Mycobacterium avium subsp. paratuberculosis (MAP) in goat milk in the semiarid region of the Brazilian Northeast - molecular characterization

Background: Goat farming has been on the rise in Brazil in recent years. Overall, 93% of the national herd is concentrated in the Northeast, with the state of Paraíba being the largest goat milk producer in the country. Considering Mycobacterium avium subsp. paratuberculosis (MAP) as a sanitary issue for the development of animal farming with risks for human health and that is a notifiable disease, this research was structured with the objective of confirming the presence and performing a molecular characterization of MAP in goat milk destined for processing plants in the semiarid region of the Brazilian Northeast. Materials, Methods & Results: Samples from 179 production units and 5 collective bulk tanks and 4 samples of pasteurized goat milk were analyzed through Real-time polymerase chain reaction (qPCR). Genetic material (DNA) for MAP was found in the goat milk sample from 1 production unit (1/179). From this positive sample, 9 lactating goats were identified in the original property, 7 of which showed MAP DNA in milk samples (77.77%). The characterization of the nucleotide sequence detected in the positive sample has 99% identity with KJ173784. Discussion: One sample (1/179), from the production units, had MAP genetic material (DNA) detected using the molecular test. Samples from these production units represent the milk from all lactating goats from each producer. Therefore, it was possible to identify from which farm the samples originated, allowing individual animals to then be tested, with milk samples collected from 9 goats and MAP DNA detected in 7 of them (77.77%) via PCR. Control and/or prevention programs need this type of surveillance in reason that it allows the tracking of possible foci from milk samples collected from dairy products or cooling stations. The use of PCR to detect MAP foci via goat milk is thus advantageous because samples are obtained in a non-invasive manner, with faster results when compared to the culture technique. The low detection via PCR in goat milk may be related to factors such as the small amount of MAP eliminated and the intermittent excretion in asymptomatic animals, as also false-positive samples. Samples from the collective bulk tanks was negative. It is possible that the combination of milk from all the properties diluted the amount of MAP. This suggests that the sensitivity of the PCR can be improved if the samples are obtained from the pooled milk from the same property. In some regions of Brazil, for example, showed the frequency of Zona da Mata region of the state of Minas Gerais, Brazil, found 1.94% of positive samples (9/464) and 9.76% (4/41) of properties with at least 1 positive sample for MAP. Different results to what were found in the semiarid region of Paraíba, where climate and production characteristics are different. Goats are susceptible to 3 strains: type "S" (Sheep), "Bison type" and type "C" (Cattle). Previous contact with this species may explain the similarity between the strain found in goat milk and those detected from bovine samples. This must also be taken into consideration during diagnosis and upon implementation of control measures for paratuberculosis in goats. Mycobacterium avium subsp. paratuberculosis was recorded for the first time in goat milk in the semiarid region, which may reveal a potential biological risk to humans and suggests the need for active surveillance of the agent.

Leptospira spp. of the urinary tract of female carrier goats in semi-arid conditions

Background: Leptospirosis is an important infectious disease in goat farming, with a worldwide distribution. It is usually transmitted by rodents and the genital route, may cause reproductive losses, negatively impacting goat farming. The diagnosis lies on serological, molecular and isolation techniques. Considering the importance of this disease for small ruminants, this work aimed to evaluate the serological, molecular findings and isolation of pathogenic leptospires in the urinary tract (kidney and bladder tissues) of goats. Materials, Methods & Results: Thirty-four adult goats were used for slaughter. Renal samples (n = 34), bladder (n = 34), were collected for isolation of the agent and molecular detection of Leptospira sp. and blood samples (n = 34) for serological testing. The polymerase chain reaction (PCR) was used as a molecular test and the microscopic serum agglutination test (MAT) was used as a serological test. Samples with DNA amplification were subjected to genetic sequencing. The presence of Leptospira DNA was found in the tissues of 8 (23.4%) goats, and of these, only 2 were positive in PCR and MAT. There was a slight agreement between the PCR and MAT techniques (k = 0.150; P = 0.436). In 6 (17.6%) samples of renal tissue and 2 (5.8%) bladder samples, Leptospira DNA was detected. The genes in a kidney tissue sample were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira sp. were detected in 6 (17.6%) of the animals tested. Discussion: Serology identified 3 predominant serogroups: Icterohaemorrhagiae, Tarassovi and Autumnalis, serogroups that are related to the presence of rodents that coexist in rural environments. Autumnalis has been reported in small ruminants, raising the hypothesis that goats are adapted, becoming chronic carriers and possible maintenance hosts. The frequency obtained (17.6%) may be the result of the mixed breed pattern and rustic characteristics inherent to the goat species. Given the characteristics of the semi-arid region, such as low rainfall and high solar incidence, it is essential to use an adapted methodology, with a lower cut-off point (1:50), as the serological titer is an established relationship between the animal species, the level of exposure throughout its evolution and the region studied. Molecular findings and bacterial isolation reveal the agent's ability to colonize the urinary tract of goats. These data show the importance that urine has in the epidemiological chain, being able to transmit the agent through direct contact with this product or through contamination of soil and water. There was no statistical agreement between the diagnostic techniques used in this study, in this case, an association between PCR and MAT is recommended to obtain data with high sensitivity and specificity. A bladder sample was sequenced and showed 99% similarity to Leptospira interrogans. In the semiarid region, the most common form of leptospirosis spread is through the sale of animals in business fairs for breeding, rearing or slaughter, as well as sharing the same property with several breeders. The introduction of chronic and asymptomatic carriers on the properties represents a serious risk for the spread of the disease. The results show the presence of Leptospira spp. in semi-arid goat herds, having as risk factors the presence of rodents and intercropping. The association of MAT and PCR is necessary for a better diagnosis of the disease.

