Equine infectious anemia virus in naturally infected horses from the Brazilian Pantanal.

Equine infectious anemia (EIA) has a worldwide distribution, and is widespread in Brazil. The Brazilian Pantanal presents with high prevalence comprising equine performance and indirectly the livestock industry, since the horses are used for cattle management. Although EIA is routinely diagnosed by the agar gel immunodiffusion test (AGID), this serological assay has some limitations, so PCR-based detection methods have the potential to overcome these limitations and act as complementary tests to those currently used. Considering the limited number of equine infectious anemia virus (EIAV) sequences which are available in public databases and the great genome variability, studies of EIAV detection and characterization molecular remain important. In this study we detected EIAV proviral DNA from 23 peripheral blood mononuclear cell (PBMCs) samples of naturally infected horses from Brazilian Pantanal using a semi-nested-PCR (sn-PCR). The serological profile of the animals was also evaluated by AGID and ELISA for gp90 and p26. Furthermore, the EIAV PCR amplified DNA was sequenced and phylogenetically analyzed. Here we describe the first EIAV sequences of the 5' LTR of the tat gene in naturally infected horses from Brazil, which presented with 91% similarity to EIAV reference sequences. The Brazilian EIAV sequences also presented variable nucleotide similarities among themselves, ranging from 93,5% to 100%. Phylogenetic analysis showed that Brazilian EIAV sequences grouped in a separate clade relative to other reference sequences. Thus this molecular detection and characterization may provide information about EIAV circulation in Brazilian territories and improve phylogenetic inferences.

Neutralising antibodies for West Nile virus in horses from Brazilian Pantanal.

Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for WNV and its most closely-related flaviviruses that circulate in Brazil to confirm the detection of specific virus-neutralising antibodies. Of the 93 (55.4%) blocking ELISA-seropositive horse serum samples, five (3%) were seropositive for WNV, nine (5.4%) were seropositive for St. Louis encephalitis virus, 18 (10.7%) were seropositive for Ilheus virus, three (1.8%) were seropositive for Cacipacore virus and none were seropositive for Rocio virus using PRNT90, with a criteria of ≥ four-fold antibody titre difference. All caimans were negative for flaviviruses-specific antibodies using the blocking ELISA. No virus genome was detected from caiman blood or mosquito samples. The present study is the first report of confirmed serological evidence of WNV activity in Brazil.

Detection of Ehrlichia sp. in Amblyomma sculptum parasitizing horses from Brazilian Pantanal wetland.

Bacteria of the genus Ehrlichia are transmitted by ticks and also are an important cause of infection in wild and domestic mammals. Infection with Ehrlichia spp. has been reported in horses, especially in the USA, Nicaragua and Brazil. In this study, we report the parasitism by Amblyomma sculptum, Rhipicephalus microplus and Dermacentor nitens ticks in horses from a ranch located in south Pantanal wetland. Molecular and serological analyzes to determine infection by Ehrlichia spp. in horses and their respective ticks were carried out. A total of 12 horses were submitted to blood collection to investigate antibodies by indirect immunofluorescence assay (IFA) using Ehrlichia canis crude antigens and to be tested by polymerase chain reaction (PCR) in order to amplify fragments of the 16S rRNA, dsb, groEL and sodB gene of Ehrlichia spp. A total of 164 tick specimens were removed from horses, stored in isopropanol and later identified as D. nitens, A. sculptum and R. microplus. DNA from ticks were extracted and subjected to the same PCR assays to detect Ehrlichia spp. Anti-Ehrlichia spp. antibodies were detected in five/12 (41.7 %) horses by IFA, with antibody titers ranging from 40 to 160. All horse DNA samples were negative for the 16S rRNA, dsb, groEL and sodB of Ehrlichia spp. One A. sculptum female was positive to all target genes of Ehrlichia. This tick was parasitizing an Ehrlichia-seropositive horse with antibody titer of 80. Nucleotide sequences of 16S rRNA, dsb, groEL and sodB genes showed close relationship with different strains of Ehrlichia detected in wild mammals, Amblyomma ticks and horses from Brazil and Argentina. Detection of anti-Ehrlichia sp. antibodies suggests that horses have been exposed to an ehrlichial agent in the Pantanal. Future studies on Ehrlichia infection should be carried out to better elucidate and to bring new information about equine ehrlichiosis, since these animals are important hosts of ticks in the Brazilian Pantanal wetlands.

