboratory for rabies and Australianbat lyssavirus.

Through the Australian Biosecurity CRC,the Molecular Diagnostics project willcontribute to increased preparednessfor exotic disease outbreaks.

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Development of assays for differentialdiagnosis of up to eight pathogens is apriority.This ability to identify differentdiseases in one assay will be extendedusing microhybridisation technology,where up to perhaps 1000 diseasemarkers may be arranged on a singletest platform.

MENANGLE VIRUS

In 1997 a new paramyxovirus was isolated from the brain, lungs and heartof stillborn piglets in a commercial piggery in New South Wales.The virus caused a drop in the farrowing(birthing) rate of sows, and an increasein stillborn and mummified piglets,which exhibited severe degenerationof the central nervous system.Twoworkers at the piggery also suffered aflu-like illness and developed antibodies.The virus was subsequently thought tohave jumped to pigs from fruit bats,which are believed to be the naturalhost of the virus in the wild.

Through the Molecular Diagnosticsproject, the genome of the Menanglevirus has been sequenced.This hasassisted its classification – identifyingwhat the virus is, and how it is relatedto other known paramyxoviruses suchas the Hendra and Nipah viruses.Thegenome sequence has also permittedthe development of rapid molecular

detection techniques such as quantitative real-time PCR, which hasimproved the diagnostic capability forthis new virus in the event of another outbreak.

The pathogenesis of the virus inweaned pigs is also being investigated.This includes studying how the virus isspread between pigs and identifyingthe target organs and tissues in whichthe virus replicates.

CAPRIPOX

Capripox viruses cause sheep and goatpox and lumpy skin disease in cattle.A threat assessment of these virusesfound that any outbreak would have asimilar impact on trade, livestock andtourism, as foot and mouth disease.The viruses are found in the Middle-East, north Africa, India, Myanmar andparts of Russia.While there is a lowrisk of introduction into Australia, theviruses still pose an economic threat,particularly to the sheep and woolindustry.There are many differentstrains of these viruses, some morepathogenic than others, and these differences are not well characterised.

As Australian quarantine forbids theimportation of capripox viruses forfurther study, we are collaborating withthe National Centre for Foreign

Animal Diseases in Canada to improveour index case (first case) diagnosticcapability and post-outbreak surveillance for sheep pox. Advanceswith this virus will assist future workon goat pox and lumpy skin disease.

FOWLPOX VACCINE

The current vaccine for fowlpox virus,which causes production losses in layerchickens, does not adequately protectolder birds or protect against differentstrains of the virus. Research in theproject identified a second virus (aretrovirus) in the genome of thefowlpox virus, which causes cancer-likediseases in animals and humans.Theretrovirus also appears to be associated with fowlpox vaccine failure.We have removed this retrovirus andconducted safety and potency testingand innocuity testing (to ensure it doesnot contain other poultry pathogens)on the resulting fowlpox virus.Vaccinemanufacturer Intervet is now conducting field trials of the retrovirus-free strain of fowlpox virus, as a vaccine against fowlpox disease.

Research at AAHL sometimes requires the use ofhigh microbiological security suits.

CHICKEN RECOMBINANTANTIBODIES

A new project aims to producerecombinant antibodies as keyreagents for the development of noveldiagnostic tests or for therapeuticapplications. Recombinant antibodiesoffer several advantages over conventional monoclonal antibodies.Highly diverse recombinant antibodylibraries can be constructed from anumber of mammalian or non-mammalian hosts and specific antibodies selected using phage-display.These antibodies can be geneticallymodified and tailored to suit particularapplications. Antibodies derived fromchickens enable the construction ofrecombinant antibodies to conservedmammalian antigens that are normallynot immunogenic. Currently we aredeveloping recombinant antibodybased reagents for diagnostic assays(competitive ELISA) to discriminatefoot and mouth disease virus in vaccinated and infected animals (cattle,pigs, sheep and others).

HIV/AIDS VACCINE

The Molecular Diagnostics project iscommitted to the development of anHIV/AIDS vaccine as part of anAustralian consortium funded by theNational Institutes of Health.The consortium will undertake Phase I andPhase IIa clinical trials of a ‘prime/boost’vaccine strategy. In double blind placebotrials, 24 volunteers will be vaccinatedwith a DNA vaccine and a recombinantfowlpox vaccine (both carrying HIVgenes expressing HIV antigens).Preliminary trials of this vaccinationstrategy in monkeys have shown significant levels of T-cell response andprotection. Anecdotal evidence suggeststhat T-cell immunity is important forthe clearance of HIV and may helpprotect against infection.The vaccineswill be studied for safety and their ability to stimulate an immuneresponse.

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ELECTRON MICROSCOPY AND EMERGING PATHOGENS

The electron microscopy and emerging pathogens projectuses advanced transmission (TEM) and scanning electronmicroscopical (SEM) techniques to diagnose, identify and characterise exotic and new viral pathogens, and to apply our knowledge and expertise to national and internationaldisease issues.

Project LeaderDr Alex HyattPh (03) 5227 5419Email alex.hyatt@csiro.au

Our achievements include identificationof the Hendra and Nipah viruses afterdisease outbreaks in livestock andhumans, identification of Australian batlyssavirus, pilchard herpes virus, theorbivirus causing kangaroo blindness,and the chytrid fungus responsible forfrog deaths worldwide.The projectalso acts as the Office Internationaledes Epizooties (OIE) reference laboratory for ranaviruses, a group oflarge DNA viruses that infect amphibians and fish.This responsibilityrequires us to supply reagents, methodsand advice to other laboratoriesaround the world.

Our diagnostic techniques are basedon immuno-electron microscopy,where gold-labelled antibodies toviruses in our reference collection areused to identify viruses in field samples.Antibodies that bind to the field sampleare seen as black spots under the TEMor SEM.

Our diagnostic capabilities will soon be enhanced by a Digital ImagingNetwork (DIN). In the event of anoutbreak of an unknown pathogen, forexample, the DIN will enable electronmicroscopy images of these pathogensto be posted on a web site for identification by other national andinternational experts. Chat and emailfacilities will also be available.

To maintain our diagnostic capabilities,techniques are continually refined andimproved, and fed by research activitiesthat boost our reference collection ofviruses and our knowledge of virusstructure and morphogenesis (how thevirus puts itself together, replicates andspreads).These activities includeresearch that will assist cane toad control, an understanding of henipaviruses and viral pathogenesis,and identification of the chytrid fungus.

Electron microscopy techniques are used to diagnose, identify and characteriseviral pathogens.

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RESEARCH ACTIVITIES

Cane ToadsIn collaboration with CSIROSustainable Ecosystems, genes specificfor cane toad (Bufo marinus) development are being sought, which,if manipulated, will interfere with metamorphosis, preventing the transition from tadpole to adult. If suchgenes are found, a means of distributingthe gene/s through the cane toad population will be required. Ranaviruses,which infect amphibians and fish, mayact as suitable distribution vehicles.Scientists in the project are now characterising a promising ranavirus,and identifying parts of the genome thatcould be removed to reduce pathogenicity. Other non-essential genes,which could be removed to provide aninsertion site for the metamorphosisgene/s, are also being sought.

Henipaviruses‘Henipavirus’ is the name given to twoparamyxoviruses that infect fruit batsor flying foxes – the Hendra and Nipahviruses.While the viruses do not appearto cause disease in fruit bats, they docause disease in other species, includinghorses, pigs and humans. However littleis known about how the viruses aretransmitted between bats, andbetween bats and susceptible species.

To study the transmission of Hendravirus, we have infected the black flyingfox (Pteropus alecto) with the virus andwill try to isolate the virus from urine,blood and tissue samples. Similar studies are planned with Nipah virus inMalaysia’s large flying fox (Pteropusvampyrus). If we can isolate the viruses,and identify the amounts needed forinfection, we will begin transmissionstudies in susceptible animals. Improveddiagnostic techniques for the viruseswill also be developed.

Ecosystem healthThe henipavirus work will feed into anew frontier of scientific research, drivenby the World Health Organisation,known as ‘ecosystem health’.Thisresearch will assess the state of theenvironment using a number of factors,including micro-organisms, and theanthropogenic (human induced) reasons for that state. In terms of henipaviruses, answers will be soughtto the reasons for the distribution offruit bats, and features of their environment that would impact on virus transmission.

Chytrid fungusSince the electron microscopical discovery of the chytrid fungus(Batrachochytrium dendrobatidis), we havecontinued to work, in collaboration withothers at AAHL, to develop a real-timePCR diagnostic assay for the fungus.Field sampling techniques have alsobeen developed, ensuring consistent sampling by frog researchersacross Australia, and subsequent PCRanalysis at AAHL. Samples are alsoexamined by electron microscopy tobuild and maintain staff competencyand standards on the equipment.

Pathogenesis of infectionIn collaboration with CSIROSustainable Ecosystems, we are investigating the use of microarrays andmicroscopy to study pathogenesis.Microarrays can be used to see whichgenes may be up or down regulatedduring infection of a host.This information will help identify likely locations of the virus in the host i.e.,infected cells or tissues. Using immuno-electron microscopy, we can thendetect the cells or tissues targeted bythe virus.

Genes specific to cane toad (Bufo marinus) development arebeing sought as a means of biological control.

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NEWCASTLE DISEASE

Newcastle disease virus (NDV) is a single stranded RNAparamyxovirus that can cause respiratory, neurological anddigestive symptoms, and death, in chickens.

Project LeaderDr Allan GouldPh (03) 5227 5119Email allan.gould@csiro.au

In Australia, most strains of the virusare avirulent (do not cause disease),but outbreaks of virulent NDVoccurred in NSW in 1998, and NSWand Victoria in 2002. Early workshowed that the 1998 outbreak wasnot caused by the incursion of anexotic strain of NDV into Australia, butby a virus already present in the country. As a result, the NDV projecthas been charged to fully characterisethe viruses responsible for virulentNDV in the Australian poultry industry,and to develop new and improveddiagnostic procedures for the disease.

CHARACTERISATION OF AUSTRALIAN NDVSTRAINS

To gain a better understanding of theviruses responsible for the NDV outbreaks, the virulent viruses isolatedfrom infected poultry tissue, and anavirulent strain that had caused respiratory symptoms in chickensaround the same time, were sequenced.Ten complete genome sequences weredetermined, and differences in the rateof mutation for certain genes werenoted.

The analysis showed that the NDVstrain responsible for the 1998 outbreak was related to the avirulentisolate, rather than an exotic incursion.A two base pair mutation in the fusionprotein (F) gene of the avirulent‘progenitor’ virus had enabled it tobecome virulent.

Newcastle disease is a highly contagious viral disease of poultry and wild birds,capable of causing up to 100% mortality. Here virulent Newcastle disease virusin the conjunctiva of a chicken is indicated by the brown stain.

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Biological studies revealed otherunique characteristics of the virulentand progenitor viruses, which becameimportant in subsequent investigations.The haemaglutinin-neuraminidase (HN)gene, for example, had an extra nineamino acids, compared to the normalamino acid sequence for this protein.Comparison with Australian and overseas isolates showed the HN protein was related to Australian NDVisolates rather than the overseas ones.

Further analysis of the HN proteinshowed that Australian NDV isolateswith a 45 amino acid extension ontheir HN protein generally replicated inthe gut of chickens, while those withHN extensions of 14, nine and sevenamino acids, were associated with ‘latesummer respiratory disease’ syndrome.

As field isolates of viruses consist of apopulation of viruses known as a‘quasi-species’, gene sequence analysisof an NDV field isolate was undertakento determine its genetic variability.Theanalysis showed that about 50% ofviruses in the quasi-species had randommutations, and that in certain field isolates up to 26% of the populationwere virulent.When laboratory purifiedvirulent and avirulent viruses wereplaced in tissue culture, only two or

three virus replication cycles wererequired to achieve the same level of genetic mutation as observed withfield isolates.

During these analyses, other viruses,such as infectious bronchitis virus (IBV)and infectious bursal disease virus(IBDV) were found to co-isolate withNDV. Could the presence of thesepathogens influence the emergence of virulent NDV from an avirulentquasi-species? Experiments in laboratorychickens showed that when the birdswere infected with IBV, and a weeklater, re-infected with an avirulent NDVstrain, there was no effect. However,when the chickens were infected withIBDV, which compromises the immunesystem, and then avirulent NDV,virulent individuals in the quasi-specieswere able to replicate to the point ofcausing disease.

These experiments are being repeatedwith other common pathogens of thechicken industry, and environmentalstressors (such as heat and nutrition),to identify other potential triggers ofvirulent virus outbreaks from avirulentstrains. Changes to industry management practices may be oneway to overcome some of these trigger mechanisms.

VIRUS MORPHOGENESIS

To gain a better understanding of theNDV life cycle and how the virus targets and infects host cells, the sub-program is establishing a reversegenetics system.This system works byconverting viral RNA into DNA, whichcan then be altered by the addition,removal or mutation of base pairs inthe bacterial plasmids into which thegenes are cloned.The modified DNA isthen turned back into RNA after theplasmids are transfected into tissue culture. After making changes to partsof the NDV genome responsible forreplication, we will infect chickens andlook for changes in how and where thevirus replicates, and its effects on thebirds’ health.

A critical component of NDV replication is the production of a‘matrix’ protein.This protein underliesthe membrane of infected cells andforms a raft to which all other proteinsare directed during viral replication inthe host cell.To study these protein-protein interactions further wewill use a ‘yeast-two-hybrid’ systemdeveloped through the Pests andDisease sub-program in Brisbane.We hope to identify protein-proteininteractions that are critical to virusassembly and which we may be able to target with a blocking agent.

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NDV VACCINES

A vaccine (V4) based on an avirulentAustralian isolate was used in combination with quarantine andslaughter to control the 1998 and2002 NDV outbreaks. Another vaccine,I-2, also based on a heat treated,avirulent, Australian NDV strain, hasbeen developed for use in developingcountries such as Asia and Africa.This heat tolerant vaccine will help villagers in these countries rise abovethe poverty line. However, given thatvirulent strains of NDV have arisenfrom avirulent strains, there was anurgent need to characterise the components of this vaccine.

To do this, both the vaccine and theoriginal virus from which it was developed were sequenced and foundto be two base pairs away from virulence. Monitoring of batches of thevaccine will now be undertaken todetect genetic drift. NDV strains inMyanmar are also being characterisedto determine which strains of the virusthe I-2 vaccine protects birds against.

The modified DNA is then turned back into RNAafter the plasmids are tansfected into tissue culture“

“NDV DIAGNOSIS

Molecular techniques (PCR and rapidsequencing) developed during thecourse of this project allow us to identify virulent strains of NDV ininfected tissue or allantoic fluid (ineggs) within 12-18 hours. Genesequences for different regions of theNDV genome, such as those for thefusion gene and the haemaglutinin neuraminidase gene, enable phylogenetic characterisation (ancestry)of the viruses, while the amino acidsequence for the fusion cleavage signalin the fusion protein enables the virulence of different strains to beassessed.

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VIROLOGY

The Virology project works with other groups at AAHL todevelop and provide diagnostic reagents and procedures forthe rapid identification of emerging pathogens including footand mouth disease virus, Hendra and Nipah viruses, andsevere acute respiratory syndrome coronavirus (SARS CoV).

Project LeaderDr Bryan EatonPh (03) 5227 5116Email bryan.eaton@csiro.au

Among the more recent projects isthe development of peptides to blockthe entry of Hendra and Nipah virusesinto cells.The peptides are derivedfrom a viral protein that facilitatesfusion of the virus and host cell membranes to permit entry of thevirus into cells or the movement ofvirus from one cell to another.We willinvestigate the therapeutic potential ofsuch peptides, which work on thesame principle as the latest HIV drugs.

Antibodies are also needed to providea positive control for SARS diagnostictests and to detect SARS CoV proteinin infected tissues and cells. A smallquantity of human antibodies derivedfrom infected human serum is currentlyavailable, but large quantities are needed to maximise Australia’s diagnostic capabilities during futureoutbreaks. Preliminary experimentshave shown that SARS CoV will replicate in some laboratory animalspecies and antibodies generated in

response to the virus infection havebeen shown to neutralise SARS CoVand to react with virus proteins in cellsinfected by SARS CoV. Rats, mice,guinea pigs and rabbits will be testedto see if the virus causes symptomssimilar to those in humans.

Some of the work of this and otherAAHL research is now expanding andevolving into projects in the new CRCfor Australian Biosecurity.

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TISSUE CULTURE AND MEDIA PRODUCTION

Our tissue culture and media production efforts ensure acontinuous supply of cell cultures and biological reagents forthe scientific programs at AAHL.

Project LeaderMs Catherine WilliamsPh (03) 5227 5000Email catherine.williams@csiro.au

Master stocks of some 120 differentcell lines are available on request,including amphibian, avian, mammalian,fish, insect and snake cell lines.TheAfrican green monkey kidney cell line,for example, is used to grow thesevere acute respiratory syndrome(SARS) virus, while bovine thyroid cellsor pig kidney cells are used to growthe foot and mouth disease virus.

The cell lines are sourced from international culture collections,research institutes, commercial companies and scientists around theworld.The Tissue Culture Unit alsoproduces a variety of primary cell linesfrom a range of animals. All the celllines are maintained by AAHL. Masterstocks are stored in liquid nitrogen,while actively growing cells are regularly prepared and replenished ingrowth media for immediate use.

The Tissue Culture Unit regularlyassists in the training of overseas scientists to AAHL in tissue culturetechniques. A collaborative project inVietnam with the national veterinarycompany on development of animproved capability for the control ofclassical swine fever on large and small pig farms, involved Vietnamesescientists visiting AAHL to improvetheir skills base in cell culture and associated quality control techniques.

The Tissue Culture Unit provides a continuous supply of cell cultures and biological reagents for AAHL’s scientific programs.

Projects include improving

retail beef yield and meat

quality without compromising

fertility and adaptation to

parasites and heat stress

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Genetics 44Livestock systems/livestockEnvironment interactions 47Stress physiology 49Ruminant nutrition 50Parasitology 54

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Program Leader Dr Bob HunterPh (07) 4923 8142Fax (07) 4923 8222Email bob.hunter@csiro.au

The northern Australian beefindustry is continually evolving to meet market demands.This evolution includes a movetowards crossbred, rather thanpurebred animals, and the re-introduction of British andtropically adapted Bos Taurusbreeds, which have better fertilityand meat and carcass qualitiesthan the Indian Bos indicus breedsthat have flourished in the harshnorthern Australian environment.To assist this transition, and toaddress various production,environmental and welfare issuesin the industry, the program conducts research in five areas:genetics, ruminant nutrition,livestock systems/livestock environment interactions, stressphysiology and parasitology.

Projects include:• Improving retail beef yield and meat

quality without compromising fertilityand adaptation to parasites and heat stress;

• Identifying new gene markers for feed efficiency, tick and worm resistance and female fertility;

• Understanding the impact of crossbreeding on different productive and adaptive traits;

• Determining the structure and productivity of northern Australian pasture communities and the northern beef industry;

• Understanding the response of cattleto heat stress and ameliorating its negative effects;

• Establishing the benefits of high molasses diets and understanding thephysiological barriers to increased molasses consumption by cattle;

• Reducing methane emissions in cattle;

• Reducing environmental degradation in southern China by establishing economically viable pasture-based beef production systems;

• Developing novel strategies for the control of ticks and other parasites that are effective, sustainable and environmentally acceptable.

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GENETICS

Genetic research aims to assist the breeding of more profitable cattle for the northern Australian beef industry.Research is being undertaken through the CRC for Cattle and Beef Quality, with funding from the Australian Centre for International Agricultural Research (ACIAR), Meat and Livestock Australia and the Northern Pastoral Group of Companies.

Sub-program LeaderDr Heather BurrowPh (07) 4923 8139Email heather.burrow@csiro.au

PERFORMANCE ANDADAPTATION

A key issue of any genetic improvement program is whether wecan change carcass and beef qualityattributes of cattle without undulycompromising key fitness traits such asreproductive performance and environmental adaptation.

To find out, a breeding program wasestablished in conjunction with themajor pastoral companies in northernAustralia, resulting in some 4800 purebred Brahman and ‘tropicallyadapted composite’ calves (a threeway cross between Sanga, Bos indicusand British breeds), all with completepedigree information.

Brahmans and tropically adapted composites were selected as parents,as they represent the extremes of performance, in tropical breeds, for the traits of interest. Brahmans, for example, are highly resistant to

environmental stressors such as heatand parasites, but are the least productive in terms of beef quality andfertility.Tropically adapted compositesin contrast, have better meat qualityand fertility, but a lower resistance toheat, ticks and worms than Brahmans.

Progeny are now being tested to see ifincreases in retail beef yield (amountof meat on the carcass), or consequentdecreases in fat cover, lead to reducedfertility and meat quality, using toolssuch as ultrasound scanning and geneticmarkers for carcass and meat quality.Early results suggest a relationshipbetween fat cover at the start of mating and subsequent fertility.Thus,tradeoffs between fertility and productivity may need to be managedin breeding programs.

Genetic markers and estimated breeding values are also being used toassess feed efficiency and heat, wormand tick resistance (adaptive traits).

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Indigenous South African cattle breeds such as this Nguni cow have beenshown to meet commercial market specifications for feed efficiency, carcassand beef quality, fertility and disease incidence.

Over the next three years, femaleprogeny from the original matings willbe assessed for all productive,reproductive and adaptive traits, asthey rear calves. Results will feed intoimproved breeding programs, andenable farmers to consider the economic tradeoffs between breedingfor improved production, reproductionor adaptation.

Research also aims to develop newgene markers for feed efficiency, wormresistance and in future, for female fertility. Fertility is regarded as a lowlyheritable trait and therefore not undergenetic control. However componentsof the trait, such as fatness, are heritable.A study is underway to evaluate therelationship between genetic markersfor fatness and ovarian activity.

BETTER BEEF FOR SOUTHAFRICA AND AUSTRALIA

In South Africa, research aims toimprove the profitability and sustainability of beef business run byresource poor farmers. Farmers arebeing taught business, marketing andteamwork skills, which will allow continuous improvement and innovation, in collaboration with localgovernment and academic organisations, beyond the project term.Indigenous southern African Sanga

breeds, such as the Nguni,Tuli andAfrikaner are also being evaluated infeedlots against commercial Bonsmaracattle for feed efficiency, carcass andbeef quality, fertility and disease incidence. Results show the indigenouscattle can meet the commercial marketspecifications for these traits.Theseresults are encouraging the commercialsector to purchase cattle from poorfarmers, and fill a 35 000 tonne shortfall in the domestic beef market.Animals from these same breeds arenow being evaluated against international market specifications,for their export potential.

Indigenous sires are also beingscreened for the presence of tendernessand marbling genes using the GeneStartests. Animals carrying favourablecopies of the genes may be used aselite sires in international markets, suchas Australia, and to improve resource-poor herds.

As South Africa experiences a climatesimilar to that of northern Australia,indigenous breeds are well adapted tothe heat and parasites that plague ournorthern beef herds. South Africancattle could therefore provide a newsource of tropically adapted geneticmaterial for use in Australia’s northernbeef industry. Calves sharing the same

parentage are being raised in eachcountry and compared for feed efficiency, carcass and meat quality andfemale fertility.This will ensure anacross-country genetic evaluation thatwill identify genetically superior lines ofcattle within the breeds of interest, inboth countries.

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CROSSBREEDING EXPERIMENT

Between 1992 and 1998, a cross-breeding experiment was undertakeninvolving Indian, African and Europeancattle breeds, to look for heteroticadvantage (hybrid vigour). Data fromthis experiment have been analysed toevaluate the breed differences in traitssuch as growth, parasite and heatresistance, temperament, fertility, andcalf mortality.

The focus of analyses has been theestimation of crossbreeding parametersfor various traits of economic importance, which can then be usedfor the prediction of performance ofuntested genotypes in similar environments. Analyses have shown, forexample, that crossbred animals have a

greater weaning weight and finalweight at 18 months than purebredanimals, that crossbred dams rear bigger calves, and that crosses betweenBritish (Bos taurus) and Zebu (Bos indicus) cattle resulted in progeny withimproved tick and worm resistance.

This information can be incorporatedinto a decision support system toallow cattle breeders to determine theoptimal breed proportions (percentageof Indian, European or African genetics)in ‘composite’ cattle, or to design acrossbreeding program to achieve production goals in a tropical environment. A novel optimisation procedure incorporating growth andtick resistance parameters, has beendeveloped and shows that in an environment of low tick challenge,

for example, the optimal breed proportions are 70% Continental and30% Zebu. In high tick challenge areas,however, the more resistant Zebu andSanga-derived breeds would comprisethe majority of the animal’s genetics.

As South Africa experiences a climate similar to that of northern Australia, indigenous breeds are welladapted to the heat and parasites that plague our northern beef herds

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Surveys of the northern beef industry by the Tropical Agri-Exports project hasled to a better understanding of the structure and productivity of the industry,and the inter-relationships between sectors and regions.(Photo: Greg Bortolussi, CSIRO Livestock Industries)

LIVESTOCK SYSTEMS/LIVESTOCK ENVIRONMENT INTERACTIONS

Research in this sub-program aims to achieve a balancebetween economic productivity and the long-term preservation of the environment.The results will aim toimprove the environmental and social impact of tropical livestock production systems.

Sub-program LeaderDr Dave SwainPh (07) 4923 8125Email dave.swain@csiro.au

ANIMAL-ENVIRONMENTINTERACTIONS

Extensive grazing systems are finelybalanced between environmentalpreservation and economic production.These livestock productionsystems have high levels of inherentvariability, for example, annual rainfalland the spatial distribution of feedresources.

Research is being undertaken toexplore the links between managementoptions and their ability to reduce riskassociated with this variability.The mainfocus of the work is exploring processesthat affect forage use. Current researchaims to determine detailed grazingbehaviour and intake within spatiallyheterogeneous environments.Workwith the nutrition group is exploringhow pasture based supplements influences growth rate and grazingbehaviour. Further work will explore

how supplements and watering pointscan be used to manipulate grazingselection behaviour.This work is supported through the developmentof a spatially explicit grazing model.The results from this work willimprove environmental impact ofextensive livestock production systems.

TROPICAL AGRI-EXPORTS

In the 1990s, a huge growth in liveexports led to industry and governmentconcerns over the capacity of thenorthern Australian beef herd to continue to meet both the live exportand box beef markets.This concernwas due in part to an absence ofknowledge about the capacity ofnorthern Australian pasture communities to sustain animal growthand reproduction.To address this concern, a survey of 375 propertiesacross 14 bioregions in northernQueensland, the Northern Territoryand the north of Western Australia(Kimberly and Pilbara), was conducted.

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The survey was supported by a varietyof industry, government and researchorganisations, and had three broadobjectives:• To define the animal production

capacity of different northern Australian rangeland communities;

• To analyse the ability of regional herds to economically and sustainably increase beef production and to meet market specifications;

• To establish relationships between the environment and the breeding performance of different female genotypes.

A face-to-face survey questionnairewas conducted with producers.Information was sought on propertydescription, pasture management anddevelopment, herd management andperformance, and information management.This was the first detailedsurvey conducted in northern Australiaexamining management practices andherd performance in a common timeframe. Results were collated for eachbioregion, and communicated to individual producers, industry, scienceand government interest groups, in 14 bioregion reports.

As a result of this work we have developed a better understanding ofthe structure and productivity of thenorthern beef industry, and the inter-relationships between sectors andregions. Agencies are better able todirect research, extension, or policyefforts. And opportunities for futureresearch will be more easily identified.The project also provides an information base to support policy and trade objectives. For example,information has been used to developpolicy to support our responsivenessto animal diseases of national importance, and to assist consultantsinvolved in the formulation of transport infrastructure developmentfor state and federal governments.

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Dr Piere Cronjé demonstrates monitoringequipment for heat stress experiments.

STRESS PHYSIOLOGY

Australia’s live export market has come under scrutinyrecently, following reports of large-scale losses of sheep andcattle due to heat stress during ocean voyages.