Molecular detection of Leishmania infantum in donkeys and mules under semiarid conditions in Brazil / Detecção molecular de Leishmania infantum em asininos e muares sob condições semiáridas

Abstract Visceral leishmaniasis is a parasitic zoonosis that mainly affects poorest and most vulnerable populations, and domestic dogs are considered to be the main source of infection to the vector and therefore humans. However, several studies have investigated the role of other vertebrate hosts in the disease cycle. In this context, the aim of the present study was to conduct a survey of Leishmania infantum infection in donkeys and mules living in a semiarid region of Brazil. Whole blood sampled from 72 equids (65 donkeys and 7 mules) was used to perform molecular diagnosis using the real-time polymerase chain reaction (qPCR) technique. A total of 25% of the samples (18/72) were positive through qPCR, but there were no significant differences between the species (donkeys or mules), sex (male or female) and abandonment situation of the animals (yes or no). Donkeys and mules living under semiarid conditions have high frequency of L. infantum infection. It is therefore worth assigning importance to these species in the epidemiological cycle of visceral leishmaniasis, either as potential reservoirs or just as an abundant food source for vectors.

Resumo A leishmaniose visceral é uma zoonose parasitária que afeta principalmente populações mais pobres e vulneráveis, e os cães domésticos são considerados as principais fontes de infecção para o vetor e, portanto, para os humanos. Porém diversos estudos têm pesquisado o papel de outros hospedeiros vertebrados no ciclo da doença. Neste contexto, objetivou-se realizar um levantamento da infecção por Leishmania infantum em asininos e muares, vivendo em região semiárida do Brasil. Foi utilizado sangue total de 72 equídeos (65 asininos e 7 muares) para a realização de diagnóstico molecular por meio da técnica de Reação em Cadeia de Polimerase em Tempo Real (qPCR). Um total de 25% das amostras (18/72) resultaram positivas na qPCR, porém não houve diferença significativa entre as espécies (asininos e muares), sexo (macho e fêmea) e situação de abandono dos animais (sim ou não). Asininos e muares, vivendo em condições semiáridas, apresentam alta frequência de infecção por L. infantum, sendo válido atribuir importância a essas espécies no ciclo epidemiológico da leishmaniose visceral, seja como um reservatório em potencial, seja apenas como uma fonte alimentar abundante para os vetores.

Leptospira interrogans infection in tegu lizard (Tupinambis merianae), Brazil / Infecção por Leptospira interrogans em lagarto teiú (Tupinambis merianae), Brasil

We used 12 tegu lizards (Tupinambis merianae) from northeastern Brazil, and we reported nine (75%) Leptospira sp. PCR-positive animals and six (50%) seropositive. Leptospira sp. DNA sequencing revealed 99% similarity with L. interrogans. Our findings indicated that this species may play a role in the transmission of human leptospirosis.(AU)

Foram utilizados 12 lagartos Teiús (Tupinambis merianae) do Nordeste do Brasil. Encontramos nove animais positivos (75%) para Leptospira sp. na PCR e seis (50%) soropositivos. O sequenciamento de DNA de Leptospira sp. revelou 99% de semelhança com L. interrogans. Os resultados indicam que esta espécie pode desempenhar um papel importante na transmissão da leptospirose humana.(AU)

Métodos diagnósticos para la detección de Leptospira spp. en muestras biológicas / Diagnostic methods for the detection of Leptospira spp. in biological samples / Métodos diagnósticos para a detecção de Leptospira spp. em amostras biológicas

Leptospirosis es una zoonosis bacteriana distribuida en todo el mundo. Puede afectar a cualquier especie animal, manteniendo el agente en la naturaleza a través de la infección renal crónica en los animales portadores, que excretan los microorganismos en el ambiente mediante la orina y pueden infectar a otros animales y el hombre mismo. Acerca de Leptospira spp., técnicas de diagnóstico han ayudado en la comprensión de los aspectos eco-epidemiológicas importantes como la distribución ambiental de los serotipos y nuevos huéspedes, y mejorar el diagnóstico clínico de la enfermedad. Esta revisión tiene como objetivo presentar las diferentes técnicas utilizadas en el diagnóstico de la leptospirosis en muestras biológicas, incluyendo algunos estudios sobre las aplicaciones prácticas de detección molecular de Leptospira spp., reafirmando la importancia del conocimiento y la elección de las técnicas adecuadas en el diagnóstico de la enfermedad.(AU)

Leptospirosis is a bacterial zoonoses distributed worldwide. It can undertake any animal species, maintaining the agent in the nature via renal chronical infection of carriers animals, which can secrete the organism through the urine into the environment, being able to infect other animals and the human. Regarding Leptospira spp., diagnostic techniques have helped understand important eco-epidemiological aspects such as environmental serovar distribution and new hosts, in addition to improve the clinical diagnosis of the disease. This review focuses on present diverse techniques used in leptospirosis diagnosis to biological samples, including some studies on practical applications for the molecular detection of Leptospira spp., reinforcing the importance of the knowledge and the choice of the correct technique in the diagnosis of the disease.(AU)

