First report of Sapajus cay naturally infected by Trypanosoma cruzi in San Pedro Department, Paraguay / Primeiro relato de infecção natural em Sapajus cay por Trypanosoma cruzi em Departamento San Pedro, Paraguai

Abstract To verify the occurrence of natural Trypanosoma cruzi infection in non-human primates from a rural endemic area of the east region of Paraguay, xenodiagnosis was performed in 35 animals belonging to two species. For genotyping and T. cruzi discrete typing unit (DTU) assignment, a combination of four markers was used, including amplification products of the small (18S) and large (24Sα) subunits of ribosomal ribonucleic acid gene, the intergenic region of mini-exon gene and the heat shock protein 60 Eco-RV polymerase chain reaction-restriction fragment length polymorphism (HSP60/EcoRV-PCR-RFLP). One specimen of Sapajus cay was found positive and infected by the DTU TcII. This result constitutes the first record of natural T. cruzi infection in a sylvatic monkey in Paraguay, harbouring a DTU associated with severe Chagas disease in humans.

Resumo Com o objetivo de verificar a infecção natural por Trypanosoma cruzi em primatas não-humanos de uma área endêmica rural da região leste do Paraguai, foi realizado o xenodiagnóstico em 35 animais pertencentes a duas espécies. Para a genotipagem foi utilizada a unidade discreta de tipagem (UDT) do T. cruzi, em uma combinação de quatro marcadores, incluindo amplificação de produtos de pequena (18S) e grande (24Sα) subunidades do gene do ácido ribonucleico ribossômico, da região intergênica de miniéxon e do gene da proteína de choque térmico 60 (HSP60/EcoRV-PCR-RFLP), pela reação em cadeia da Polimerase. Um espécime de Sapajus cay se mostrou positivo pelo UDT TcII. Este resultado constitui o primeiro relato da infecção natural pelo T. cruzi em um macaco silvestre no Paraguai, abrigando um UDT associado com a doença de Chagas grave em humanos.

SEROLOGICAL DETECTION OF HEPATITIS A VIRUS IN FREE-RANGING NEOTROPICAL PRIMATES (Sapajus spp., Alouatta caraya) FROM THE PARANÁ RIVER BASIN, BRAZIL

Nonhuman primates are considered as the natural hosts of Hepatitis A virus (HAV), as well as other pathogens, and can serve as natural sentinels to investigate epizootics and endemic diseases that are of public health importance. During this study, blood samples were collected from 112 Neotropical primates (NTPs) (Sapajus nigritus and S. cay, n = 75; Alouatta caraya, n = 37) trap-captured at the Paraná River basin, Brazil, located between the States of Paraná and Mato Grosso do Sul. Anti-HAV IgG antibodies were detected in 4.5% (5/112) of NTPs, specifically in 6.7% (5/75) of Sapajus spp. and 0% (0/37) of A. caraya. In addition, all samples were negative for the presence of IgM anti-HAV antibodies. These results suggest that free-ranging NTPs were exposed to HAV within the geographical regions evaluated.

Serological evidence for Saint Louis encephalitis virus in free-ranging New World monkeys and horses within the upper Paraná River basin region, Southern Brazil

Introduction Saint Louis encephalitis virus (SLEV) primarily occurs in the Americas and produces disease predominantly in humans. This study investigated the serological presence of SLEV in nonhuman primates and horses from southern Brazil. Methods From June 2004 to December 2005, sera from 133 monkeys (Alouatta caraya, n=43; Sapajus nigritus, n=64; Sapajus cay, n=26) trap-captured at the Paraná River basin region and 23 blood samples from farm horses were obtained and used for the serological detection of a panel of 19 arboviruses. All samples were analyzed in a hemagglutination inhibition (HI) assay; positive monkey samples were confirmed in a mouse neutralization test (MNT). Additionally, all blood samples were inoculated into C6/36 cell culture for viral isolation. Results Positive seroreactivity was only observed for SLEV. A prevalence of SLEV antibodies in sera was detected in Alouatta caraya (11.6%; 5/43), Sapajus nigritus (12.5%; 8/64), and S. cay (30.8%; 8/26) monkeys with the HI assay. Of the monkeys, 2.3% (1/42) of A. caraya, 6.3% 94/64) of S. nigritus, and 15.4% (4/26) of S. cay were positive for SLEV in the MNT. Additionally, SLEV antibodies were detected by HI in 39.1% (9/23) of the horses evaluated in this study. Arboviruses were not isolated from any blood sample. Conclusions These results confirmed the presence of SLEV in nonhuman primates and horses from southern Brazil. These findings most likely represent the first detection of this virus in nonhuman primates beyond the Amazon region. The detection of SLEV in animals within a geographical region distant from the Amazon basin suggests that there may be widespread and undiagnosed dissemination of this disease in Brazil. .

