Detection of Pathogenic Leptospira in Captive Chelonians (Kinosternon scorpioides-Linnaeus, 1766) in the Brazilian Amazon.

Leptospirosis is a zoonosis of great importance for One Health. In this context, the Amazonian biome may harbor numerous hosts for Leptospira spp. that contribute to the maintenance of the pathogen in the environment. Some reptiles, such as chelonians, have been little studied in terms of their involvement with leptospires. The objective of this study was to detect Leptospira spp. DNA in Kinosternon scorpioides turtles kept in captivity in a region of the Brazilian Amazon. A total of 147 samples of blood (n = 40), cloacal fluid (n = 27), cloacal lavage (n = 40), and stomach (n = 40) were collected from 40 chelonians. After DNA extraction, the samples were subjected to amplification of a 331 base pair product of the 16S rRNA gene using the Lep1 and Lep2 primers. PCR products were Sanger sequenced, assembled, and subjected to online blast search and phylogenetic analysis. Of the animals tested, 40% (16/40, 95% confidence interval [CI]: 25-55) had at least one or two samples positive for Leptospira spp. Considering the total number of samples collected, 12.93% (19/147) were positive, being blood clots (27.5%; 11/40), followed by cloacal washings (10%; 4/40), cloacal fluid (11.11%; 3/27) and gastric washings (2.5%; 1/40). Of these, 11 samples were sequenced and showed 99% to 100% identity with Leptospira interrogans sequences, which was confirmed by phylogenetic analysis. This is the first study to detect pathogenic Leptospira DNA in chelonians in a region of the Brazilian Amazon. It has been concluded that K. scorpioides turtles in captivity have been exposed to pathogenic Leptospira.

Leptospira strains isolated from cattle in the Amazon region, Brazil, evidence of a variety of species and serogroups with a high frequency of the Sejroe serogroup.

In Brazil, there have been few leptospires isolated from cattle, especially in the Amazon, implying that the epidemiology of the disease in this region is still largely unclear. In a previous study, 52 Leptospira isolates were obtained from urine of cattle raised in the Brazilian Amazon and, to achieve a greater understanding of Leptospira infection in cattle of this region, the present study aimed to serologically and molecularly characterizes all these isolates. The laboratory assays used were the microscopic agglutination test (MAT) adopting a panel of polyclonal antisera against Leptospira spp. for serogrouping the isolates, DNA sequencing (secY) and multiple locus variable number tandem repeat analysis (MLVA). The isolates belonged to five species: 20/52 were identified as L. borgpetersenii (38.5 %); 18/52 as L. kirschneri (34.6 %); 9/52 as L. santarosai (17.3 %); 3/52 as L. noguchii (5.8 %) and 2/52 as L. interrogans (3.8 %). With serogrouping, nine different serogroups were detected, with a high frequency of the Sejroe serogroup. MLVA showed that all L. borgpetersenii isolates had a profile compatible with serovar Hardjo; moreover, the other isolates demonstrated a diversity of patterns, and some of them may represent strains not yet characterized. In the Brazilian Amazon, the leptospires circulating in cattle revealed the unique aspects of infections in this area which, in addition to a variety of strains, were characterized by a high frequency of the Sejroe serogroup, highlighting the serovar Hardjo, which has not been reported in other regions of Brazil.

Serological prevalence of Toxoplasma gondii infection in cats (Belém, Pará, Brazil).

We evaluated the prevalence of anti-Toxoplasma gondii antibodies in the serum samples collected from domestic cats in Belém, Pará, Brazil. We also correlated the presence of T. gondii antibodies with environmental variables and cat-owner habits. Four-hundred and forty-seven serum samples from domestic cats were analyzed. The sera were tested using an indirect immunofluorescence assay. Among the animals analyzed, 21.92% (98/447) were seropositive. A statistically significant association was found in relation to age and serology among the animals over 1 year old (p<0.01): in the group up to 1 year old, 12.82% (20/156) of the animals were positive, and in the group over 1 year old, 26.80% (78/291) were positive. Our results show that the cats in Belém, Pará region have anti-T. gondii antibodies, and their owners are not aware of toxoplasmosis or how to prevent its transmission.