Identification of large genetic variations in the equine infectious anemia virus tat-gag genomic region.

The aetiological agent of equine infectious anaemia (EIA) is the retrovirus equine infectious anemia virus (EIAV) that infects all members of the Equidae family. The EIA is widely disseminated in the Brazilian territory with a high seroprevalence in the Brazilian Pantanal and is mainly diagnosed using agar gel immunodiffusion (AGID). There are few complete EIAV genome sequences available in GenBank, which had an impact on molecular detection studies. In this study, we conducted molecular detection and sequencing of EIAV proviral DNA from Brazilian horses. We analysed the genomic region from exon 1 of tat to gag (tat-gag). Comparative serological tests, comprising AGID and two enzyme-linked immunosorbent assays (ELISAs), were also conducted. Of the 133 samples, 58 were positive in the tat-gag PCR, and 49 nucleotide sequences of 272 bp were obtained. Using this developed tat-gag PCR EIAV proviral DNA was detected in 7% of the AGID-negative samples and 26% of the AGID-negative samples were positive in at least one of the ELISA tests used. Using phylogenetic analysis, the Brazilian Pantanal EIAV sequences grouped in a different clade of EIAV sequences from other countries. Thus, the EIAV sequences can contribute to the knowledge of the tat-gag genomic region in the circulating viruses in the Brazilian Pantanal, in addition to providing new information about the genetic diversity. In addition, the serological results demonstrate the greater sensitivity of the ELISAs used in this study compared to AGID for EIA diagnosis.

Serologic evidence of the recent circulation of Saint Louis encephalitis virus and high prevalence of equine encephalitis viruses in horses in the Nhecolândia sub-region in South Pantanal, Central-West Brazil.

As in humans, sub-clinical infection by arboviruses in domestic animals is common; however, its detection only occurs during epizootics and the silent circulation of some arboviruses may remain undetected. The objective of the present paper was to assess the current circulation of arboviruses in the Nhecolândia sub-region of South Pantanal, Brazil. Sera from a total of 135 horses, of which 75 were immunized with bivalent vaccine composed of inactive Eastern equine encephalitis virus (EEEV) and Western equine encephalitis virus(WEEV) and 60 were unvaccinated, were submitted to thorough viral isolation, reverse transcriptase polymerase chain reaction (RT-PCR) and neutralization tests for Saint Louis encephalitis virus (SLEV), EEEV, WEEV and Mayaro virus (MAYV). No virus was isolated and viral nucleic-acid detection by RT-PCR was also negative. Nevertheless, the prevalence of neutralizing antibodies in horses older than seven months was 43.7% for SLEV in equines regardless of vaccine status, and 36.4% for WEEV and 47.7% for EEEV in unvaccinated horses. There was no evidence of MAYV infections. The serologic evidence of circulation of arboviruses responsible for equine and human encephalitis, without recent official reports of clinical infections in the area, suggests that the Nhecolândia sub-region in South Pantanal is an important area for detection of silent activity of arboviruses in Brazil.

High Genomic Variability in Equine Infectious Anemia Virus Obtained from Naturally Infected Horses in Pantanal, Brazil: An Endemic Region Case.

