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Acta Tropica 107 (2008) 168–173

Contents lists available at ScienceDirect

Acta Tropica

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nfection rates and genotypes of Trypanosoma rangeli and T. cruzi infectingree-ranging Saguinus bicolor (Callitrichidae), a critically endangered primatef the Amazon Rainforest

. Maia da Silvaa, R.D. Naiffb, A. Marcili a, M. Gordoc, J.A. D’Affonseca Netoc, M.F. Naiffb, A.M.R. Francob,. Campanera, V. Valented, S.A. Valented, E.P. Camargoa, M.M.G. Teixeiraa,∗, M.A. Milese

Departamento de Parasitologia, Instituto de Ciencias Biomedicas, Universidade de Sao Paulo (USP), Sao Paulo, Sao Paulo, BrazilCoordenacao de Pesquisas em Ciencias da Saude, Instituto Nacional de Pesquisas da Amazonia (INPA), Manaus, Amazonas, BrazilDepartamento de Biologia, Projeto Sauim-de-Coleira, Universidade Federal do Amazonas (UFAM), Manaus, Amazonas, Brazil

Instituto Evandro Chagas, Belem, Para, BrazilDepartment of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, UK

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Article history:Received 27 February 2008Received in revised form 2 April 2008Accepted 22 May 2008Available online 29 May 2008

Keywords:Trypanosoma cruziTrypanosoma rangeliNeotropical primatesSaguinus bicolorAmazoniaEmergent disease

a b s t r a c t

Parasites of wild primatepotential to infect humanpanosoma cruzi and T. ransurvey of trypanosomiasifrom the Brazilian Amazoyielded blood smears pos(39.6%) whereas T. cruzi wgenus Megatrypanum. Hewere morphologically diacultures of these two specand two of T. rangeli, withremained as established c

and 54 to lineage B (87.1%). TeTransmission of both trypanosowild primates as reservoirs for

. Introduction

Some vector-borne zoonoses such as trypanosomiasis andeishmaniasis pose considerable risk to humans and wildlifend may have deleterious effects that trigger or accelerate theecline of threatened species. Saguinus bicolor (bare-faced tamarin)elongs to the family Callitrichidae with a geographical distributionestricted to the vicinities of the cities of Manaus, Rio Preto da Eva,nd Itacoatiara, State of Amazonas (Egler, 1983). This species wasncluded in the Red List of Threatened Species of the Internationalnion for the Conservation of Nature and Natural Resources (IUCN),nd is considered, with Cebus kaapori, the most endangered primatepecies of the Amazonian region (Rylands et al., 2003).

∗ Corresponding author. Fax: +55 11 30917417.E-mail address: mmgteix@icb.usp.br (M.M.G. Teixeira).

001-706X/$ – see front matter © 2008 Elsevier B.V. All rights reserved.oi:10.1016/j.actatropica.2008.05.015

important for conservation biology and human health due to their highthe Amazon region, non-human primates are commonly infected by Try-which are also infective to man and several mammals. This is the firstcritically endangered species of tamarin, Saguinus bicolor (Callitrichidae),forest. Of the 96 free-ranging specimens of S. bicolor examined 45 (46.8%)

for trypanosomes. T. rangeli was detected in blood smears of 38 monkeysever detected. Seven animals (7.3%) presented trypanosomes of the sub-ltures detected 84 positive tamarins (87.5%). Seventy-two of 84 (85.7%)d as T. rangeli and 3 (3.1%) as T. cruzi. Nine tamarins (9.4%) yielded mixedhich after successive passages generated six cultures exclusively of T. cruzione culture remaining mixed. Of the 72 cultures positive for T. rangeli, 62

es and were genotyped: 8 were assigned to phylogenetic lineage A (12.9%)n established cultures of T. cruzi were genotyped as TCI lineage (100%).me species, their potential risk to this endangered species and the role oftrypanosomes infective to humans are discussed.

© 2008 Elsevier B.V. All rights reserved.

S. bicolor is a diurnal arboreal monkey that uses a mosaic of veg-etation, including primary and secondary forests, “campinarana”forests, and forest edges. Tamarins are primarily insectivores andfrugivorous but also feed on nectar, exudates and small vertebrates(Egler, 1993; Rylands and Faria, 1993; Vidal and Cintra, 2006). Asother tamarins, this species lives in groups and uses tangles of twigsand lianas in trees, holes in trunks, and palms as nocturnal restingsites. The areas inhabited by S. bicolor have abundant palm trees,which provide the nocturnal refuges for the tamarins and are thepreferential ecotopes of hematophagous triatomine bugs, notablyR. pictipes and R. robustus, which are the most important vectorsof Trypanosoma cruzi and T. rangeli all over Amazonia (Miles et al.,1983a; Coura et al., 2002; Maia da Silva et al., 2007; Aguilar et al.,2007).