Serological prevalence of Toxoplasma gondii infection in cats (Belém, Pará, Brazil) / Prevalência sorológica de infecção por Toxoplasma gondii em gatos (Belém, Pará, Brasil)

Abstract We evaluated the prevalence of anti-Toxoplasma gondii antibodies in the serum samples collected from domestic cats in Belém, Pará, Brazil. We also correlated the presence of T. gondii antibodies with environmental variables and cat-owner habits. Four-hundred and forty-seven serum samples from domestic cats were analyzed. The sera were tested using an indirect immunofluorescence assay. Among the animals analyzed, 21.92% (98/447) were seropositive. A statistically significant association was found in relation to age and serology among the animals over 1 year old (p<0.01): in the group up to 1 year old, 12.82% (20/156) of the animals were positive, and in the group over 1 year old, 26.80% (78/291) were positive. Our results show that the cats in Belém, Pará region have anti-T. gondii antibodies, and their owners are not aware of toxoplasmosis or how to prevent its transmission.

Resumo Foi avaliada a prevalência de anticorpos anti-Toxoplasma gondii em amostras de soro, coletadas de gatos domésticos, em Belém, Pará, Brasil. Também correlacionou-se a presença de anticorpos contra T. gondii com variáveis ambientais e hábitos de proprietários. Quatrocentas e quarenta e sete amostras de soro de gatos domésticos foram analisadas. Os soros foram testados por ensaio de imunofluorescência indireta, e as amostras de fezes foram analisadas por meio de reação em cadeia da polimerase. Entre os animais analisados, 21,92% (98/447) foram soropositivos. Foi encontrada associação estatística significante em relação à idade e à sorologia entre os animais acima de 1 ano (p <0,01): no grupo de até 1 ano, 12,82% (20/156) dos animais positivos e, no grupo acima de 1 ano, 26,80% (78/291) positivos. Os resultados mostram que os gatos daquela região possuem anticorpos anti-T. gondii, e seus donos não estão cientes da toxoplasmose ou de como evitar sua transmissão.

Detection of antibodies against Leptospira spp in free-living marsupials caught in the Eastern Amazon.

INTRODUCTION: Serological surveys are important to assess the health status of wild animals. In this study, antibodies against Leptospira spp, causal agents of leptospirosis, were detected in free-living marsupials in the State of Pará, Brazil. METHODS: Nineteen blood samples collected from marsupials in the municipalities of Peixe-Boi, Viseu, and Castanhal were subjected to microscopic agglutination tests. RESULTS: In total, 36.8% (7/19) of samples were positive, and two exhibited co-agglutination. The most frequent serovars were Icterohaemorrhagiae (60%; 3/5), Panama (20%; 1/5), and Nupezo (20%; 1/5). CONCLUSIONS: Anti-Leptospira spp antibodies currently circulate in free-living marsupials in Northeastern Pará.

Detection of antibodies against Leptospira spp in free-living marsupials caught in the Eastern Amazon

Abstract INTRODUCTION: Serological surveys are important to assess the health status of wild animals. In this study, antibodies against Leptospira spp, causal agents of leptospirosis, were detected in free-living marsupials in the State of Pará, Brazil. METHODS: Nineteen blood samples collected from marsupials in the municipalities of Peixe-Boi, Viseu, and Castanhal were subjected to microscopic agglutination tests. RESULTS: In total, 36.8% (7/19) of samples were positive, and two exhibited co-agglutination. The most frequent serovars were Icterohaemorrhagiae (60%; 3/5), Panama (20%; 1/5), and Nupezo (20%; 1/5). CONCLUSIONS: Anti-Leptospira spp antibodies currently circulate in free-living marsupials in Northeastern Pará.