Equine infectious anemia virus (EIAV) is a persistent lentivirus that causes equine infectious anemia (EIA). In Brazil, EIAV is endemic in the Pantanal region, and euthanasia is not mandatory in this area. All of the complete genomic sequences from field viruses are from North America, Asia, and Europe, and only proviral genomic sequences are available. Sequences from Brazilian EIAV are currently available only for gag and LTR regions. Thus, the present study aimed for the first time to sequence the entire EIAV genomic RNA in naturally infected horses from an endemic area in Brazil. RNA in plasma from naturally infected horses was used for next-generation sequencing (NGS), and gaps were filled using Sanger sequencing methodology. Complete viral genomes of EIAV from two horses were obtained and annotated (Access Number: MN560970 and MN560971). Putative genes were analyzed and compared with previously described genes, showing conservation in gag and pol genes and high variations in LTR and env sequences. Amino acid changes were identified in the p26 protein, one of the most common targets used for diagnosis, and p26 molecular modelling showed surface amino acid alterations in some epitopes. Brazilian genome sequences presented 88.6% nucleotide identity with one another and 75.8 to 77.3% with main field strains, such as EIAV Liaoning, Wyoming, Ireland, and Italy isolates. Furthermore, phylogenetic analysis suggested that this Brazilian strain comprises a separate monophyletic group. These results may help to better characterize EIAV and to overcome the challenges of diagnosing and controlling EIA in endemic regions.

Toxoplasmose ovina e equina: ocorrência de anticorpos em animais criados em centro de pesquisa na região do pantanal Sul Mato-Grossense, Brasil / Ovine and equine toxoplasmosis: occurrence of antibodies in animals raised in a research center in the pantanal region of Mato Grosso do Sul, Brazil / Toxoplasmosis ovina y equina: ocurrencia de anticuerpos en animales criados en un centro de investigación en la región del pantanal en el Mato Grosso del Sur, Brasil

A toxoplasmose é uma zoonose de distribuição mundial que acomete vertebrados homeotérmicos, incluindo o ser humano. Toxoplasma gondii, seu agente etiológico, é capaz de infectar seus hospedeiros por diversas vias de transmissão, dentre elas a ingestão de cistos teciduais, presente na carne crua ou malcozida, e a ingestão de oocistos, presentes na água e alimentos contaminados, sendo essas as principais rotas de infecção na cadeia epidemiológica da toxoplasmose. Neste sentido, a contaminação ambiental com oocistos esporulados apresenta-se como um importante fator de risco para a infecção toxoplásmica em animais herbívoros, como os equinos e ovinos. Enquanto os equinos mostram resistência à infecção e a manifestações de sinais clínicos da doença, em ovinos, a toxoplasmose pode ser responsável por importantes distúrbios reprodutivos como aborto e morte neonatal. Sendo assim, este estudo teve como objetivo pesquisar a presença de anticorpos específicos anti- T. gondii em equinos e ovinos criados na Fazenda Experimental Nhumirim, propriedade da Embrapa Pantanal, Mato Grosso do Sul, Brasil. Para tanto foram coletadas amostras de soro de 101 equinos e 99 ovinos entre março de 2011 e maio de 2015. As amostras de soro foram enviadas para o Laboratório de Toxoplasmose e outras Protozooses, do Instituto Oswaldo Cruz, na Fundação Oswaldo Cruz/RJ. Os soros foram submetidos à reação de imunofluorescência indireta, sendo consideradas positivas as amostras sororreagentes a partir da diluição de 1:64 para ambas as espécies. Anticorpos anti-T. gondii foram detectados em 15% (15/99) dos ovinos e 10% (10/101) dos equinos. A ocorrência de anticorpos específicos anti-T. gondii nas amostras de soro desses animais, aponta para uma contaminação ambiental da propriedade, onde possivelmente, felídeos silvestres estariam atuando como dispersores do parasito...(AU)