Some species of trypanosomes are infective to human and non-human primates around the world (Marinkelle, 1976). Neotropicalmonkeys in addition to harbouring T. cruzi and T. rangeli are also

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infected by T. saimiri and T. mycetae, species allied to T. rangeli,and by T. devei, T. minasense and T. lambrechti, which belong tothe subgenus Megatrypanum. Except for T. cruzi and T. rangeli, noother species infecting monkeys have been reported as infectiveto humans. Trypanosomes were commonly reported in monkeysof the families Callitrichidae (tamarins and marmosets), Cebidae(squirrel monkeys and capuchins), Aotidae (night monkeys) andAtelidae (spider monkeys and howler monkeys) (Deane et al.,1970; Sousa et al., 1974; Marinkelle, 1976; Lanham et al., 1984;D’Alessandro et al., 1986; Ziccardi and Lourenco-de-Oliveira, 1997;Ziccardi et al., 2000; Lisboa et al., 2004a,b, 2006, 2007; Maia daSilva et al., 2004a,b, 2007).

Although the available data indicate that trypanosomiasisshould be very common in some Amazonian primates in thewild, few species of wild primates have so far been examined.The largest survey on Amazonian primates was done using bloodsmear microscopy, hemoculture and xenodiagnosis and revealed ahigh prevalence of trypanosomes (67.9%) in 165 squirrel monkeys(Saimiri sciureus and Saimiri ustus), with the following trypanosomespecies: T. rangeli (35.2%), T. saimirii (32.1%), T. minasense (33.3%)and T. cruzi (10.3%). Mixed infections were present in 52.7% of theinfected animals (Ziccardi and Lourenco-de-Oliveira, 1997). Highrates of T. rangeli were also reported for Saguinus midas (51%),whereas a lower prevalence was found in Alouatta macconelli (16%)in French Guiana (De Thoisy et al., 2001). T. rangeli was also themost common species (30.4%) infecting tamarins and marmosetscaptured in Amazonia and maintained in the National Centre ofPrimates, Para, Brazil (Ziccardi et al., 2000). T. rangeli has alsobeen recovered from several monkey species from the Amazo-nian States of Rondonia, Acre, Amazonas and Para (Miles et al.,1983b; Maia da Silva et al., 2004a,b, 2007). This trypanosome wasalso highly prevalent in squirrel monkeys, tamarins and marmosetsfrom Colombia, Bolivia and Panama (Sousa et al., 1974; Marinkelle,1976; D’Alessandro et al., 1986). In Brazil, there are no reports ofT. rangeli in wild primates outside Amazonia, although this specieswas reported in sylvatic rodent, the common opossum, R. neglectusand R. nasutus (Steindel et al., 1991; Ramirez et al., 2002; Dias et al.,2007).

The infection rates with T. cruzi in Amazonian monkeys rangedfrom 10.3% using parasitological methods (Ziccardi and Lourenco-de-Oliveira, 1997; Ziccardi et al., 2000) to 46% detected by serology(Lisboa et al., 2006). Seroprevalence for T. cruzi was high (46%)for specimens of golden lion tamarins living in a broader areaof the Atlantic Forest, and also in a captive primate unit located

inside the Atlantic Forest (26.5%) (Lisboa et al., 2004b, 2006). InAmazonian monkeys lower prevalences have been reported for T.cruzi compared to T. rangeli. T. rangeli is considered non-pathogenicfor man, but its pathogenicity for non-human primates has notbeen investigated. In contrast, T. cruzi is pathogenic in humansand also in non-human primates from both the Old and NewWorlds. In experimentally infected monkeys, T. cruzi infection isgenerally asymptomatic but can also present symptoms similarto those of acute and chronic human Chagas disease (Miles et al.,1979; Rosner et al., 1988; Carvalho et al., 2003). Naturally infectedgolden lion tamarins displayed typical signs of disease, with 45%of the animals presenting cardiac abnormalities (Monteiro et al.,2006).