First report of yellow fever virus in non-human primates in the State of Parana, Brazil

Sylvatic yellow fever is a zoonosis associated mainly with wild animals, especially those in the genus Alouatta, that act as the source of infection. Once infected, these animals pass the disease on to humans by way of an infected mosquito belonging to the genera Aedes, Haemagogus, or Sabethes. The present study is the first report of a case of yellow fever in non-human primates (NHP) in the State of Paraná, Brazil. After the case was diagnosed, several prophylactic measures were adopted to prevent outbreaks of the disease in humans.

Reemergence of yellow fever: detection of transmission in the State of São Paulo, Brazil, 2008 / Reemergência de febre amarela: detecção de transmissão no Estado de São Paulo, Brasil, 2008

INTRODUCTION: Following yellow fever virus (YFV) isolation in monkeys from the São José do Rio Preto region and two fatal human autochthonous cases from the Ribeirão Preto region, State of São Paulo, Brazil, two expeditions for entomological research and eco-epidemiological evaluation were conducted. METHODS: A total of 577 samples from humans, 108 from monkeys and 3,049 mosquitoes were analyzed by one or more methods: virus isolation, ELISA-IgM, RT-PCR, histopathology and immunohistochemical. RESULTS: Of the 577 human samples, 531 were tested by ELISA-IgM, with 3 positives, and 235 were inoculated into mice and 199 in cell culture, resulting in one virus isolation. One sample was positive by histopathology and immunohistochemical. Using RT-PCR, 25 samples were processed with 4 positive reactions. A total of 108 specimens of monkeys were examined, 108 were inoculated into mice and 45 in cell culture. Four virus strains were isolated from Alouattacaraya. A total of 931 mosquitoes were captured in Sao Jose do Rio Preto and 2,118 in Ribeirão Preto and separated into batches. A single isolation of YFV was derived from a batch of 9 mosquitoes Psorophoraferox, collected in Urupês, Ribeirão Preto region. A serological survey was conducted with 128 samples from the municipalities of São Carlos, Rincão and Ribeirão Preto and 10 samples from contacts of patients from Ribeirão Preto. All samples were negative by ELISA-IgM for YFV. CONCLUSIONS: The results confirm the circulation of yellow fever, even though sporadic, in the Sao Paulo State and reinforce the importance of vaccination against yellow fever in areas considered at risk.

INTRODUÇÃO: A partir do isolamento do vírus febre amarela (VFA), de macacos, da região de São José do Rio Preto e de dois casos humanos autóctones fatais, da região de Ribeirão Preto, Estado de São Paulo, foram realizadas duas expedições para pesquisa entomológica e avaliação ecoepidemiológica. MÉTODOS: Um total de 577 amostras de humanos, 108 de macacos e 3.049 mosquitos foram analisados por um ou mais métodos: isolamento viral, ELISA-IgM, RT-PCR, histopatologia e imunohistoquímica. RESULTADOS: De 577 amostras humanas, 531 foram testadas por ELISA-IgM, sendo 3 positivas, 235 foram inoculadas em camundongos, 199 em cultura de células, obtendo-se 1 isolamento viral. Uma amostra foi positiva por histopatologia e imunohistoquímica. Por RT-PCR foram processadas 25 amostras com 4 reações positivas. Os 108 espécimes de macacos foram inoculados em camundongos, 45 em cultura de células, obtendo-se 4 isolamentos de VFA, de Alouatta caraya. Um total de 931 mosquitos foram capturados em São José do Rio Preto e 2.118 em Ribeirão Preto e separados em lotes. Um único isolamento de VFA foi derivado de um lote de 9 mosquitos Psorophora ferox, coletados em Urupês, região de Ribeirão Preto. Um inquérito sorológico foi realizado com 128 amostras dos municípios de São Carlos, Rincão e Ribeirão Preto e mais 10 amostras de contactantes de pacientes de Ribeirão Preto. Todas as amostras foram negativas por ELISA-IgM para VFA. CONCLUSÕES: Os resultados confirmam a circulação, mesmo que esporádica, do VFA no Estado de São Paulo e reforça a importância da vacinação antiamarílica nas áreas consideradas de risco.