Toxoplasmosis is a worldwide zoonosis that affects homeothermic vertebrates, including humans. Toxoplasma gondii, its etiological agent, is capable of infecting its hosts through several transmission routes, including the ingestion of tissue cysts, present in raw or undercooked meat, and the ingestion of oocysts, present in contaminated water and food, being these are the main routes of infection in the epidemiological chain of toxoplasmosis. In this sense, environmental contamination with sporulated oocysts is an important risk factor for toxoplasmic infection in herbivorous animals, such as horses and sheep. While horses show resistance to infection and manifestations of clinical signs of the disease, in sheep, toxoplasmosis may be responsible for important reproductive disorders such as abortion and neonatal death. Therefore, this study aimed to investigate the presence of specific anti-T. gondii antibodies in horses and sheep raised on the Experimental Farm Nhumirim, property of Embrapa Pantanal, Mato Grosso do Sul, Brazil. For this purpose, serum samples were collected from 101 horses and 99 sheep between March 2011 and May 2015. The serum samples were sent to the Toxoplasmosis and other Protozoal Diseases Laboratory, at Oswaldo Cruz Institute, at Oswaldo Cruz Foundation/RJ. Sera were subjected to indirect immunofluorescence test, and seroreactive samples were considered positive from a 1:64 dilution for both species. Anti-T. gondii were detected in 15% (15/99) of sheep and 10% (10/101) of horses. The occurrence of specific anti-T. gondii in the serum samples of these animals, points to an environmental contamination of the property, where possibly wild felids would be acting as dispersers of the parasite...(AU)

La toxoplasmosis es una zoonosis de distribución mundial que afecta a los vertebrados homeotérmicos, incluidos los humanos. Toxoplasma gondii, su agente etiológico, es capaz de infectar a sus huéspedes a través de varias vías de transmisión, entre ellas la ingestión de quistes tisulares, presentes en carnes crudas o poco cocidas, y la ingestión de ooquistes, presentes en agua y alimentos contaminados, siendo estos las principales vías de infección en la cadena epidemiológica de la toxoplasmosis. En este sentido, la contaminación ambiental con ooquistes esporulados es un factor de riesgo importante para la infección por toxoplasma en animales herbívoros, como caballos y ovejas. Mientras que los caballos muestran resistencia a la infección y las manifestaciones de los signos clínicos de la enfermedad, en las ovejas, la toxoplasmosis puede ser responsable de importantes trastornos reproductivos como el aborto y la muerte neonatal. Por lo tanto, este estudio tuvo como objetivo investigar la presencia de anticuerpos específicos anti-T. gondii en caballos y ovejas criados en la Granja Experimental Nhumirim, propiedad de Embrapa Pantanal, Mato Grosso do Sul, Brasil. Para ello, se recolectaron muestras de suero de 101 caballos y 99 ovejas entre marzo de 2011 y mayo de 2015. Las muestras de suero se enviaron al Laboratorio de Toxoplasmosis y otros Protozoos, del Instituto Oswaldo Cruz, en la Fundação Oswaldo Cruz / RJ. Los sueros se sometieron a una reacción de inmunofluorescencia indirecta y las muestras serorreactivas se consideraron positivas a partir de una dilución 1:64 para ambas especies. Anti-T. gondii se detectaron en el 15% (15/99) de las ovejas y el 10% (10/101) de los caballos...(AU)

Toxoplasmose ovina e equina: ocorrência de anticorpos em animais criados em centro de pesquisa na região do pantanal Sul Mato-Grossense, Brasil / Ovine and equine toxoplasmosis: occurrence of antibodies in animals raised in a research center in the pantanal region of Mato Grosso do Sul, Brazil / Toxoplasmosis ovina y equina: ocurrencia de anticuerpos en animales criados en un centro de investigación en la región del pantanal en el Mato Grosso del Sur, Brasil