Leptospirose é uma zoonose bacteriana distribuída mundialmente. Pode acometer qualquer espécie animal, mantendo o agente na natureza via infecção crônica renal de animais carreadores, os quais excretam os organismos pela urina no meio ambiente, podendo infectar outros animais e humanos. Em relação à Leptospira spp., as técnicas diagnósticas têm auxiliado na compreensão de aspectos eco-epidemiológicos importantes tais como a distribuição ambiental dos sorovares e novos hospedeiros, além de aprimorar o diagnóstico na clínica da doença. A revisão tem como objetivo apresentar as diversas técnicas utilizadas no diagnóstico da leptospirose em amostras biológicas, incluindo alguns estudos sobre aplicações práticas da detecção molecular de Leptospira spp., reafirmando assim a importância do conhecimento e da escolha das técnicas corretas no diagnóstico da doença.(AU)

Métodos diagnósticos para la detección de Leptospira spp. en muestras biológicas / Diagnostic methods for the detection of Leptospira spp. in biological samples / Métodos diagnósticos para a detecção de Leptospira spp. em amostras biológicas

Leptospirosis es una zoonosis bacteriana distribuida en todo el mundo. Puede afectar a cualquier especie animal, manteniendo el agente en la naturaleza a través de la infección renal crónica en los animales portadores, que excretan los microorganismos en el ambiente mediante la orina y pueden infectar a otros animales y el hombre mismo. Acerca de Leptospira spp., técnicas de diagnóstico han ayudado en la comprensión de los aspectos eco-epidemiológicas importantes como la distribución ambiental de los serotipos y nuevos huéspedes, y mejorar el diagnóstico clínico de la enfermedad. Esta revisión tiene como objetivo presentar las diferentes técnicas utilizadas en el diagnóstico de la leptospirosis en muestras biológicas, incluyendo algunos estudios sobre las aplicaciones prácticas de detección molecular de Leptospira spp., reafirmando la importancia del conocimiento y la elección de las técnicas adecuadas en el diagnóstico de la enfermedad.

Leptospirosis is a bacterial zoonoses distributed worldwide. It can undertake any animal species, maintaining the agent in the nature via renal chronical infection of carriers animals, which can secrete the organism through the urine into the environment, being able to infect other animals and the human. Regarding Leptospira spp., diagnostic techniques have helped understand important eco-epidemiological aspects such as environmental serovar distribution and new hosts, in addition to improve the clinical diagnosis of the disease. This review focuses on present diverse techniques used in leptospirosis diagnosis to biological samples, including some studies on practical applications for the molecular detection of Leptospira spp., reinforcing the importance of the knowledge and the choice of the correct technique in the diagnosis of the disease.

Leptospirose é uma zoonose bacteriana distribuída mundialmente. Pode acometer qualquer espécie animal, mantendo o agente na natureza via infecção crônica renal de animais carreadores, os quais excretam os organismos pela urina no meio ambiente, podendo infectar outros animais e humanos. Em relação à Leptospira spp., as técnicas diagnósticas têm auxiliado na compreensão de aspectos eco-epidemiológicos importantes tais como a distribuição ambiental dos sorovares e novos hospedeiros, além de aprimorar o diagnóstico na clínica da doença. A revisão tem como objetivo apresentar as diversas técnicas utilizadas no diagnóstico da leptospirose em amostras biológicas, incluindo alguns estudos sobre aplicações práticas da detecção molecular de Leptospira spp., reafirmando assim a importância do conhecimento e da escolha das técnicas corretas no diagnóstico da doença.

Toxoplasma gondii antibodies and related risk factors in mammals at Sorocaba zoo, São Paulo, BrazilToxoplasma gondii antibodies and related risk factors in mammals at Sorocaba zoo, São Paulo, Brazil / Anticorpos anti-Toxoplasma gondii e fatores de risco em mamíferos do Zoológico de Sorocaba, São Paulo, Brasil

Toxoplasmosis is a parasitic zoonosis with worldwide distribution and different wild species either are involved in maintenance of the pathogen in the environment as definitive or intermediate hosts serving as font of feeding or prey to definitive hosts. The present study aimed to investigate Toxoplasma gondii antibodies and the risk factors relating to infection in mammals at Sorocaba Zoo, state of São Paulo, Brazil. Serum samples collected from 153 animals were analyzed using the modified agglutination test (MAT; cutoff ? 25). Seropositivity was found in 62 animals (40.5%; 95% CI: 33.1-48.5%), with different titers. Significant differences were observed in relation to the ages of the animals, origin, presence of free-ranging animals in the enclosure and feeding habits, through univariate analysis (p ? 0.05). Multivariate logistic regression analysis showed that only age (p = 0.03) had significance for the study. Adult animals were 3.5 more likely to become infected than were young ones. In relation to feeding habits, herbivores (80%) and carnivores (46.5%) were the animals most affected. These results highlight the presence of T. gondii in animals at Brazilian zoos, and suggest that continuous transmission is occurring at zoos.

Toxoplasmose é uma zoonose parasitária com distribuição mundial e diversas espécies silvestres estão envolvidas na manutenção do patógeno no ambiente, seja como hospedeiro definitivo ou intermediário. O objetivo do estudo foi pesquisar anticorpos de Toxoplasma gondii e os fatores de risco relacionados com a infecção de mamíferos do Zoológico de Sorocaba, Estado de São Paulo, Brasil. Foram coletadas 153 amostras de soro dos animais e realizado teste de aglutinação modificada (MAD, ponto de corte 16). Sessenta e dois (40,5%; IC95% 33,1-48,5%) animais foram soropositivos, com diferentes títulos. Diferenças significativas foram observadas com relação a idade dos animais, origem, presença de animais de vida livre dentro dos recintos e hábitos alimentares, utilizando análise univariada (p ? 0,05). A análise pela regressão logística multivariada mostrou que apenas idade (p = 0.03) foi significativo no estudo. Animais adultos apresentaram 3,5 vezes mais chance de se tornarem infectados que os jovens. Com relação aos hábitos alimentares, animais herbívoros (80%) e carnívoros (46,5%) foram os mais afetados. Esses resultados ressaltam a presença do T. gondii nos animais dos zoológicos brasileiros e sugerem contínua transmissão no zoológico.