Risco de transmissão do vírus da raiva oriundo de sagui (Callithrix jacchus), domiciliado e semidomiciliado, para o homem na região metropolitana de Fortaleza, estado do Ceará / Risks of transmitting rabies virus from captive domiciliary common marmoset (Callithrix jacchus) to human beings, in the metropolitan region of Fortaleza, state of Ceará, Brazil

INTRODUÇÃO: Uma variante do vírus da raivafoi identificadaem associação a casos de raiva humanos, no Estado do Ceará, transmitidos por saguis (Callithrix jacchus), primatas frequentemente criados como animais de estimação. Essa variante não apresenta proximidade antigênica ou relação genética com as variantes do vírus encontradas em morcegos e mamíferos terrestres das Américas. O objetivo do estudo foi avaliar os fatores de risco de transmissão do vírus da raiva oriundo de sagui (C. jacchus), criado como animal de estimação, para o homem na região metropolitana de Fortaleza, Ceará. MÉTODOS: Foi aplicado um questionário estruturado aos criadores de saguis, residentes nos municípios de Aquiraz e Maranguape, Ceará, enfocando o manejo e a interação desses primatas com humanos. Para avaliação da ocorrência de antígenos rábicos, através do teste de imunofluorescência direta (IFD), foram coletadas amostras de saliva dos saguis domiciliados e semidomiciliados. Com base nos resultados obtidos desses espécimes, foram analisadas amostras de sistema nervoso central (SNC). RESULTADOS: Na análise dos questionários, observou-se a proximidade dos criadores de saguis durante o manejo desses animais nos domicílios, bem como, seus conhecimentos limitados sobre a raiva, demonstrando haver risco quanto à transmissão do vírus. De 29 amostras de saliva de saguis reavaliadas, uma (3,4 por cento) apresentou reação de IFD positiva. De 11 amostras de SNC, três (27,3 por cento) apresentaram positividade. CONCLUSÕES: Os dados laboratoriais estão de acordo com os achados dos questionários, confirmando haver risco da transmissão do vírus da raiva devido à convivência de humanos com saguis (C. jacchus).

INTRODUCTION: In the State of Ceará, a new variant of the rabies virus was identified associated with cases of human rabies transmitted by common marmosets (Callithrix jacchus), which are frequently kept as pets. This new variant does not present antigenic proximity or genetic relationship to variants of the virus isolated from bats and terrestrial mammals from the American continent. The present study aimed to evaluate the risk factors of rabies virus transmission from common marmosets (C. jacchus) maintained as pets in the metropolitan region of Fortaleza, State of Ceará, Brazil, to human beings. METHODS: A questionnaire focusing on animal management and interaction between humans and primates was applied to individuals who had marmosets in the municipalities of Aquiraz and Maranguape. In order to evaluate the presence of rabies antigens by direct immunofluorescence test (DIF), samples of saliva were collected from domiciliary captive marmosets. Based on the detection of rabies antigens, biopsy samples of central nervous system (CNS) were analyzed. RESULTS: Analysis of questionnaire data verified that a close relation exists between humans and their pet marmosets, especially during management practices. Additionally, these people showed minimal knowledge regarding rabies, which represents a greater risk of infection. Of the 29 saliva samples evaluated, one (3.4 percent) was positive for DIF reaction and of the 11 CNS samples, three (27.3 percent) were positive. CONCLUSIONS: Laboratory data are in agreement with the questionnaire findings, which confirm an increased risk of rabies virus transmission due to the close relation between humans and marmosets.