A toxoplasmose é uma zoonose de distribuição mundial que acomete vertebrados homeotérmicos, incluindo o ser humano. Toxoplasma gondii, seu agente etiológico, é capaz de infectar seus hospedeiros por diversas vias de transmissão, dentre elas a ingestão de cistos teciduais, presente na carne crua ou malcozida, e a ingestão de oocistos, presentes na água e alimentos contaminados, sendo essas as principais rotas de infecção na cadeia epidemiológica da toxoplasmose. Neste sentido, a contaminação ambiental com oocistos esporulados apresenta-se como um importante fator de risco para a infecção toxoplásmica em animais herbívoros, como os equinos e ovinos. Enquanto os equinos mostram resistência à infecção e a manifestações de sinais clínicos da doença, em ovinos, a toxoplasmose pode ser responsável por importantes distúrbios reprodutivos como aborto e morte neonatal. Sendo assim, este estudo teve como objetivo pesquisar a presença de anticorpos específicos anti- T. gondii em equinos e ovinos criados na Fazenda Experimental Nhumirim, propriedade da Embrapa Pantanal, Mato Grosso do Sul, Brasil. Para tanto foram coletadas amostras de soro de 101 equinos e 99 ovinos entre março de 2011 e maio de 2015. As amostras de soro foram enviadas para o Laboratório de Toxoplasmose e outras Protozooses, do Instituto Oswaldo Cruz, na Fundação Oswaldo Cruz/RJ. Os soros foram submetidos à reação de imunofluorescência indireta, sendo consideradas positivas as amostras sororreagentes a partir da diluição de 1:64 para ambas as espécies. Anticorpos anti-T. gondii foram detectados em 15% (15/99) dos ovinos e 10% (10/101) dos equinos. A ocorrência de anticorpos específicos anti-T. gondii nas amostras de soro desses animais, aponta para uma contaminação ambiental da propriedade, onde possivelmente, felídeos silvestres estariam atuando como dispersores do parasito...

Toxoplasmosis is a worldwide zoonosis that affects homeothermic vertebrates, including humans. Toxoplasma gondii, its etiological agent, is capable of infecting its hosts through several transmission routes, including the ingestion of tissue cysts, present in raw or undercooked meat, and the ingestion of oocysts, present in contaminated water and food, being these are the main routes of infection in the epidemiological chain of toxoplasmosis. In this sense, environmental contamination with sporulated oocysts is an important risk factor for toxoplasmic infection in herbivorous animals, such as horses and sheep. While horses show resistance to infection and manifestations of clinical signs of the disease, in sheep, toxoplasmosis may be responsible for important reproductive disorders such as abortion and neonatal death. Therefore, this study aimed to investigate the presence of specific anti-T. gondii antibodies in horses and sheep raised on the Experimental Farm Nhumirim, property of Embrapa Pantanal, Mato Grosso do Sul, Brazil. For this purpose, serum samples were collected from 101 horses and 99 sheep between March 2011 and May 2015. The serum samples were sent to the Toxoplasmosis and other Protozoal Diseases Laboratory, at Oswaldo Cruz Institute, at Oswaldo Cruz Foundation/RJ. Sera were subjected to indirect immunofluorescence test, and seroreactive samples were considered positive from a 1:64 dilution for both species. Anti-T. gondii were detected in 15% (15/99) of sheep and 10% (10/101) of horses. The occurrence of specific anti-T. gondii in the serum samples of these animals, points to an environmental contamination of the property, where possibly wild felids would be acting as dispersers of the parasite...

La toxoplasmosis es una zoonosis de distribución mundial que afecta a los vertebrados homeotérmicos, incluidos los humanos. Toxoplasma gondii, su agente etiológico, es capaz de infectar a sus huéspedes a través de varias vías de transmisión, entre ellas la ingestión de quistes tisulares, presentes en carnes crudas o poco cocidas, y la ingestión de ooquistes, presentes en agua y alimentos contaminados, siendo estos las principales vías de infección en la cadena epidemiológica de la toxoplasmosis. En este sentido, la contaminación ambiental con ooquistes esporulados es un factor de riesgo importante para la infección por toxoplasma en animales herbívoros, como caballos y ovejas. Mientras que los caballos muestran resistencia a la infección y las manifestaciones de los signos clínicos de la enfermedad, en las ovejas, la toxoplasmosis puede ser responsable de importantes trastornos reproductivos como el aborto y la muerte neonatal. Por lo tanto, este estudio tuvo como objetivo investigar la presencia de anticuerpos específicos anti-T. gondii en caballos y ovejas criados en la Granja Experimental Nhumirim, propiedad de Embrapa Pantanal, Mato Grosso do Sul, Brasil. Para ello, se recolectaron muestras de suero de 101 caballos y 99 ovejas entre marzo de 2011 y mayo de 2015. Las muestras de suero se enviaron al Laboratorio de Toxoplasmosis y otros Protozoos, del Instituto Oswaldo Cruz, en la Fundação Oswaldo Cruz / RJ. Los sueros se sometieron a una reacción de inmunofluorescencia indirecta y las muestras serorreactivas se consideraron positivas a partir de una dilución 1:64 para ambas especies. Anti-T. gondii se detectaron en el 15% (15/99) de las ovejas y el 10% (10/101) de los caballos...