Sub-program LeaderDr Pierre CronjéPh (07) 4923 8144Email pierre.cronje@csiro.au

Heat waves in the northern part ofAustralia have also claimed the lives ofmany feedlot cattle. As the number ofhot days will increase over the next 50 years, the plight of these animalswill worsen unless solutions to thisproblem are found.

High temperatures also impact on avariety of animal production traits suchas growth, fertility and milk yield.Thissubprogram aims to develop strategiesto ameliorate the negative effects ofheat stress on animal performance andwelfare in the cattle feedlot and liveexport industries, in a number of ways.

Firstly, a scientific description of howhigh and how long an elevated bodytemperature can be sustained in cattlebefore tissue injury occurs is required.Prolonged or excessive heat stresscauses a breakdown of the gut liningresulting in the entry of endotoxinsfrom the gut contents into the blood,and an acute phase immune response.This leads to a sudden fall in bloodpressure, and death. Heat shock proteins (HSP) are synthesised in damaged gut cells during this process,and are used to assess the extent oftissue damage due to heat stress.

In our climate simulators, HSP concentrations, heart rates, core bodytemperatures, respiration rates andfeed intakes will be monitored in aseries of experiments that exposegroups of cattle to different maximumtemperatures for different lengths of time. Results will enable us to characterise the pathophysiology ofhyperthermia, and to identify the besttemperature regime (causing aresponse but not excessive stress) for future experiments. Definition ofthe ‘critical thermal maximum’ for tissue damage, feed intake, growth,metabolism, morbidity and mortalitywill enable the structuring of clear,practical guidelines for use by the feedlot and live export industry onwhen to intervene during hot weather.

Practical diets and feeding strategies forfeedlots and the live export industryare also being investigated in collaboration with feed companies.Feed additives that protect the gut cellsfrom damage during heat stress andcompounds that induce the secretionof HSP and thus maintain gut integrityduring heat stress are currently beingdeveloped and evaluated.

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RUMINANT NUTRITION

Projects in this sub-program are investigating the benefits ofincreased molasses in the diets of feedlot and pasture-basedcattle, and methods to increase dietary intake.

Sub-program LeaderDr Peter KennedyPh (07) 4923 8182Email peter.kennedy@csiro.au

HIGH MOLASSES DIETS

Molasses has traditionally been seen asa drought supplement and a by-productwith little inherent value. But researchsuggests there are both economic andproductivity benefits when cattle arefed high molasses diets. Early trials haveshown that cattle fed diets containingup to 60% molasses can achieveliveweight gains of up to 1.6 kg/day.

Trials conducted at a collaborativeindustry site have shown that cattle feddiets containing 25% and 50%molasses, had similar carcass and meatquality characteristics to grain-fed animals, including colour, moisture, pH,tenderness and fat content. Animalsalso achieved liveweight gains of 1.3kg/day and 1.7 kg/day respectively,compared to 2 kg/day for grain-fedcontrols.

If 500 000t of molasses were availableannually, and represented 50% of thediet of intensively fed cattle, it wouldbe possible to add 100kg of liveweightto about 1.3 million head.This gaincould be pivotal to continuing the supply of northern cattle and beef into

Asian markets, especially during thedry season and drought. A continueddemand for molasses from the live-stock industry could also generate anew domestic market for Queenslandsugar producers.

A second feed trial is now underway.Endorsement of the results by Meatand Livestock Australia is expected toincrease industry acceptance ofmolasses.

MOLASSES FOR PASTUREFINISHING OF CATTLE

Increasing the molasses content in thediet of grazing animals could overcomeweight loss due to poor nutrition during the dry season in northernAustralia, and produce more marketable animals at a younger age.

In a series of pen-fed studies, steerswere given chopped roughages of differing nutritive value, with or without molasses. Liveweight gains ofup to 900g/day were achieved whenmolasses comprised 32% of the diet.To improve this gain, supplements are

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Cattle fed diets containing 25% and 50% molasses have similar carcass andmeat quality characteristics to grain-fed animals, including colour, tendernessand fat content.

being investigated, which provide additional protein for absorption in thesmall intestine, when cattle consumelarge quantities of molasses.The behaviour of grazing animals whengiven a choice between molasses andpasture is also being studied. Resultswill inform decision support systemsthat aim to predict the feed intake andgrowth rate of cattle fed molasses-based diets, when molasses is offeredto grazing animals.

RUMINANT NUTRITION

As molasses offers a cheap alternativeto other traditional energy rich feedssuch as grains, for growth and finishingin the northern beef industry, anunderstanding of the factors influencingmolasses intake is necessary. Currently,we have little information on patternsand amounts of molasses eaten by animals grazing northern Australianpastures throughout the day, andwhether rapid molasses consumptionreduces intake and utilization of theenergy from pasture.

To find out, different amounts ofmolasses were pumped directly intothe rumen of fistulated cattle eachmorning.The amount of time it tookfor animals to begin to eat hay, andhow much they ate, was then measured. Osmolality, pH, digesta flow,and fermentation end products in therumen, were also measured.

Results to date show that the moremolasses given, the greater the changein rumen osmolality, and the longer ittakes for osmolality to return to normal. Similarly, large amounts ofmolasses cause big drops in pH (leading to an acidic rumen), and alonger lag time to return to normal.This drop in pH in the rumen accompanies an increase in butyricacid, one of the three main acidsresulting from rumen fermentation.

The types and numbers of microbespresent in the rumen at different timesof the day are also being investigated,as different organisms are responsiblefor processing fibre, grain andmolasses. Changes in microbial floracould affect how soon an animal feelslike eating after a molasses meal.

Results from this study will feed intoother molasses research for pasture-based animals, to identify the best timeof day or feed combination in which tooffer molasses, in order to increaseintake. Alternatively, a supplement thatregulates osmolality or alters the pathof fermentation may be identified.

ENERGY AND PROTEINFROM MOLASSES

The energy value of molasses, on itsown or mixed with hay, is beingassessed by measuring the energy balance (energy input and output) incattle fed molasses.The amount ofenergy available to animal tissues(metabolisable energy intake) frommolasses is reduced by microbialmethane production in the rumen.Thiscan be measured by analysing thegasses expired by cattle.

Microbes, which grow on the energy inmolasses, provide an important sourceof protein for cattle, and measurementssuggest that improvements could bemade in the efficiency of microbialprotein synthesis. Studies are inprogress to confirm this inefficiencyand identify its causes.

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GROWTH PATH BIOLOGY

Calves in northern Australia are routinely weaned at the beginning ofthe dry season and undergo a periodof nutrient deprivation resulting in prolonged weight stasis or loss.Alterations in nutrient supply cancause significant variations in growthand development, and previous studiesindicate that the timing of nutrient supply affects carcass composition,marbling and meat quality. However,studies also suggest that animalsdeprived of feed could experiencerapid compensatory weight gains during feedlot finishing, overcomingnegative effects on muscle developmentand meat toughness.

Research in the sub-program aims todocument these compensatory weightgains and the developmental aspects of muscle that impact on carcass composition and meat quality.Such information will underpin recommendations for achieving high eating quality beef at less cost.

To do this, groups of Belmont Redweaner steers were placed on diets to achieve either ; a liveweight gain of0.6 kg/day (super maintenance),maintenance, or a 10% liveweight loss(sub-maintenance), over four months.Some animals were then slaughteredand their carcass characteristics determined. Others were left to graze

or fed a feedlot diet until they reached400 kg, when they were slaughtered.Remaining steers were grazed untilgrowth rates fell to less than 0.6kg/day, and were then returned to a‘growth’ diet, before slaughter.

Results to date show that animalsdeprived of sufficient food exhibitaccelerated growth rates once full feedis offered. Intramuscular fat and musclearea also increases rapidly. Histologicalexamination of muscle tissue indicatesthat nutritionally restricted animals losemuscle mass, although connective tissuemass remains constant. However, imageanalysis and histology showed no significant difference in muscle massafter nutritional compensation.

REDUCING METHANEEMISSIONS

A patented antimethanogen in acyclodextrin complex, produced by theAustralian Animal Health Laboratory,could prevent the production ofmethane by rumen methanogens. Initialexperiments in respiration chambershave shown that cattle fed the compound had a reduction in methaneemissions. A larger trial involving animalsfed the antimethanogen twice daily, for85 days, has demonstrated slightincreases in live weight gain and anassociated decrease in methane emissions.Tissue samples are now

being examined for chemical residues.If the samples are cleared, the case foractively pursuing commercialisation isstrong.Trials on dairy cattle are imminent.The knock-on effect of thecompound on other rumen microbes,such as acetogens and cellulolytic bacteria, will also be investigated.

RUMINANT PRODUCTIONIN CHINA

In the degraded red and yellow soilsregions of southern China, an ACIAR-funded project to develop economicallyviable pasture-based beef productionsystems for smallholder farmers isunderway.The project aims to stabilisethe soil in deforested areas by encouraging farmers to plant fodderspecies for their cattle.This will reducerunoff and erosion, and improve thecapacity of farmers to supply thedomestic beef market by improvinganimal nutrition and growth.

Because of the harsh climatic conditionsand distinct seasonality of foragegrowth in the region, year-round provision of fresh and conserved,nutritious cattle feed has been difficult.However, previous ACIAR projectshave identified suitable forage typesadapted to these environmental conditions.The challenge now is todevelop an economically-viable

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Dr Nigel Tomkins monitors microbial methane production in cattle in nearbyrespiration chambers. Cattle fed an antimethanogen product twice daily, for 85 days, demonstrated increases in live weight gain and decreases in methane emissions.

forage-based ruminant production system that integrates with other activities of small-holder farmers. ‘Feedyear plans’ will be developed whichidentify forage options for each monthof the year, and integrate into plantingand harvesting activities.These plansaim to provide a high nutritive valuediet, resulting in liveweight gainsapproaching 0.5 kg/day.

Strategies to utilise forages, cropresidues and by-products, supportedby computer software to predict animalperformance, will also be developedfor China and tropical Australia.Thisinformation will help formulate supplements, predict animal performance, and aid transferability ofinnovations beyond this project.

If 500 000t of molasses were available annually, andrepresented 50% of the diet of intensively fed cattle,it would be possible to add 100kg of liveweight toabout 1.3 million head

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PARASITOLOGY

With the conversion of the northern beef herd to highBrahman (Bos indicus) content over the last 40 years, researchinto parasitism and its effect on animal productivity has waned.

Sub-program LeaderDr Ian SutherlandPh (07) 4923 8187Email ian.sutherland@csiro.au

The introduction of more parasite-susceptible Bos taurus genesfor improved carcass and meat quality,however, has revived the need for parasite control. As existing controlmethods, based on chemical treatmentand increased Bos indicus content, areno longer appropriate, novel strategiesfor the control of endo- and ecto-parasites that are effective,sustainable and environmentally acceptable are required.

As ticks and tick-borne diseases are amajor cause of lost productivity in Bos taurus breeds, the first priority ofthe recently formed parasitology groupis the development of a tick infectionmodel.To do this, groups of previouslyuninfected Bos indicus and Bos tauruscattle will be gradually challenged withsmall numbers of ticks (trickle challenge).The number of tick larvae,nymphs and adults present will bemonitored throughout the experiment.Blood samples, and attached ticks, willalso be taken at regular intervals andstored until the animals appear tobecome immune (based on ticknumbers and immunological indicators). Once immunity appears, a

variety of molecular techniques will beused to investigate changes in both thehost and parasite at critical points during the host-parasite relationship.

In terms of the host response, cytokineproduction and gene expression in theblood of infected and uninfected (control) animals will be investigatedusing real time PCR and microarraysrespectively. In the ticks, a combinationof proteomics and molecular techniques will be used to detect anddefine changes during attachment andfeeding on different cattle genotypeswith differing immune capability.Thecombination of these approaches willenable a direct comparison of changesin gene expression in a host and parasite simultaneously.

There are a number of potential outcomes of this work such as the discovery of major genes for tick-resistance and anti-parasite immunity; the identification of potentialvaccine targets to prevent tick attachment; the discovery of noveldrug targets and possibly novel molecules with medical or veterinary applications.

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Novel control methods for endo- and ecto-parasites, that are effective, sustain-able and environmentally acceptable to the northern beef industry are beingsought by the Parasitology group. (Photo: Dr Greg Bortolussi, CSIRO LivestockIndustries)

While the tick infection model formsthe basis of strategic research for theparasitology group in the immediatefuture, several other areas of interestwill be included when appropriate.These include:• Determining the prevalence of

tick-borne disease agents, Anaplasmaand Babesia, in ticks in Queensland.

• Identifying specific targets in buffalo fly (Haematobia irritans exigua) larvaeand delivering a relevant control agent to cattle faeces.

• Understanding the biology and epidemiology of the filarial nematode,Stephanofilaria stilesi, transmitted by buffalo fly;

Once immunity appears, a variety of molecular techniques will be used to investigate changes in both the host and parasite at critical points during the host-parasite relationship

“ “• Examining the effect of parasites on

methane production. As parasites such as gastrointestinal nematodes reduce feed intake for significant periods, methane production over the life of the animal may be significantly over-estimated when extrapo lating from indoor measurements.

• Investigate the epidemiology of the protozoan parasite Neospora caninum(a major cause of abortion in cattle),and the relatedness of parasites in different parts of Australia.

• Develop a diagnostic test for acaricide resistance in ticks using Random Amplified Polymorphic DNA-PCR.

The Infectious Diseases and

Food Safety program aims to

enhance Australia's disease

control capacity by improving

our understanding of viral and

bacterial diseases and

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Avian virology 58Johne’s disease 60Protein biochemistryand proteomics 62Plant-associated toxins 65Molecular detection 68

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Program Leader Dr Marion AndrewPh (03) 5227 5745Fax (03) 5227 5555Email marion.andrew@csiro.au

The Infectious Diseases and FoodSafety program aims to enhanceAustralia's disease control capacityby improving our understandingof viral and bacterial diseases andplant-associated poisonings, anddeveloping technologies toimprove diagnostic tests and vaccine performance for betterdisease control in livestock.Thereare five projects within the program, which are conducted atthe Australian Animal HealthLaboratories (AAHL) in Geelong.

The avian virology project is investigatingthe characterisation of variant infectiousbursal disease viruses and the development of vaccines and improveddiagnostic tests. Similarly, work on thebacterial disease, Johne’s disease, focuseson developing more sensitive diagnostictests that can be used to detect infectedanimals at an earlier stage of disease.The work is also identifying bacterialproteins that could be used for a safer,more efficacious vaccine.

The protein biochemistry group isdeveloping and implementing newtechnologies in protein chemistry andproteomics, and applying these technologies to the characterisationand purification of viral and bacterialproteins, including paramyxovirusreceptor proteins and antigens forJohne’s disease.They have also developed a sensitive diagnostic test

for lice detection, which will help todecrease the amount of pesticideresidue in wool, and a vaccine for pink eye in cattle.

The plant-associated toxins group isinvolved in the identification of suspected plant-associated poisoning ofanimals and the development of strategies for managing such poisonings.The group also works on the development of accurate and sensitivemethods for the detection of thesetoxins, including corynetoxin, responsiblefor annual rye-grass toxicity, andpyrrolizidine alkaloids, which cause liverdisease in humans.The group is working to determine the NoObserved Effect Level for corynetoxin,which will establish allowable levels ofthe toxin in grains for animal andhuman consumption.

The molecular detection sub-programapplies and develops molecular technologies for the characterisation ofemerging paramyxoviruses, found infruit bats. New diagnostic tests andvaccine strategies for foot and mouthdisease (FMD) will enable rapid diagnosis and improved control of thedisease, without the need for massslaughter. And the development of antibodies to diagnose severe acuterespiratory syndrome (SARS) inhumans, and tests for SARS antibodiesin different animal species, will contribute to strategic internationalresearch on the virus.

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AVIAN VIROLOGY

Infectious bursal disease virus (IBDV) causes one of the mosteconomically important diseases in the intensive poultryindustry. A large number of IBDV strains have been identifiedthroughout the world and grouped according to their pathogenicity (ability to cause disease) and antigenicity (abilityto stimulate an immune response).Three major groups havebeen identified: classical virulent, very virulent (vvIBDV) andantigenic variants.

Project LeaderJagoda IgnjatovicPh (03) 5227 5769Email jagoda.ignjatovic@csiro.au

All groups of IBDV are immuno-supressive and impede the successfulcontrol of all other poultry pathogens.The viruses act by destroying B cells inthe bursa (an immunological organnear the tail), which are essential for antibody-mediated immunity, leavingbirds vulnerable to other viral and bacterial infections.While the classicalvirulent and antigenic variants cause noor low mortality, vvIBDV strains causeup to 70% mortality in susceptiblepoultry flocks.

Very virulent IBDV emerged in 1987 inEurope, and has since spread to manycountries including South East Asia. InIndonesia, vv IBDV is widespread insmall poultry holdings and causes mortalities of up to 40%. As availablevaccines to control vvIBDV are tooexpensive for small poultry holdings,we are working to develop a moresuitable vaccine.

To do this we have passaged threeIndonesian isolates of vvIBDV throughcell culture to reduce virulence.Theresulting viruses will be cloned andmonitored for virulence by DNAsequencing.Two DNA mutations havebeen shown to be associated with lossof virulence but not loss of antigenicity,and it is these two mutations we will look for. Suitableclones will then be used to infectchickens and their effect on B cells and bird health monitored.

Only mild strains of IBDV have beendetected in Australia, but previouscharacterisation of Australian IBDV isolates by the project team has indicated that they are changing, withthe emergence of antigenic variantsand strains showing greater virulence.Australian IBDV strains differ geneticallyfrom IBDV strains of other countries.

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For that reason it is unknown if thesame type of genetic changes associatedwith vvIBDV would also change local,low pathogenic IBDV into vvIBDV.

To determine the degree of mutationlocal IBDV strains must undergobefore they become vvIBDV-likestrains, we are developing a reversegenetics system.This system works byconverting viral RNA into cDNA,which can then be altered by the addition, deletion or mutation of DNAbase pairs in genes involved in virulence.This altered DNA is then convertedback into RNA and inserted into the

viral structure.The effect of thesemutations on virulence can then bestudied by infecting chickens with thealtered virus.

The incursion or appearance ofvvIBDV strains would pose a seriousthreat to the Australian poultry industryin terms of economic losses and itssustainability.Thus, we must ensure thattechniques are available to rapidlydetect the incursion or appearance ofsuch strains.We have developed chicken recombinant antibodies thatcan differentiate vvIBDV from all otherstrains.These antibodies were isolated

The resulting viruses will be cloned and monitoredfor virulence by DNA sequencing“

“from chickens infected with differentIBDV strains, including vvIBDV, andthen cloned into bacteriophages (viruses that infect bacteria).The resultis a collection of bacteriophage withantibodies specific to different strainsof the virus displayed on the bacteriophage surface.Those with diagnostic potential, such as specificityfor vvIBDV, have been selected.

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JOHNE’S DISEASE

Johne's disease (JD) is a widespread, contagious and chronicintestinal inflammation of ruminants, particularly cattle andsheep, caused by Mycobacterium avium subspecies paratuberculosis.

Project LeaderDr Mark TizardPh (03) 5227 5753Email mark.tizard@csiro.au

The bacterium, a relative of M. bovis,which causes bovine tuberculosis, istransmitted mainly by the faecal-oralroute to young animals.The disease hasa long and variable incubation period(from a few months to 2-4 years insheep and up to 10 years in cattle)before clinical symptoms appear.Whenthey do, decline and death follows relatively rapidly. Clinical symptomsinclude wasting and scouring, due todisruption of the gut lining, and shedding of bacteria in the faeces.

JD is increasing in prevalence and hassignificant livestock trade implications.The disease results in restrictions oninterstate livestock movement and theoverseas export of live animals.Through a voluntary national MarketAssurance Program, herds and flocksare classified according to JD status tominimize the risk of introducing thedisease into clean herds or flocks.However, for businesses affected by JD,there are no guaranteed ways toregain a disease free status and this hasled to the introduction of the use of avaccine to control the disease in sheepflocks.This reduces the impact of clinical

disease but does not eradicate it fromthe flock. Policy to control and minimisethe impact of JD on the farming community is now being developed.

The focus of the JD project is to provide a better understanding of thedisease and generate new tools toimprove management and reduce itseconomic and social impacts on theAustralian sheep and cattle industries.To do this, JD scientists are taking anintegrated approach to address threekey issues: pathogenesis, bacterium-macrophage interactions, and bacterialmetabolism and biochemistry. Duringthe course of these studies,opportunities will be taken to exploitthe research findings to improve diagnostic tests and the current vaccine.

PATHOGENESIS

This new research initiative aims touncover how the bacterium establishesinfection in the animal and how infection progresses towards clinicalexpression.To do this we will use animal models (laboratory based goatsand cattle), and infected animals from

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the field, to look at the spread of disease within the animals, and correlate this with their immunologicalresponse at different stages of infection.The leading edge technologies ofgenomics and proteomics will be usedto compare M. paratuberculosis in thelaboratory (in ‘couch potato’ mode)and the diseased animal (in ‘field soldier’mode) to look for differences in geneand protein expression.This will involvestudies of samples retrieved from naturally and experimentally infectedanimals and also the establishment oflaboratory based experimental systems.

BACTERIUM-MACROPHAGEINTERACTIONS

A central feature of disease caused bymycobacteria (like JD and TB) is thatthe infectious agent makes its homewithin host macrophages, the cells thatthe animal’s immune system normallyuses to capture and destroy invadingbacteria.The bacterium does this byevading the mechanisms that themacrophage uses to destroy foreignbodies, and then drawing the nutrientsit needs to survive and grow from themacrophage. Details of how the bacteria achieve this feat are not clear.A simple way of replicating what happens in the disease state is to mixthe bacterium with host macrophagesin laboratory tissue culture. Again, bothproteomic and genomic methods will

be employed to study changes in protein and gene expression in boththe bacterium and macrophages.

BACTERIAL METABOLISMAND BIOCHEMISTRYUnderstanding bacterial metabolismand biochemistry is essential to understanding the processes of pathogenesis and bacterium-macrophage interactions.This disciplineoverlaps with proteomic analysis beingundertaken by members of the ProteinBiochemistry and Proteomics Group,integrated as part of the Johne’s disease research team (see below).

IMPROVING THE CONTROLOF JD

By defining the disease process, we aimto identify infection mechanisms andpathways that bacteria use, and thegenes responsible.These basic studieswill improve our understanding of theprocesses underlying this disease, andthis knowledge will provide moredefined and better targeted reagentsthat can significantly improve the control of JD.

The current vaccine for JD, Gudair™,has had some success in Australia,slowing the rate of progression of clinical disease and decreasing mortality.But it does not stop all infection andultimately does not stop infected animals from shedding the bacteria

Johne’s disease, caused by Mycobacterium avium subspecies paratuberculosis,is a chronic intestinal inflammation of ruminants, particularly sheep and cattle.CSIRO Livestock Industries aims to develop new tools to improve managementof the disease and reduce its economic and social impacts.

and infecting others.The vaccine is also relatively crude.Thus, benefits may beachieved from a better-defined reagent– including reduced site lesions,improved animal welfare, reduced carcass damage, and opportunities todistinguish infected and vaccinated animals.

During 2003/2004, the group will hostexperts from the United StatesDepartment of Agriculture NationalAnimal Disease Center, and theUniversity of Minnesota, to collaborateon the production of DNA microarraysof M. paratuberculosis.This work willcomplement the work conducted bythe Protein Biochemistry andProteomics team members.

For many years there has been the suggestion of a link between JD andCrohn’s disease (an inflammatory bowldisease in humans).Though the link isweak, it remains a threat to livestockindustries and their production systems.Our vision for JD research is to providethe knowledge and tools to assistindustry to minimize the presentimpacts on production, and to helpindustry position itself to deal effectivelywith any future issues of market access and assurance of product integrity and safety.

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PROTEIN BIOCHEMISTRY AND PROTEOMICS

The Protein Biochemistry and Proteomics Group use cuttingedge technology to identify, purify and characterise native andrecombinant proteins, peptides and antigens of biological significance, and to solve complex problems relevant to livestock industries.

Project LeaderDr Wojtek MichalskiPh (03) 5227 5772Email wojtek.michalski@csiro.au

Research spans five main objectives:Johne’s disease diagnosis, characterisingemerging paramyxoviruses, developinga vaccine for Pinkeye and a lice detection test for sheep, characterisingbioactive peptides in milk, and application of proteomics to ryegrasstoxicity.The group also provides adviceand protein analysis expertise to othersat AAHL and supports a strong studenttraining program in conjunction withmajor Australian universities and thosein the Netherlands and Poland.

JOHNE’S DISEASE PATHOGENESIS ANDDIAGNOSIS

Johne’s disease (JD) is a chronic intestinal inflammation of ruminants,particularly cattle and sheep, caused byMycobacterium avium subspeciesparatuberculosis (see Johne’s diseaseproject). Using proteomics (analysis ofthe protein complement of the bacterium) we aim to identify M.paratuberculosis antigens with diagnosticpotential. It is hoped these antigens willenable diagnosis of the disease beforeclinical symptoms appear. In collaboration with the University ofSydney and Meat Livestock Australiawe will also study the pathogenesis ofJD in sheep and cattle using in vitro andin vivo systems.This will involve studyingthe interaction of the bacterium in itshost cells – macrophages - in tissue culture.

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A member of the Protein Biochemistry and Proteomics group identifies structural proteins of a newly emergent Nipah virus on an Edman degradationamino acid sequencer.

NEWLY EMERGEDPARAMYXOVIRUSES

Paramyxoviruses such as the Hendraand Nipah viruses are genetically similar, but have many significant differences at the protein level. Byidentifying these protein differences wecan design diagnostic tests that willcomplement genetic tests.We also aimto identify the host cell receptors forthe Hendra and Nipah viruses. Bystudying the interaction between proteins on the virus surfaces, andreceptors on host cells, we may gaininsights into the evolution of theseviruses and the processes involved inviral infectivity.

PINK EYE

Pink eye or infectious bovine kerato-conjunctivitis (IBK) is a painful,infectious eye disease of cattle causedby Moraxella bovis.The bacterium causes ulceration of the cornea, whilethe subsequent pain leads to weightloss and/or reduced milk production.Treatment is by topical application ofantibiotic cream.While this is effective,it is also labour intensive.

Initially a vaccine against M. bovis pili(hair-like projections from the bacterium’s surface) was proposed.However, as there are seven serotypesof M. bovis, a vaccine would need to

target the seven different pili. Instead,the protein biochemistry group isolatedthree haemolysin proteins - toxic proteins that degrade the cornea andthat are common to all serotypes.These antigens could by used singly orin combination, in a recombinant vaccine formulation. Commercial partnerships are now being sought forvaccine trials planned for 2003/2004.

LICE

Lice infestation of sheep is a majorproblem for the wool industry. As nosimple detection method is available toconfirm the presence of lice, farmersend up chemically treating flocks thataren’t infested. It is estimated that inAustralia, 80% of flocks are treated butonly 25% are infested with lice.Toovercome this problem we have developed a hand-held test based onantigens specific to the louse egg,juvenile and adult. During shearing,grease from the shearing comb ismixed with a solution and spotted ona pregnancy-type test strip. Antibodiesbound to the test strip then pick uplouse antigens, and a colour reactionresults. Development of the lice detection test was done over 6 yearsin close collaboration with NSWAgriculture’s Elizabeth McArthurAgricultural Institute and AustralianWool Innovation. On-farm trials of thetest will be performed in 2003/2004 inNew South Wales.

BIOACTIVE PEPTIDES IN MILK

When milk and milk-based productsare digested, enzymes in the digestivetract break the milk proteins into smaller peptides. Studies have shownthat many of these smaller peptidesare bioactive, having antibacterial,antiviral and even anticancer properties.This bioactivity can be increased byincreasing the positive charge on thepeptide, through amidation and otherchemical modifications. In collaborationwith Food Science Australia, we haveidentified two new bioactive peptides.The effect of amidation on their activityis now being studied. Such peptidescould be used as therapeutics in food products.