Circulation of antibodies against yellow fever virus in a simian population in the area of Porto Primavera Hydroelectric Plant, São Paulo, Brazil / Circulação de anticorpos contra o vírus amarílico em população simiana da região da usina hidrelétrica de Porto Primavera, São Paulo, Brasil

Yellow fever (YF) is an acute viral infectious disease transmitted by mosquitoes which occurs in two distinct epidemiological cycles: sylvatic and urban. In the sylvatic cycle, the virus is maintained by monkey's infection and transovarian transmission in vectors. Surveillance of non-human primates is required for the detection of viral circulation during epizootics, and for the identification of unaffected or transition areas. An ELISA (enzyme-linked immunosorbent assay) was standardized for estimation of the prevalence of IgG antibodies against yellow fever virus in monkey sera (Alouatta caraya) from the reservoir area of Porto Primavera Hydroelectric Plant, in the state of São Paulo, Brazil. A total of 570 monkey sera samples were tested and none was reactive to antibodies against yellow fever virus. The results corroborate the epidemiology of yellow fever in the area. Even though it is considered a transition area, there were no reports to date of epizootics or yellow fever outbreaks in humans. Also, entomological investigations did not detect the presence of vectors of this arbovirus infection. ELISA proved to be fast, sensitive, an adequate assay, and an instrument for active search in the epidemiological surveillance of yellow fever allowing the implementation of prevention actions, even before the occurrence of epizootics.

A febre amarela (FA) é doença infecciosa aguda de origem viral transmitida por mosquitos. No ciclo silvestre, o vírus é mantido por meio da infecção de macacos e da transmissão transovariana nos vetores. A vigilância sobre populações de primatas não humanos torna-se necessária para detectar a circulação viral, quando ainda está restrito a epizootias, e para determinar sua presença em regiões indenes ou de transição para a doença. Padronizou-se a técnica ELISA (Enzyme Linked Immunosorbent Assay) para determinar a prevalência de anticorpos da classe IgG contra o vírus da FA em soros de bugios (Alouatta caraya) da região do reservatório da Usina Hidrelétrica de Porto Primavera, SP. Foram testados soros de 570 macacos sendo que nenhuma amostra mostrou-se reativa para a presença de anticorpos contra o vírus da FA. Os resultados são coerentes com a epidemiologia da FA na região. Mesmo sendo área de transição, não se conhece, até o momento, ocorrência de epizootia ou surto de FA em humanos e investigações entomológicas não apontaram a presença de vetores para esta arbovirose. A técnica mostrou-se sensível, rápida e útil à vigilância epidemiológica como instrumento de busca ativa permitindo desencadear ações preventivas, como vacinação, antes mesmo do surgimento de epizootias.

Circulation of Campylobacter spp. in rhesus monkeys (Macaca mulatta) held in captivity: a longitudinal study

Campylobacteriosis is an extremely important zoonosis, circulating freely in the environment. In nonhuman primates kept in open facilities and bred for experimental purposes, the presence of Campylobacter spp. could cause severe damage to the production and interfere with the results of scientific research. In this paper, we assessed the circulation of Campylobacter spp. in a colony of clinically healthy rhesus monkeys (Macaca mulatta) destined to research. The analysis was carried out during seven non-consecutive years. Data showed that despite several changes made in animal management along the studied years in order to control this zoonosis, reduction of bacterial charge did not occur. Significant differences among the age groups and sex were observed. Infants showed higher susceptibility than adult animals. In general males were more infected than females. Modifications adopted in the handling techniques need to be reviewed with the intent of improving the production, reducing bacterial infection of the stock and avoiding undesirable cross reactions in the research carried out with these animals. Therefore, this paper alerts professionals that work directly with captive rhesus monkeys about the risks of Campylobacter spp. infection and possible interference on the experimental procedures.

Leptospira spp detection by Polymerase Chain Reaction (PCR) in clinical samples of captive black-capped Capuchin monkey (Cebus apella)

Leptospirosis is a widely distributed zoonosis that affects domestic and wild animals, and that has the man as the end point of its epidemiological chain. Leptospirosis diagnosis in primates is more difficult than in other animal species, as clinical signs and lesions are less evident and antibody response is detected only for short periods. The aim of this article was to describe the detection of Leptospira spp using polymerase chain reaction (PCR), in clinical samples from one captive black-capped Capuchin monkey (Cebus apella), which presented characteristics compatible with leptospirosis (jaundice and haemorrhagic kdney) in the macroscopic post-mortem examination. A friable kidney fragment and urine sample were cultured and submitted to experimental inoculation in guinea pigs and PCR using genus specific primer pair targeting the 16S rRNA region from Leptospira interrogans serovar canicola. Isolation of the agent was negative both in culture and experimental inoculation. The PCR amplification of the clinical samples showed a 330 pb amplified fragment that corresponds to the Leptospira genus. Based on these results PCR was considered an important tool for leptospira detection in nonhumam primates, more sensitive and specific than other techniques, especially considering that the viability of the pathogen was not possible. These advantages enable the detection of the leptospiras in urine and kidney, even when autolysed, frozen or badly conserved, which prevented the isolation and experimental inoculation from positive results.