Factors affecting the performance of Pantaneiro horses

This study aimed to assess the physical performance of Pantaneiro horses with and without equine infectious anemia (EIA) under functional conditions of cattle management. The horses were subjected to a performance test and split into two groups according to a completely randomized design: animals were chosen from populations testing positive and negative for EIA. Performance was measured as a function of a data envelopment analysis (DEA) model considering four outputs and one unitary input. The output measures were the distance achieved in the performance test, hematocrit as a weighted average over the test duration, respiratory rate as weighted average over the test duration, and the level of lactic acid at the test termination. Weights for the hematocrit and the respiratory rate output variables were determined by means of factor analysis. The performance score was a weighted average of the output variables with the weights defined by the averages of the optimum individual multipliers in the DEA analysis. Contextual variables of interest were age, horse weight, room temperature, and corporal temperature. Only groups and room temperature were statistically significant effects, as indicated by a bootstrap analysis. The performance of group positive for EIA is significantly lower than that of the group negative for EIA and room temperature has a negative effect.(AU)

Neutralising antibodies for West Nile virus in horses from Brazilian Pantanal

Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for WNV and its most closely-related flaviviruses that circulate in Brazil to confirm the detection of specific virus-neutralising antibodies. Of the 93 (55.4 percent) blocking ELISA-seropositive horse serum samples, five (3 percent) were seropositive for WNV, nine (5.4 percent) were seropositive for St. Louis encephalitis virus, 18 (10.7 percent) were seropositive for Ilheus virus, three (1.8 percent) were seropositive for Cacipacore virus and none were seropositive for Rocio virus using PRNT90, with a criteria of > four-fold antibody titre difference. All caimans were negative for flaviviruses-specific antibodies using the blocking ELISA. No virus genome was detected from caiman blood or mosquito samples. The present study is the first report of confirmed serological evidence of WNV activity in Brazil.

Serologic evidence of the recent circulation of Saint Louis encephalitis virus and high prevalence of equine encephalitis viruses in horses in the Nhecolândia sub-region in South Pantanal, Central-West Brazil

As in humans, sub-clinical infection by arboviruses in domestic animals is common; however, its detection only occurs during epizootics and the silent circulation of some arboviruses may remain undetected. The objective of the present paper was to assess the current circulation of arboviruses in the Nhecolândia sub-region of South Pantanal, Brazil. Sera from a total of 135 horses, of which 75 were immunized with bivalent vaccine composed of inactive Eastern equine encephalitis virus (EEEV) and Western equine encephalitis virus(WEEV) and 60 were unvaccinated, were submitted to thorough viral isolation, reverse transcriptase polymerase chain reaction (RT-PCR) and neutralization tests for Saint Louis encephalitis virus (SLEV), EEEV, WEEV and Mayaro virus (MAYV). No virus was isolated and viral nucleic-acid detection by RT-PCR was also negative. Nevertheless, the prevalence of neutralizing antibodies in horses older than seven months was 43.7 percent for SLEV in equines regardless of vaccine status, and 36.4 percent for WEEV and 47.7 percent for EEEV in unvaccinated horses. There was no evidence of MAYV infections. The serologic evidence of circulation of arboviruses responsible for equine and human encephalitis, without recent official reports of clinical infections in the area, suggests that the Nhecolândia sub-region in South Pantanal is an important area for detection of silent activity of arboviruses in Brazil.