Toxoplasma gondii antibodies and related risk factors in mammals at Sorocaba zoo, São Paulo, BrazilToxoplasma gondii antibodies and related risk factors in mammals at Sorocaba zoo, São Paulo, Brazil / Anticorpos anti-Toxoplasma gondii e fatores de risco em mamíferos do Zoológico de Sorocaba, São Paulo, Brasil

Toxoplasmosis is a parasitic zoonosis with worldwide distribution and different wild species either are involved in maintenance of the pathogen in the environment as definitive or intermediate hosts serving as font of feeding or prey to definitive hosts. The present study aimed to investigate Toxoplasma gondii antibodies and the risk factors relating to infection in mammals at Sorocaba Zoo, state of São Paulo, Brazil. Serum samples collected from 153 animals were analyzed using the modified agglutination test (MAT; cutoff ? 25). Seropositivity was found in 62 animals (40.5%; 95% CI: 33.1-48.5%), with different titers. Significant differences were observed in relation to the ages of the animals, origin, presence of free-ranging animals in the enclosure and feeding habits, through univariate analysis (p ? 0.05). Multivariate logistic regression analysis showed that only age (p = 0.03) had significance for the study. Adult animals were 3.5 more likely to become infected than were young ones. In relation to feeding habits, herbivores (80%) and carnivores (46.5%) were the animals most affected. These results highlight the presence of T. gondii in animals at Brazilian zoos, and suggest that continuous transmission is occurring at zoos.(AU)

Toxoplasmose é uma zoonose parasitária com distribuição mundial e diversas espécies silvestres estão envolvidas na manutenção do patógeno no ambiente, seja como hospedeiro definitivo ou intermediário. O objetivo do estudo foi pesquisar anticorpos de Toxoplasma gondii e os fatores de risco relacionados com a infecção de mamíferos do Zoológico de Sorocaba, Estado de São Paulo, Brasil. Foram coletadas 153 amostras de soro dos animais e realizado teste de aglutinação modificada (MAD, ponto de corte 16). Sessenta e dois (40,5%; IC95% 33,1-48,5%) animais foram soropositivos, com diferentes títulos. Diferenças significativas foram observadas com relação a idade dos animais, origem, presença de animais de vida livre dentro dos recintos e hábitos alimentares, utilizando análise univariada (p ? 0,05). A análise pela regressão logística multivariada mostrou que apenas idade (p = 0.03) foi significativo no estudo. Animais adultos apresentaram 3,5 vezes mais chance de se tornarem infectados que os jovens. Com relação aos hábitos alimentares, animais herbívoros (80%) e carnívoros (46,5%) foram os mais afetados. Esses resultados ressaltam a presença do T. gondii nos animais dos zoológicos brasileiros e sugerem contínua transmissão no zoológico.(AU)

Anti-leptospirosis agglutinins in Brazilian capybaras (hydrochoerus hydrochaeris)

The interest in commercial use of wild animals is increasing, especially regarding raising of capybaras. Although this wild species is potentially lucrative for the production of meat, oil and leather, it is suggested as a probable reservoir of leptospires. Methods Due to the economic importance of this species and the lack of studies concerning leptospirosis, the presence of anti-leptospirosis agglutinins was assayed in 55 serum samples of capybaras (Hydrochoerus hydrochaeris) from commercial and experimental breeding flocks located in São Paulo state, Paraná state, and Rio Grande do Sul state, Brazil. Samples were obtained through cephalic or femoral venipunction (5 to 10 mL). Microscopic agglutination test was used according to the Brazilian Health Ministry considering as cut-off titer of 100. Results Out of the 55 samples analyzed, 23 (41.82 %) tested positive. The most prevalent serovar was Icterohaemorrhagiae (56.52 %) in 13 samples, followed by Copenhageni in nine samples (39.13 %), Pomona in four samples (17.39 %), Djasiman and Castellonis in three samples each (13.04 %), Grippotyphosa, Hardjo, Canicola, and Cynopteri in two samples each (8.7 %), and Andamana and Bratislava in one sample each (4.34 %). Conclusions These results suggest the evidence of exposure toLeptospira spp. and the need of new studies to evaluate a higher number of capybaras from different regions to better understand the importance of leptospirosis infection in these animals and verify the zoonotic role of this species as a possible source of infection to humans and other animals.(AU)

Anti-leptospirosis agglutinins in Brazilian capybaras (hydrochoerus hydrochaeris)

The interest in commercial use of wild animals is increasing, especially regarding raising of capybaras. Although this wild species is potentially lucrative for the production of meat, oil and leather, it is suggested as a probable reservoir of leptospires. Methods Due to the economic importance of this species and the lack of studies concerning leptospirosis, the presence of anti-leptospirosis agglutinins was assayed in 55 serum samples of capybaras (Hydrochoerus hydrochaeris) from commercial and experimental breeding flocks located in São Paulo state, Paraná state, and Rio Grande do Sul state, Brazil. Samples were obtained through cephalic or femoral venipunction (5 to 10 mL). Microscopic agglutination test was used according to the Brazilian Health Ministry considering as cut-off titer of 100. Results Out of the 55 samples analyzed, 23 (41.82 %) tested positive. The most prevalent serovar was Icterohaemorrhagiae (56.52 %) in 13 samples, followed by Copenhageni in nine samples (39.13 %), Pomona in four samples (17.39 %), Djasiman and Castellonis in three samples each (13.04 %), Grippotyphosa, Hardjo, Canicola, and Cynopteri in two samples each (8.7 %), and Andamana and Bratislava in one sample each (4.34 %). Conclusions These results suggest the evidence of exposure toLeptospira spp. and the need of new studies to evaluate a higher number of capybaras from different regions to better understand the importance of leptospirosis infection in these animals and verify the zoonotic role of this species as a possible source of infection to humans and other animals.