T. cruzi and T. rangeli are both generalist parasites infective for awide range of mammals including man and several species of wildmonkeys, and both species are transmitted by the same vectors inoverlapping areas of Amazonia. However, knowledge on the epi-demiology of these trypanosomes in both human and non-humanprimates is hindered by diagnostic methods of limited sensitivityand specificity. Genotyping and phylogenetic analysis of 13 isolatesof T. rangeli from Amazonian wild monkeys were reported in our

pica 107 (2008) 168–173 169

previous studies (Maia da Silva et al., 2004b, 2007) whereas only7 isolates of T. cruzi from captive Amazonian monkeys have beengenotyped (Lisboa et al., 2007). The small number of isolates char-acterized prevents a correct appraisal of the genotypes of T. cruziand T. rangeli circulating in free-ranging primates, their transmis-sion dynamics and pathogenicity.

Diseases transmitted among human and wild and domestic ani-mals, especially those caused by generalist pathogens pose risksto both wildlife preservation and human health. Parasites of non-human primates are also very relevant to human health since nearlyone-third of the protozoan parasites of wild monkeys have alsobeen reported in humans (Nunn et al., 2005; Brooks and Ferrao,2005; Lebarbenchon et al., 2008). Knowledge of the health status ofwildlife and monitoring of the parasitic infections are prerequisitesfor conservation strategies. The main objective of this study was toassess trypanosomiasis in the endangered tamarin S. bicolor aimingto (a) determine the rates of infection by T. cruzi and T. rangeli byblood smear microscopy and hemoculture; (b) diagnose culturedtrypanosomes by molecular methods; (c) characterize the geno-types of the trypanosomes isolated from S. bicolor; (d) provide dataabout transmission and potential risk of trypanosome infections fortamarins and humans.

2. Materials and methods

2.1. Area of study, capture and handling of monkeys

The blood samples from S. bicolor examined in this study werecollected during the fieldwork of “Projeto Sauim-de-Coleira” exe-cuted by the Universidade Federal do Amazonas. The animals usedin this study were captured using Tomahawk traps baited withbanana, between the years 2001 and 2005, in several forest frag-ments in Manaus, Amazonas State, Brazil, and in two neighbouringareas, the Reserve Adolfo Ducke of INPA (Instituto Nacional dePesquisa da Amazonia), Road AM-010, km 26, and a farm on thesame road, km 43. Ninety-six males and females of all ages, infantsexcluded, were captured, kept in a relatively cool place in coveredtraps in order to minimize their stress, and anesthetized (using0.1 ml intramuscular, 1% ketamine hydrochloride) to permit minorprocedures including clinical examination and blood sampling.After complete recuperation, about 4 h after capture, the tamarinswere set free at the site of capture. The predominant vegetationin all localities is secondary and/or primary “terra-firme” forests,

with borders around the fragments, and in some places (ReservaDucke) there are a few hectares of “campinarana” (Fig. 1). A groupcomposed of biologists and veterinarians was responsible for cap-turing and handling the tamarins, according to the permit of IBAMA(Instituto Brasileiro do Meio Ambiente).

2.2. Detection of trypanosomes by microscopy and hemocultureof tamarin blood samples

Blood of tamarins was collected (0.3–0.6 ml) from the femoralvein and examined for the presence of trypanosomes by hemo-culture and by microscopy of blood smears. Two Giemsa-stainedsmears on glass slides were prepared from each tamarin. Morphol-ogy of blood trypomastigotes was analysed in a first attempt atspecies identification according to the features described by Hoare(1972). Blood samples were inoculated into hemoculture tubes (3–5for each animal) containing NNN medium with a 0.9% saline overlay(Miles et al., 1981a) and incubated at 28 ◦C. Giemsa-stained epi-mastigotes of cultured trypanosomes were also analysed aiming atmorphological identification of trypanosome species according toHoare (1972).

170 F. Maia da Silva et al. / Acta Tropica 107 (2008) 168–173

of cap

Fig. 1. Geographical area

2.3. DNA preparation, molecular diagnosis and genotyping oftrypanosome isolates from S. bicolor

Trypanosomes cultured in LIT medium were used for DNAextraction and tested by two PCR assays for species-specific diag-nosis: (a) PCR based on ribosomal sequences for simultaneousdetection of T. cruzi and T. rangeli (Souto et al., 1999) and (b) theT. rangeli-specific assay TraSL-PCR (Maia da Silva et al., 2007). Afterconfirmation of the specific diagnosis, isolates from all cultureswere genotyped through the following methods: TraSL-PCR forgenotyping of T. rangeli lineages (Maia da Silva et al., 2007); PCRbased on mini-exon markers for genotyping of T. cruzi lineages(Fernandes et al., 2001). Reference-isolates/strains of the differentphylogenetic lineages of T. rangeli (A, San Augustin; B, AM80; C, PG;D, SC58) and T. cruzi (TCI, G; TCIIa, CanIII; TCIIb, Y; TCIIc, Tc3663)were used for comparison with the isolates from S. bicolor.