Health problems of Callithrix jacchus in captivity

A 20-year retrospective study was carried out to determine the prevalence of diseases occurring in 265 (155 males and 110 females) common marmosets (Callithrix jacchus), kept at the Säo Paulo Zoo. A total of 564 clinical disorders were detected: digestive problems, 50.2percent (protozoa, 44.3 percent; helminths, 26.3 percent; bacteria, 7.0 percent; non-especific causes, 22.4 percent); respiratory problems (pneumonia), 16.3 percent; injuries, 13.8 percent; nutritional deficiency, 4.4 percent; reproductive problems (obstetrical) 2.5 percent; septicemia, 1.6 percent; circulatory problems, 1.6 percent; sporadic cases, 4.0 percent; inconclusive cases, 5.5 percent. Little attention has been paid to the sanitary care of marmosets in Brazil. Since most of the available information comes from the international literature, the occurrence of major diseases and their etiological agents in Brazil are relevant data

Infecçäo experimental de macacos Cebus apella sp pelo Trypanosoma cruzi: avaliaçäo clínica, eletrocardiográfica e anatomopatológica / The experimental infection of Cebus apella sp by Trypanosoma cruzi: its clinical electrocardiography and anatomopathological assessment

Dos trinta e dois macacos capturados no interior do Estado de Säo Paulo e mantidos em laboratório em gaiolas individuais (24 a 25 Cº e 70 por cento de umidificaçäo) após vários xenodiagnósticos negativos, 12 foram infectados por via intraperitoneal com diferentes cepas de Trypanosoma cruzi, cujas formas tripomastigotas injetadas variaram de 1.10**5 a 5.10**6. Os 20 macacos restantes foram mantidos como controle. No período de 1 a 6 anos tanto animais inoculados como os näo inoculados, foram submetidos a xenodiagnóstico e teste sorológico de aglutinaçäo direta, exame clínico e o eletrocardiograma. Posteriormente os macacos foram necropsiados e todos os órgäos submetidos a exame macro e microscópico. O exame clínicos e o eletrocardiograma näo revelaram alteraçöes. Dos 12 macaços infectados, 4 apresentaram evidências de infecçäo ao exame histopatológico: um com formas amastigotas nos tecidos e 3 com miocardite crônica de grau leve. A parasitemia foi comprovada em 66,66 por cento dos animais na fase aguda e a sorologia em 91,66 por cento na fase crônica. Conclui-se que os macacos Cebus näo expressaram suscepitibilidade ao desenvolvimento das lesöes que caracterizam a fase crônica da doença de Chagas mas poderiam ser usados para manter as cepas de T. cruzi e estudos de pesquisa sorológico a longo termo

Seroepidemiological survey on dengue and Japanese encephalitis virus infections in Asian monkeys.

To investigate the ecology of dengue and Japanese encephalitis (JE) viruses in the forest in Asia, a seroepidemiological survey was carried out on 358 Southeast Asian cynomolgus (Macaca iris), 33 Indian bonnet (Macaca radiata) and 37 Japanese (Macaca fuscata) monkey sera by a plaque reduction neutralization test. The results indicated that Southeast Asian monkeys were naturally infected with these viruses but the frequency of antibody to them varied considerably according to the geographical origin of the monkeys. The frequency of antibody to one or more types of dengue virus were 87.2, 49.5, 34.3, 34.2 and 14.9% in Malaysian, Vietnamese, Cambodian, Indonesian and Filipino cynomolgus monkey sera, respectively. None of the Indian bonnet monkey sera neutralized type 1 dengue virus which was the only virus type examined with this monkey species. Monkey sera collected in Japan where dengue virus infection had not been known since 1944 did not significantly neutralize dengue viruses. JE virus antibody was detected at 29.7, 9.0, 8.6, 2.7, 1.4 and 0% in Japanese, Cambodian, Vietnamese, Indonesian, Filipino and Malaysian monkey sera respectively.