TOXICOPROTEOMICS OFPLANT-ASSOCIATED TOXINS

In collaboration with the Plant-Associated Toxins (PAT) group, we aretaking a toxicoproteomic approach tounderstanding the effects that corynetoxins have on the proteins ofexposed animals. Corynetoxins areproduced by the bacterium,Rathayibacter toxicus, which is carriedby nematodes (Anguina funesta) to theseedheads of several plants includingannual ryegrass (Lolium rigidum).

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Ingestion of corynetoxin by sheep andcattle, leads to the fatal neurologicaldisease, annual ryegrass toxicity (ARGT).

Corynetoxins and their relatives,tunicamycins, inhibit the action of a glucosamine transferase enzyme, whichis involved in the first step in the formation of N-glycosylated proteins(proteins containing sugars on theiramino (N) terminals).This results inunder-glycosylation of proteins, whichprevents proper protein function.

Toxicoproteomics provides an excitingnew avenue for identifying associationsbetween corynetoxins/tunicamycins,proteins, and the pathological effectsexhibited by intoxicated livestock.Proteomic platform technologies,

including two-dimensional electrophoresis, mass-spectrometryand bioinformatics, will be employed toanalyse tissue samples taken from ratsand sheep exposed to various doses ofthe toxins.We hope to define potentialprotein biomarkers that may help diagnose exposure to this class of toxins.We also hope to identify thelevels of corynetoxins or tunicamycinsin the diet of rats and sheep that donot have any effect on proteins thatmay be critical to health in the long orshort terms.This will feed into the NoObserved Effect Limit (NOEL)research being undertaken by the PAT group.

The group also provides advice and protein analysis expertise to others at AAHL and supports a strong student training program in conjunction with major Australian universities and those in theNetherlands and Poland

“ “

OTHER EXPERTISE

The protein biochemistry laboratorycontains a range of protein analysisequipment, including chromatography,mass spectrometry and amino acidanalysis and sequencing instruments,and associated staff expertise.Thisallows us to assist with or collaborateon other projects at AAHL and otherCSIRO Livestock Industries sites.Computer aided molecular modellingof proteins also helps identify the mostlikely protein structure.

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PLANT-ASSOCIATED TOXINS

Many plants, and bacteria or fungi associated with plants,produce chemicals that are toxic to animals and humans.Animals are exposed to these plant-associated toxins (PATs)through grazing or via contaminated feed, while humans caningest foods contaminated with these toxins, such as grains orhoney, or contaminant residues in animal-derived food such asmeat, dairy products and eggs.

Project LeaderDr Steven ColegatePh (03) 5227 5739Email steve.colgate@csiro.au

The Plant-Associated Toxins (PATs)research group aims to:• reduce the impact of PATs on

farmers, animals, livestock feed industries and human food-related industries;

• contribute to the safeguarding of domestic and international markets for Australian agricultural produce byhelping to control contamination by PATs, and ;

• contribute to human food safety issues by helping to control contamination by PATs.

To do this the group has three majorresearch projects.The first focuses oninvestigating new PATs, their mechanisms of action, pathogenesis ofthe poisoning disease and developmentof management strategies.The secondis developing sensitive analytical methodsfor the detection of PATs in the foodsupplies of animals and humans, such ascorynetoxins and pyrrolizidine alkaloids.The third toxicology project aims to

identify the highest level of corynetoxins- responsible for annual ryegrass toxicity - that can be tolerated withoutadverse health effects in animals.ThisNo Observed Effect Level (NOEL) forcorynetoxins may then be extrapolatedto humans, leading to the regulation ofsafe levels in food.

PLANT-ASSOCIATEDTOXIN INVESTIGATIONS

Our expertise in toxin isolation andidentification enables us to investigateoutbreaks of poisonings around thecountry. New toxins identified throughthis work can provide leads for futurework.Various techniques such as highpressure liquid chromatography, thinlayer chromatography and mass spectrometry are used to separate andidentify these small bioactive molecules.The pathological effects of the purifiedtoxins on animals can then be studiedin order to completely define the poisoning syndrome.

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Animals are exposed to plant-associated toxins through grazingor via contaminated feed.The Plant-Associated Toxins (PAT)group undertakes a range of projects to reduce the impact ofPATs on animal and human health, and help safeguard Australia’sagricultural markets.

The work also involves developingassays for animal feed and human food,identifying management measures toprevent animal poisoning and feed contamination, and developing antidotesand/or vaccines.We have developedexperimental, systemic vaccines thatprotect sheep against lupinosis andannual ryegrass toxicity.The lattershows promise in pen trials butrequires substantial funding to progressthe development.Work in conjunctionwith CSIRO Livestock Industries inArmidale is also investigating thepotential of mucosal vaccination.

One of the toxins being studied indetail is that produced by the introduced pasture-improvement plant,Phalaris. In collaboration with CSIROPlant Industry and Agriculture NSW,the group is aiming to identify thetoxin causing ‘sudden death syndrome’in sheep grazing Phalaris pasture. If wecan identify Phalaris plants with lowlevels of toxin, traditional crossbreedingmethods may be employed to breednon-toxic pasture.

DETECTING PLANT-ASSOCIATED TOXINS

Annual ryegrass toxicity (ARGT) arosein South Australia and WesternAustralia about 50 years ago, and isnow a huge problem for Australia’slivestock and grain and fodder exportindustries.The toxins are produced bya bacterium, Rathayibacter toxicus,which is carried by nematodes(Anguina spp.) to the seedheads of several plants including annual ryegrass(Lolium rigidum). Ingestion of contaminated ryegrass by sheep andcattle leads to neurological disease.The toxins may also contaminate grain destined for the human food chain.As little is known about the triggers for corynetoxin production by the bacterium, work in the group is lookingfor changes in gene expression in theplant and the bacterium at differentstages of ryegrass growth.

We have also developed a detectiontest for corynetoxins, funded by theGrains Research and DevelopmentCorporation (GRDC) and the RuralIndustries Research and DevelopmentCorporation (RIRDC).To do this,antibodies to the toxins were raised insheep. As the toxins are too small tostimulate the immune system naturally,a ‘hapten’ derived from the toxins, wasattached to a larger carrier protein.Serum from the sheep was then testedfor cross reactivity with the toxins and

other potential biomolecular interferences.The antibodies were thenincorporated into an Enzyme-LinkedImmunosorbent Assay (ELISA), whichcan quantitatively detect the presenceof toxins in grain down to levels ofabout 25 micrograms per kilogram.

The test was recently validated byAgriculture Western Australia and theSouth Australian Research andDevelopment Institute. Reagents weresent to these laboratories, along withsamples of wheat and barely grainextracts. Analysis of the samplesshowed the test was suitably robustand accurate for use in other laboratories.The test will now be optimised for use with hay/fodder.These tests will assure export marketsof the quality of our fodder and grainfor livestock and human use.Commercial partners will be sought todevelop and market these tests further.

Inter-laboratory validation of an ELISAtest for pyrrolizidine alkaloids (PAs)from Heliotropium europaeum is alsounderway.This introduced plant contaminates pasture and produces arange of DNA-damaging and potentialcancer-promoting toxins. In humans,PAs in herbal medicines can cause liverdisease. PAs can also be found in grainscontaminated by Heliotrope seeds, andin honey (from the nectar of PA-producing plants). It is not known

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whether animals poisoned by PAsdirect the toxins into tissues thathumans eat.The ELISA is currentlybeing tested on wheat and barley grain samples.

NO OBSERVED EFFECTLEVEL (NOEL)

The major focus of the PAT group is todetermine the level of corynetoxinsthat animals can ingest, without anynegative health effects, especially following long term, low level exposureto the toxins.The project, funded by theGRDC, is based on international guide-lines for the investigation of chronictoxicity, and will run in three phases.

The first phase has involved a six monthtrial with rats and sheep, developingstandard operating procedures andidentifying a range of corynetoxindoses that would produce an effect,based on studies in the literature.

A 12 month study followed, where ratsreceived purified toxin mixed in ratpellets ad lib, and post mortem tissuesamples were taken from sacrificed animals at three, six and nine months,to test for biochemical changes. Bodyweight and feed intake was also measured and neurological assessmentsmade. Similarly, sheep received dosesof the toxin three times a week.

Analysis of the tissue samples andother data collected for the rats andsheep is now underway.

Ingestion of corynetoxins at a toxicdose of 1mg/kg body weight, leads todeath of sheep, cattle and pigs in 4-5days. However, it is not known whetheringestion of small amounts of toxinover time, has a cumulative effect thateventually results in adverse healtheffects. Preliminary results from our 12 month study show that sheep fed8-10 times more than the lethal dose,in 12 months, are clinically healthy.This suggests a balance between ingestionof the toxin, detoxification by the liverand excretion from the body, may be possible.

A collaborative project with theProtein Biochemistry and Proteomicsgroup will also help define potentialprotein biomakers that may be ofvalue in diagnosing exposure tocorynetoxins and their relatives.

The Plant-Associated Toxins group is determining the levelof corynetoxins that animals can ingest, without any negativehealth effects, especially following long term, low level exposure to the toxins.

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MOLECULAR DETECTION

This technology-driven project applies and develops moleculartechnologies for the rapid characterisation of emerging viruses.The current focus is on emerging paramyxoviruses - a family of negative-stranded RNA viruses that can infect animals and humans.

Project LeaderDr Linfa WangPh (03) 5227 5121Email linfa.wang@csiro.au

Recently, novel paramyxoviruses havebeen found in fruit bats or flying foxes(Megachiroptera). In 1994, for example,the laboratory identified the Hendravirus, which spread from bats to horsesand then to humans.When a similarvirus emerged in pigs and humans inMalaysia, serology and genomesequencing showed it was related tothe Hendra virus.The Nipah virus wassubsequently traced back to fruit bats.

Paramyxoviruses from fruit bats, pigs,rodents, shrews and other animals havealso been isolated, and their molecularstructures (genome, proteins, antigens)characterised.While some of theseviruses have not yet been associatedwith any known disease, they will beavailable for future reference shouldnew infections arise in animals orhumans from these viruses or closedrelated viruses.

Work in the project also aims todevelop tests for better disease surveillance.Through the CRC forAustralian Biosecurity, for example, weare developing a test to rapidly identifyan outbreak of foot and mouth disease(FMD) in Australia. As the current policy to slaughter animals infectedwith the disease has animal welfare andpublic perception issues, new vaccinationstrategies are being considered. ‘Ring’vaccination, for example, involves containing infected animals and vaccinating those within a certainradius of the infected property. Suchstrategies will also require a test to differentiate between animals that havebeen infected and those that havebeen vaccinated.

Through the CRC we are also developing ‘multiplex’ diagnostic assaysthat will provide more accurate resultsfor diagnosing diseases caused by

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Novel paramyxoviruses including the Hendraand Nipah viruses, have been found in Australianand Malaysian flying foxes, respectively.

closely related viruses. In the past, mostassays relied on one antibody or antigenfor disease diagnosis. Using multiplextechnologies, we will be able to conductassays at the same time and in thesame sample.This will not only increasesensitivity and specificity, but will alsoprovide intra-assay comparisons thatare needed to differentiate betweendiseases caused by closely relatedviruses, such as those caused byHendra and Nipah viruses, MurrayValley and Japanese encephalitis viruses,and West Nile and Kunjin viruses.

The sub-program is also involved indeveloping diagnostic reagents for

Paramyxoviruses from fruit bats, pigs, rodents, shrews andother animals have also been isolated, and their molecularstructures (genome, proteins, antigens) characterised“ “

severe acute respiratory syndrome(SARS) in Australia.We are also playinga leading role in an international collaboration to develop an antibodytest for SARS-CoV (coronavirus) infection in different animal species.Thenatural host of SARS has not yet beenfound, but this test will allow us toidentify the different species that thevirus can infect. In a recent WorldHealth Organisation sponsored missionto China, the project leader joined agroup of international experts to out-line strategic plans for future researchon animal reservoirs of SARS-CoV andits transmission to humans.

In 2002, the MLS program

published the first information

on field trials of genetically

modified sheep, revealing the

benefits and limitations of this

new technology

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Reducing environmentalimpact 73Greater productivity 76New/improved products 78

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Program Leader Dr Rob Kelly

Ph (07) 9333 6685Fax (07) 9333 7688Email rob.kelly@csiro.au

The Mediterranean LivestockSystems Program (MLS) based atFloreat Park in Perth, has threekey research areas: environmentalissues in agriculture, greater livestock and pasture productivity,and new products from sheep and cattle.

Through the reducing environmentalimpacts sub-program, we aim to helpgraziers sustainably manage and adaptto saline and water logged environments.To do this, innovative animal production methods are beingdeveloped in collaboration with theCRC for Plant Based Management ofDryland Salinity, and others. Projectactivities planned over the next fouryears include:•Screening existing salt tolerant plants

for nutritive value and secondary compounds;

•Understanding the impact of grazing on biodiversity, plant persistence and water use, and the movement of salt,sediment and water ;

•Measuring the performance of animalsfed salt tolerant plants, and the effect of combining saline plants with other feed types.

The sub-program also aims to reducegreenhouse gas emissions from ruminant livestock through a vaccinetargeting methane-generating micro-organisms in the rumen.Vaccinepreparations are currently being studied in sheep. Development of ananti-methanogen vaccine for cattle, anda vaccine against protozoa in sheep -which consume rumen bacteria andthus remove a valuable source of nitrogen from ruminants’ diets – arealso underway. Reducing the activity ofmethanogens and protozoa is expected to increase productivity insheep and cattle.

Through the greater productivity sub-program, a website has been launchedthat provides farmers in southernAustralia with estimates of pasture biomass and growth rates.This information (based on satellite images),allows farmers to better manage theirpastures and boost profitability.Wehave reliably and accurately deliveredthis information at the shire level, andhave piloted delivery at the farm andpaddock level. Near infrared reflectancespectroscopy, is also being used tomeasure hay quality, allowing farmersto predict feed intake and animal performance – particularly in dairy cattle. A ruminant model and decisionsupport software package (AusBeef),is being developed to assist consultants/managers of cattle feedlots

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in formulating rations, and for devisingfeeding strategies and other management options.

The new and improved productssub-program aims to improve meatflavour and composition, milk production and quality, and enhancewool processing and garment quality.Some of the projects, for example, lookat enriching the nutritional value ofmeat and milk by introducing omega-3fatty acids into the diets of ruminants.A commercial plant has been established in India to produce a protected feed supplement that boosts

Through the reducing environmental impacts sub-program, we aim to help graziers sustainably manage and adapt to saline and water logged environments“ “

milk production in Indian dairy herds.And by selectively breeding for sheepwith low felting wool, we expect toreduce shrinkage and pilling of garments.

In 2002, the MLS program publishedthe first information on field trials ofgenetically modified sheep, revealingthe benefits and limitations of this new technology.

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REDUCING ENVIRONMENTAL IMPACT

Projects in this sub-program focus on two of the major environmental threats in Australia – salinity and greenhousegas production.

Sub-program LeaderDr David MastersPh (08) 9333 6691Email david.masters@csiro.au

The sub-program aims to improve theeconomic viability of saltland pasturefor livestock production, while increasingenvironmental sustainability and managing the effects of salinity andwaterlogging. In a separate project,greenhouse gas reductions are alsobeing sought using vaccines againstmethane-producing microbes in sheepand cattle. A vaccine against rumenprotozoa aiming to increase wool production is also a major researchactivity.

ANIMAL PRODUCTIONFROM SALINE LANDS(APSL)Dryland salinity is the most devastatingthreat to our agricultural environment.Across Australia, 5.7 million hectares -predominantly in the wheat-sheepzones - are threatened, with this figureset to triple to 17.1 million hectares by2050. In Western Australia alone, 16percent of agricultural land is at risk.

As land reclamation is not a short-term option in many areas, new landuses are needed, which provide an economic return and help control risingwatertables.Through the AnimalProduction from Saline Lands umbrellaproject, a variety of projects are investigating land use and animal production issues.

Sustainable grazing on saline landsWith the support of the CRC forPlant-based Management of DrylandSalinity and the national Land,Waterand Wool program, the APSL groupare involved in a number of projectson grazing salt-tolerant pastures.Saltland pasture in Western Australia isdominated by saltbush (Atriplex spp),bluebush (Maireana spp), and ‘volunteer’understorey species made up of nativeand introduced grasses, weeds andlegumes. Studies have shown that saltland pastures can maintain low tomoderate growth rates and wool production in young sheep duringautumn. However these weight gainshave been linked to low salt understorey pasture species, ratherthan saltbush or bluebush.

Dryland salinity is a major threat to Australia’s agricultural systems. CSIROLivestock Industries aims to improve the economic viability of saltland pasturefor livestock production while increasing environmental sustainability.

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The project aims to modify this understorey to increase the feedingvalue of saltland pasture, and improveliveweight gains and wool quality andquantity.Two five-hectare plots atTammin, east of Perth, have been sownwith recommended understoreyspecies.These will be allowed to flowerand set seed in spring.The pasture willbe grazed the following autumn.Understorey plants will then be examined for minerals (salts), proteincontent, fibre content and digestibility,before, during and after grazing, andwill be compared to understoreyplants in two unmodified plots. In thegrazing sheep, liveweight change, woolquality and wool quantity will be measured. Results from this project will allow farmers to assess the economic value of modifying their salt-affected land.

A second project within SustainableGrazing on Saline Lands, being undertaken at Yealering (south-east ofPerth), looks at how to optimise supplementary feeding on saltland pasture. Farmers typically supplementtheir flock once saltbush availability islow. However, animal house studiessuggests there may be a liveweightbenefit in feeding small amounts ofgrain or straw as soon as animals areput out to pasture.

To test this hypothesis in the field, onegroup of sheep were supplementedwith barley grain or barley straw onthey day they went into the saltbushpaddock.Two other groups were supplemented once 75% of the saltbushhad been grazed.The liveweight gainand wool growth of the different feeding strategy groups were compared.

At a third saltbush site at Lake Grace,south-east of Perth, set stocking androtational grazing will be compared.With set stocking, where a certainamount of sheep per hectare are lefton the same pasture for weeks ormonths, liveweights are expected toincrease initially, but then decrease asthe higher nutritive value foodbecomes less abundant. By using rotational grazing around paddocks toreduce the amount of time thesesheep are left to graze one area wemay be able to increase the overallproductivity from the area, and encourage the growth on new highernutritive value plant material.

Nutritive value and secondary compoundsPlants growing in hostile environments,such as saline land, have mechanisms todeal with environmental challenges.These mechanisms may help the plantswithstand the osmotic stress within thetissues or in other cases may bedesigned to discourage grazing andgive the plant a competitive advantage.The result is often the accumulation of

secondary compounds, such asoxalates, nitrates, betaine, tannins oreven salt, which may be beneficial orharmful to animals. Research in thesubprogram aims to identify such compounds in salt-tolerant pastureplants.The effect of different salt concentrations on secondary compounds and the nutritive value of salt-tolerant plants are also being studied.

Nutritional studies have shown thatsaltbush can provide crude protein (9-17%) to grazing ruminants duringsummer and autumn, but is low indigestible energy. In addition, mineralcontent (salt) can be as high as 30%.The high salt acts as a limiter to intakeand is probably the main reason sheepdo not gain weight when only high saltplants are available for grazing.

Under some circumstances, high levelsof salt have benefits. A high salt intakeresults in higher water consumptionand a faster flow of nutrient throughthe gastrointestinal tract (particularlythe rumen).The rapid passage mayreduce the breakdown of protein bythe rumen microbes and increase theprotein available for absorption by theanimal. More protein means morewool growth for sheep.This beneficialeffect is usually dependent on theother components of the diet (ie thosein addition to high salt plants).

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Saltbush and carcass and meat qualityAnecdotal evidence suggests sheep fedon saltbush have meat that tastes better and has less fat.To scientificallyassess the effect of grazing saltbush onsheep carcass and meat quality, we arestudying various physiological and biochemical responses, in the animalhouse and field.

In one animal house study, hoggetwethers were fed either saltbush withbarley (field simulation); lupins, barleyand hay (control); or lupins, barley andhay with added salt (control plus salt),for 70 days. Animal performance wasmeasured by liveweight gain, woolgrowth and body fatness.

Results showed that animals fed thesaltbush and barley diet gained thesame amount of weight as controls(60-80 g/day). However fat content inthe meat of animals fed saltbush wassignificantly lower than the control andcontrol plus salt groups.This may berelated to lower levels of insulin andIGF (insulin-like growth factor), whichare involved in the fat metabolismpathway. Further studies are underwayto examine this.

To assess any impact on carcass qualityfollowing slaughter, carcass weight, pH,and colour were measured in the animal house experiment, and musclesamples taken for glycogen analysis. Nosignificant difference in carcass qualitywas found.

Following slaughter a sensory panelassessed meat samples for tenderness,juiciness, flavour and aroma. No significant difference in these attributeswas observed between the diets,indicating that saltbush has no detrimental effect on eating quality.

Field trials are now underway, withsheep grazing either saltbush or stubbleplots. Early results suggest the meat ofanimals grazing saltbush has a higherwater content.This may be associatedwith more tender and juicy meat.

REDUCING METHANE BYVACCINE TECHNOLOGYSheep and cattle contribute 14% oftotal Australian greenhouse emissions,due to the activity of methane producing microorganisms known as‘methanogens’ in the rumen. As agreenhouse gas, methane is 23 timesmore potent than carbon dioxide.Methane production also accounts forabout 12% of the animal’s gross energy intake.

To try and reduce methane productionby methanogens in the rumen, we arecomparing two vaccine formulations insheep.The first formulation targets awide range of methanogens and thesecond formulation targets threemethanogens that dominate themethanogen population.The project istargeting a reduction in methane emission of 7-20% with a concurrent

increase in liveweight gain.Trials on cattle are anticipated in the future.Related projects are currently beingnegotiated with the AustralianGreenhouse Office.

Boosting wool productionA vaccine targeting protozoa in therumen is being developed and testedwith funds from Australian WoolInnovation. Inhibition of the protozoaleads to improved supply to the animalof microbial protein from the rumen.Early research suggests that by reducingthe numbers and activity of protozoa,wool production and liveweight gainsshould increase.

Saltbush (Atriplex spp) plays an important role in sustainable agricultural production on saline land.

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GREATER PRODUCTIVITY

The Greater Productivity sub-program aims to provide precision land management information and tools to assistproducers, land managers and agribusiness to predict animalperformance and product output, and to manage their businesses profitably and sustainably.

Sub-program LeaderDr David HenryPh (08) 9333 6689Email dave.henry@csiro.au

PASTURES FROM SPACE This project aims to provide accurateestimates of pasture production andpasture quality using remote sensing.For pasture production, satellites thatmeasure the reflectance of visible lightare used to show the amount of‘greenness’ across a target region, andthis is calibrated to estimate pasturebiomass in kg/ha.This is then combinedwith climate data to produce real timeestimates of pasture growth rates(PGR) in kg/ha/day.

The technology is being trialed byWestern Australian farmers, and subsequently modified to improve itsaccuracy, frequency and accessibility.PGR and biomass information is broadcast on ABC Radio, signposted inregional areas, and available on threewebsites developed in collaborationwith Agriculture Western Australia, theDepartment of Land Administration,and the FarmShed. Currently, PGR arepresented as averages for each shireand are available weekly, while biomassis shown within individual farms and is

available monthly. In future years theweb sites will deliver whole farm information at the paddock level, atintervals tailored to individuals.

This technology will assist producerswith management decisions such asgrazing rotations, feed budgeting,fertiliser application and other precisionagriculture techniques. Already, strategicgrazing and feed budgeting in theWestern Australian wool industry hasimproved the quantity and quality(fibre diameter and staple strength) ofwool produced. Analogous benefits arepossible in the meat, beef and dairyindustries. Application of the technologyin other states is now being assessed.

Recent work aims to develop pasturequality measurement techniques, andinvolves the use of airborne and satellite-mounted near infraredreflectance spectroscopy (NIRS) equipment.The technique measuresthe wavelengths of infrared lightreflected by pasture, which variesdepending on the amount of protein,fibre, lignin, oil, starch and other

mediterraneanlivestock systems

The pastures from space project provides accurate estimates of pasture production and pasture quality using remote sensing. Dr David Henry andMarion Barnes examine a map of pasture growth rates across Western Australia.

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components in the pasture. It is hopedthat in the future, PGR, biomass andquality measurements from satelliteswill be tailored for individual users andavailable as required all year round.

More information is available atwww.pgr.csiro.au

LIFETIME WOOLIn Mediterranean climates, the seasonalfluctuation in pasture generally meansthat at some stage during pregnancy, agrazing ewe will become undernourished.This affects wool follicle development in the unborn foetus, leading to reduced wool qualityand quantity in later life. It also affectswool quality and quantity in the ewe.

The lifetime wool project - a collabora-tion between the Department ofPrimary Industries,Victoria,Department of Agriculture,WesternAustralia, CSIRO, and wool producers -aims to manage pasture biomass toimprove ewe nutrition during pregnancyand achieve predictable wool qualityand quantity in ewes and their offspring,over their lifetime.

Wool quantity and quality will be monitored in Merino ewes fed on pastures carrying 800, 1100, 1400,2000 or more than 3000kg of dry matter/ha, over three years. Progenywill also be monitored. By identifyingwhen and by how much nutritionshould be improved during pregnancy

to optimise wool characteristics, arange of practical guidelines will bedeveloped.These will enable wool producers to choose their optimumfeeding solution during pregnancy and lactation to increase profits. An economic analysis on the potentialbenefits of improved nutrition predictsgains of $5-$6 per ewe per year.

PREMIUM GRAINS FORLIVESTOCK PROJECT(PGLP) AND AUSBEEF Through the PGLP, scientists havefound that the nutritional value of arange of cereal grains (wheat, barley,sorghum, triticale and oats) when fedto poultry, pigs and ruminants, varies byabout 30%. Some of the largest variations have been observed inexperiments involving cattle. Many ofthe chemical and physical characteristicsof these grains that are responsible forthe variations in nutritional value havebeen identified. Rapid and cheap methods based on near infrared (NIR)technologies have been developed tomeasure these characteristics, yielding alarge amount of information that couldbe directly transferred to a laptop orpersonal computer.

To make this nutritional informationaccessible and useful to both buyersand sellers of feed grains, the PGLP hasalso developed a detailed mathematicalmodel of a ruminant.The ruminantmodel consists of sub-models of

voluntary intake, the rumen, the lowergut, and body growth.The model simulates observed feeding behaviourin cattle feedlots, ad libitum feeding,restricted feeding, and other morecomplex feeding strategies.The ruminant model forms the basis ofAusBeef, a decision support softwarepackage designed to assist consultantsor managers of cattle feedlots to formulate rations, devise feeding strategies and identify other manage-ment options. AusBeef may also beused by the feed grains industry toassess the value of their products foruse in cattle feedlots.

The ultimate aim of the package is toallow users to send feed informationobtained using a NIR device, directlyinto AusBeef, where it can be combined with other feed databaseinformation to evaluate the economicvalue of different grains and feeding options.

Other capabilities to be incorporatedinto AusBeef in the future include:• management options to avoid

heat stress;• the ability to evaluate economic

differences between feedlot sites;• the ability to examine management

options for their capacity to reduce greenhouse gas emissions, such as methane production.

Satellites such as this Japanese owned satellite, areused to measure the reflectance of solar radiationfrom the Earth’s surface. CSIRO Livestock Industrieshas developed an accurate method for measuring pasture biomass from this data.

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NEW/IMPROVED PRODUCTS

To meet consumer demand and future markets for newproducts and services, this sub-program is developing technologies to produce milk and sheep meat products that improve human health.

Sub-program LeaderDr Norm AdamsPh (08) 9333 6687Email norm.adams@csiro.au

Improvements in wool processing andgarment quality are also being pursued.The sub-program will develop cost-effective ways to produce these products by determining their mostprofitable place within current production systems.

MEAT FROM MERINOS The Merino industry is under pressureto increase meat production and, atthe same time, grow finer, high qualitywool. As meat and wool both consistof protein, there is competitionbetween the two for amino acids (protein building blocks).