3. Results

3.1. Diagnosis and prevalence rates of trypanosomiasis in S.bicolor by microscopic examination of blood smears andhemoculture

Microscopic examination (ME) of blood smears of wild-caught

specimens of S. bicolor revealed trypanosomes in 45 (46.8%) ofthe 96 individuals examined. Of these positive smears, 38 (84.4%)presented slender trypomastigotes morphologically resemblingthose of T. rangeli, whereas 7 (15.6%) exhibited large trypomastig-otes similar to those of species of the subgenus Megatrypanum.Small trypomastigotes typical of T. cruzi were not detected. Pri-mary hemocultures revealed that 71 tamarins were positive fortrypanosomes. Most T. rangeli cultures were derived from micro-scopically positive animals whereas T. cruzi cultures were frommicroscopically negative or T. rangeli positive animals. Morpholog-ical analysis of Giemsa-stained epimastigotes from recent culturesrevealed 72 cultures exclusively of T. rangeli (75% of tamarins) and3 of T. cruzi (3.2%), whereas 9 were observed to be mixed cultures(9.45%) of both these species. After successive passages in culture,the nine mixed cultures generated six pure cultures of T. cruzi, T.rangeli became the sole species in two cultures and only one cultureremained mixed with these two species. These species were all con-firmed by molecular diagnosis of established cultures, which are allcryopreserved. Only one tamarin generated separate cultures of T.rangeli or T. cruzi from different hemoculture tubes. Ten tamarinsproduced positive hemocultures that were not established after

tures of Saguinus bicolor.

successive cultures. No other trypanosome species besides T. cruziand T. rangeli could be cultured for more than 2 weeks. Culturesof Megatrypanum trypanosomes, including some similar to T. deveidescribed in S. midas niger (Lanham et al., 1984) were never estab-lished, dying after 7–10 days.

3.2. Molecular diagnosis and prevalence rates, and genotyping ofT. rangeli and T. cruzi infecting S. bicolour

Results of molecular diagnosis for all trypanosome cultures fromS. bicolor corroborated data obtained by morphological analysis,confirming higher prevalence rates of trypanosomiasis caused byT. rangeli, detected in 72 cultures (75%), than for T. cruzi, detectedin 10 cultures (10.4%) from the 96 tamarins examined. All cultureswere tested by a duplex PCR based on ribosomal sequences, whichdistinguishes T. rangeli from T. cruzi (Souto et al., 1999) (Fig. 2A).T. rangeli identity was confirmed by a species-specific PCR assay(TraSL-PCR) capable of detecting all lineages of this species (Maiada Silva et al., 2007) (Fig. 2B).

Of a total of 72 primary cultures 62 isolates of T. rangeli remainedestablished cultures, 8 isolates were ascribed to phylogenetic lin-eages A (12.9%) and 54 to lineage B (87.1%) (Fig. 2B). All 10 culturesof T. cruzi, including one mixed with T. rangeli, were genotyped aslineage TCI (Fig. 2C). Genotyping was done using two PCR assays

based on spliced leader (SL or mini-exon) sequences developed togenotype T. rangeli (Maia da Silva et al., 2007) or T. cruzi (Fernandeset al., 2001).

4. Discussion

Because wild primates are reservoirs of several parasites infec-tious to humans, parasitological surveys of non-human primatesprovides an important opportunity to better understand the epi-demiology, transmission dynamics and emergence risk of variousanthropozoonoses. Although Brazil, especially Amazonia, presentsa vast diversity of wild primates, viral, bacterial and protozoaninfections of these primates are poorly studied compared to OldWorld primates (Hugot, 1998; Nunn et al., 2005).