Mycobacterium genavense infection in two species of captive snakes

Background: Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.(AU)

Mycobacterium genavense infection in two species of captive snakes

Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.(AU)

Seroprevalencia de leptospirosis en ovinos de la región centro oeste de la provincia de São Paulo / Serological profile of leptospirosis in sheep from midlle western region of São Paulo State, Brazil / Soroprevalência da leptospirose em ovinos da região centro-oeste do Estado de São Paulo

Los últimos años hubo un aumento significativo en el rebaño de ovinos en São Paulo. Con el fin de contribuir a la salud de esta creciente actividad agrícola, se estudió la seroprevalencia de leptospirosis en 400 ovejas de ocho fincas ubicadas en la región centro oeste de São Paulo. El suero de estos ovinos se sometió a una prueba de aglutinación microscópica (MAT), la principal técnica serológica para el diagnóstico de la leptospirosis. En la bateria de antígenos utilizada en la prueba de aglutinación microscópica se encontraban representantes de los serogrupos de todos los serotipos del país. De los ovinos evaluados, el 3% (12 animales) tuvieron resultados seropositivos para la leptospirosis. En cuanto a la prevalencia de diferentes serotipos de animales seropositivos destacado Leptospira Pomona (75%), L. hardjo (16,6%) y L. castellonis (8,4%). La vacunación de los ovinos y el control de los roedores son las formas principales de evitar las pérdidas económicas de la leptospirosis, y para evitar la propagación de enfermedades zoonóticas entre los trabajadores agrícolas.(AU)

Over the past years, there have been a significant increase in herd sheep in the state of São Paulo. Aiming to contribute to the health of these growing farming activities, we studied the seroprevalence of leptospirosis in 400 sheep from eight farms placed in the midlle western region of São Paulo state. The serum of these sheep was submitted to the microscopic agglutination test (MAT), the main technique for serological diagnosis of leptospirosis. The antigens used in the microscopic agglutination test included serogroups of all serovars described in the country. 3% (12 animals) had seropositive results for leptospirosis. Regarding the prevalence of different serotypes in positive animals highlight was Leptospira pomona (75%), followed by L. hardjo (16.6%) and L. castellonis (8.4%). Vaccination of sheep and control of rodents are the major means to prevent economic losses arising from leptospirosis and the spread of this zoonotic disease among rural workers.(AU)

Nos últimos anos, houve aumento significativo do rebanho ovino no estado de São Paulo. Com o objetivo de contribuir com a sanidade desta crescente atividade agropecuária, estudouse a soroprevalência da leptospirose em 400 ovinos, provenientes de oito criatórios situados na região centro-oeste do estado de São Paulo. Realizou-se o teste de soroaglutinação microscópica (SAM), principal técnica sorológica para o diagnóstico de leptospirose. A bateria de antígenos empregada na prova de aglutinação microscópica incluiu representantes de sorogrupos de todos os sorovares existentes no país. Dos ovinos avaliados, 3% (12 animais) apresentaram resultado soropositivo para leptospirose. Quanto à prevalência dos diferentes sorotipos, destacou-se Leptospira pomona (75%), L. hardjo (16,6%) e L. castellonis (8,4%). A vacinação dos ovinos e o controle dos roedores são os principais meios de evitar prejuízos econômicos decorrentes da leptospirose, além de prevenir a propagação desta zoonose entre os trabalhadores rurais.(AU)

Mycobacterium genavense infection in two species of captive snakes

Mycobacterium is an important zoonotic agent with companion, livestock and wildlife animals reportedly playing a role as reservoirs. Although its association with reptiles has been described, the disease cycle remains to be fully established, particularly in snakes. Accordingly, this study aimed to report the occurrence of mycobacteriosis with clinical pneumonia in one exotic python snake (Python molurus) and one native green snake (Philodryas olfersii) from the Sorocaba Zoo, São Paulo state, Brazil. Methods: Diagnosis was based on necropsy, histopathological examination, Ziehl-Neelsen stain and immunohistochemistry. Results: Using a nested PCR followed by DNA sequencing and bioinformatics analysis, the causative Mycobacterium species was identified as Mycobacterium genavense. Conclusion: Mycobacterium genavense is an infectious zoonotic agent of animal and public health concerns.