Studies through the Australian SheepIndustry CRC, and in collaborationwith the Department of Agriculture inWestern Australia, indicate that sheepwith higher fleece weights (and there-fore higher protein turnover) havereduced amounts of energy (glucose)available for liveweight gain, fat deposition and reproduction.Thus,selection of sheep for greater woolgrowth may be detrimental toincreased growth and reproduction.

To try and understand the movement(‘partitioning’) of nutrients from feedto wool, meat and fat, metabolic studieslooking at protein turnover, hormonelevels and glucose synthesis, in sheepselected for genetic extremes of fibrediameter, fleece weight and liveweight,will be set up in Western Australia and Armidale.

In Western Australia, the impact offleece weight and fibre diameter onthe nutrients available for growth andreproduction are being examined.Protein synthesis in skin and muscle willbe compared in individual sheep, andthe efficiency of wool growth as a fraction of skin protein turnover will bedetermined at different fibre diameters.Hormones and metabolites will bemeasured to determine the way inwhich amino acid metabolism affectsthe energy balance of sheep. Proteinexpression profiles on muscle fromsheep differing in wool characteristicswill allow estimation of the impact onenergy metabolism in muscle.Theseanalyses will enable us to predictimprovements in meat characteristicsof sheep resulting from changes inwool growth.

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In Armidale, work will consider the efficiency of amino acid use for wooland muscle in sheep differing in fleeceweight and fibre diameter. Measures ofgrowth rate, skin characteristics andfeed intake under field conditions, willbe made.

These studies will provide biologicalmarkers that predict the impact ofwool growth on the flow of nutrientsto meat quality and quantity, and recommendations on feeding regimesto optimise nutrient partitioning to thedesired products in specific genotypes.

EFFECT OF PARASITES ONNUTRIENT REQUIREMENTSSheep affected by nematode parasitessuch as Trichostrongylus colubriformis andOsertagia circumcincta, have anincreased requirement for nutrients,particularly protein, in order to mounta strong immune response. Growinglambs, pregnant and lactating ewes, andwool-producing sheep, also have anincreased requirement for protein, andare susceptible to parasite infectionduring periods of nutrient deficiency.

This project is part of a broader effortin the Australian Sheep Industry CRCthat aims to answer three questions:• How much extra protein do infected

sheep require, what type of protein (amino acids) do they require, and when do they require it;

• How do you manipulate nutrition to improve production in infected sheep, and;

• What is the metabolic mechanism bywhich sheep become resistant to parasites?

The nutritional requirements of parasiteresistant Rylington Merinos, bred bythe Department of Agriculture,Western Australia, are being comparedwith susceptible sheep, in animal houseexperiments. Early studies in which halfthe resistant and susceptible sheepwere infected with T. colubriformis andO. circumcinta, showed no difference involuntary feed intake and body weightgains as a result of parasitism. However,resistant, infected sheep were found tohave lower concentrations of theantioxidant glutathione, which is thoughtto be involved in immune function.A decrease in some amino acids in infected resistant and susceptible sheepwas also observed, suggesting theamino acids could be involved in host-parasite interactions.These sameamino acids have been implicated inimmune function in human health.

A long-term field experiment will beconducted by the Department ofAgriculture to continue this work andto look at the partitioning of nutrientsin infected sheep.

GENETICALLY MODIFIEDSHEEPMerino, Border Leister and Poll Dorsetsheep, genetically modified with anextra copy of sheep growth hormonegene were tested in field conditionsover three years to assess how themodification influenced growth, andwool and milk production. Growthhormone affects important characteris-tics of an animal's development,including growth rate and fatness.Thefirst research of its type in Australiademonstrated a range of genetic interactions.Wool growth, for example,increased in Merinos, but decreased inPoll Dorsets. In general, however, GMsheep had bigger bones, similaramounts of muscle, and less fat, andthey grew faster and produced doublethe amount of milk. However, theyrequired more care, with overgrowntoes and retained testes being commonproblems.The work has highlightedpossible side-effects that traditionalbreeding programs, which select animals for large, lean lambs, may need to consider.

Research on sheep genetically modified with an extra copy of the sheep growthhormone gene found that while sheep grew bigger and ewes produced moremilk, there were some side-effects for the animals.

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FISH OILS IN MEAT ANDMILK Consumption of omega-3 polyunsatu-rated fatty acids, found in fish oil,reduces the risk of heart disease by upto 30%. As the Western diet rarelyincludes the recommended two fishmeals per week, improving the omega-3 content in meat and milk maybe a way to improve human health indeveloped countries.

As polyunsaturated fatty acids interferewith digestion in the rumen, we haveused CSIRO technology (Rumentek) toencapsulate omega-3 oils from tuna ina protein-aldehyde matrix.This matrixis stable at rumen pH, enabling the oilto by-pass the rumen and go straightto the small intestine. Here it breaksopen, releasing the omega-3 oils forabsorption directly into the blood andsubsequent incorporation into meat ormilk.While omega-3 fatty acids arealready present in low amounts in

meat, we have been able to triple meatconcentrations, without affecting feedintake or growth rate. Similarly, milkconcentrations of omega-3 have risenfrom zero to half the recommendeddaily intake, with no effect on milkyield. Experiments were conducted ondairy and meat sheep, dairy goats anddairy cows.This project is now part ofCSIRO’s Agrifood flagship project.

BOOSTING CONJUGATEDLINOLEIC ACID IN MEATAND MILKConjugated linoleic acids (CLA) are afamily of partly hydrogenated fattyacids with potential anti-cancer properties.They are found naturally inthe meat and milk of ruminants, andare higher in animals fed on pasturethan on grain or conserved feed.A study of the components of cowmilk in Western Australia, for example,showed CLA concentrations dropped

from 3% in winter, when pasture wasfresh and abundant, to about 0.4% during the dry summer months.

CLA concentrations also vary betweenbreeds. A comparison of differentbreeds of dairy sheep, for example, hasshown that there are more CLAs inMerino milk, than in the milk of theAwassi or East Friesian.

This project aims to boost the CLAcontent of meat and milk by combiningthe right breed of animal with the rightdietary supplement. CLA’s are notfound naturally in livestock feed, butare produced during bacterial fermentation of precursor molecules indietary ingredients (such as canola,soybean or cotton oil) in the rumen.We are now looking to see how heritable the trait is in Merino sheep,in order to select for animals with naturally high CLA concentrations intheir meat or milk; and whethercracked canola seed will provide a

Consumption of omega-3 polyunsaturated fatty acids,found in fish oil, reduces the risk of heart disease by up to 30%“ “

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suitable CLA precursor. Other benefitsof feeding precursors, such asimproved growth on low quality feeds,or higher milk yields, will also be investigated. Such benefits could helpbalance the cost of oilseed processing.

IMPROVING DAIRY MILKPROTEIN LEVELS The Australia New Zealand FoodAuthority has stipulated that fresh milkmust contain 3.1% crude protein.However, dairies in Queensland andWestern Australia have been unable tomeet this target over the summermonths.

To find out why, a group of representatives from the Departmentsof Agriculture in Western Australia andQueensland, the QueenslandDepartment of Primary Industries, theDairy Research and DevelopmentCorporation (DRDC) and local dairycompanies, was formed.Their projectaimed to understand the cause of lowmilk protein, develop cost-effectivestrategies to correct it, and providefarmers with the knowledge and toolsto manipulate protein concentrations.

A group of ‘monitor farms’ consisting ofeight high protein and eight low proteinfarms were selected in WesternAustralia and Queensland, and information on feed quality and quantity,and the results of herd tests (milk production, milk quality and calving

patterns) were examined. Informationfrom the Australian Dairy HerdImprovement database, which collatesherd information and test data fromfarms throughout Australia, was also analysed.

The project found that the mostimportant factor affecting protein concentration was the interactionbetween the season (feed quality) andthe stage of lactation in the cow. InQueensland and Western Australia,most cows were found to be calving insummer – in order to meet thedemand for fresh milk year round andfor the premium price paid for summermilk.This meant that cows were in theearly stages of lactation at the worsttime of year for nutrition. In early lactation, the cow’s demand for proteinand energy is high, and cannot be fullymet by summer pasture and conservedfeed, resulting in low milk protein.

In contrast, dairies in Victoria (whichrun 60% of the Australian dairy herd),and other states, had most cows calvingoutside summer, and those beingmilked during summer were in the laterstages of lactation, when milk proteincontent is highest.

The type of supplement fed to cowswas also found to be important. Highstarch supplements such as maize, pushmilk protein up, while lupins, whichincrease the amount of milk produced,tend to push down the protein

percentage. Breed also plays an important role in protein concentration,with Jersey cows having 5 g/L moreprotein than Holsteins or Freisians.

To boost milk production, farmers inWestern Australia and Queenslandwere advised to consider changingtheir calving patterns, and to keep latestage lactating cows in milk for longer,rather than drying them out too soon.They were also advised to replacesome lupins with starchy cereals suchas triticale and wheat. And selecting forhigh milk yield and high protein concentrations within a breed, or bycrossbreeding was also an option.Implementation of this advice overthree years, led to an increase in thestates’ average milk protein content of 1.5 g/L.

With deregulation of the dairy industry,farmers now have less incentive toproduce milk in summer. However,farmers still need strategies to manipulate protein content.To test different management strategies andevaluate their impact on protein levelsand farm income, CSIRO, the DRDCand the Australian Dairy HerdImprovement Scheme developed a‘Dairy Ready Reckoner’ computer program.The program incorporatesfour years of milk production and composition data for 10 000 farms and41 dairying regions around Australia. Itcan review historical data for individual

Wool felting (shrinkage) is a heritable trait which can be manipulated throughthe selective breeding of sheep.

CSIRO Livestock Industries research has tripled theamount of healthy omega-3 polyunsaturated fatty acidsin lamb, and boosted concentrations in the milk ofsheep, goats and cows.

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and district dairy farm performance topredict milk income through changingcalving pattern, breed, milk yield, milkcomposition and factory pricing schedules.

The ability to calculate feed costs isnow being incorporated and field validation of the software on four farms,is underway. A checklist for farmers, forraising milk protein levels during summer, has also been developed.

SHRINK-PROOF WOOL Wool felting or shrinkage causes significant problems in the manufactureand care of knitted and woven woolproducts, particularly fine wool garments. However, recent workthrough the Australian Sheep IndustryCRC, and in collaboration with theDepartment of Agriculture,Western

Australia, has shown that felting is aheritable trait that can be manipulatedthrough selective breeding of sheep.

Over 2000 wool samples from theKatanning Merino resource flocks weretested for various wool fibre traits,including fibre diameter, staple strengthand staple length. ‘Feltball density' wasalso tested by placing one gram ofclean, hand-carded wool in a containerwith water.This was then placed in afront-loading tumble drier at 20degrees for 30 minutes.The smaller theresulting feltballs, the greater the felting.

Using pedigree information and removing the effects of fibre diameter,fibre curvature and yield, a moderateheritability for felting of 0.39 wasdetermined.This figure can now beused to help woolgrowers designbreeding programs for low felting wool.

Breeding also offers the potential toreduce chemical treatment of woolproducts to prevent shrinkage andother undesirable characteristics.Preliminary processing studies conducted with CSIRO Textile andFibre Technology have shown that knitted fabric made from low-feltingwool has reduced pilling and shrinkage.Low-felting wool also produced longer‘hauter’ (length) and less entanglementduring the scouring (cleaning) process,resulting in fewer breakages duringspinning.We are now investigating theunderlying causes of felting, which arethought to be due to variations in thesize, shape and/or structure of wool fibres.

Over 2000 wool samples from the Katanning Merinoresource flocks were tested for various wool fibre traits,including fibre diameter, staple strength and staple length“ “

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Dr Tony Schlink and Melanie Ladyman are investigating theheritability of dust accumulation in wool. Genetic selection of sheep for fleeces that attract less dust, could see improvements in economically desirable fleece traits.

DUSTY WOOL In Mediterranean environments, dustyconditions reduce the value of finewool, as the dust accumulates on andin the fleece, reducing clean wool yieldand style grades - a visual score of thewool.Traditionally, growers and sheepclassers have estimated dust contentby measuring how far dust has penetrated the fleece. However,research in collaboration with theDepartment of Agriculture,WesternAustralia, and the University ofWestern Australia shows dust penetration is not an accurate indicator of dust content.

Wool containing more wax, for example, tends to accumulate largeamounts of dust at the tips of the fibre.With less wax, dirt is able to penetratethe fleece further, but the amount ofdirt is less than that in a waxy fleece.

Analysis of the wax, suint (sweat) anddust content of some 1050 fleeces,showed all traits were heritable andcorrelated with wool yield.Thus, geneticselection of sheep for fleeces thatattract less dust, could lead to modestimprovements in clean fleece weightand other economically desirable traits.

Studies are underway to determine aheritability figure for dust content andidentify other heritable traits, such as

wax content, which could be used asan indirect measure of dust content.A correlation between wax contentand ultraviolet light damage is alsounder investigation.

ANGORA RABBITSA feasibility study into the potential foran angora rabbit fibre industry inAustralia has been conducted.Thereport at http://www.rirdc.gov.au/rep-orts/NAP/03-014.pdf, advises potentialinvestors to conduct careful marketresearch before proceeding.While animal husbandry skills in Australia arecompetitive, current world prices andthe dominant market hold of a fewcountries, suggests only limited scopefor an Australian industry.

Our program is applying and

developing cutting edge

science to produce vaccines

and therapeutics against

diseases of economic

significance to livestock

industries worldwide

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Development of noveltherapeutics 86Gene technologies 88Viral delivery 90

vaccinesand therapeutics

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Program Leader Dr Chris PrideauxPh (03) 5227 5791Fax (03) 5227 5555Email chris.prideaux@csiro.au

The Vaccines and Therapeuticsprogram aims to develop andcommercialise products that willenhance the health, welfare andproduction of AustralianLivestock.These products mustbe compatible with a move awayfrom the use of antibiotics inlivestock industries as recommended by the JETACAR(Joint Expert Technical AdvisoryCommittee on AntibioticResistance) report.

Implementation of policies furtherrestricting chemical and antibiotic usein production animals, changes in husbandry and farm management practices, continuing emergence ofhyper-virulent pathogens and advancesin vaccine technologies will see newdemands and opportunities for vaccinesto control disease and promote welfare.With this end in mind, our program isapplying and developing cutting edgescience to produce vaccines and therapeutics against diseases of economic significance to livestockindustries worldwide.

Through closer examination of animalspost-vaccination, and examininghost/pathogen interactions duringinfections, we will use micro-array technologies to gain an insight intowhat constitutes a protective immuneresponse.This information will then beapplied to design more effective

vaccines, and to generate genetic markers for selective breeding.

The program also has extensive experience in the use of cytokines -natural modulators or regulators of theimmune system - to control diseasesand promote the welfare and growthof production animals. For example,delivery of a fowl adenovirus to chickens, which had been modified tocarry the cytokine gene for chickeninterferon gamma (ChIFN-γ), boostedthe birds’ immunity and improved theirgrowth rate by 5-10%.This workreceived a CSIRO Medal in 2002.Similar work is being undertaken inpigs and will be extended to other livestock species in the future.

The search for ‘natural’ replacementsfor chemical usage in livestock industrieshas also led us to investigate the therapeutic potential of bacteriocins,small bioactive polypeptides producedby bacteria.This work is expected toexpand to look at polypeptides produced by the host to protectagainst microbial invasion, and theirexploitation as therapeutics and potentially as genetic markers forincreased innate immunity.

Novel technologies, such as RNA interference (RNAi), for controllingviral, microbial and parasitic infection,provide opportunities for generatingIntellectual Property.The program isnow exploring its potential.

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DEVELOPMENT OF NOVEL THERAPEUTICS

Over the past few decades, antibiotics, chemicals and vaccineshave been used in intensively farmed livestock, such as pigs,poultry and feed-lot cattle, to control infectious diseases,promote growth and improve feed efficiency.

Project LeaderPorcine ImmunologyDr David StromPh (03) 5227 5740Email david.strom@csiro.au

Project LeaderAvian ImmunologyDr Andrew BeanPh (03) 5227 5792Email andrew.bean@csiro.au

Program ManagerPoultry CRCDr John LowenthalPh (03) 5227 5759Email john.lowenthal@csiro.au

However, the emergence of drugresistant organisms in human medicinehas fuelled concern that antibiotic usein animals may speed the developmentof resistance in both human and animalpathogens. Livestock industries aroundthe world are now facing restrictionson the use of antibiotics and chemicalsfor food production animals.This maylead to an increased incidence of infections and reduced productivity.Industry is also seeing the emergenceof hyper-virulent pathogens that aredifficult to control with current vaccines.Thus, safe, alternative vaccinationstrategies and therapeutics are needed.

The Therapeutics team encompassestwo parallel projects, porcine and avianimmunology as well as involvement inthe Poultry CRC.The general aims are to:• better understand immune function

in chickens and pigs;• characterise natural immune

modulators, known as cytokines;• assess their therapeutic potential in

enhancing disease resistance and productivity, and vaccine effectiveness;

• collaborate on the development of simple, effective delivery mechanismsfor such therapeutics and;

• develop the National Facility for Poultry Immuno-Genomics

IMMUNE FUNCTION

The Therapeutics team aims to gain abetter understanding of immune function, in particular, the innate (firstline of defence) immune response.They are studying a group of innateimmune molecules known as ‘toll-like’and ‘TREM’ (triggering receptorexpressed on myeloid cells) receptors.These receptors recognise potentiallypathogenic material (such as virusesand bacteria or their products) and signal the immune system to attack.By learning more about how these receptors work, the team hope tomodulate the response to infection,thereby enhancing production andimproving animal welfare. Otherimmune system molecules such asporcine bone marrow stem cells anddendritic cells are also under study.

Dr John Lowenthal (left) and Dr Mike Johnson are part of a teamthat developed a way to deliver cytokines to chickens, boostingtheir immunity and improving growth rates by 5-10%. Such newtherapeutics could provide an alternative to antibiotics.

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Cytokine therapy trials in commercial piggeries showed pigs given purifiedporcine cytokines had less disease and gained up to 10% more weight than pigs given antibiotics.

CYTOKINE THERAPEUTICSCytokines are proteins produced bythe immune system, in response toinfection by bacteria, viruses and parasites.They help the innate immunesystem identify the type of invadingorganism, and to mount an adaptiveimmune response. Generally, this willbe an antibody response to bacteriaand parasites, and a ‘cell-mediated’(such as T cells) response to viruses.Thus, cytokines could provide a sourceof natural therapeutics, especially foryoung, intensively housed animals,whose immune systems are immatureand regularly challenged.

In 1990 the Avian Cytokine Projectwas established to clone, express andcharacterise chicken cytokines andassess their therapeutic potential. Oneof the cytokines developed - chickeninterferon gamma (ChIFN-γ) - boostedthe immunity of chickens and improvedtheir growth rate. In collaboration withscientists from the Viral Vector Project,the genes coding for ChIFN-γ wereinserted into a harmless, species-specific fowl adenovirus.Trials showedthat delivery of the virus to broilerchickens via their drinking water resulted in improved growth rates of 5-10%, possibly due to the immuno-enhancing properties ofChIFN-γ. This technology has beenlicensed to a spin-off company,VectoGen Limited, and a leading pharmaceutical company is evaluatingits commercial application.The workreceived a CSIRO Medal in 2002.

A parallel project for pigs was initiatedin 1993 and a similar strategy is underway for the development ofcytokine therapy in pigs.Trials in commercial piggeries showed that pigsgiven purified porcine cytokines and antibiotics had less disease and gainedup to 10% more weight than thosegiven antibiotics. Additional trials incommercial piggeries showed that pigsgiven purified porcine cytokines hadsubstantial health and productivityimprovements over those medicatedwith or without antibiotics (i.e. naturalalternatives to antibiotics).The conceptof enhancing the immune competenceof pigs and increasing the resistance toinfection has been further tested byspecific infectious disease challengemodels. Specific porcine cytokines were shown to decrease the incidenceand pathology of infectious disease.It is anticipated that this strategy could be adapted to other animals,including humans.

FUTURE DIRECTIONS

As there are many different types ofcytokines, much of the work in pigsand chickens now involves identifyingthe function of each cytokine andassessing its biological activity in tissueculture. Biologically active cytokines can then be inserted into suitable viral vectors for large scale delivery.

This work could lead to the development of cytokine therapeuticstargeted to particular pathogens or for the initiation of a particular immune

response (antibody versus cell mediated) as well as increasing innateor mucosal immunity.The use ofcytokines as ‘adjuvants’ which, whencombined with a vaccine, increase theeffectiveness of that vaccine, is alsobeing studied. Such adjuvants couldovercome the need to develop newvaccines for different and/or more virulent strains of the same virus, orenable administration of lower dosesof expensive vaccine.

The Australian poultry industry is supporting the search for alternativetherapeutics through funding by theRIRDC Chicken Meat program,Australian Egg Corporation Limitedand the Poultry CRC. Research isaimed at countering the rise of hypervirulent pathogens such as veryvirulent Marek’s disease virus; reducingthe need for chemicals to control coccidiosis; and reducing reliance onthe use of antibiotics for other common infections and growth promotion.

Similarly, the pig cytokine work, fundedthrough the Australian Pork Limited, andan international pharmaceutical company, may help overcome the needfor antibiotics to control the commonrespiratory disease, Actinobacillus pleuropneumoniae; side effects of harmfulimmune responses, such as inflammation;and the problem of the internal wormparasite, Ascaris suum in pigs.

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GENE TECHNOLOGIES

The major focus of the gene technologies group is to identifyantigens from bacterial pathogens that can be used to producerecombinant vaccines for animal diseases, and assist the pushto reduce antibiotic use.

Project LeaderRecombinant Vaccines and BacteriocinsDr Rob MoorePh (03) 5227 5760Email rob.moore@csiro.au

Project LeaderRNAiDr Tim DoranPh (03) 5227 5788Email timothy.doran@csiro.au

Bacteriocins - small bioactive polypeptides produced by some bacteria to kill others - are also beinginvestigated as potential therapeutics,with particular focus on necroticenteritis in chickens.Through the newlyestablished Australian Poultry CRC, theproject will use microarrays to look athost-pathogen interactions during thenecrotic enteritis disease process.Theapplication of RNAi technology toimprove disease resistance and productivity is also progressing.

RECOMBINANT VACCINES

The bacterium Mycoplasma hyopneumoniae causes pneumonia inpigs, is endemic in pig production systems, and reduces productivity.Vaccines for the disease are producedcommercially and they generally conferabout 80% protection. However, as thevaccines are difficult and expensive toproduce, the development of a recombinant vaccine is desirable.

To do this, the genome of M. hyopneu-moniae was chopped into small DNAfragments and inserted into a plasmid

vector in Escherichia coli. Using this‘expression library’ we developedmethods to then remove ‘junk’ clonesand screened the remaining clones forantigenic activity. DNA sequencing wasalso performed on some clones toidentify genes likely to code for usefulantigens.These two approaches overcome problems commonlyencountered using more traditionalscreening methods that identify antigensby screening expression libraries usingserum from animals exposed to disease.As pathogens often express antigensthat act as a smoke screen to theimmune system, animals develop animmune response to a range of decoyantigens, rather than the antigens thatcould be protective.

By targeting the whole genome, ratherthan serological antigens, we have identified a number of potentially protective antigens to M. hyopneumoniae.These antigens have been tested asrecombinant antigens in conventionalvaccine formulations and by DNA injection.

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Dr Tim Doran and his team are investigating the use of RNAi to target genesfor viruses causing disease in sheep, cows, chickens and pigs.

Early trials of the recombinant vaccinein ‘clean’ pigs – an isolated populationbred by the Victorian Department ofAgriculture – gave about 95% protection. However, field trials onclean pigs in the wider pig communityresulted in about 70% protection.Thisdecrease in protection is likely due togenetic differences in the pigs.We arenow aiming to increase protectionusing cytokines, such as interleukin-3,to boost animals’ responses to the vaccine.

The techniques developed to identifyprotective M. hyopneumoniae antigensare now being applied to Brachyspirahyodysentariae, which causes swine dysentery.

BACTERIOCINS

Another approach to identifying antibiotic alternatives is to look forantibacterial compounds that aren’tconventional antibiotics. Bacteriocins,for example, are produced by somebacteria to compete against others inecological niches.We are investigatingways to use these small antimicrobialpolypeptides to target pathogenic gutbacteria, such as Clostridium perfringens, which causes necroticenteritis in chickens.

As necrotic enteritis is a difficult diseasemodel to reproduce, further study ofthe disease induction is needed and suchwork is being supported by the RIRDC.

Through the Australian Poultry CRC amicroarray facility has been established,which will allow us to look at changesin gene expression in the host andpathogen during different stages ofinfection. Changes in gene expressionin host and pathogen, during manipulation of the host immune system with cytokines or vaccines, forexample, will also be investigated.

RNAI TECHNOLOGIES

RNA interference (RNAi) is thought tobe an ancient cellular defence mechanism, developed by plants andanimals to protect against viral infection.Since its discovery, its mechanism ofaction has been determined and usedto silence or knock out genes of interest in eukaryotic organisms.The function of these genes can then be determined.

The system relies on double strandedRNA (dsRNA) molecules targeting thegene of interest.When a constructcontaining a short DNA sequencefrom the target gene is introduced intohost cells, via a virus, bacteria or plasmid,the DNA is transcribed to producesmall dsRNA molecules that activatethe cell’s RNAi mechanism. Expressionof the target gene is then prevented.

We have developed a range of plasmidvectors that will be used to targetgenes for viruses that affect sheep,cows, chickens and pigs. In collaboration

with CSIRO Division of MolecularScience we are also trialing delivery ofRNAi constructs through an adenovirus.Future work will focus on bacterial vectors.

Applications of the technology includedeveloping transgenic chickens containing a plasmid targeting genes forMarek’s disease virus (a herpes virus),for example. Such chickens would beresistant to the disease. So far we have successfully used RNAi to blockthe replication of a herpes virus in continuous cell lines.

Also on the agenda are genetic constructs targeting essential genes inintestinal nematodes or fly strike larvae,which could be introduced into gutbacteria of livestock. Nematodes eatingthese bacteria, or fly larvae grazing onthese bacteria in the animals’ faeces,would then die.The technology couldalso be used to silence important production genes in livestock animals,for example, the myostatin gene, whichnegatively regulates muscle growth andthe DMRT1 gene involved in sex differentiation in chickens.

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VIRAL DELIVERY

The Viral Delivery project has developed a number of fowland porcine adenovirus recombinant vectors that have beenused, in collaboration with the Novel Therapeutics project, todeliver cytokines and vaccine antigens into chickens and pigs.

Project LeaderDr Mike JohnsonPh (03) 5227 5768Email michael.johnson@csiro.au

Technology license agreements fortheir commercialisation through a newcompany,Vectogen Ltd, are in place.This project aims to assist in the further development and registrationof these commercial products for theAustralian and international markets, specifically:• fowl adenovirus expressing the

spike antigen from infectious bronchitis virus,

• fowl adenovirus expressing chicken interferon gamma,

• porcine adenovirus expressing the gp55 antigen from classical swine fever virus and

• the development of new products utilising the adenovirus vector systems

FOWL ADENOVIRUS

Adenoviruses are a group of virusesthat infect almost all species, but varyin their ability to cause disease. Some,such as the fowl adenovirus (FAV), areinnocuous and easy to grow in tissueculture, making them useful for geneticengineering purposes.We have usedFAV as a vector to transport genes for

therapeutic molecules (cytokines), andprotective antigens (vaccines) into cells.

In terms of vaccines, we have targetedinfectious bronchitis virus (IBV) ofchickens.When a bird is vaccinatedwith live attenuated IBV, it developsantibodies against a range of ‘smokescreen’ antigens, rather than the antigensthat will protect it.The recombinantFAV vaccine, however, delivers only theantigen that will protect the birds – the S1 subunit of the spike protein.Importantly, the vaccine can be delivered in water.