Wild primates living in the Atlantic Forest presented trypanoso-miasis caused by T. cruzi, malaria and toxoplasmosis, which areall diseases caused by hemoprotozoa that share human and wildhosts (Lisboa et al., 2004a,b, 2006; Deane, 1992; Duarte et al., 2006;Garcia et al., 2005). Data about T. cruzi in naturally infected non-human primates focused primarily on the golden lion tamarin, anendangered species of the Atlantic Forest that may get severely sick

F. Maia da Silva et al. / Acta Tropica 107 (2008) 168–173 171

ide gecificl., 200

Fig. 2. Agarose gel (2%) showing amplified fragments stained with ethidium bromsimultaneous detection of T. cruzi and T. rangeli (Souto et al., 1999). (B) T. rangeli-spbased on mini-exon gene markers for genotyping of T. cruzi lineages (Fernandes et a

when infected by T. cruzi (Monteiro et al., 2006). There are few stud-ies about protozoa of free-ranging primates in the Amazon region,with reports about malaria and trypanosomiasis (Deane et al., 1970,1972; Miles et al., 1983b; Ziccardi and Lourenco-de-Oliveira, 1997;Ziccardi et al., 2000).

Here, we described a large survey of trypanosomes in theendangered wild primate from Amazonia, the tamarin S. bicolor.Morphological and molecular diagnosis of cultures from 96 spec-imens of S. bicolor revealed prevalence rates higher for T. rangeli(75%) than for T. cruzi (10.4%). High prevalences of T. rangeli havealready been described in other wild primates from Brazilian Ama-zonia, besides the description of T. saimirii and T. minasense, whichare synonymies of T. rangeli (Ziccardi and Lourenco-de-Oliveira,1997; Ziccardi et al., 2000; Maia da Silva et al., 2004a,b, 2007).

T. rangeli isolates were ascribed to phylogenetic lineages A orB confirming previous studies that demonstrated the existence ofthese lineages in wild primates and triatomine bugs from north-ern (Amazonas State), eastern (Para State) and western (Rondoniaand Acre States) Brazilian Amazonia (Maia da Silva et al., 2004b,2007). In Amazonia, Rhodnius species are also infected by T. cruziand T. rangeli, frequently in mixed infections (Miles et al., 1983b;

enerated by the following PCR assays. (A) PCR based on ribosomal sequences forTraSL-PCR for genotyping of T. rangeli lineages (Maia da Silva et al., 2007). (C) PCR1).

Coura et al., 2002; Maia da Silva et al., 2004b, 2007). Comparativephylogeographical analysis permitted the association of T. rangelilineages with complexes of Rhodnius species. Lineage A occurs inhumans and triatomines in Central America and northwest SouthAmerica, and is transmitted by species of the complex R. prolixus,represented by R. robustus in Amazonia. Lineage B, the only lineageso far found infecting man in Brazilian Amazonia, was associatedwith R. brethesi, which belongs to the complex that includes R. pic-tipes and appears to be restricted to Amazonia. Lineages A and B,besides primates, also circulate in other wild mammals includingopossum, sloth and anteater. The lineages C and D were never foundin Amazonia, being restricted to Central America and Colombia (lin-eage C) and Southern Brazil (lineage D) (Maia da Silva et al., 2004b,2007).

Before this study, only seven T. cruzi isolates from Amazonianprimates held in captivity had been genotyped as lineage TCI,including two isolates of S. bicolor (Lisboa et al., 2007). In this study,10 T. cruzi isolates from wild-caught S. bicolor were assigned to TCI.The finding that TCI, known to be associated with the sylvatic cycleof T. cruzi in all Latin America, circulates in Amazonian S. bicoloragrees with reports on the presence of T. cruzi belonging to TCI or,

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less frequently to TCIIa (formerly known as zymodeme 3) in othersylvatic mammals and in humans in this region (Miles et al., 1981a;Coura et al., 2002). TCI is associated with the sylvatic cycle in Brazil,responsible for acute and chronic Chagas disease in Amazonia andin other rural areas (Miles et al., 1981a; Coura et al., 2002; Teixeiraet al., 2006; Aguilar et al., 2007). In contrast, TCI is associated withthe domestic cycle and with acute and chronic myocardiopathy inVenezuela, Colombia and Panama (Miles et al., 1981b; Anez et al.,2004; Sousa et al., 2006). In the Atlantic Forest, however, wild pri-mates were found infected with TCI and/or TCII (Lisboa et al., 2004a,2006). While specimens of free-living S. bicolor harbour TCI in Ama-zonia, a specimen of S. bicolor from the Primatology Center of Rio deJaneiro in the Atlantic Forest was found infected with TCII, whichis associated with domestic cycle and human disease in this area(Lisboa et al., 2007).