Seroprevalencia de leptospirosis en ovinos de la región centro oeste de la provincia de São Paulo / Serological profile of leptospirosis in sheep from midlle western region of São Paulo State, Brazil / Soroprevalência da leptospirose em ovinos da região centro-oeste do Estado de São Paulo

Los últimos años hubo un aumento significativo en el rebaño de ovinos en São Paulo. Con el fin de contribuir a la salud de esta creciente actividad agrícola, se estudió la seroprevalencia de leptospirosis en 400 ovejas de ocho fincas ubicadas en la región centro oeste de São Paulo. El suero de estos ovinos se sometió a una prueba de aglutinación microscópica (MAT), la principal técnica serológica para el diagnóstico de la leptospirosis. En la bateria de antígenos utilizada en la prueba de aglutinación microscópica se encontraban representantes de los serogrupos de todos los serotipos del país. De los ovinos evaluados, el 3% (12 animales) tuvieron resultados seropositivos para la leptospirosis. En cuanto a la prevalencia de diferentes serotipos de animales seropositivos destacado Leptospira Pomona (75%), L. hardjo (16,6%) y L. castellonis (8,4%). La vacunación de los ovinos y el control de los roedores son las formas principales de evitar las pérdidas económicas de la leptospirosis, y para evitar la propagación de enfermedades zoonóticas entre los trabajadores agrícolas.

Over the past years, there have been a significant increase in herd sheep in the state of São Paulo. Aiming to contribute to the health of these growing farming activities, we studied the seroprevalence of leptospirosis in 400 sheep from eight farms placed in the midlle western region of São Paulo state. The serum of these sheep was submitted to the microscopic agglutination test (MAT), the main technique for serological diagnosis of leptospirosis. The antigens used in the microscopic agglutination test included serogroups of all serovars described in the country. 3% (12 animals) had seropositive results for leptospirosis. Regarding the prevalence of different serotypes in positive animals highlight was Leptospira pomona (75%), followed by L. hardjo (16.6%) and L. castellonis (8.4%). Vaccination of sheep and control of rodents are the major means to prevent economic losses arising from leptospirosis and the spread of this zoonotic disease among rural workers.

Nos últimos anos, houve aumento significativo do rebanho ovino no estado de São Paulo. Com o objetivo de contribuir com a sanidade desta crescente atividade agropecuária, estudouse a soroprevalência da leptospirose em 400 ovinos, provenientes de oito criatórios situados na região centro-oeste do estado de São Paulo. Realizou-se o teste de soroaglutinação microscópica (SAM), principal técnica sorológica para o diagnóstico de leptospirose. A bateria de antígenos empregada na prova de aglutinação microscópica incluiu representantes de sorogrupos de todos os sorovares existentes no país. Dos ovinos avaliados, 3% (12 animais) apresentaram resultado soropositivo para leptospirose. Quanto à prevalência dos diferentes sorotipos, destacou-se Leptospira pomona (75%), L. hardjo (16,6%) e L. castellonis (8,4%). A vacinação dos ovinos e o controle dos roedores são os principais meios de evitar prejuízos econômicos decorrentes da leptospirose, além de prevenir a propagação desta zoonose entre os trabalhadores rurais.

PCR and qPCR for detection of Porcine Circovirus type 2 (PCV2) in captive white-lipped (Tayassu pecari) and collared (Tayassu tajacu) peccaries from Southern Brazil / PCR e qPCR para detecção de Circovírus Suíno tipo 2 (PCV2) em catetos (Pecari tajacu) e queixadas (Tayassu pecari) de cativeiro do Sul do Brasil

Porcine circovirus type 2 (PCV2) is an emergent virus found in commercial pig farms and may cause clinical or subclinical infection. The collared peccary (Pecari tajacu) and white-lipped peccary (Tayassu pecari) may also be infected by PCV2. Accordingly, the aim of this study was to identify PCV2 in whole blood samples of captive peccaries (16 collared and 6 white-lipped) by conventional and quantitative PCR (qPCR). Although the housekeeping gene (c-myc) DNA was successfully amplified, all 22 peccaries tested were negative for PCV2 by both molecular methods. In conclusion, although PCV2 may be endemic in free ranging wild pigs of Central and Northern Brazil, lack of serological and molecular PCV2 evidence (in whole blood) of both captive and free-range wild pigs may indicate low risk of disease in Southern Brazil.

O circovírus suíno tipo 2 (PCV2) é um vírus emergente encontrado em granjas comerciais de suínos que pode causar infecção clínica ou subclínica. Os catetos (Pecari tajacu) e as queixadas (Tayassu pecari) também podem se infectar com PCV2. Deste modo, o objetivo deste estudo foi identificar o PCV2 em amostras de sangue total de pecaris de cativeiro (16 catetos e seis queixadas) por PCR convencional e quantitativo (qPCR). Embora o gene constitutivo (c-myc) tenha sido amplificado com sucesso, todas as 22 amostras de pecaris avaliadas foram negativas para PCV2 em ambos os métodos moleculares. Em conclusão, embora o PCV2 seja endêmico em suínos selvagens de vida livre do Centro e Norte do Brasil, a ausência de evidência sorológica e molecular de PCV2 (em sangue total) de ambos pecaris de cativeiro e de vida livre no Sul do Brasil podem indicar baixo risco de doença no Sul do Brasil.