Similarly FAV has been used to deliverthe cytokine gene, chicken interferongamma (ChIFN-γ) into host cells,boosting the chickens’ immunity andimproving their growth rate by 5-10%.

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PORCINE ADENOVIRUS

Innocuous adenoviruses that infect pigs(PAV) can be used in the same way, todeliver cytokines and vaccine antigens.Classical swine fever, which causes diarrhoea and death in pigs, is a bigproblem in Europe and Asia.Trials withPAV containing the gene for the gp55(glycoprotein) antigen, delivered byinjection or oral dose, have demonstrated protection of pigs fromthe disease. Recent trials of the recombinant vaccine in Vietnam haveresulted in more than 60% protection.This should improve once suitable vaccine production and storage facilities are available in Vietnam.

Adenoviruses can be used to deliver cytokinesand vaccine antigens to pigs and chickens.

When a bird is vaccinated with live attenuated IBV, itdevelops antibodies against a range of ‘smoke screen’antigens, rather than the antigens that will protect it“ “

PAV has also been used to deliver thegene for the cytokine, granulocytecolony stimulating factor (GCSF), topigs. A single dose of the therapeuticcan boost the natural immunity of pigsto Actinobacillus pleuropneumoniae, acommon respiratory pathogen. Smallscale trials have shown GCSF confersmore than 90% protection against infection.

In the future we aim to characterisehost responses that typify a successfulvaccination or therapeutic delivery,using microarrays to detect changes ingene expression when animals aregiven a cytokine or vaccine, followedby a disease challenge.

The recombinant adenovirus technologywill also be extended to develop vaccines and therapeutics for other diseases of pigs and chickens, and other animals.

The Temperate Livestock

Systems program has three

main research areas; disease

resistance and animal

welfare, productivity and

enterprise diversification,

and information systems

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Disease resistance andanimal welfare 94Productivity and enterprisediversification 98Information Project Systems 101

temperatelivestock systems

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Program Leader Dr Laurie PiperPh (02) 6776 1349Fax (02) 6776 1334Email laurie.piper@csiro.au

The Temperate LivestockSystems program, located at theF.D. McMaster Laboratory atChiswick, near Armidale, hasthree main research areas;disease resistance and animalwelfare, productivity and enterprise diversification, andinformation systems.Throughthese subprograms we aim to:• Improve production efficiency • Improve product quality, and

develop new products and production options for temperate livestock systems;

• Reduce reliance on chemicals to control parasitic disease; and

• Develop information systems to improve livestock management and breeding programs, and facilitate technology transfer of our research.

The disease resistance and animalwelfare sub-program looks at a rangeof parasite resistance, management,diagnosis and control issues.Theseinclude improving the delivery and lifeof anthelmintic drugs, understandinghost-parasite interactions, biologicalcontrol of nematodes, and developingrace-side diagnostic tests for variousworm species. On the host front, weare breeding animals for parasite resistance and developing more effectivedelivery systems for vaccination.

As animal welfare issues influenceexport market access, the subprogramis looking at the response of animals to

Australian production systems, with theaim of modifying them if necessary.Weare also developing electronic devicesto remotely record and monitor animalphysiology, deliver drugs for parasitecontrol, or to use in electronic livestockmanagement applications.

The productivity and enterprise diversification subprogram aims toincrease profitability in the sheep andemerging industries (yabbies, meat rabbits), by developing improvedbreeding, nutritional and managementtechnologies.The recently constructedReproductive Technology Centre willhouse much of the cloning, IVF andgene modification work.We are alsolooking at improving ultrafine woolproduction with our Towards 13Micron project. And, using the naturalvariation in wool fibre attributes, weaim to identify new products.

The information systems sub-programcomprises projects with a strong customer focus, which specialise in the use of computer modelling, data management and statistical analysis forresearch and technology transfer.AUSPIG for example, is a decision support tool that looks at factorsaffecting production profitability in thepig industry, such as interactionsbetween pig type, feed, environment,market prices, labour and capital.Developments in e-commerce willallow customers to access our toolsover the Internet.

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DISEASE RESISTANCE AND ANIMAL WELFARE

The major cause of lost production and animal deaths in temperate livestock production systems is internal parasitism of sheep.

Sub-program LeaderDr Ian ColditzPh (02) 6776 1460Email ian.colditz@csiro.au

The Barber’s pole worm (Haemonchus contortus) has developedresistance to some of the major chemicals used to control it.

Control measures and production losses cost approximately $220 millionannually. Current measures for controlling internal parasites rely heavilyon anthelmintic drugs. However,resistance of parasites has beenreported for all the anthelmintics incommercial use.Within ten years thelevel of resistance in worms (nema-todes) is expected to leave currentanthelmintics ineffective, by which timelosses due to parasitism could spiral tomore than $700 million annually.Accentuating this threat is the knowledge that no new anthelminticcompounds are expected to becomeavailable in the near future.

To address this problem, the diseaseresistance part of the subprogramlooks at managing and understandingaspects of host and parasite resistance,improving drug delivery and diseasediagnosis, and biological control. Otherprojects aim to understand the geneticbasis of host resistance to mastitis andsheep blowfly strike, and to measureand improve animal welfare.

OVERCOMING PARASITERESISTANCE

Barber’s pole worm (Haemonchus contortus) has developed resistance tosome of the major chemicals used tocontrol it, including Ivermectin. Studieshave shown that resistance toIvermectin is caused by a single gene,which we are now trying to identify.Using genes that confer Ivermectinresistance on the freeliving soil nematode, Caenorhabditis elegans, PCRprimers have been designed thatamplify related sequences in H contortus. If the gene involved isidentified, a DNA assay will be developed to detect resistance in parasites on-farm.This will allow farmersto implement alternative managementmeasures before the levels of resistance get out of hand.

In collaboration with the University ofNew England, we are looking at theheritability of nematode egg counts insheep and fibre goats in the Philippines,and the correlation between circulatingantibodies to parasites and faecal egg

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The nematode-eating fungus, Duddingtonia flagrans, is being investigated for its biological control potential. Here the fungal mycelium has ensnared a nematode larva.

counts. A breeding project will bedeveloped to select for nematoderesistant animals.The use of medicatedblocks (molasses/urea blocks containinganthelminthics), in combination withgrazing rotation methods, is also beinginvestigated, in order to reduce thenematode egg burden in paddocks.

IMPROVING HOST RESISTANCE

We aim to boost the mucosal immunityof young lambs to the three main gutparasites, Haemonchus contortus,Trichostrongylus (black scour worm) andOstertagia (small brown stomachworm).This will ensure a high level ofimmunity throughout life, boosting theanimals’ health and productivity andreducing the need for drenches.

As poor nutrition is known to reduceimmunity, dietary factors that influenceresistance to nematodes are beingstudied. Sheep selected for resistanceto H. contortus, are also being studied,to see if their immunity is related totheir nutritional intake and/or moreefficient digestion. Methods of deliveringnematode vaccines to mucosal lymphoid tissues in order to promote a more effective immune response arebeing investigated.

Vaccines to ingested plant toxins, suchas those found in rye grass andheliotrope, are being introduced

mucosally. If these vaccines stimulateimmunity, sheep may be able to extractnutritional value from poisonous plants,and assist weed control.

Through the Nemesis program (page 102) and other applied research,farmers can select sheep for nematode resistance.We have also bred our ownlines of sheep, resistant to Haemonchus.Using these ‘demonstration’ flocks, theeffects of host resistance on productivity, scouring, managementpractices, and drenching requirementsare being studied. Results to date showthat sheep selected for resistance toone nematode are generally crossresistant to others, and pass fewernematode eggs in their droppings.Weaner sheep selected for nematoderesistance remain resistant as adults,even during lambing and lactation. Andmodelling predictions show that selection for resistant animals willreduce the need for drenches in 9-13years.This work allows farmers to seethe economic value of becominginvolved in selection programs. Moreinformation on breeding for resistanceis available at: www.csiro.au/nemesis

By studying feed intake, diet selection,nutritional and immunological parameters in resistant and susceptiblelines of sheep, we hope to determinethe mechanisms contributing to nematode resistance.

To understand the genetic basis of hostresistance to nematodes, gene expression in sheep bred for resistanceto Haemonchus or Trichostrongylus(‘high lines’), will be compared withgene expression in sheep sensitive tothese parasites (‘low lines’).We will useour knowledge of genes involved inimmune response to parasites to discover new genes using microarrays.In other experiments, gene expressionin ‘normal’ sheep, which have developedimmunity to these parasites after anumber of infections given under controlled conditions, will be examined.

DISEASE DIAGNOSIS

A chemical field test is being developedto detect the number and species ofnematode eggs in sheep faeces.Thetest will allow farmers to detect thenumber of Haemonchus, Trichostrongylusor Ostertagia eggs in a sample in minutes, allowing decisions aboutwhether a drench is needed, or thetype of drench needed.

A new project aims to develop a realtime PCR test to diagnose the level ofparasitic nematode larvae on pasture.The test would aim to distinguishbetween different nematode speciesand ultimately determine if they carry genes for resistance to different drenches.

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Peter Bradley, coordinator of parasitological services, preparesnematodes for internal and external research clients.

PARASITE MANAGEMENT

Sustainable parasite control requiresthe integration of several nematodecontrol methods, such as drenching,pasture spelling, improved nutrition, andbiological control. However, testing thebenefits of integrated methods is difficult. A model has been developed -for use by agricultural and veterinaryadvisors - which combines informationon climate, farm management and parasite control practices, and thedynamics of different parasite lifecycles. It then predicts the success ofcontrol strategies in terms of wormpopulations, sheep mortalities andselection for drug resistance.

Current development work on‘WormWorld’ (page 101) aims to combine the three separate models foreach of the major nematode species ofeconomic importance to the Australiansheep industry (Haemonchus contortus,Trichostrongylus colubriformis andOstertagia circumcincta), into a singlemodel.The model will be tested underdifferent conditions across Australia.Eventually, the model will be combinedwith information on feed intake andherbage density to provide estimatesof production benefits and play a rolein whole farm management.

BIOCONTROL

With a commercial partner, we aredeveloping a biocontrol agent forworms of sheep and other livestock.Duddingtonia flagrans, a nematode-eating fungus, has beenshown to reduce the number of nematode larvae emerging from thefaeces of young sheep.The fungus canboost sheep productivity throughimproved nematode control and hasno detectable impacts on other soil and pasture organisms. If commercialisation of the fungus provessuccessful, D. flagrans could assist farmers battling the rising tide ofnematodes resistant to anthelminticchemicals. It will also help fulfil anincreasing demand for agricultural commodities free of chemical inputs.

DRUG DELIVERY

Controlled release devices foranthelmintics are being designed, whichremain in the rumen and deliver continuous or pulsed doses of a drug.Such devices will improve the efficacyof dosed drugs, by ensuring optimumdosing conditions are met – directedto the right tissue at the right time, inthe right concentration and chemicalform, for the right period of time. Atthe same time new coatings for drug

tablets are being developed, for fastand slow release.These studies willensure a more responsible and controlled approach to drug administration.

PHYSIOLOGICAL TELEMETRY

Encapsulated electronic devices arebeing developed to provide real-timeinformation on animal health, welfareor location. For example, devices delivered to the rumen could measuretemperature, pressure and pH, anddetect heat stress, bloat or acidosis.Similar devices may be used intravaginally to detect oestrus andimprove the timing of artificial insemination. Spatial devices couldmonitor stock movement or theft,or determine the ‘tilt’ of a bull to trackmating.These platform technologies arebeing developed through Smart Spaces- Emerging Sciences Initiative.

BLOWFLY STRIKE

Fleece rot and blowfly strike are aninterrelated disease complex of sheep.Previous work has shown that sheepthat develop fleece rot are predisposedto blowfly strike. However, animalsresistant to blowfly strike can be selected.Through the Australian Sheep

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Dr Keith Ellis holds one of his encapsulated electronic devices, which could provide real time information on animal health and physiology.

Industry CRC we are investigating thiscapacity for resistance, by taking fleeceout of the equation, and implanting flylarvae on the skin of resistant and sensitive animals. Any differences in theresponse of these sheep will provide astarting point for identifying genesinvolved in resistance, and/or hostdefence mechanisms that could beused to prime immunity.

MASTITIS

Mastitis is an infection of the mammary(udder) tissue in cattle, sheep andgoats, caused by a range of bacteria,yeast and fungi.Through the CRC forInnovative Dairy Products, the hostdefence response to infection is beinginvestigated. Using a microarray ofudder tissue and leucocyte cDNA,gene expression is being studied insheep and cattle with mastitis causedby Staphylococcus aureus or Escherichiacoli. Differences in gene expressionbetween different infections will allowidentification of genes involved in hostresistance. Protective gene expressionprofiles could later become a target forvaccination strategies or immunotherapy.

ANIMAL WELFARE

We expect to see a move towards the accreditation of animal welfare standards in livestock production systems in the future, to satisfy consumer demands and concerns.Scientific validation of what constitutesgood or bad animal welfare is needed,however, to underpin any objectivewelfare assessment.

The stress-responsive systems of sheepand cattle, and their interactions withimmunity and disease, are being studied.By measuring animals’ responses toexperimental and normal productionstressors (such as shearing, live transport and pre-slaughter management), we hope to identifygenes that confer greater adaptabilityand tolerance to production systems.We are also aiming to improve or provide alternatives to managementpractices that are problematic, such as mulesing.

Early research suggests that selection ofcattle with more tolerant temperamentswill lead to improvements in meatquality and tenderness. If a link betweentemperament and key productivitytraits is proven, adoption of temperament testing in geneticimprovement programs should increase.

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At the new Centre for Advanced Reproductive Technologies,researchers led by Dr Jon Hill are developing or improving anumber of advanced reproductive techniques, including cloningand embryo transfer.

PRODUCTIVITY AND ENTERPRISE PRODUCTION

The focus of the Productivity and Enterprise Diversificationsub-program is to increase profitability in the sheep andemerging industries (yabbies, rabbits), by developing improvedbreeding, nutritional and management technologies andimproved strains of animals.

Sub-program LeaderDr Ian PurvisPh (02) 6776 1373Email ian.purvis@csiro.au

BREEDING ULTRAFINESHEEP

The ultrafine wool market (below 16microns) is a small but lucrative market,which is under threat from syntheticfibres. High quality ultrafine wool is currently produced by expensive‘shedded’ sheep systems.The challengeis to produce sheep that are geneticallycapable of growing high quality, ultrafinewool in the paddock. In 1998, theToward 13 Micron (T13) selection program was initiated to develop astrain of Merino with a mean fibrediameter averaging less than 14microns, while maintaining or improvingother wool quality attributes. SelectGene software was used (page 101) toobjectively select the best animals foruse in the breeding program and ourrams regularly win or place in the premier sire evaluation contests. Sixram breeders, who will eventually owna large part of the genetically improvedflock, are funding the project, togetherwith CSIRO.Through these breeders

the delivery of high quality ultrafineMerinos to the industry is guaranteed.

NOVEL FIBRE ATTRIBUTES

To fill new or niche markets, new products will be developed based onthe natural variation in wool fibreattributes, such as crimp, felting andpilling propensity, and pigmentation.Ultrafine wool is another such product,and a project is underway to understand the attributes of ultrafinewool fibres that have a propensity tofelt.This may lead the development ofwool that is naturally resistant to felting(shrink-proofed).

Development of new practical measurement techniques is requiredfor the evaluation of some of thesenovel attributes. For example, we hopeto develop a test that can measure thecylindricity of fibres. More cylindricalfibres can be packed closer together toproduce a thinner yarn and higherquality fabric.

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The genetic improvement of yabbies (Cherax destructor) is enabling the Australianyabby industry to increasingly supply European and Asian markets.

PRECISION WOOL

Through the Australian Sheep IndustryCRC management benchmarks arebeing developed to help producersgrow better quality wool, in terms ofstaple strength and fibre diameter profile (changes to fibre diameter alongthe length of a fibre). As nutrition andgenetics are known to influence thesetraits, management strategies will bedeveloped based on our study of howthese attributes vary between sheepon farms, between farms in regions,and between regions.

Fine wool is sometimes discountedbecause the staple is too long. A short-term solution may be to reduceshearing intervals from once every 12months, to every nine months.Thiscould mean that sheep are shorn inwinter, rather than spring. As staplestrength is generally weaker in winter,this change in shearing interval willalter the point at which the fibresbreak and therefore the staple length.Preliminary studies are underway todetermine if such a change is feasible interms of wool quality, processing andanimal welfare.

MOLECULAR MARKERSFOR GENETIC GAIN

Molecular techniques are being developed to complement traditionalbreeding strategies and identify markersfor genes that have a major influence

on commercially important traits insheep.These markers will be used toidentify genes in young animals, forimproved growth, reproduction or disease resistance, for example.Younganimals with superior genotypes canthen be selected for breeding programsand/or introduced into our advancedreproduction program.

ADVANCED REPRODUCTIVE TECHNOLOGIES

Through the CRC for Innovative DairyProducts we aim to improve cloning(nuclear transfer) techniques in farmanimals. Currently 33% of clones areborn with an abnormality, usually in thelungs or cardiovascular system. As theplacenta is not properly developed inmost of these clones, there is likely tobe a link between the abnormal placenta and abnormal clones.To findout, molecular genetic microarray technology will be used to look atchanges in the expression of genes inthe placenta and other cloned tissues.As we identify critical genes that areincorrectly expressed in the earlystages of the cloning process, we canbegin to rectify the problem. Animproved cloning technique will provide a useful tool for animal production and research.

To efficiently incorporate commerciallyimportant genes – such as those

identified in the molecular markersproject - into livestock, we are developing or improving a number ofother advanced reproductive techniques:• in vitro embryo production• embryo transfer• artificial insemination • gamete cryopreservation, and• cell culture

These techniques will enable farmersto improve their flocks or herds andincrease production. For example, ifeggs and sperm from sexually immatureanimals with superior genotypes arecombined in a test tube (in vitroembryo production), the resultingembryo can be implanted into amature animal to speed genetic gains.

FRESHWATER CRAYFISH

World demand for freshwater crayfishis high and Australian yabbies are of aconsistently higher quality and largersize than their Northern Hemispherecounterparts. Consequently, theAustralian yabby industry has thepotential to expand to supply traditional European crayfish markets,as well as export into Asia wherethere has been a significant increase infreshwater crayfish consumption.Thereis also an opportunity to more efficiently use the water in our farmdams by farming freshwater crayfishmore extensively.

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Meat rabbits are providing a useful means of income diversificationfor farmers in temperate Australia.The Crusader project hasdeveloped improved breeding stock, technologies and manage-ment strategies, to optimise farmers’ productivity and profit.

At the freshwater aquaculture researchfacility in Armidale, a program of genetic improvement of yabbies(Cherax destructor) is underway.Strains of yabbies have been collectedfrom river catchments in easternAustralia, and mated to produce a newstrain with superior traits, includinggreater size and growth rate. Selectedyabbies grow about 24% faster thanunselected animals.We are now lookingat ways to produce single sex and/orsterile yabbies, to prevent matings withwild animals and to allow farmers tocontrol stocking densities in theirponds. Because yabbies shed theirexoskeleton as they grow, internalultraviolet identification tags have beendeveloped.We are now evaluating thefeasibility of growing fish and crayfishtogether, and are developing a cagesystem for the fish, to stop them eatingyoung yabbies.

RABBITS

Meat rabbit farming has the potentialto provide a useful means of incomediversification for farmers in the temperate regions of Australia. OurCrusader Project provides researchand development to support profitableand sustainable Australian meat rabbitproduction.The focus of the project ison identifying and implementing methods to improve key profit drivers.Individual areas of work that have been

identified by industry as a high priority are management of disease,improved survival of kittens andgrowers, and providing optimumnutrition and benchmarking performance.The project is deliveringinformation systems, feed formulations and improved breedingstock to optimise productivity and profit.

The resulting gains in productivityand genetic improvement will ensurea regular supply of product to theprocessing industry, enabling securemarkets to be established. Farmersare now using breeding stock, andtechnology and management strategies developed by theCrusader Project, including estimatedbreeding values, restricted antibioticuse, and improved kit boxes.

Crusader has joined forces with theFrench rabbit breeding group inINRA to explore the opportunitiesof breeding for improved diseaseresistance in rabbits.This has been animportant component of theCrusader research program since itsinception and is of particular interestto the industry in Europe as theywork to reduce reliance on antibiotics for disease control.More information is available atwww.csiro.au/crusader

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INFORMATION SYSTEMS

This sub-program comprises projects that specialise in the useof computer modelling, data management and statistical analysis for research and technology transfer.The projects aim to build business relationships with internal and external customers in the Australian livestock industries.

Sub-program LeaderDr Ian PurvisPh (02) 6776 1373Email ian.purvis@csiro.au

AUSPIG

AUSPIG is a decision support systemthat applies research to pig farm management, to assist users to put inplace more profitable pig managementstrategies.The software looks at factorswhich effect profitability in pig production by accounting for theeffects and interactions of factors suchas pig type, feed, environment, marketprices, labour, capital and otherresources. Developed in collaborationwith Australian Pork Limited, AUSPIG is used in pig enterprises throughout Australia.

WORMWORLD

WormWorld models the epidemiologyof nematodes. It combines informationon climate, farm management, parasitecontrol practices, and the dynamics ofdifferent parasite life cycles, and predictsthe success of control strategies interms of worm populations, sheepmortalities and selection for drugresistance.The model may be incorporated into CSIRO’s integratedfarm management tool, FarmWi$e.

SELECTGENE

This software tool assists fine wool rambreeders to optimise their breedingprograms and improve profitability.Thetool draws on information from theFine Wool Project, supported by TheWoolmark Company and completed in2000.This project determined theinheritance of fine wool traits inMerinos, and generated informationthat enhanced the competitive positionof Australian fine and superfine woolson the global apparel textile market.

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Select Breeding Services helps Merino breeders identify breedingobjectives, while the sire evaluation database maintains a list ofbreeding values for Australian rams.

Using this information, SelectGenehelps ram breeders set breeding goalsand strategies to achieve these.Wehave used the software to breed ourown prize-winning rams through theTowards 13 Micron project (page 98).The tool is available to breedersthrough Select Breeding Services.

SELECT BREEDINGSERVICES

This consultancy business was established in 1998 to transfer genetictechnologies developed by the division,to the fine wool industry.The servicehelps Merino breeders identify abreeding objective, based on heritabletraits for fine wool (fibre diameter, staplelength, staple strength and fleece weight,for example), correlations betweenthese traits, and economic values forwool production, wool quality and processing performance.The serviceworks closely with the Nemesis program to incorporate nematoderesistance into fine wool breedingobjectives.The principles behind ourservice have also been used to designbreeding programs in the forestry andaquaculture industries.

SIRE EVALUATION DATABASE

In collaboration with the AustralianMerino Sire Evaluation Association,CSIRO maintains a database of breedingvalues for Australian rams.Thesebreeding values are determined at 13Central Test Sire Evaluation sites acrossthe country. Ewes are mated to siresinvolved in the program, and thebreeding values of their progeny areevaluated.These values are based oncommercially important traits such asgreasy and clean fleece weights, fibrediameter, staple strength and length,faecal egg count, and body weight.Breeders can then compare siresacross regions and decide which wouldprove most useful in their breedingprograms.The interactive database isavailable at http://mss.anprod.csiro.au

NEMESIS

This technology transfer tool, fundedby Australian Wool Innovation, helpseducate industry about the importanceof breeding Merino sheep for nematode resistance. For breedersusing the service, we estimate breedingvalues for faecal egg counts in youngrams.These values are then used todetermine suitable sires for a breedingprogram. Nemesis works closely withSelect Breeding Services to includenematode resistance traits in fine woolbreeding objectives.

E-COMMERCE

We aim to bring together many of thesoftware tools developed by CSIROLivestock Industries, and build a framework that allows access and subscription to these tools via the internet.

E-SHEEP

Through the Australian Sheep IndustryCRC we are developing technologythat will allow industry to move fromflock management towards individualanimal management.This will involvelinking new or existing hardware products, such as ear tags, readers anddrafting equipment, with softwareproducts such as SelectGene.The project will enable management decisions to be made in near-real time,based on electronic data captured onindividual animals as they move aboutthe paddock.

GENETICS DATABASE

The Information Systems sub-programmanages a database of sheep and cattle breeding experiments from the1940s to the present.The database isan integral component of the dailymanagement and statistical analysis of current genetics experiments.

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The e-sheep project is developing technology that will allow industry to movefrom flock management towards individual animal management.

WOOL SALE DATABASE

This database contains records of allwool sold through the auction system,from the early 1970s to the present.The data is used for research purposes,through an agreement with AuswoolDirect Pty Ltd.

COMPUTER SUPPORT

We provide statistical support to staffon site, and basic and applied researchin bioinformatics and mathematicalgenetics.

The service helps Merino breeders identify a breedingobjective, based on heritable traits for fine wool,correlations between these traits, and economic values for wool production, wool quality and processing performance

“ “

CSIRO Livestock Industries

conducts research projects

through a number of

Cooperative Research

Centres (CRCs)

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CRC for Australian biosecurity 105CRC for Australian sheepindustry 105CRC for cattle and beef quality 105

CRC for innovative dairyproducts 106CRC for plant-basedmanagement of dryland salinity 106CRC for poultry 106CRC for vaccine technology 106

crcinvolvement

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CSIRO Livestock Industries conducts research projectsthrough a number ofCooperative Research Centres (CRCs).

CRC FOR AUSTRALIANBIOSECURITY

Emerging infectious diseases are amajor threat to trade and tourism inAustralia.The new AustralianBiosecurity CRC aims to address thisthreat through research that willenhance our country’s diagnostic andsurveillance capabilities for emerginginfectious diseases. Given that the goalsof the CRC parallel those of CSIROLivestock Industries’ Australian AnimalHealth Laboratory (AAHL) in Geelong,AAHL will play a major role in the CRC.

Projects originating in AAHL’sDiagnostic Sciences and InfectiousDiseases and Food Safety programswill expand and evolve into three mainresearch areas.The first will extend ourdiagnostic and surveillance capabilitieswith known diseases, such as foot andmouth disease and Hendra and Nipahviruses, towards development of laboratory and field-based technology,that will rapidly detect and identify newpathogens before an outbreakbecomes catastrophic. Rather than creating new technologies, those

already on the crest of developmentwill be investigated.These include DNAchips, where thousands of DNA fragments from different pathogens can be arranged on a microchip andhybridised to a sample, and ‘luminex’technology, which allows multiple serological reactions to be performedin one assay. Hand-held tests that canbe used in the field will also be developed, to speed diagnosis and toprovide user-friendly tests for untrainedfield officers in developing countries.This work will tap into platform technology developed by Brisbanebiotechnology company, PanBio.

The second research area for AAHLaims to understand basic biologicalquestions about virus transmission andpathogenesis. Japanese encephalitis,Nipah virus and West Nile virus are onour doorstep, but little is known abouthow these viruses could survive inAustralia and their effect on nativefauna. Monitoring the movement ofanimals that carry such viruses, such asthe flight path of Australian flying foxesthat could carry Nipah virus, or thesusceptibility of Australian avifauna toWest Nile virus, and other ecologicalinformation, will assist our understandingof these diseases and enhance ourcapacity to carry out surveillance andrespond appropriately should the viruses arrive in Australia.

The third main area of researchinvolves development of diagnosticreagents (recombinant antibodies) forthe severe acute respiratory syndrome(SARS) coronavirus, and determininghow the virus kills and where it comes from.

CRC FOR AUSTRALIANSHEEP INDUSTRY

www.sheep.crc.org.au

Through the Australian Sheep IndustryCRC, the Livestock Applications ofBiotechnology program is developing asheep tissue resource bank, identifyinggenes for production traits and developing tests to screen for sheepwith genes for pigmented wool.

The Temperate Livestock Systems e-sheep project, aims to develop precision sheep production technologiesthat will allow industry to move fromflock management towards individualanimal management.