Estudo de Paramyxovirus, Mycoplasma e de bacilos Gram-negativos no trato respiratório de serpentes Crotalus durissus terrificus

Serpentes Crotalus durissus terrificus foram avaliadas em três ocasiões: assim que retiradas da natureza e entregues ao Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP) da UNESP, Campus de Botucatu, após um período de quarentena e depois da permanência por aproximadamente 90 dias em uma baia externa. Foram colhidas amostras de sangue para realização de hemograma e sorologia para Paramyxovirus Ofídico (OPMV), swab de glote e lavado traqueopulmonar, para avaliação citológica e pesquisa de OPMV, Mycoplasma e bacilos Gram-negativos. Nos animais que foram a óbito realizaram-se necropsias e pesquisas dos microrganismos citados em traquéia e pulmão. Cepas de Salmonella, Pseudomonas e Aeromonas isoladas foram submetidas a testes de susceptibilidade a drogas. Na primeira colheita (n = 51), os títulos ao teste de inibição da hemaglutinação foram considerados inespecíficos e duas serpentes foram positivas para o OPMV, ao nested PCR. Na quarentena, nove exemplares vieram a óbito, duas das quais mostraram-se positivas para o OPMV. Na segunda colheita (n = 42), os títulos mantiveram-se, porém nove serpentes foram positivas para o OPMV, duas das quais ao teste de hemaglutinação da cultura de células VERO, tornando-se este o primeiro relato do isolamento do OPMV a partir de um animal vivo. Na baia externa, 24 animais foram a óbito, dos quais 23 foram positivos para o OPMV. Na terceira colheita (n = 18), os títulos variaram de 160 a 5.120, e amostras de todas as serpentes foram positivas para o OPMV. Na avaliação citológica, estes lavados apresentaram elevada celularidade com características de processo inflamatório de misto a macrofágico. Salmonella e Pseudomonas foram isoladas a partir de todas as colheitas, porém Aeromonas apenas de amostras de órgãos. Dentre as drogas testadas, os três gêneros de interesse...

Crotalus durissus terrificus snakes were evaluated in three occasions: as soon as they were captured from nature and delivered at the Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP) - UNESP, Botucatu, SP, Brazil; after quarantine; and after spent nearly 90 days in an outside enclosure. Blood samples were collected for hematology and Ophidian Paramyxovirus (OPMV) serology, and glottis swabs and tracheal washings for cytological evaluation, OPMV, Mycoplasma and Gramnegative bacilli search. Dead animals were necropsied and the abovementioned microorganisms investigated in trachea and lung. Salmonella, Pseudomonas and Aeromonas isolated strains were submitted to antimicrobial susceptibility tests. In the first sampling (n = 51), the titles to the hemagglutination inhibition test were considered negative and two snakes were nested PCR positives to OPMV. During quarantine, nine specimens died, and two of them were positives to OPMV. In the second sampling (n = 42), titles had no change, but nine snakes turned positive to OPMV, two of them in the hemagglutination test of the VERO cells culture, what is the first report of the OPMV isolation from a live animal. In the external enclosure, 24 animals died, with 23 revealing to be OPMV positive. In the third sampling (n = 18), serological titles ranged from 160 to 5,120, and samples from all snakes were OPMV positives. In the cytological evaluation, these washings showed to be highly cellular with features of mixed cell to macrophagic inflammation. Salmonella and Pseudomonas strains were isolated in all three samplings, but Aeromonas only from trachea and lung samples. Among all the tested drugs, the three bacterial genera were susceptible to amikacin, gentamicin and enrofloxacin. The isolation of Mycoplasma was not obtained. Nematodes of the genus Rhabdias were observed in the tracheal washings. OPMV was the microorganism isolated in...