PCR and qPCR for detection of Porcine Circovirus type 2 (PCV2) in captive white-lipped (Tayassu pecari) and collared (Tayassu tajacu) peccaries from Southern Brazil / PCR e qPCR para detecção de Circovírus Suíno tipo 2 (PCV2) em catetos (Pecari tajacu) e queixadas (Tayassu pecari) de cativeiro do Sul do Brasil

Porcine circovirus type 2 (PCV2) is an emergent virus found in commercial pig farms and may cause clinical or subclinical infection. The collared peccary (Pecari tajacu) and white-lipped peccary (Tayassu pecari) may also be infected by PCV2. Accordingly, the aim of this study was to identify PCV2 in whole blood samples of captive peccaries (16 collared and 6 white-lipped) by conventional and quantitative PCR (qPCR). Although the housekeeping gene (c-myc) DNA was successfully amplified, all 22 peccaries tested were negative for PCV2 by both molecular methods. In conclusion, although PCV2 may be endemic in free ranging wild pigs of Central and Northern Brazil, lack of serological and molecular PCV2 evidence (in whole blood) of both captive and free-range wild pigs may indicate low risk of disease in Southern Brazil.(AU)

O circovírus suíno tipo 2 (PCV2) é um vírus emergente encontrado em granjas comerciais de suínos que pode causar infecção clínica ou subclínica. Os catetos (Pecari tajacu) e as queixadas (Tayassu pecari) também podem se infectar com PCV2. Deste modo, o objetivo deste estudo foi identificar o PCV2 em amostras de sangue total de pecaris de cativeiro (16 catetos e seis queixadas) por PCR convencional e quantitativo (qPCR). Embora o gene constitutivo (c-myc) tenha sido amplificado com sucesso, todas as 22 amostras de pecaris avaliadas foram negativas para PCV2 em ambos os métodos moleculares. Em conclusão, embora o PCV2 seja endêmico em suínos selvagens de vida livre do Centro e Norte do Brasil, a ausência de evidência sorológica e molecular de PCV2 (em sangue total) de ambos pecaris de cativeiro e de vida livre no Sul do Brasil podem indicar baixo risco de doença no Sul do Brasil.(AU)

Detección molecular de Leishmania spp. en material de hemocultivo, y diagnostico sorologico para leishmaniasis em perros del Barrio de la Conquista, São Manuel-SP, Brasil / Molecular detection of Leishmania spp. in blood culture material, and leishmaniasis serological diagnosis in dogs from Bairro da Conquista, Sao Manuel, Brazil / Detecção molecular de Leishmania spp. em material de hemocultura, e diagnóstico sorológico para leishmaniose em cães procedentes do Bairro da Conquista, São Manuel-SP, Brasil

Leishmaniasis es una enfermedad tropical desatendida causada por protozoos del género Leishmania, y transmitida a hospederos mamíferos por flebótomos infectados. Esta zoonosis incluye un amplio espectro de manifestaciones como lesiones cutáneas en el sitio de la picadura de los flebótomos hasta la leishmaniasis visceral sistémica, y pueden presentarse como leishmaniasis tegumentaria americana (LTA) y leishmaniasis visceral (LV). En las zonas urbanas, los perros (Canis familiaris) son la principal fuente de infección de LV, además son de gran importancia en la transmisión de la enfermedad, probablemente debido al mayor parasitismo cutáneo y por estar infectados en casi todos los sitios brasileños de leishmaniasis visceral humana. Fueron evaluadas 197 muestras de sangre de perros del barrio de la Conquista, en el municipio de São Manuel-SP, Brasil. Las pruebas serológicas de inmunofluorescencia indirecta (IFI) y ELISA fueran realizadas, y todos los animales fueron no reactivos, así como en los exámenes parasitológicos de punción de nódulos linfáticos. En el hemocultivo, 48/197 (24,3%) de las muestras presentaron crecimiento de tripanosomas, incluso tres muestras fueron positivas en la técnica de reacción en cadena de la polimerasa (PCR) para Leishmania spp. Solamente un animal (0,5%) con sintomatología compatible se mostró positivo en la PCR de Leishmania spp. El estudio [...](AU)

Leishmaniasis is a neglected tropical disease caused by Leishmania genus protozoa, and transmitted to mammals by infected phlebotomines. This zoonosis determines a high spectrum of clinical manifestations, like skin lesions at the local of phlebotomines bite, or more severe signs in case of visceral systemic leishmaniasis. According to the Leishmanias specie infection, the disease can be clinically presented as American Tegumentary Leishmaniasis (ATL) or Visceral Leishmaniasis (VL). Dog (Canis familiaris) is the main VL infection source in the urban area, and has high importance in the disease transmission, probably due to its high cutaneous parasitism, and also because dogs are frequently diagnosed infected in all Brazilian regions where human visceral leishmaniasis cases are related. 197 blood samples were collected from dogs in Bairro da Conquista, in São ManuelSP Brazil. Serum samples were evaluated by Indirect Immunofluorescent Antibody Test (IFAT) and ELISA for Leishmania spp. diagnosis, and all resulted negative. The cytology of dogs lymph nodes was also negative for Leishmania spp. 48/197 (24.3%) blood samples were positive to culture, with tripanosomatides visualization. Three of these positive samples (1.5%) were also positive by PCR for Leishmania spp. One blood sample collected from a VL symptomatic dog (0.5%) that was positive to culture was also positive by [...](AU)

A leishmaniose é uma doença tropical negligenciada causada por protozoários do gênero Leishmania, transmitidos para os hospedeiros mamíferos por flebotomíneos infectados. Esta zoonose inclui um amplo espectro de manifestações clínicas, desde lesões cutâneas no local da picada dos flebotomíneos até a leishmaniose visceral sistêmica, podendo se apresentar como Leishmaniose Tegumentar Americana (LTA) e Leishmaniose Visceral (LV). Na área urbana, o cão (Canis familiaris) é a principal fonte de infecção da LV, pois assume grande importância na transmissão da doença, provavelmente devido ao seu maior parasitismo cutâneo e por apresentar-se infectado em quase todos os focos brasileiros de calazar humano. Foram analisadas 197 amostras de sangue de cães procedentes do bairro da Conquista, no município de São ManuelSP. Exames sorológicos pela Reação de Imunofluorescência Indireta (RIFI) e ELISA foram realizados, sendo todos os animais não reagentes, bem como negativos ao exame parasitológico de punção de linfonodos. À hemocultura, 48/197 (24,3%) amostras apresentaram crescimento de tripanossomatídeos, sendo que, destas, três amostras (1,5%) foram positivas à técnica de Reação em Cadeia de Polimerase (PCR) para Leishmania spp. Apenas um animal (0,5%) com sintomatologia compatível foi positivo à PCR para Leishmania spp. O estudo ressalta a importância de esforços contínuos e [...](AU)