CRC FOR CATTLE ANDBEEF QUALITY

www.beef.crc.org.au

Through the CRC for Cattle and BeefQuality, the Livestock Applications ofBiotechnology program in Brisbane hasdeveloped the GeneSTAR Marblingand GeneSTAR Tenderness commercial

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DNA tests.These tests help beef producers to breed animals containinggenes for desirable fat distribution inmuscle tissue, and meat tenderness,respectively. Other projects includestudying environmental and predisposition factors that influencemarbling in Japanese Black steers,identifying and regulating genes formeat marbling, and developing bovinemicroarrays to identify genes important for beef quality.

CRC FOR INNOVATIVEDAIRY PRODUCTS

www.dairycrc.com

The CRC for Innovative DairyProducts is applying cutting-edgegenetic research to create health-enhancing new milk productsworldwide and essential herd efficiencies.Through the Genes and Proteins sub-program of the Livestock Applicationsof the Biotechnology program, genesinvolved in mastitis resistance are beingsought using microarray technology.

CRC FOR PLANT-BASEDMANAGEMENT OFDRYLAND SALINITY

www.crcsalinity.com

Through an improved understanding ofthe way natural and agricultural ecosystems work, the Salinity CRC willprovide new plant-based land use systems that lessen the economic,environmental and social impacts ofdryland salinity and help to sustainrural communities.The MediterraneanLivestock Systems program, through itsAnimal Production from Saline Landsproject, is involved in a number of projects looking at increasing the feedingvalue of saltland pastures to improveliveweight gains and wool quality and quantity.

CRC FOR POULTRY

The Poultry CRC aims to build a poultryindustry with enhanced bird health,welfare standards and environmentalmanagement, producing high quality'clean and green' products.TheAustralian Animal Health Laboratorywill be a major participant in the CRC,aiming for sustainable production ofchicken meat without reliance onantibiotics by applying new technologies such as genomics andRNA interference.

CRC FOR VACCINETECHNOLOGY

www.crc-vt.qimr.edu.au

The CRC for Vaccine Technology isresponsible for the development andcommercialisation of new vaccines,therapies and platform technologies forthe benefit of human and veterinaryhealth.Through the CRC, the LivestockApplications of Biotechnology Programhas identified proteins that may formthe basis of a tick fever vaccine.

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This list includes books, book chapters and journal articles by staff of CSIRO Livestock Industries for the period 2000-July 2003.It does not include conference or newsletter articles or extension materials.

Abraham, G., Hooper, P.T.,Williamson, M.M. and Daniels, P.W. (2001) Investigations of emerging zoonotic diseases.In: Anthology of Biosafety IV: Issues in Public Health. (Ed. Richmond, J.Y.), 261-279. (American Biological Safety Association:Mundelein, IL).

Abraham, G., Muschialli, J. and Middleton, D. (2002) Animal experimentation in level 4 facilities. In: Anthology of Biosafety V: BSL-4 Laboratories. (Ed. Richmond, J.Y.), 343-359. (American Biological Safety Association: Mundelein, IL.).

Accioly, J.M., Beatty, D.T., Barnes,A.L., Pethick, D.W.,Taylor, E.G.,Tudor, G.D.,White, C.L., Maloney, S.K., McCarthy, M.R.,Pluske, J.R. and Costa, N.D. (2003) Nutrition during live export of cattle. Recent Advances in Animal Nutrition in Australia, 14, 49-56.

Adams, N.R. and Kelly, R.W. (2000) Staple strength. - Overview. Asian-Australasian Journal of Animal Sciences,13, (Suppl. C), 20-21.

Adams, N.R. and Wiese, S.C. (2000) Variability of wool fibre diameter and carcass characteristics in sheep. Proceedings of the Nutrition Society of Australia, 24, 134.

Adams, N.R., Liu, S. and Masters, D.G. (2000) Regulation of protein synthesis for wool growth. In: Ruminant Physiology:Digestion, Metabolism, Growth, and Reproduction. (Ed. Cronjé, P.B.), 255-272. (CABI:Wallingford, UK).

Adams, N.R., Briegel, J.R.,Thompson, M.J. and Sammels, L.M. (2000) Metabolic hormones and tissue concentrations of mRNAfor IGF-I in lines of sheep that differ in their protein synthesis response to feed intake. Journal of Endocrinology,167, 315-320.

Adams, N.R., Hewett, L.J., Schlink,A.C. and Briegel, J.R. (2000) Phenotypic correlates of staple strength. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. A), 289-292.

Adams, N.R., Liu, S.M., Briegel, J.R. and Greeff, J.C. (2000) Protein metabolism in skin and muscle of sheep selected for or against staple strength. Australian Journal of Agricultural Research, 51, 541-546.

Adams, N.R. and Briegel, J.R. (2002) Variation in fibre diameter along the wool staple in sire progeny groups. Proceedings of the Australian Society of Animal Production, 24, 5-8.

Adams, N.R., Briegel, J.R. and Blache, D. (2002) Feed intake, liveweight and wool growth rate in Merino sheep with different responsiveness to low- and high-quality feed. Australian Journal of Experimental Agriculture, 42, 399-405.

Adams, N.R., Briegel, J.R. and Robertson, I.R.D. (2002) Wool characteristics affecting differences among sire progeny groups instaple strength. Proceedings of the Australian Society of Animal Production, 24, 1-4.

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Adams, N.R., Briegel, J.R. and Ward, K.A. (2002) The impact of a transgene for ovine growth hormone on the performance oftwo breeds of sheep. Journal of Animal Science, 80, 2325-2333.

Adams, N.R., Lea, J.M., Briegel, J.R. and Schlink,A.C. (2002) Effect of a 3-day fast on stress hormones in sheep with low or high staple strength. Proceedings of the Australian Society of Animal Production, 24, 9-12.

Adams, N.R. and Cronjé, P.B. (2003) A review of the biology linking fibre diameter with fleece weight, liveweight, and reproduction in Merino sheep. Australian Journal of Agricultural Research, 54, 1-10.

Adelson, D.L., Hollis, D.E. and Brown, G.H. (2002) Wool fibre diameter and follicle density are not specified simultaneously during wool follicle initiation. Australian Journal of Agricultural Research, 53, 1003-1009.

Alitheen, N., McClure, S.J. and McCullagh, P. (2001) Segration of B lymphocytes into stationary apoptotic and migratory proliferating subpopulations in agglomerate culture with ileal epithelium. European Journal of Immunology, 31, 2558-2565.

Alitheen, N., McClure, S. and McCullagh, P. (2003) Development of B cells in the gut-associated lymphoid tissue of mid-gestational fetal lambs. Developmental and Comparative Immunology, 27, 639-646.

Allingham, P.G., Harper, G.S., Hennessy, D.W. and Oddy,V.H. (2001) The influence of pre-weaning nutrition on biochemical and myofibre characteristics of bovine semitendinosus muscle. Australian Journal of Agricultural Research, 52, 891-902.

Altmann, K.G. and Gorman, G.E. (2000) The use of gifts and exchanges in a university library serials collection. Serials Librarian, 39, 23-38.

Anderson, N. (2000) Parasitic infections of sheep and cattle in the winter rainfall regions of Australia. In: Of Vets, Viruses and Vaccines:The Story of CSIRO's Animal Health Research Laboratory, Parkville. (Ed. Butcher, B.W.), 236-246. (CSIRO Publishing: Collingwood,Vic).

Anderton, N.A., Cockrum, P.A., Colegate, S.M., Edgar, J.A. and Flower, K. (2000) New alkaloids from Phalaris spp.: a cause for concern? In: Natural and Selected Synthetic Toxins: Biological Implications. (Eds.Tu, A.T. and Gaffield,W.), 140-153. (American Chemical Society:Washington, D.C.).

Anderton, N.A., Cockrum, P.A., Colegate, S.M., Edgar, J.A. and Flower, K. (2000) Transfer of pyrrolizidine alkaloids into eggs:food safety implications. In: Natural and Selected Synthetic Toxins: Biological Implications. (Eds.Tu, A.T. and Gaffield,W.),118-128. (American Chemical Society:Washington, D.C.).

Andrew, M.E., Morrissy, C.J., Lenghaus, C., Oke, P.G., Sproat, K.W., Hodgson,A.L.M., Johnson, M.A. and Coupar, B.E.H. (2000)Protection of pigs against classical swine fever with DNA-delivered gp55. Vaccine, 18, 1932-1938.

Ariza, F., Harrison, B. and Drinkwater, R.D. (2001) The assignment by linkage mapping of four genes from human chromosome 22 to bovine chromosome 5 and 17. Animal Genetics, 32, 371-374.

Ashes, J.R., Gulati, S.K., Kitessa, S.M., Fleck, E. and Scott,T.W. (2000) Utilisation of rumen protected N-3 fatty acids by ruminants. In: Recent Advances in Animal Nutrition 2000. (Eds. Garnsworthy, P.C. and Wiseman, J.), 129-140. (Nottingham University Press: Nottingham, UK).

Bagust,T.J. (2000) Diseases of poultry. In: Of Vets, Viruses and Vaccines:The Story of CSIRO's Animal Health Research Laboratory,Parkville. (Ed. Butcher, B.W.), 226-235. (CSIRO Publishing: Collingwood,Vic).

Bagust,T.J. (2000) Specialised Laboratory Services - Specified pathogen-free poultry. In: Of Vets, Viruses and Vaccines:The Story of CSIRO's Animal Health Research Laboratory, Parkville. (Ed. Butcher, B.W.), 298-302. (CSIRO Publishing: Collingwood,Vic).

Baker, S.K. (2000) Reducing methane from livestock. In: Greenhouse Gases - Meeting our Bottom Line. 20-21. (Australian Academy of Science: Canberra, ACT).

Baker, S.K., Klein, L. and Williams, J.R. (2000) Selection by sheep grazing a grass-dominant annual pasture. Proceedings of the Nutrition Society of Australia, 24, 75.

Baker, S.K., D'Antuono, M.F., Edwards, N.J., Klein, L. and Williams,A.J. (2000) Variance components in estimation of methane emissions. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. B), 162.

Baker, S.K., Edwards, N.J., Holloway, P.E. and Wright,A.D.G. (2001) Opportunities for manipulation of rumen fermentation.Microbiology Australia, 22, A34.

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Baker, S.K., Henry, D.C., Klein, L., MILls, H.R., Pimm, C.L. and Store, L.A. (2001) Degradation of canola and lupin meals in the rumen. Proceedings of the Nutrition Society of Australia, 25, S37.

Baker, S.K. and Kilminster,T.F. (2002) Methanogens in Zebu-cross cattle, identified presumptively with immunofluorescence microscopy. Proceedings of the Australian Society of Animal Production, 24, 272.

Baker, S.K., Purser, D.B., Barnes, M.J. and Williams, J.R. (2002) Capeweed (Arctotheca calendula), a component of pasture.Proceedings of the Australian Society of Animal Production, 24, 271.

Baker, S.K., Schoep,T., Edwards, N.J. and Wright,A.-D.G. (2002) Methanogens in kangaroos. Reproduction Nutrition Development, 42, (Suppl. 1), S77.

Barendse,W. and Armitage S.M. (2001) The single strand conformational analysis of cattle and human single nucleotide polymorphisms may be biased towards specific sequence motifs that minimize local secondary structure of single strand DNA.Animal Biotechnology, 12, 21-28.

Barker, J.S.F.,Tan, S.G., Moore, S.S., Mukherjee,T.K., Matheson, J.-L. and Selvara, O.S. (2001) Genetic variation within and relationships among populations of Asian goats (Capra bircus). Journal of Animal Breeding and Genetics, 118, 213-233.

Barnes, E.H., Dobson, R.J., Stein, P.A., Le Jambre, L.F. and Lenane, I.J. (2001) Selection of different genotype larvae and adult worms for anthelmintic resistance by persistent and short-acting avermectin/milbemycins. International Journal for Parasitology, 31, 720-727.

Barton, S.A. and Brewer, H.G. (2000) Are skin and follicle characteristics associated with wool quality and production throughout the life of the animal within the CSIRO Fine Wool Project flock? In: Finewool 2000 - Breeding for Customer Needs., Armidale, NSW. (Eds. Swan, A.A. and Piper, L.R.), 53-54. (CSIRO Livestock Industries and The Woolmark Company:Armidale, NSW).

Barton, S.A., Purvis, I.W. and Brewer, H.G. (2001) Are wool follicle characteristics associated with wool quality and productionin hogget and adult sheep? Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 289-292.

Beck, C., Middleton, D., Maclean,A. and Lavelle, R. (2002) Osteochondrosis of the second cervical vertebra of a horse.Equine Veterinary Journal, 34, 210-212.

Beh, K.J., Riffkin, C.D., Davies, K.P., Diienno, K. and Maddox, J.F. (2000) Dinucleotide repeat polymorphism at the ovine McMA7, McMA10, McMA13, McMA16, McMA17, McMA27, McMA29, McMA42, McMA47 and McMA49 loci. Animal Genetics, 31, 228-229

Beh, K.J., Callaghan, M.J., Leish, Z., Hulme, D.J., Lenane, I. and Maddox, J.F. (2001) A genome scan for QTL affecting fleeceand wool traits in Merino sheep. Wool Technology and Sheep Breeding, , 88-97.

Beh, K.J., Hulme, D.J., Callaghan, M.J., Leish, Z., Lenane, I.,Windon, R.G. and Maddox, J. (2002) A whole genome scan for quantitative trait loci affecting resistance to Trichostrongylus colubriformis in sheep. Animal Genetics, 33, 97-106.

Bell,A.M. and Eady, S.J. (2001) Improvements in worm resistance can be passed from studs to client flocks. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 199-202.

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Dynes, R.A. and Henry, D.A. (2002) Biomechanical properties of plant species in animal pastures grazed to different levels of feed on offer. Proceedings of the Australian Society of Animal Production, 24, 292.

Dynes, R.A., Henry, D.A. and Masters, D.G. (2003) Characterising forages for ruminant feeding. Asian-Australasian Journal of Animal Sciences, 16, 116-123.

Eady, S.J. (2001) Sensible approaches to new animal industry development in Australia. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 39-42.

Eady, S.J. and Smith, J.L. (2001) Production costs in sheep bred for worm resistance during Haemonchus challenge and infection. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 191-194.

Eady, S.J.,Woolaston, R.R. and Barger, I.A. (2003) Comparison of genetic and nongenetic strategies for control of gastrointestinal nematodes of sheep. Livestock Production Science, 81, 11-23.

Eaton, B.T. (2001) Introduction to current focus on Hendra and Nipah viruses. Microbes and Infection, 3, 277-278.

Eaton, B.T. (2003) Henipaviruses: transcriptional regulation and cell fusion. In: Emergence and Control of Zoonotic Viral Encephalitis, (Fondation Merieux: Lyon, France).

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Edgar, J.A., Roeder, E. and Molyneux, R.J. (2002) Honey from plants containing pyrrolizidine alkaloids: a potential threat to health. Journal of Agricultural and Food Chemistry, 50, 2719-2730.

Edirisinghe,A., Chapman, G.E. and Louis, J.P. (2001) Radiometric corrections for multispectral airborne video imagery.Photogrammetric Engineering and Remote Sensing, 67, 915-922.

Edirisinghe,A., Chapman, G.E. and Louis, J.P. (2001) A simplified method for retrieval of ground level reflectance of targets from airborne video imagery. International Journal of Remote Sensing, 22, 1127-1141.

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Edwards, N.J. and Baker, S.K. (2000) The population of rumen methanogens in both pen-fed and grazing animals is reduced by2-bromoethanesulphonic acid. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. A), 479.

Edwards, N.J.,Ansotegui, R.P., Leadbetter, E. and Costa, N.D. (2000) Straw supplementation of cattle browsing tagasaste during summer/autumn reduces sand accumulation in the rumen. Asian-Australasian Journal of Animal Sciences, 13,(Suppl. B), 138.

Edwards, N.J., McNeill, D.M.,Allen, G.M. and Oldham, C.M. (2000) Increasing the level of superphosphate application to tagasaste increases cattle performance. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. B), 136.

Edwards, S.F. (2002) The condition of high containment laboratory HEPA filters after 13 years of service. Applied Biosafety:Journal of the American Biological Safety Association, 7, 64-73.

Edwards, S.F., Lamb, B. and Maurer, D. (2002) Design and operation of a high containment sewage treatment facility. In:Anthology of Biosafety V: BSL-4 Laboratories. (Ed. Richmond, J.Y.), 319-342. (American Biological Safety Association:Mundelein, IL.).

Eisemann, C.H.,Wijffels, G.L. and Tellam, R.L. (2001) Secretion of the type 2 peritrophic matrix protein, peritrophin-15, from the cardia. Archives of Insect Biochemistry and Physiology, 47, 76-85.

Ellis, P.M., Daniels, P.W. and Banks, D.J.D. (2000) Japanese encephalitis. Veterinary Clinics of North America: Equine Practice,16, 565-578.

Elvin, C.M., Liyou, N.E., Pearson, R., Kemp, D.H. and Dixon, N.E. (2003) Molecular cloning and expression of the dihydrofolatereductase (DHFR ) gene from adult buffalo fly (Haematobia irritans exigua): effects of antifolates. Insect Molecular Biology,12, 173-183.

Emery, D.L., McClure, S.J. and Davey, R.J. (2000) Protection of Merino lambs against Haemonchus contortus by trickle infection of neonates. Parasitology International, 49, 165-170.

Engelke, C.F., McSweeneyl, C.S.,Vercoe, P.E. and Gregg, K. (2002) MALDI TOF mass spectrometry can be used to identify changes in the protein composition of rumen bacteria:The response of the novel ruminal bacterium lplr3-7 when grown in the presence of diaminopropionic acid. Proceedings of the Australian Society of Animal Production, 24, 61-64.

Faichney, G.J., Gordon, G.L.R. and Welch, R.J. (2000) Effect of free lipid on rumen microbes and fiber digestibility in sheep.Proceedings of the Comparative Nutrition Society, 3, 56-59.

Faichney, G.J., Rintoul,A.J. and Gordon, G.L.R. (2000) Effect of dietary free lipid on anaerobic fungi in the rumen of sheep.Proceedings of the Nutrition Society of Australia, 24, 132.

Faichney, G.J., Gordon, G.L.R.,Welch, R.J. and Rintoul,A.J. (2002) Effect of dietary free lipid on anaerobic fungi and digestion in the rumen of sheep. Australian Journal of Agricultural Research, 53, 519-527.

Farn, J.L., Strugnell, R.A., Hoyne, P.A., Michalski,W.P. and Tennent, J.M. (2001) Molecular characterisation of a secreted enzymewith phospholipase B activity from Moraxella bovis. Journal of Bacteriology, 183, 6717-6720.

Feng, C.G., Britton,W.J., Palendira, U., Groat, N.L., Briscoe, H. and Bean,A.G.D. (2000) Up-regulation of VCAM-1 and differential expansion of beta integrin-expressing T lymphocytes are associated with immunity to pulmonary Mycobacterium tuberculosis infection. Journal of Immunology, 164, 4853-4860.

Field, H., Daniels, P.W., Ong, B.L.,Aziz, J., and Bunning, M. (2002) Manual on the Diagnosis of Nipah Virus Infection in Pigs,(FAO: Bangkok).

Firth, D.J., Jones, R.M., Mcfadyen, L.M., Cook, B.G. and Whalley, R.D.B. (2002) Selection of pasture species for groundcover suited to shade in mature macadamia orchards in subtropical Australia. Tropical Grasslands, 36, 1-12.

Ford, J.T.,Wong, C.W. and Colditz, I.G. (2001) Effects of dietary protein types on immune responses and levels of infection with Eimeria vermiformis in mice. Immunology and Cell Biology, 79, 23-28.

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Franklin, I.,Adelson, D.L., Cam, G.R., Sutton, R. and Van Hest, B. (2000) Biotechnology - genetic engineering and wool production: discovering quality related genes. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 43.

Frisch, J.E., O’Neill, C.J. and Kelly, M.J. (2000) Using genetics to control cattle parasites - the Rockhampton experience.International Journal for Parasitology, 30, 253-264.

Fulloon, O.A., Piper, L.R. and Swan,A.A. (2000) Comparison of subjective assessment and objective measurement for classing fine wool Merinos. In: Finewool 2000 - Breeding for Customer Needs. (Eds. Swan, A.A. and Piper, L.R.), 55-56. (CSIRO Livestock Industries and The Woolmark Company: Armidale, NSW).

Fulloon, O.A., Swan,A.A., Piper, L.R. and Van der Werf, J. (2001) Comparison of subjective assessment and objective measurement for the classing of fine wool Merinos. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 514-516.

Fuqing, Z., Khounsy, S., Nianzu, Z. and Blacksell, S.D. (2000) Considerations regarding the transport of samples and development of diagnostic protocols for the detection of classical swine fever virus under endemic conditions. In: Classical Swine Fever and Emerging Diseases in Southeast Asia. (Ed. Blacksell, S.D.), (ACIAR: Canberra).

Galea, C.A., Dalrymple, B.P., Kuypers, R. and Blakeley, R. (2000) Modification of the substrate specificity of porcine pepsin for the enzymatic production of bovine hide gelatin. Protein Science, 9, 1947-1459.

Gardener, C.J.,Whiteman, L.V. and Jones, R.M. (2001) Patterns of seedling emergence over 5 years from seed of 38 species placed on the soil surface under shade and full sunlight in the seasonally dry tropics. Tropical Grasslands, 35, 218-225.

Garner, M.G., Sawarkar, S.D., Brett, E.K., Edwards, J.R., Kulkarni,V.B., Boyle, D.B. and Singh, S.N. (2000) The extent and impactof sheep pox and goat pox in the State of Maharastra, India. Tropical Animal Health and Production, 32, 205-223.

Gazzola, C., Hill, R.A., O’Neill, C.J., Jeffery, M.R., Reid, D.A.,White, D.H. and Burns, B.M. (2000) Scrotal circumference at puberty and its relationship to testes mass in three genotypes of beef cattle. Asian-Australasian Journal of Animal Sciences,13, (Suppl. B), 146.

Gazzola, C. and Spiers,W.G. (2002) Effects of the alpha(2)-adrenoceptor agonist, guanfacine, on growth rate, glucose,corticosterone, insulin and energy partitioning in rats. Animal Science, 74, 455-459.

Gill, H.S.,Altmann, K.G., Cross, M.L. and Husband,A.J. (2000) Induction of T helper 1- and T helper 2-type immune responsesduring Haemonchus contortus infection in sheep. Immunology, 99, 458-463.

Gleeson, L.J. (2002) A review of the status of FMD in Southeast Asia and approaches to control and eradication. Revue Scientifique et Technique de l’Office International des Epizooties, 21, 465-472.

Gobius, K.S., Xue, G.P.,Aylward, J.H., Dalrymple, B.P., Swadling,Y.J., McSweeney, C.S. and Krause, D.O. (2002) Transformation and expression of an anaerobic fungal xylanase in several strains of the rumen bacterium Butyrivibrio fibrisolvens. Journal of Applied Microbiology, 93, 122-133.

Gordon, G.L.R., Phillips, M.W., Rintoul,A.J. and White, S.W. (2000) Increased intake of fibrous feed by sheep orally dosed witha culture of an elite non-indigenous anaerobic gut fungus. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 143.

Gordon, G.L.R., Phillips, M.W.,White, S.W., Rintoul,A.R. and Mitchell, P.A. (2001) Can anaerobic fungi be manipulated in the sheep rumen to improve the utilisation of poor-quality feed? Recent Advances in Animal Nutrition in Australia, 13, 43-48.

Gough, J.M.,Akhurst, R.J., Ellar, D.J., Kemp, D.H. and Wijffels, G.L. (2002) New isolates of Bacillus thuringiensis for control of livestock ectoparasites. Biological Control, 23, 179-189.

Gould,A.R., Kattenbelt, J.A., Selleck, P.W., Hansson, E., Della-Porta,A.J. and Westbury, H.A. (2001) Virulent Newcastle diseasein Australia: molecular epidemiological analysis of viruses isolated prior to and during the outbreaks of 1998-2000. Virus Research, 77, 51-60.

Gray, G.D.,Ancheta, P.A.,Arguzon, J.A., Barcelo, P.M., Casis, C., Cerbito,W.A., Dobson, R.J., Dumilon, R., Cruz, E.M., Knox,M.R., Le Jambre, L.F., Mateo, K.S., Suba, M.S. and Villar, E.C. (2000) Better worm control for goats and sheep in the Philippines. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. A), 548.

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Greeff, J.C. and Schlink,A.C. (2001) The inheritance of felting of Merino wool. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 497-500.

Greeff, J.C. and Schlink,A.C. (2002) The inheritance of felting of Merino wool. Wool Technology and Sheep Breeding, 50, 6-10.

Greenwood, P.L., Hearnshaw, H., Hennessy, D.W. and Harper, G.S. (2001) Early-life nutrition and growth and development of beef cattle. Australian Rural Science Annual, 12, 62-63.

Greenwood, P.L., Hearnshaw, H., Hennessy, D.W.,Thompson, J.M. and Harper, G.S. (2001) Body and carcass characteristics of newborn Piedmontese x Hereford and Wagyu x Hereford female calves. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 473-476.

Greenwood, P.L., Hearnshaw, H., Hennessy, D.W.,Thompson, J.M. and Harper, G.S. (2001) Body composition of Piedmontese x Hereford and Wagyu x Hereford newborn calves. Journal of Animal Science, 79, (Suppl. 1), 434.

Greenwood, P.L., Newby, K., Hearnshaw, H., Hennessy, D.W.,Thompson, J.M. and Harper, G.S. (2002) Nutrition of Hereford cows from ~100 days of gestation to near term influences body and carcass characteristics of Piedmontese and Wagyu sired newborn calves. Proceedings of the Australian Society of Animal Production, 24, 305.

Greenwood, P.L.,Wolcott, M., Hearnshaw, H., Hennessy, D.W., Morris, S.G. and Harper, G.S. (2002) Fetal growth capacity influences nutritional status of Hereford cows during pregnancy. Proceedings of the Australian Society of Animal Production, 24, 304.

Gulati, S.K.,Ashes, J.R. and Scott,T.W. (2000) Healthier butter fat spreads better. Feed Mix, 8, (6), 20-22.

Gulati, S.K., Kitessa, S.M.,Ashes, J.R., Fleck, E., Byers, E.B., Byers,Y.G. and Scott,T.W. (2000) Protection of conjugated linoleic acids from ruminal hydrogenation and their incorporation into milk fat. Animal Feed Science and Technology, 86, 139-148.

Gulati, S.K., Kitessa, S.M.,Ashes, J.R., Simos, G.C.,Wynn, P.C., Fleck, E. and Scott,T.W. (2000) Designing milk fat for the new millennium by dietary strategies. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. A), 538-541.

Gulati, S.K., Kitessa, S.M. and Scott,T.W. (2001) Manipulating milk on farm by supplementary feeding for better human nutrition. Australian Journal of Dairy Technology, 56, 150.

Gulati, S.K., McGrath, S.,Wynn, P.C. and Scott,T.W. (2001) Rumen protected conjugated linoleic acids: effects on milk composition in dairy cows. Proceedings of the Nutrition Society of Australia, 25, S85.

Gulati, S.K., May, C.,Wynn, P.C. and Scott,T.W. (2002) Milk fat enriched in n-3 fatty acids. Animal Feed Science and Technology, 98, 143-152.

Gulati, S.K., McGrath, S.,Wynn, P.C. and Scott,T.W. (2003) Preliminary results on the relative incorporation of docosahexaenoic and eicosapentaenoic acids into cows milk from two types of rumen protected fish oil.International Dairy Journal, 13, 339-343.

Hacker, J.B. and Waite, R.B. (2001) Selecting buffel grass (Cenchrus ciliaris) with improved spring yield in subtropical Australia.Tropcial Grasslands, 35, 205-210.

Hacker, J.B.,Wen, S.L.,Ying, Z.Y. and Pengelly, B.C. (2001) Selecting Chamaecrista spp. for soil stabilisation and forage in southern China. Tropical Grasslands, 35, 96-113.