The observed predominance of T. rangeli over T. cruzi in non-human primates in Amazonia deserves some consideration. If onlyblood smears were considered, this finding could reflect a method-ological bias since acute T. cruzi infection evolves rapidly to chronicinfection with parasitemias subpatent to microscopy even in rein-fected animals. In contrast, T. rangeli infection yields lasting patentparasitemias, as evidenced in this study by the high positivity ofblood smears. However, the 6:1 predominance of T. rangeli over T.cruzi in hemocultures confirms the higher prevalence of T. rangeliinfection, since hemoculture is considered to be more effective forT. cruzi than T. rangeli. Unfortunately, the small blood samples per-mitted to be collected from S. bicolor precluded serology that mightclarify the real prevalence of T. cruzi chronic infection in these pri-mates.

It is very likely that transmission peculiarities are responsible forthe predominance of T. rangeli in S. bicolor. Both trypanosomes aretransmitted by the same triatomines, but through distinct routes ofinfection. T. cruzi is transmitted by contamination of injured skin orintact mucosa with infective forms present in bug faeces, whereasT. rangeli is transmitted by inoculation, during biting or probing,of infective forms present in the salivary glands of Rhodnius spp.Inoculation is a far more efficient means of transmission than con-tamination and this may well contribute to the predominance of T.rangeli infections.

The possibility of oral infection of tamarins cannot be dis-counted. Insects are very important in the diet of S. bicolor that couldbe infected by ingestion of trypanosome-infected triatomines.However, there is no proof that S. bicolor actually eats triatomines.Oral transmission is known to be a very efficient route for T. cruzi

infection, responsible for outbreaks of acute Chagas disease inBrazilian Amazonia and elsewhere (Coura et al., 2002; Pinto et al.,2004). Although there are several outbreaks of orally acquired Cha-gas disease in Amazonia, there are no reports of human infectionswith T. rangeli attributable to oral transmission. If oral infections oftamarins by both trypanosomes were equally efficient this wouldnot accord with the predominance of T. rangeli in wild tamarinsor of T. cruzi in humans. Blood-to-blood exposure during fightingand transplacentary transmission are other possible transmissionroutes (Eberhard and D’Alessandro, 1982; Lisboa et al., 2004a,b),but these routes would be expected to favour T. cruzi over T.rangeli. Regardless of the mechanisms underlying the transmis-sion of trypanosomes to tamarins, the fact remains that T. rangeliunquestionably prevails over T. cruzi infections.

As to the health of trypanosome-infected animals, despite thesuperficial clinical evaluation, it did not seem to be impaired, butthis does not preclude the existence of undergoing pathogenic pro-cesses. T. cruzi infection in primates is known to remain subclinicalfor years, but long-term follow ups revealed severe complica-tions. Tamarins physically debilitated by T. cruzi infection may havereduced lifetimes and reduced ability to get food and reproduce

pica 107 (2008) 168–173

(Monteiro et al., 2006). Despite there are no reports of pathogenicityfor mammals, the very high prevalences of T. rangeli in Amazo-nian wild primates indicate that the course of infections shouldbe carefully assessed in these animals.

Besides helping to design strategies to protect the species, theunderstanding of the transmission dynamics of the trypanosomesinfecting S. bicolor could help to evaluate the role of primates asreservoirs of human trypanosomes in the Amazon region. Theirtransmission cycles take place in palm trees and other animalrefuges inhabited by triatomine bugs in close proximity of humandwellings. Deforestation and other anthropogenic alterations ofwildlife habitats can change population dynamics, the behaviourand the geographic range of reservoirs and vectors of parasites,allowing emergence of human diseases of wildlife origin (Brooksand Ferrao, 2005; Lebarbenchon et al., 2008). Occurrence of Cha-gas disease in Amazonia is a current source of public health concernbecause of the wide dispersion of infected vectors, intensive humanmigration into the region and increasing number of human cases(Coura et al., 2002; Aguilar et al., 2007). Further epidemiologicalstudies are necessary to establish the real impact of trypanosomeinfections on wildlife preservation and the presumed risks for thehealth of humans and wild primates.

Acknowledgements

This work was funding by the Brazilian agency CNPq (ConselhoNacional de Pesquisa) and by the Wellcome Trust, UK. We are deeplyindebted to several collaborators for their inestimable help in thefieldwork (Projeto Sauim-de-Coleira) supported by PROBIO/MMA,FNMA/MMA, WCS, Apenheul Primate Conservation Trust, Shal-don Wildlife Trust, and Philadelphia Zoo. Flavia Maia da Silva andArlei Marcili are sponsored by CAPES (PRODOC) and CNPq, respec-tively.

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