Detection of anti-Toxoplasma gondii antibodies in carthorses in the metropolitan region of Curitiba, Paraná, Brazil / Detecção de anticorpos anti-Toxoplasma gondii em cavalos carroceiros da região metropolitana de Curitiba, Paraná, Brasil

Toxoplasma gondii, the agent for toxoplasmosis, has worldwide distribution. Horses normally play a secondary role in its life cycle, but movement around urban areas, feeding on grass and the increasing use of carthorses for gathering recyclable material in some urban areas of Brazil may increase their exposure to T. gondii infection. The aim of the present study was to investigate the frequency of anti-T. gondii antibodies in carthorses in the metropolitan region of Curitiba, PR. IgG antibodies against T. gondii were detected using the indirect fluorescence antibody test (IFAT) (titers ≥ 64). Seventeen (17.0%) of the 100 horses sampled were seropositive. There were no statistical differences in relation to sex (p = 0.28) or age (p = 0.15). Our findings suggest that carthorses are exposed to T. gondii infections and that no associations with age or sex exist.

Toxoplasma gondii, agente da toxoplasmose, tem distribuição mundial. Geralmente cavalos desempenham papel secundário no ciclo de vida do T. gondii. Entretanto, a circulação em meio urbano, a alimentação com gramíneas e o aumento no uso de cavalos carroceiros para recolhimento de material reciclável em algumas áreas urbanas do Brasil podem aumentar a exposição desses cavalos ao T. gondii. O objetivo deste estudo foi investigar a frequência de anticorpos anti-T. gondii em cavalos carroceiros da região metropolitana de Curitiba, PR. Os anticorpos da classe IgG contra T. gondii foram detectados pela reação de imunofluorescência indireta (RIFI) (títulos ≥ 64). Dezessete (17%) dos 100 cavalos avaliados foram positivos e não houve diferença entre sexos (p = 0,28) ou idade (p = 0,15). Esses achados sugerem que cavalos carroceiros estão expostos a infecções por T. gondii, não existindo associação com a idade ou o sexo dos animais.

Spatial distribution of seropositive dogs to Leptospira spp. and evaluation of leptospirosis risk factors using a decision tree

Background: Leptospirosis is an important zoonotic disease associated with poor areas of urban settings of developing countries and early diagnosis and prompt treatment may prevent disease. Although rodents are reportedly considered the main reservoirs of leptospirosis, dogs may develop the disease, may become asymptomatic carriers and may be used as sentinels for disease epidemiology. The use of Geographical Information Systems (GIS) combined with spatial analysis techniques allows the mapping of the disease and the identification and assessment of health risk factors. Besides the use of GIS and spatial analysis, the technique of data mining, decision tree, can provide a great potential to find a pattern in the behavior of the variables that determine the occurrence of leptospirosis. The objective of the present study was to apply Geographical Information Systems and data prospection (decision tree) to evaluate the risk factors for canine leptospirosis in an area of Curitiba, PR. Materials, Methods & Results: The present study was performed on the Vila Pantanal, a urban poor community in the city of Curitiba. A total of 287 dog blood samples were randomly obtained house-by-house in a two-day sampling on January 2010. In addition, a questionnaire was applied to owners at the time of sampling. Geographical coordinates related to each household of tested dog were obtained using a Global Positioning System (GPS) for mapping the spatial distribution of reagent and non-reagent dogs to leptospirosis. For the decision tree, risk factors included results of microagglutination test (MAT) from the serum of dogs, previous disease on the household, contact with rats or other dogs, dog breed, outdoors access, feeding, trash around house or backyard, open sewer proximity and flooding. A total of 189 samples (about 2/3 of overall samples) were randomly selected for the training file and consequent decision rules. The remained 98 samples were used for the testing file. The seroprevalence showed a pattern of spatial distribution that involved all the Pantanal area, without agglomeration of reagent animals. In relation to data mining, from 189 samples used in decision tree, a total of 165 (87.3%) animal samples were correctly classified, generating a Kappa index of 0.413. A total of 154 out of 159 (96.8%) samples were considered non-reagent and were correctly classified and only 5/159 (3.2%) were wrongly identified. On the other hand, only 11 (36.7%) reagent samples were correctly classified, with 19 (63.3%) samples failing diagnosis. Discussion: The spatial distribution that involved all the Pantanal area showed that all the animals in the area are at risk of contamination by Leptospira spp. Although most samples had been classified correctly by the decision tree, a degree of difficulty of separability related to seropositive animals was observed, with only 36.7% of the samples classified correctly. This can occur due to the fact of seronegative animals number is superior to the number of seropositive ones, taking the differences in the pattern of variable behavior. The data mining helped to evaluate the most important risk factors for leptospirosis in an urban poor community of Curitiba. The variables selected by decision tree reflected the important factors about the existence of the disease (default of sewer, presence of rats and rubbish and dogs with free access to street). The analyses showed the multifactorial character of the epidemiology of canine leptospirosis.