Hammond, J.M., McCoy, R.J., Jansen, E.J., Morrissy, C.J., Hodgson,A.L.M. and Johnson, M.A. (2000) Vaccination with a single dose of a recombinant porcine adenovirus expressing the classical swine fever virus gp55 (E2) gene protects pigs against classical swine fever. Vaccine, 18, 1040-1050.

Hammond, J.M., Jansen, E.S., Morrissy, C.M.,Williamson, M., Hodgson,A.L.M. and Johnson, M.A. (2001) Oral and sub-cutaneous vaccination of commercial pigs with a recombinant porcine adenovirus expressing the classical swinefever virus gp55 gene. Archives of Virology, 146, 1787-1793.

Hammond, J.M., Jansen, E.S., Morrissy, C.J., van der Heide, B., Goff,W.V.,Williamson, M.M., Hooper, P.T., Babiuk, L.A.,Tikoo,S.K. and Johnson, M.A. (2001) Vaccination of pigs with a recombinant porcine adenovirus expressing the gD gene from pseudorabies virus. Vaccine, 19, 3752-3758.

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Hammond, J.M. (2002) Porcine adenovirus. In: Animal Health and Production Compendium. (CABI:Wallingford, UK).

Hammond, J.M., Jansen, E.S., Hodgson,A.L.M. and Johnson, M.A. (2002) Development of porcine adenovurus as a vaccine andtherapeutic delivery vector for swine. In: The World of Microbes 29-V-198 p.67. (EDK Medical and Scientific International Publisher : Paris).

Hammond, S.A.,Tsonis, C.G., Sellins, K., Rushlow, K., Scharton-Kersten,T., Colditz, I.G. and Glenn, G.M. (2000) Transcutaneous immunization of domestic animals: opportunities and challenges. Advanced Drug Delivery Reviews, 43, 45-55.

Hanbury, C.D.,White, C.L., Mullan, B.P. and Siddique, K.H.M. (2000) A review of the potential of Lathyrus sativus L. and L. cicera L. grain for use as animal feed. Animal Feed Science and Technology, 87, 1-27.

Hansen, R.W. (2000) Starting out - developing an occupational health, safety and environment role in a medium-sized enterprise. In: Towards a Safe and Civil Society: Occupational and Environmental Health and Safety in History,Theory and Practice. (Ed. Eddington, I.), 78-83. (International Commission on Occupational Health:Toowoomba, Qld).

Harper, G.S.,Allingham, P.G. and Landsberg, M.J. (2000) Age and gender effects on two biochemical markers of muscle development in cattle. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. B), 318-321.

Harper, G.S., Le Feuvre, R.P. and Oddy,V.H. (2000) Liveweight data as predictors of objective measures of meat quality.Asian-Australasian Journal of Animal Sciences, 13, (Suppl. B), 115-118.

Hatcher, S., Lightfoot, R.J. and Purvis, I.W. (2000) Mating Awassi rams to Merino ewes causes an initial level of fibre contamination which decreases to an insignificant level eight weeks post-mating. Australian Journal of Experimental Agriculture, 40, 363-369.

Hatcher, S., Lightfoot, R.J. and Purvis, I.W. (2000) Previous Awassi grazing on a paddock or grazing in the same or adjoining paddock, does not cause fibre contamination of fleeces of Merino sheep. Australian Journal of Experimental Agriculture,40, 379-385.

Hatcher, S., Lightfoot, R.J. and Purvis, I.W. (2000) Transfer of contaminant lamb fibres to their dam's fleece, and loss within four weeks of weaning. Australian Journal of Experimental Agriculture, 40, 371-378.

Hayes, B., Shepherd, R.K. and Newman, S. (2002) Look ahead mate selection schemes for multi-breed beef populations.Animal Science, 74, 13-23.

Hearnshaw, H., Hennessy, D.W., Greenwood, P.L., Harper, G.S. and Morris, S. (2001) Gestation length, birth traits and preweaning growth of Wagyu-, Piedmontese- and Angus-sired calves. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 337-340.

Hennessy, D.R., Page, S.W. and Gottschall, D. (2000) The behaviour of doramectin in the gastrointestinal tract, its secretion in bile and pharmacokinetic disposition in the peripheral circulation after oral and intravenous administration to sheep.Journal of Veterinary Pharmacology and Therapeutics, 23, 203-213.

Hennessy, D.R., Praslicka, J. and Bjorn, H. (2000) The disposition of pyrantel in the gastrointestinal tract and effect of digesta flow rate on the kinetic behaviour of pyrantel in the pig. Veterinary Parasitology, 92, 277-285.

Hennessy, D.R. and Alvinerie, M.R. (2002) Pharmacokinetics of the macrocyclic lactones: Conventional wisdom and new paradigms. In: Macrocyclic Lactones in Antiparasitic Therapy. (Ed.Vercruysse, J. and Rew, R.S.), 97-123. (CABI:Wallingford, UK).

Hennessy, D.W., Morris, S.G. and Allingham, P.G. (2001) Improving the pre-weaning nutrition of calves by supplementation of the cow and/or the calf while grazing low quality pastures - 2. Calf growth, carcass yield and eating quality. Australian Journal of Agricultural Research, 41, 715-724.

Hennessy, D.W., Hearnshaw, H., Greenwood, P.L., Harper, G.S. and Morris, S.G. (2002) The effects of low or high quality pastures on the live weight of cows at calving and on the birth weight of calves sired by Wagyu or Piedmontese.Proceedings of the Australian Society of Animal Production, 24, 311.

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Henry, D.A., Baker, S.K. and Dynes, R.A. (2000) Additional characters of Australian hays measured using near infrared reflectance spectroscopy. Asian-Australasian Journal of Animal Sciences , 13, (Suppl. B), 161.

Henry, D.A., Dynes, R.A. and Hulm, E.L. (2000) Carbohydrate content of hays changes with time of cutting and nitrogen applications. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. B), 186.

Henry, D.A., Moore, G. and Dynes, R.A. (2002) Variability in nutritive value allows better selection of perennial pasture legumes. Proceedings of the Australian Society of Animal Production, 24, 312.

Henry, D.A., Edirisinghe,A., Donald, G. and Hill, M. (2002) Monitoring pastures using satellite remote sensing. Proceedings of the Australian Society of Animal Production, 24, 81-84.

Henryon, M. and Purvis, I.W. (2000) Eggs and hatchlings of the freshwater crayfish, marron (Cherax tenuimanus), can be successfully incubated artificially. Aquaculture, 184, 247-254.

Henshall, J.M., Burrow, H.M. and Tier, B. (2001) Segregation analysis for major genes affecting adaptive traits in cattle grazed in the tropics. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 167-170.

Henshall, J.M.,Tier, B. and Kerr, R.J. (2001) Estimating genotypes with independently sampled descent graphs. Genetical Research, 78, 281-288.

Henshall, J.M.,Tier, B. and Kerr, R.J. (2001) Marker assisted segregation analysis in complex pedigrees. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 301-304.

Heriveau, C., Dimier-Poisson, I., Lowenthal, J.W., Naciri, M. and Quere, P. (2000) Inhibition of Eimeria tenella replication after recombinant IFN-gamma activation in chicken macrophages, fibroblasts and epithelial cells. Journal of Parasitology, 92, 37-49.

Hill, J., Fisher, P., Linke, R., Cain, D., Roman, H. and Schlafer, D. (2003) Trophoblast MHC-1 expression and apoptosis in bovine nuclear transfer embryos. Theriogenology, 59, 258.

Hill, M.J. (2000) Applications for spatial data in grassland monitoring and management. Spatial Information for Land Use Management,113-128.

Hill, M.J., Donald, G.E., Donnelly, J.R. and Moore,A.D. (2000) Integrating spatial data with a grazing system model: assessing variability of pasture and animal production at a regional scale. Asian-Australasian Journal of Animal Sciences, 13,(Suppl. C), 128-131.

Hill, M.J. and Donald, G.E. (2003) Estimating spatio-temporal patterns of agricultural productivity in fragmented landscapes using AVHRR NDVI time series. Remote Sensing of Environment, 84, 367-384.

Hilton, L.S., Bean,A.G.D., Kimpton,W.G. and Lowenthal, J.W. (2002) Interleukin-2 directly induces activation and proliferation of chicken T cells in vivo. Journal of Interferon and Cytokine Research, 22, 755-763.

Hilton, L.S., Bean,A.G.D. and Lowenthal, J.W. (2002) The emerging role of avian cytokines as immunotherapeutics and vaccineadjuvants. Veterinary Immunology and Immunopathology, 85, 119-128.

Hilton, L.S., Bean,A.G.D. and Lowenthal, J.W. (2002) Recent advances in avian cytokines. Veterinary Immunology and Immunopathology, 85, 119-128.

Hodgson,A.L.M. (2000) Vaccine Vectors - Bacterial Vaccine Vectors. In: Of Vets, Viruses and Vaccines:The Story of CSIRO's Animal Health Research Laboratory, Parkville. (Ed. Butcher, B.W.), 286-289. (CSIRO Publishing: Collingwood,Vic).

Holloway, P.E. and Baker, S.K. (2000) Development of antibodies to rumen methanogens in sheep. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 264.

Holloway, P.E. and Baker, S.K. (2002) Natural immune responses in sheep to methanogenic archaea. Reproduction Nutrition Development, 42, (Suppl. 1), S13-14.

Hooper, P.T. (2000) New fruit bat viruses affecting horses, pigs, and humans. In: Emerging Diseases of Animals. (Eds. Brown, C.and Bolin, C.), 85-99. (ASM Press:Washington, DC).

Hooper, P.T. (2001) Identification and eradication of Menangle virus from pigs. Australian Veterinary Journal, 79, 190-191.

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Hooper, P.T., Gould,A.R., Hyatt,A.D., Braun, M.A., Kattenbelt, J.A., Hengstberger, S.G. and Westbury, H.A. (2000) Identification and molecular characterisation of Hendra virus in a horse in Queensland. Australian Veterinary Journal,78, 281-282.

Hooper, P.T. and Williamson, M.M. (2000) Hendra and Nipah virus infections. Veterinary Clinics of North America: Equine Practice, 16, 597-603, xi.

Hooper, P.T., Zaki, S.R., Daniels, P.W. and Middleton, D. (2001) Comparative pathology of the diseases caused by Hendra and Nipah viruses. Microbes and Infection, 3, 315-322.

Huang,A., Scougall, C.A., Lowenthal, J.W., JILbert,A.R. and Kotlarski, I. (2001) Structural and functional homology between duck and chicken interferon-gamma. Developmental and Comparative Immunology, 25, 55-68.

Hung, B.S.,Wang, X.Q., Cam, G.R. and Rothnagel, J.A. (2001) Characterization of mouse Frizzled-3 expression in hair follicle development and identification of the human homolog in keratinocytes. Journal of Investigative Dermatology, 116, 940-946.

Hunter, R.A. (2000) High molasses diets for intensive finishing of steers. Asian-Australasian Journal of Animal Sciences, 13,(Suppl. B), 112.

Hunter, R.A., Magner,T. and Allingham, P.G. (2000) Sustained growth promotion, carcass characteristics, and meat quality ofsteers treated with oestradiol-17b. Australian Journal of Agricultural Research, 51, 133-138.

Hunter, R.A., Burrow, H.M. and McCrabb, G.J. (2001) Sustained growth promotion, carcass and meat quality of steers slaughtered at three liveweights. Australian Journal of Experimental Agriculture, 41, 1033-1040.

Hunter, R.A., Day,A. and Blakeley, S.K. (2002) Role of high molasses diets in the live exports supply chain. Proceedings of the Australian Society of Animal Production, 24, 109-112.

Hunter, R.A., Harper, G.S. and McCrabb, G.J. (2002) The effect of ingestion of coal mine pit water on the productivity of pregnant and lactating beef cows. Proceedings of the Australian Society of Animal Production, 24, 105-108.

Hyatt,A.D., Gould,A.R., Zupanovic, Z., Cunningham,A.A., Hengstberger, S.,Whittington, R.J., Kattenbelt, J.A. and Coupar,B.E.H. (2000) Comparative studies of piscine and amphibian iridoviruses. Archives of Virology, 145, 301-331.

Hyatt,A.D., Zaki, S.R., Goldsmith, C.S.,Wise,T.G. and Hengstberger, S.G. (2001) Ultrastructure of Hendra virus and Nipah virus within cultured cells and host animals. Microbes and Infection, 3, 297-306.

Hyatt,A.D.,Williamson, M.M., Coupar, B.E.H., Middleton, D., Hengstberger, S.G. , Gould,A.R., Selleck, P.,Wise,T.G.,Kattenbelt, J., Cunningham,A.A. and Lee, J. (2002) First identification of a ranavirus from the green tree python (Chondrapython virdis). Journal of Wildlife Diseases, 38, 239-252.

Hynd, P.I. and Masters, D.G. (2002) Nutrition and wool growth. In: Sheep Nutrition. (Eds. Freer, M. and Dove, H.), 165-188.(CSIRO Publishing: Collingwood,Vic).

Ignjatovic, J. and Sapats, S.I. (2000) Avian infectious bronchitis virus. Revue Scientifique et Technique de l'Office International des Epizooties, 19, 493-508.

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Ignjatovic, J. and Sapats, S. (2002 ) Confirmation of the existence of two distinct genetic groups of infectious bursal disease virus in Australia. Australian Veterinary Journal, 80, 689-694.

Ignjatovic, J., Reece, R. and Ashton, F. (2002) Susceptibility of three genetic lines of chicks to infection with a nephropathogenicT strain of avian infectious bronchitis virus. Journal of Comparative Pathology, 128, 92-98.

Ignjatovic, J., Sapats, S. and Gould, G. (2002) Detection of vvIBDV strains and Australian variants in poultry. Poultry Digest,17, 14-15.

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Ingham,A., Sproat, K.,Tizard, M. and Moore, R. (2001) Recombinant antimicrobial peptides active against veterinary pathogens. Microbiology Australia, 22, A96.

Ingham,A., Zhang,Y. and Prideaux, C. (2002) Attenuation of Actinobacillus pleuropneumoniae by inactivation of aroQ.Veterinary Microbiology, 84, 263-273.

Ingham,A., Ford, M., Moore, R.J. and Tizard, M. (2003) The bacteriocin, Pisciolin 126, retains anti-listerial activity in vivo. Journal of Antimicrobial Chemotherapy, 51, 1365-1371.

Islam.A.F.M.F.,Wong, C.W.,Walkden-Brown, S.W., Colditz, I.G.,Arzey, K.E. and Groves, P.J. (2002) Immunosuppressive effects of Marek's disease virus (MDV) and herpesvirus of turkeys (HVT) in broiler chickens and the protective effect of HVT vaccination against MDV challenge. Avian Pathology, 31, 449-461.

Jacob, R., Pethick, D.W., Masters, D.G. and Milton, J. (2001) Changes in muscle glycogen in sheep with low levels of basal glycogen fed two levels of metabolisable energy. Recent Advances in Animal Nutrition in Australia, 13, 8A.

Jerry, D.R. (2000) Smart farming - animals. Growing better yabbies. In: The Biotechnology Revolution. 14. (University of Technology Sydney: Sydney, NSW).

Jerry D.R. (2001) Electrical stimulation of spermatophore extrusion in the freshwater yabby (Cherax destructor).Aquaculture, , 317-322.

Jerry, D.R., Elphinstone, M.S. and Baverstock, P.R. (2001) Phylogenetic relationships of Australian members of the family Percichthyidae inferred from mitochondrial 12S rRNA sequence data. Molecular Phylogenetics and Evolution, 18, 335-347.

Jerry, D.R., Piper, L.R. and Purvis, I.W. (2001) Differences in growth parameters among populations of the yabby Cherax destructor (Clark). Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 51-54.

Jerry, D.R., Purvis, I.W. and Piper, L.R. (2001) Opportunities for genetic improvement in crustacean species. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 55-59.

Jerry, D.R., Stewart,T., Purvis, I.W. and Piper, L.R. (2001) Evaluation of visual implant elastomer and alphanumeric internal tags as a method to identify juveniles of the freshwater crayfish, Cherax destructor. Aquaculture, 193, 149-154.

Jerry, D.R., Purvis, I.W. and Piper, L.R. (2002) Genetic differences in growth among wild populations of yabby, Cherax destructor (Clark). Aquaculture Research, 33, 917-923.

Jitradakdee, S., Unajak, S., SittidILokratna, N., Hodgson, R.A.J., Cowley, J.A.,Walker, P.J., Panyim, S. and Boonsaeng,V. (2003) Identification and analysis of Gp116 and Gp64 structural glycoproteins of Yellow Head Nidovirus of Penaeus monodonshrimp. Journal of General Virology, 84, 863-873.

Johara, M.Y., Field, H.,Azmin Mohd Rashdi,A.M., Morrissy, C., van der Heide, B., Rota, P.,Azri bin Adzhar,White, J., Daniels,P.W., Jamaluddin,A. and Ksiazek,T. (2001) Nipah virus infection in bats (order Chiroptera) in peninsular Malaysia. Emerging Infectious Diseases, 7, 439-441.

Johnson, M.A., Pooley, C. and Lowenthal, J.W. (2000) Delivery of avian cytokines by adenovirus vectors. Developmental and Comparative Immunology, 24, 354.

Johnson, M.A., Pooley, C., Ignjatovic, J. and Tyack, S.G. (2003) A recombinant fowl adenovirus expressing the S1 gene of infectious bronchitis virus protects against challenge with infectious bronchitis virus. Vaccine, 21, 2730-2736.

Johnston, D.J., Henshall, J.M. and Tier, B. (2001) Estimate of the genetic correlation between calving success and days to calving in Angus females. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 353-356.

Johnston, D.J., Herd, R.M., Reverter,A. and Oddy,V.H. (2001) Heritability of IGF-1 in beef cattle and in association with growthand carcase traits. Proceedings of the Association for Advancement of Animal Breeding and Genetics, 14, 163-164.

Johnston, D.J., Herd, R.M., Reverter,A. and Oddy,V.H. (2001) Use of the two-allele model in half-sib designs. Proceedings of the Association for Advancement of Animal Breeding and Genetics, 14, 409-412.

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Johnston, D.J., Reverter,A., Burrow, H.M., Oddy,V.H. and Robinson, D.L. (2003) Genetic and phenotypic characterisation of animal, carcass, and meat quality traits from temperate and tropically adapted beef breeds. 4. Correlations among animal,carcass and meat quality traits. Australian Journal of Agricultural Research, 54, 107-118.

Johnston, D.J., Reverter,A., Ferguson, D.M.,Thompson, J.M. and Burrow, H.M. (2003) Genetic and phenotypic characterisationof animal, carcass, and meat quality traits from temperate and tropically adapted beef breeds. 3. Meat quality traits.Australian Journal of Agricultural Research, 54, 135-147.

Jones, R.M. (2001) Evaluation of legumes and grasses in coastal south-east Queensland. Tropical Grasslands, 35, 85-95.

Jung, H.G. and Baker, S.K. (2002) Particle size distribution in feeds and boli. Journal of Animal Science, 80, (Suppl. 1), 361.

Kahn, L.P., Leng, R.A. and Piper, L.R. (2000) Rumen microbial yield from sheep genetically different for fleece weight.Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 137.

Kahn, L.P., Knox, M.R.,Walkden-Brown, S.W. and Lea, J.M. (2001) Enhancing life-long resistance to nematode parasites:possibilities from nutritional supplementation of recently weaned sheep. Recent Advances in Animal Nutrition in Australia,13, 87-95.

Kahn, L.P.,Walkden-Brown, S.W., Lea, J.M. and McClure, S.J. (2002) Protein supplementation but not oral vaccination with infective larvae accelerates the development of resistance to Trichostrongylus colubriformis of grazing weaners.Proceedings of the Australian Society of Animal Production, 24, 113-116.

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Kahn, L.P., Knox, M.R.,Walkden-Brown, S.W. and Lea, J.M. (2003) Regulation of the resistance to nematode parasites of single-and twin-bearing Merino ewes through nutrition and genetic selection. Veterinary Parasitology, 114, 15-31.

Kamath,A.T., Groat, N.L., Bean,A.G.D. and Britton,W.J. (2000) Protective effect of DNA immunization against mycobacterial infection is associated with the early emergence of interferon-gamma (IFN-gamma)-secreting lymphocytes. Clinical and Experimental Immunology, 120, 476-482.

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Kennedy, P.M. and Lowry, J.B. (2002) Do tree leaves promote digestion of grass by cattle? Proceedings of the Australian Society of Animal Production, 24, 121-124.

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Kirkland, P.D., Daniels, P.W., Nor, M.N.M., Love, R.J., Philbey,A.W. and Ross,A.D. (2002) Menangle and Nipah virus infections of pigs. Veterinary Clinics of North America - Food Animal Practice, 18, 557-571.

Kitessa, S.M., Gulati, S.K.,Ashes, J.R., Scott,T.W., Fleck, E. and Bode, O. (2000) Fatty acid composition of meat from lambs fedprotected tuna oil supplements. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 201.

Kitessa, S.M. and Nicol,A.M. (2001) The effect of continuous or rotational stocking on the intake and live-weight gain of cattleco-grazing with sheep on temperate pastures. Animal Science, 72, 199-208.

Kitessa, S.M., Gulati, S.K.,Ashes, J.R., Scott,T.W. and Fleck, E. (2001) Effect of feeding tuna oil supplement protected against hydrogenation in the rumen on growth and n-3 fatty acid content of lamb fat and muscle. Australian Journal of Agricultural Research, 52, 433-437.

Kitessa, S.M., Simos, G.C., Gulati, S.K., Scott,T.W. and Wynn, P.C. (2001) Healthier milkfat from n-3 enriched milk. Australian Journal of Dairy Technology, 56, 96.

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Kitessa, S.M., Gulati, S.K.,Ashes, J.R., Fleck, E., Scott,T.W. and Nichols, P.D. (2001) Utilisation of fish oil in ruminants - II.Transfer of fish oil fatty acids into goats' milk. Animal Feed Science and Technology, 89, 201-208.

Kitessa, S.M. and Gulati, S.K. (2002) Manipulation of the fatty acid profiles of meat and milk using protected lipid supplements - a brief review. Proceedings of the Australian Society of Animal Production, 24, 125-128.

Knox, M.R. (2000) Nutritional approaches to nematode parasite control in sheep. Feed Mix, 8, 12-15.

Knox, M.R. and Faedo, M. (2001) Biological control of field infections of nematode parasites of young sheep with Duddingtonia flagrans and effects of spore intake on efficacy. Veterinary Parasitology, 101, 155-160.

Knox, M.R. (2002) Effectiveness of copper oxide wire particles for Haemonchus contortus control in sheep. Australian Veterinary Journal, 80, 224-227.

Knox, M.R., Josh, P.F. and Anderson, L.J. (2002) Deployment of Duddingtonia flagrans in an improved pasture system: dispersal,persistence and effects on free-living soil nematodes and microarthropods. Biological Control, 24, 178-182.

Kongsuwan, K., Dalrymple, B.P.,Wijffels, G.L. and Jennings, P.A. (2002) Cellular localisation of the clamp protein during DNA replication. FEMS Microbiology Letters, 216, 255-262.

Kongsuwan, K. and Jennings, P.A. (2001) Biological function and cellular localisation of GFP-ß fusion protein in Escherichia coli.Microbiology Australia, 22, A120.

Kotze,A.C. (2000) Oxidase activities in macrocyclic-resistant and -susceptible Haemonchus contortus. Journal of Parasitology,86, 873-876.

Kotze,A.C. and McClure, S.J. (2001) Haemonchus contortus utilises catalase in defence against exogenous hydrogen peroxide in vitro. International Journal for Parasitology, 31, 1563-1571.

Kotze,A.C. and Sales, N. (2001) Inheritance of diflubenzuron resistance and monooxygenase activities in a laboratory-selectedstrain of Lucilia cuprina (Diptera: Calliphoridae). Journal of Economic Entomology, 94, 1243-1248.

Kotze,A.C., Dobson, R.J.,Tyrrell, K.L. and Stein, P.A. (2002) High-level ivermectin resistance in a field isolate of Haemonchus contortus associated with a low level of resistance in a larval stage: implications for resistance. Veterinary Parasitology,108, 255-263.

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Krause, D.O., Smith,W.J. and McSweeney, C.S. (2001) Extraction of microbial DNA from rumen contents containing plant tannins. Biotechniques, 31, 294-298.

Krause, D.O., Bunch, R.J., Conlan, L.L., Kennedy, P.M., Smith,W.J., Mackie, R.I. and McSweeney, C.S. (2001) Repeated ruminal dosing of Ruminococcus spp. does not result in persistence, but changes in other microbial populations occur that can be measured with quantitative 16S-rRNA-based probes. Microbiology-SGM, 147, 1719-1729.

Krause, D.O., Bunch, R.J., Dalrymple, B.P., Gobius, K.S., Smith,W.J.M., Xue, G.P. and McSweeney, C.S. (2001) Expression of a modified Neocallimastix patriciarum xylanase in Butyrivibrio fibrisolvens digests more fibre but cannot effectively compete with highly fibrolytic bacteria in the rumen. Journal of Applied Microbiology, 90, (3), 388-96.

Krause, D.O., Smith,W.J.M., Conlan, L.L., Gough, J.M.,Williamson, M.A. and McSweeney, C.S. (2003) Diet influences the ecology of lactic acid bacteria and Escherichia coli along the digestive tract of cattle: neural networks and 16S rDNA.Microbiological-SGM, 149, 57-65.

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Lamb, P.R., Purvis, I.W. and Robinson, G.A. (2000) Genetic improvement of wool quality processing implications of finer,longer, lower crimp wool. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 16-19.

Lanigan, M.D., Kalman, K., Lefievre,Y., Pennington, M.W. and Chy, K.G. (2002) Mutating a critical lysine in ShK toxin alters its binding configuration in the pore-vestibule region of the voltage-gated potassium channel, Kv1.3. Biochemistry, 41,11963-11971.

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Le Blanc Smith, P.M., Nguyen, S. and Edwards, S.F. (2002) Biological testing of a laboratory pathological waste incinerator.Applied Biosafety: Journal of the American Biological Safety Association, 7, 52-63.

Le Jambre, L.F., Gill, J.H., Lenane, I.J. and Baker, P. (2000) Inheritance of avermectin resistance in Haemonchus contortus.International Journal for Parasitology, 30, 105-111.

Lee, G.J.,Atkins, K.D. and Swan,A.A. (2000) Genetic variation in feed efficiency. In: Finewool 2000 - Breeding for Customer Needs (Eds. Swan, A.A. and Piper, L.R.), 59-60. (CSIRO Livestock Industries and The Woolmark Company: Armidale, NSW).

Lee, G.J.,Atkins, K.D. and Swan,A.A. (2001) Genetic parameters for pasture intake and wool growth efficiency in Merino sheep. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 505-508.

Lee, S.T., Schoch,T.K., Stegelmeier, B.L., Gardner, D.R.,Than, K.A. and Molyneux, R.J. (2001) Development of enzyme-linked immunosorbent assays for the hepatoxic alkaloids riddelliine and riddelliine N-oxide. Journal of Agricultural and Food Chemistry, 49, 4144-4151.

Lehnert, S.A. and Johnson, S.E. (2002) Expression of hemocyanin and digestive enzyme messenger RNAs in the hepatopancreas of the Black Tiger Shrimp Penaeus monodon. Comparative Biochemistry and Physiology B - Biochemistry and Molecular Biology, 133, 163-171.

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Leyhe-horn, B., Ziron, C., Prinzenberg, E.M., Barendse,W., Hiendleder, S. and Erhardt, G. (2001) Mapping of Oxytocin (OXT)to the central region of bovine chromosome 13 by linkage analysis using SSCP. Journal of Animal Science, 79, 777-778.

Li,Y., Kerr, R., Henshall, J.M., Harrison, B., Goddard, M. and Barendse,W. (2001) Comparison of QTL detection methods usingmarker data on beef quality traits - Single-trait single-sire family versus two-allele multi-trait and multi-family model.Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 413-416.

Li,Y., Byrne, K., Miggiano, E.,Whan,V., Moore, S.S., Keys, S., Crocos, P., Preston, N. and Lehnert, S.A. (2003) Genetic mapping of the kuruma prawn Penaeus japonicus using AFLP markers. Aquaculture, 219, 143-156.

Lin, C.-L., Chung, C.-S., Heine, H.G. and Chang,W. (2000) Vaccinia virus envelope H3L protein binds to cell surface heparan sulfate and is important for intracellular mature virion morphogenesis and virus infection in vitro and in vivo. Journal of Virology, 74, 3353-3365.

Linn, M.L., Gardner, J.,WarrILow, D., Darnell, G.A., McMahon, C.R., Field, I., Hyatt,A.D., Slade, R.W. and Suhrbier,A. (2001) Arbovirus of marine mammals: a new alphavirus isolated from the elephant seal louse, Lepidophthirus macrorhini. Journal of Virology, 75, 4103-4109.

Liu, S.M. and Masters, D.G. (2000) Quantitative analysis of methionine and cysteine requirements for wool production of sheep. Animal Science, 71, 175-185.

Liu, S.M., Blache, D., Mata, G. and Masters, D.G. (2000) Plasma leptin and body fat in grazing Merinos fed protected canola meal or methionine. Proceedings of the Nutrition Society of Australia, 24, 135.

Liu, S.M., Masters, D.G., Mata, G.,Wielinga, C. and Gulati, S.K. (2000) Responses to protected canola meal and methionine in grazing Merino weaners - Glutathione, methionine and cysteine. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 189.

Liu, S.M., Sammuels, L.M., Fitzgerald, N., Mata, G. and Adams, N.R. (2000) Insulin-like factor-1 mRNA and fractional protein synthesis rate in the skin of Merino lambs. Proceedings of the Nutrition Society of Australia, 24, 255.

Liu, S.M., Mata, G., Figliomeni, S., Powell, B.C., Nesci,A. and Masters, D.G. (2000) Transsulfuration, protein synthesis rate and follicle mRNA in the skin of young Merino lambs in response to infusions of methionine and serine. British Journal of Nutrition, 83, 401-409.

Liu, S.M. (2002) Determination of 2H2O at very low enrichment by gas chromatography mass spectrometry for measuring body water space. Proceedings of the Australian Society of Animal Production, 24, 323.

Liu, S.M., Murray,A., Schlink,A.C., Mata, G. and Masters, D.G. (2002) The effects of intradermal injections of spermidine on the growth rate of fibres and mitosis of wool follicles in Merino lambs. Animal Science, 75, 33-40.

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Lynn, D.H., Gransden, S.G.,Wright,A.D.G. and Josephson, G. (2000) Characterization of a new species of the ciliate Tetrahymena (Ciliophora : Oligohymenophorea) isolated from the urine of a dog: First report of tetrahymena from a mammal. Acta Protozoologica, 39, 289-294.

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Ward, K.A. (2001) Phosphorus-friendly transgenics. Nature Biotechnology, 19, 415-416.

Westbury, H.A. (2000) Emerging viruses of animals in Australia and Southeast Asia. In: Classical Swine Fever and EmergingDiseases in Southeast Asia. (Ed. Blacksell, S.D.), 15-20. (ACIAR: Canberra).

Westbury, H.A. (2000) Hendra virus: a highly lethal zoonotic agent. Veterinary Journal, 160, 165-6.

Westbury, H.A. (2001) Newcastle disease virus: an evolving pathogen? Avian Pathology, 30, 5-11.

White, C.L.,Young, P., Phillips, N. and Rodehutscord, M. (2000) The effect of dietary protein source and protected methionine(Lactet) on wool growth and microbial protein synthesis in Merino wethers. Australian Journal of Agricultural Research, 51,173-183.

White, C.L., van Houtert, M.F.J., Phillips, N.,Young, P.,Taylor, R., Coupar, F.,Ashes, J.R. and Gulati, S.K. (2000) Protected canolameal increases milk protein concentration in dairy cows fed a silage-based diet. Asian-Australasian Journal of Animal Sciences,13, (Suppl. A), 162.

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White, C.L. (2001) Factors affecting milk protein concentration in Australian Diary cows. Australian Journal of DairyTechnology, 56, 153.

White, C.L.,Tabe, L.M., Dove, H., Hamblin, J.,Young, P., Phillips, N.,Taylor, R., Gulati, S.K.,Ashes, J.R. and Higgins,T.J.V. (2001) Increased efficiency of wool growth and live weight gain in Merino sheep fed transgenic lupin seed containing sunflower albumin. Journal of the Science of Food and Agriculture, 81, 147-154.

White, C.L., Hanbury, C.D.,Young, P., Phillips, N.,Wiese, S.C., Milton, J.B., Davidson, R.H., Siddique, K.H.M. and Harris, D.(2002) The nutritional value of Lathyrus cicera and Lupinus angustifolius grain for sheep. Animal Feed Science and Technology,99, 45-64.

Wiese, S.C.,White, C.L., Masters, D.G., Davidson, R.H. and Milton, J.T.B. (2000) The performance of prime lambs fed diets with urea, lupins or canola meal as a source of protein. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 138.

Wiese, S.C.,White, C.L., Masters, D.G., Milton, J.T.B. and Davidson, R.H. (2000) Performance and carcase attributes of lambs fed protected methionine. Asian-Australasian Journal of Animal Sciences, 13, (Suppl. C), 281.

Wiese, S.C.,White, C.L., Masters, D.G., Milton, J.T.B. and Davidson, R.H. (2003) The growth performance and carcass attributes of Merino and Poll Dorset x Merino lambs fed rumen-protected methionine (Smartamine (TM)-M). Australian Journal of Agricultural Research, 54, 507-513.

Wijffels, G.L., Eisemann, C.H., Riding, G.A., Pearson, R.D., Jones,A.,Willadsen, P. and Tellam, R.L. (2001) A novel Family of chitin binding proteins from insect type II peritrophic matrix: cDNA sequences, chitin binding activity and cellular localisation. Journal of Biological Chemistry, 18, 15527-15536.

Willadsen, P. (2001) The molecular revolution in the development of vaccines against ectoparasites. Veterinary Parasitology,101, 353-367.

Williams, D.T.,Wang, L.-F., Daniels, P.W. and Mackenzie, J.S. (2000) Molecular characterisation of the first Australian isolate of Japanese encephalitis virus, the Fu strain. Journal of General Virology, 81, 2471-2480.

Williams, D.T., Daniels, P.W., Lunt, R.A.,Wang, L.F., Newberry, K.M. and Mackenzie, J.S. (2001) Experimental infections of pigs with Japanese encephalitis virus and closely related Australian flaviviruses. American Journal of Tropical Medicine and Hygiene,65, 379-387.

Williamson, M.M. and Daniels, P.W. (2000) Nipah virus. In: Animal Health and Production Compendium. (CABI:Wallingford, UK).

Williamson, M.M., Hooper, P.T., Selleck, P.W.,Westbury, H.A. and Slocombe, R.F. (2000) Experimental Hendra virus infection in pregnant guinea-pigs and fruit bats (Pteropus poliocephalus). Journal of Comparative Pathology, 122, 201-207.

Williamson, M.M., Hooper, P.T., Selleck, P.W.,Westbury, H.A. and Slocombe, R.F.S. (2001) A Guinea-pig model of Hendra virusencephalitis. Journal of Comparative Pathology, 124, 273-279.

Wilson, K., Li,Y.,Whan,V., Lehnert, S.A., Byrne, K.A., Moore, S., Pongsomboon, S.,Tassanakajon,A., Rosenberg, G., Ballment,E., Fayazi, Z., Swan, J., Kenway, M. and Benzie, J. (2002) Genetic mapping of the black tiger shrimp Penaeus monodon with amplified fragment length polymorphism. Aquaculture, 204, 297-309.

Wilson, S.C., Fell, L.R., Colditz, I.G. and Coliins, D.P. (2002) An examination of some physiological variables for assessing the welfare of beef cattle in feedlots. Journal of Animal Welfare, 11, 305-316.

Windon, R.G., Chaplin, P.J., Beezum, L., Coulter,A., CaHill, R., Kimpton,W., Drane, D., Pearse, M., Sjolander,A.,Tennent, J.M.and Scheerlinck, J.P.Y. (2000) Induction of lymphocyte recruitment in the absence of a detectable immune response.Vaccine, 19, 572-578.

Windon, R.G., Chaplin, P.J., McWaters, P.G.,Tavarnasi, M.,Tzatzaris, M., Kimpton,W.G., CaHill, R.N.P., Beezum, L., Coulter,A.,Drane, D., Sjolander,A., Pearse, M., Scheerlinck, J.-P.Y. and Tennent, J.M. (2001) Local immune responses to influenza antigen are synergistically enhanced by the adjuvant ISCOMATRIX. Vaccine, 20, 490-497.

Winter, P. and Colditz, I.G. (2001) Studien zur Produktion von Zytokinen nach Experimentell Induzierter Mastitis mit einem S.Epidermidis Stamm beim Schaf. (Studies on Production of Cytokines after experimental induced mastitis with S.Epidermidis-strain in ewes). In: Innere Medizin und Klinische Laboratoriumsdiagnostik, 27-29. (Deutschen VeterinarmedizinschenGesellschaft: Gießen, Germany).

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Winter, P. and Colditz, I.G. (2002) Immunological responses of the lactating ovine udder following experimental challenge withStaphylococcus epidermidis. Veterinary Immunology and Immunopathology, 89, 57-65.

Winter, P., Colditz, I.G., Fuchs, K. and Walshe, K. (2003) Serum amyloid A in serum and milk of ewes with mastitis induced experimental Staphylococcus epidermidis. Veterinary Record, 152, 558-562.

Winter, P., SchILcher, F., Fuchs, K. and Colditz, I.G. (2003) Dynamics of experimentally induced Staphylococcus epidermidismastitis in East Friesian ewes. Journal of Dairy Research, 70, 157-164.

Wood, P.R. (2000) Modulation of the response - ovine cytokines. In: Of Vets, Viruses and Vaccines:The Story of CSIRO's Animal Health Research Laboratory, Parkville. (Ed. Butcher, B.W.), 279-281. (CSIRO Publishing: Collingwood,Vic).

Wood, P.R. (2000) Mycobacterial diseases - renewed investigations of tuberculosis diagnosis. In: Of Vets, Viruses and Vaccines:The Story of CSIRO's Animal Health Research Laboratory, Parkville. (Ed. Butcher, B.W.), 167-169. (CSIRO Publishing: Collingwood,Vic).

Woolaston, R.R. and Elwin, R.L. (2001) No adaptation of roundworms to resistant hosts. Proceedings of the Association for the Advancement of Animal Breeding and Genetics, 14, 187-190.

Woolaston, R.R. and Windon, R.G. (2001) Selection of sheep for response to Trichostrongylus colubriformis larvae: genetic parameters. Animal Science, 73, 41-48.

Wright,A.D.G., Rogers, S., Baker, S.K. and Colorni,A. (2001) Marine fish parasite Cryptocaryon irritans finds taxonomic home. Microbiology Australia, 22, A120.

Wright,A.-D.G. and Colorni,A. (2002) Taxonomic re-assignment of Cryptocaryon irritans, a marine fish parasite. European Journal of Protistology, 37, 375-378.

Wright,A.D.-G.,Williams,A.J. and Baker, S.K. (2002) Novel group of archaea recovered from 16S rRNA clone libraries prepared from ovine rumen contents. Reproduction Nutrition Development, 42, (Suppl. 1), S77.

Wright, K., Berger, L., Nichols, D.K., Speare, R., Sredl, M.J., Pessier,A. and Johnson, B. (2001) Amphibian population decline.Journal of Herpetological Medicine and Surgery, 11, 14-27.

Yashiro, K., Lowenthal, J.W., O’Neil,T.E., Ebisu, S.,Takagi, H. and Moore, R.J. (2001) High-level production of recombinant chicken interferon-gamma by Brevibacillus choshinensis. Protein Expression and Purification, 23, 113-120.

Yob, J.M., Field, H.E., Rashdi,A.M., Morrissy, C.,Van der Heide, B., Rota, P.A., Binadzhar,A.,White, J.R., Daniels, P.W.,Aziz,A.J.and Ksiazek,T.G. (2001) Nipah virus infection in bats (order Chiroptera) in peninsular Malaysia. Emerging Infectious Diseases, 7, 439-41.

Zhang,Y.,Tennent, J.M., Ingham,A., Beddome, G., Prideaux, C.T. and Michalski,W.P. (2000) Identification of type 4 fimbriae on Actinobacillus pleuropneumoniae, FEMS Microbiology Letters, 189, 15-18.

Zinn, R.A., Gulati, S.K., Plascencia,A. and Salinas, J. (2000) Influence of ruminal biohydrogenation on the feeding value of fat infinishing diets for feedlot cattle. Journal of Animal Science, 78, 1738-1746.

Chief of DivisionShaun Coffey SL

Research ManagementMarion Andrew GABernard Bindon ARShaun Coffey SLPeter Daniels GARobert Hunter RHRobert Kelly FRLaurie Piper ARChristopher Prideaux GAStephen Prowse GAMichael Rickard GAJames Rowe ARPeter Willadsen SLRobert Woolaston SL

Research ScientistsNorman Adams FRSuzanne Baker FRWilliam Barendse SLAndrew Bean GAEmma Bermingham FRSharon Bishop-Hurley RHMaree Bowen RHDavid Boyle GAAlan Brownlee SLHeather Burrow RHGraham Cam SLIan Colditz ARSteven Colegate GAAxel Colling GABarbara Coupar GAJeff Cowley SLMark Crane GAPierre Cronje RHBrian Dalrymple SLJoyMaree Dempsey ARStuart Denman SLThomas Dixon SLRobert Dobson ARSonja Dominik ARTimothy Doran GARobyn Dynes FRSandra Eady ARBryan Eaton GAJohn Edgar GAAsoka Edirisinghe FRKeith Ellis AR

Christopher Elvin SLAndrew Fisher ARRosalind Gilbert SLLaurence Gleeson GAJeffrey Gorman SLAllan Gould GAChristian Gray SLAndrew Hall ARThomas Hall SLKim Halpin GAJeffrey Hammond GAVolker Haring GAGregory Harper SLRachel Hawken SLHans Heine GADavid Henry FRJohn Henshall ARJonathan Hill ARMichelle Hope SLPeter Hunt ARAlexander Hyatt GAJagodina Ignjatovic GAAaron Ingham SLMichael Johnson GAMatthew Kelly ARDavid Kemp SLPeter Kennedy RHCaroline Kerr SLJames Kijas SLSoressa Kitessa FRAndrew Knill GAMalcolm Knox ARKritaya Kongsuwan SLRajendra Kooloth Valappil SLAndrew Kotze SLMark Lanigan GACaroline Lee ARLisa Leeton SLSigrid Lehnert SLLeo Lejambre ARJoanne Lello ARYutao Li SLShimin Liu FRJohn Lowenthal GARussell Lyons SLDavid Masters FROliver Mayo SLSusan McClure ARKenneth McColl GASandra McKean GA

Christopher McSweeney SLWojciech Michalski GADeborah Middleton GADavid Miron ARRobert Moore GARalf Moser SLBarry Nagorcka CAHayley Norman FRBelinda Norris SLJacqueline Pallister GALaurelea Pickering SLSam Popovski FRKishore Prayaga RHCharles Press ARNorman Purdie RHIan Purvis ARSuzanne Rea FRAntonio Reverter-Gomez SLBevan Robertson RHAshok Santra FRSandra Sapats GAAnthony Schlink FRRobert Seymour SLVittoria Stevens GADavid Stewart GAYlva Strandberg SLAlanDavid Strom GAIan Sutherland RHJuliet Sutherland RHDavid Swain RHAndrew Swan ARRoss Tellam SLKhin Than GAMark Tizard GANigel Tomkins RHMaria VanHulten SLPeter Walker SLLinfa Wang GAYonghong Wang SLColin White FRGene Wijffels SLYvette Williams FRRoss Windon ARAndre-Denis Wright FRMeng Yu GAYamei Zhang GA

CODEAR–ArmidaleBH-BakersHillWABL–BelmontCA–BlackMountain(Canberra)FR–FloreatGA–AAHLRH–RockhamptonSA-SamfordSL–StLuciaND–LongPocketLaboratories

stafflist

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Research ProjectsJeisane Accioly FRDawood Al-Ajmi SLKrista Alley RHPeter Allingham SLRachel Amos GALester Anderson ARMichael Anderson ARMary-Ann Anderson GANeil Anderton GASarah Andrews FRManija Asif GAWanchai Assavalapsakul SLSteven Atkinson ARNeil Bagnall SLDenise Barendse SLMarion Barnes FRWesley Barris SLSally Barton ARKerrie Beales GAGary Beddome GAAmy Bell ARLuke Bell ARSusan Belson ARJohn Bingham GAAndrew Blakely ARGregory Bortolussi RHTimothy Bowden GANeil Bower SLVictoria Boyd GADonna Boyle GAAdam Brahim RHHeather Brewer ARJanice Briegel FRSusan Briscoe SLMary Broadway GAMatthew Bruce GAKerri Bruce GARowan Bunch SLKeren Byrne SLLee Cadogan SLDavid Callan ARYu Cao GAMaria Cardoso GAAnne Carlyle ARAndrew Carr SLAndrew Chalmers ARTamasine Chamberlain GAAmanda Choice ARClaus Christophersen FRGemma Clark GACarolyn Cluderay SLJohn Cole ARErin Collins SLJames Conlan GALaurence Conlan SLSerge Corbeil GANicholas Corbet RHGary Crameri GASandra Crameri GABrian Cross ARDavid Cummins GASousanna Daglas GAJane Dale GAChristopher Darcy GARhonda Davey ARCassandra David GACarl Davis SLAlan Day RH

Christine Dennis ARBrian Dennison ARLeanne Dierens SLChristine Dimmock SLGraham Donald FRWilliam Doughty GAEmma Doyle ARKelly Drake ARChristine Duch GATimothy Dyall ARLauretta Eaton GASharon Egan GASamuel Ellis GAClare Engelke FRJohn Exley ARElizabeth Farrell ARDrewe Ferguson ARRobert Flack ARRhys Fogarty GAAdam Foord GAMark Ford GAOlivia Fulloon ARDean Gibson RHMelinda Goga GASarah Goldie GADonna Gonzales GAGregory Good ARJohn Gorham ARJoanne Gough SLGaylene Gould GAMicaela Grandison FRKathleen Granger GADiane Green GAJae Greenwood GANicole Griffin RHNicholas Gudkovs GAJulia Hammond GAAnna Hansson AREric Hansson GAJanine Harrison ARDavid Harrison GABlair Harrison SLDebbie Hickey ARMarlene Hillard ARBradley Hine ARBarney Hines SLNicola Hogan AREdward Holland ARPeter Hooper GABiserka Horvatic GAMarie Hosking GARobert Hudson ARElizabeth Hulm FRDonna Jackson GAVijayalakshmi Janardhana GAElisa Jansen GAKristie Jenkins GADayna Johnson GASamuel Johnson SLHaydn Jones GARichard Jones SLPeter Josh ARTroy Kalinowski ARMatthew Kane SLAdam Karpala GAJacqueline Kattenbelt GABronwyn Kelley SLGregory Kemp SLJudith Kenny AR

Louis Klein FRMelissa Kowalski GALuke Lambeth GAShea Larkin GADaniel Layton GAChristine Leger ARIan Lenane SLQingmin Li FRYvette Lieschke-Mercer ARSuzanne Lowther GARoss Lunt GACallum Mack ARBahar Maghsoudi ARDanielle Magoffin GARobyn Malcolm ARSimone Martin FRGonzalo Mata FRSandra Maxwell ARDianne Mayberry FRNadia Mayfield GARussell McCulloch SLJennifer McEachern GAKatherine McMaster ARKelly McNicol RHPeter McWaters GASean McWilliam SLJennifer Meadows SLGreer Meehan GAMoira Menzies SLAgnieszka Michalewicz GAPhilippa Miller GAMakoto Mitsumori SLKirsten Morris GAChristopher Morrissy GAAnne Murray FRDamien Nankervis RHDharani Narayanan SLRobert Nethery ARKim Newberry GADominic Niemeyer ARDangThiHoang Oanh SLMargaret O'Grady SLJohn O'Grady SLPamela Oke GABrian Oldfield GAJeanette Olejnik RHVeronica Olsen GATerri O'Neil GAChristopher O'Neill RHRalph Ophuis GAJanine Ortenburg GADilok Ounpomma GARoslyn Owen FRJagadish Padmanabha SLDavid Paull ARChristina Pavlov SLKelly Pearce FRRoger Pearson SLThuy Phan SLNathan Phillips FRMarc Picault SLGary Pickard GALindsay Pritchard GALeah Protyniak GAAnthony Pye GAPeter Quigley ARMatthew Reed ARMegan Retallick GA

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Robert Rich ARGeorge Riding SLAllan Rintoul FRJordan Robinson ARTanya Robinson RHChristina Rootes GAGail Russell GAYoichi Sakaguchi SLAbdi Salah FRDarren Schafer GAKaren Schutt RHPaul Selleck GAKaylene Selleck GASarah Shaw RHScott Sheedy GABrian Shiell GAMuhammad Siddiqi FRWarren Sim RHJoanne Slater GALysandra Slocombe ARAlbert Smith ARJennifer Smith ARKellie Smith FRClarence Smith GAWendy Smith SLPatricia Sollars GAChumporn Soowannayan SLSomsrong Spiess GAKathryn Sproat GAKelly Steeper GAGarth Stephenson GAPhilip Stewart GAPaula Stiles GAPeter Stockwell SLSally Stockwell SLManuela Stolic SLLesley Store FRIvan Stratov GAMyles Stritzke RHJianguo Su SLHadi Sulaxono GAJane Swingler GARichard Sydenham GAAndrew Sydenham GAKim Szalnowski GAMary Tachedjian GASiokHwee Tan SLKarina Tane RHMasaaki Taniguchi SLTrevor Taylor GAMichael Taylor SLJesse Thomas GAMerle Thomas SLJames Thyer GAMohamad Tiong FRAndrew Toovey FRAndrea Trebbin SLLee Trinidad GALily Tsen GADamien Turner SLScott Tyack GAKerri Tyrrell ARAytac Unal GAGrant Uphill ARBrenda VanDerHeide GARosemary VanDriel GAJill Vaughan GAJohn Vercoe RHSomasuntharam Vignarajan AR

Rhonda Voysey GATony Vuocolo SLVicki Whan SLJohn White GAPriyanja WijegoonawardaneSLMichelle Wilkins GAAndrew Williams FRJudith Williams FRLynette Williams GAPaul Williams RHSamuel Williams RHMatthew Wilmot FRTerry Wise GAChinChingDarrenWong SLLynda Wright GAPeter Wright GAPaul Young FRJohn Young GAKeryn Zirkler AREric Zurcher CA

Technical ServicesJoshua Allan GARichard Amstutz GAColin Arklay GAMichael Balfour NDMatthew Balfour NDMalcolm Beaton GAGiovanni Bianchi GATerrence Blain GAMatko Bojmic GARick Bowman NDPeter Bradley ARMegan Braun GAPhilip Bullock BHGraham Burke ARDonald Carlson GAMichael Carthew FRLaurence Clark GAGerald Clune BHNoel Collins GALuis Costoso GALester Couchman SALindsay Cranston BLDarryl Crowe GARobyn Daniel SLLindsay Donahoo GASteven Edwards GAAlan Edwards GAAndrew Eichorn ARMichael Fanning GAHilton Faulmann GAAlexander Fierka GAFrank Filippi GAAiden Gebbie GADennisJohn Gladman GAIan Harrison GALindy Hart SLRobert Hensel GARoss Hodge GARaymond Honnery ARGary Hourticolon GAMilton Howard GAJohn Hutchinson GAPeter Kavenagh GARaymond Kempton GARobert Kerslake SLJoel Kingshott GASteven Kunjka GAPeter LeBlancSmith GA

Jun Liang GARonald Lockwood ARAlexander Matheson GADavid Maurer GAPhillip McCabe GAChester McCombe GANeil Michell GAGarry Monkivitch GARonald Murphy GAJohn Muschialli GARobert Nicholls GAColin Northwood GAIan Norton GAGregory Pepyat GACarolyn Pimm FRMark Rechenberg GADavid Reid GABruce Riches SLKurts Rode-Bramanis SLCraig Rooke GAColin Russell NDRichard Ryan GAJennifer Ryan SLBen Salmetti GAJonathan Salmon GATrevor Shell GANeil Slater GATerry Smith GAColin Taylor GAWilliam Vandenheuvel RHDavid Walker GABill Wells GACatherine Williams GASusanne Wilson GAAnthony Winiecki GABrett Wood NDCarol Wykes GA

Communication & InformationKlaus Altmann ARJustin Baiocchi ARMichael Batson GADaniel Benz SLArthur Berry GAPaul Bladen GAJohn Brumby GAOliver Dempsey ARJennifer Dunn RHKaye Guidetti SLGerald Haaima SLAnnette Hill SLEmma Homes GAAndrew Hooley SLRonald Horst SLPatrick Ledwith SLSue Mahoney GAJudith Maunders GAJane McIntyre SLPompeo Mignone GARobyn Mills SLHoward Philpott GAMargaret Puls FRWendy Pyper SLChristopher Short SLNeil Silver GAVeronica Toohey SLChristopher Weber ARMaria Welsh GAKerri White SLCatherine Young SL

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General ManagementAlan Chambers SLGregory Davies SLMartyn Jeggo GAJack Malecki GAPeter Rodgers GAAnne Tuppack SL

Administrative ServicesDallas Blakely ARGraham Frost ARCatherine Holmes ARDebra Lane ARFrances Nicholls ARSally Reed ARDavid Simmons ARCarolyn Adams SLYvette Baglieri GAKenneth Barker GAIrwin Barrett-Lennard FRHelen Chapman FRAnthony Coppens FRJoanna Cray SLLesley Cundy RHAnne Edwards GASandra Elson GAKristi-Ann Fenech SLMatthew Fenech SLCindy Fraser GARoslyn Gaydon SLRobert Gilham FRGarry Gould GAAnne Grace SLColin Hamilton RHKathleen Hamilton SLJulie Hughan GATrevor Kelly GANelly Kremko SLDianne Kunz SLCatherine Leather SLDebbie Logan GAPhillipa Loveridge SLGlennis McDonald FRTrent McKenzie SLLeanne Moloney GAChristopher Morrissy GAPhilip O'Dowd GASuzanne O'Dowd GABernadette O'Keefe GADianne O'Keefe SLOlalla Parker GAAnne Partridge GASuzie Perilli GAMichelle Primmer GALyn Read SLNicole Roberts SLKerrie Ryan GARobyn Sallaway SLManjit Sanghera SLMaree Schafer SLCarole Scott GAIan Southey FRMatthew Taylor SLNeil Templeman SLSonya Watson SLCatrin Waye RHPhilip White RHDenise Williams RHKenneth Woodward SLKathleen Zahnleiter SL

General ServicesDayna Anderson ARJanice Baran GAJackie Boyle GAJanice Brockman BHJason Burville GAJennifer Clarke GARoss Cook GAElaine Cranny GARobert Dandy GAAllan Farmer GAAileen Fitzgerald GAMary Giuffre GAWinsome Goff GAKeith Harder BLPatricia Hutchinson GACraig Jones BLSandra Kahle GAJill King GAPatricia Lockwood ARDiane Lodge GAKevin Lodge GAHerbert Marell GACarol Michell GATanya O'Neill GAScott Prewett BLElizabeth Regan GAPauline Robinson ARGary Rowe GAAnna-Maria Russell GAMargaret Ryan GALuke Schafer SLKathleen Snowden GAWayne Spalding GAShane Speer ARJeffrey Spence GAGraham Vick BLAdele Wheeler BLDavid Wilkinson ARAnn-Louise Williams GA