Negative results for anti-Leptospira sp. agglutinins in free-living Iguanas (Iguana iguana) from the Amazon region of Brazil / Resultados negativos para aglutininas anti-Leptospira sp. em iguanas de vida livre (Iguana iguana) da região amazônica do Brasil

ABSTRACT: Although, serological studies for leptospirosis in iguanas have already been performed, there is no report in the Amazon region. Therefore, the present study aimed to investigate the presence of anti-Leptospira sp. in free living Iguanas-verdes (Iguana iguana) of Mangal das Garças Park, metropolitan region of Belém, Pará, Northern region of Brazil. Twenty-nine blood samples of Iguana-verde (I. iguana) were collected from Mangal das Garças Park. For the serological diagnosis of Leptospira sp. it was employed microscopic agglutination test (MAT) with 25 live antigens of Leptospira sp. as antigens. It was observed that all samples were non-reactive, suggesting no exposure to the agent.

RESUMO: Embora estudos sorológicos para leptospirose em iguanas já tenham sido realizados, não a relatos na região amazônica. Portanto, o presente trabalho teve como objetivo pesquisar a presença de anticorpos anti-Leptospira sp. em Iguanas-verdes (Iguana iguana) de vida livre do Parque Mangal das Garças, região metropolitana de Belém, PA, e Norte do Brasil. Foram coletadas 29 amostras de sangue de Iguana-verde (I. iguana) que frequentaram o Parque Mangal das Garças. Para o diagnóstico sorológico da infecção por Leptospira sp. foi empregado o teste de aglutinação microscópica (MAT) com 25 antígenos vivos de Leptospira sp. como antígenos. Observou-se que todas as amostras foram não reagentes, o que sugere não exposição ao agente.

Seroprevalence for brucellosis and leptospirosis in dogs from Belém and Castanhal, State of Pará, Brazil / Soroprevalência para brucelose e leptospirose em cães de Belém e Castanhal, Pará, Brasil

Brucellosis and leptospirosis are widely spread bacterial infections and dogs are the most important source of infection and reservoir for diseases. Dogs can disseminate the agents in the environment and transmit them to humans and/or other animals. The objective of this study was assess the occurrence of reactive to antibodies anti-Leptospira spp., Brucella canis and B. abortus in Belém and Castanhal, State of Pará, Amazon, Brazil. A total of 156 samples were randomly collected in the city of Belém and 158 samples in Castanhal. The anti-B. canis antibodies research was performed by Agar Gel Immunodiffusion (AGID) with and without 2-mercaptoethanol serum treatment (AGID-2ME). To assess the anti-B. abortus antibodies, the technique of Fast Seroagglutination with buffered acidified plate antigen (BAPAT) was used. For anti-Leptospira spp. antibodies research, the Microscopic Agglutination Technique (MAT) was used. No animal reacted to Brucella abortus and one animal was reactive to B. canis at the AGID, but it was negative to the AGID-2ME test. Seventeen percent of dogs (47/274) presented anti-Leptospira spp. antibodies, with prevalence of serovar Canicola. The dogs from Belém and Castanhal are not source of infection for B. abortus and B. canis, however, they are reservoirs for different serovars of Leptospira spp.

Brucelose e leptospirose são enfermidades bacterianas amplamente disseminadas e o cão é importante fonte de infecção e reservatório para ambas, podendo eliminar o agente no meio ambiente, e transmiti-lo para humanos e/ou outros animais. O presente estudo teve como objetivo pesquisar a ocorrência de cães reagentes para anticorpos contra Leptospira spp., Brucella canis e B. abortus em Belém e Castanhal, Pará, Amazônia, Brasil. Foram colhidas de forma aleatória 156 amostras no município de Belém e 158 amostras em Castanhal. A pesquisa de anticorpos anti-B. canis foi realizada pela técnica de Imunodifusão em Gel de Ágar (IDGA) com e sem tratamento do soro com 2-mercaptoetanol (IDGA-2ME) e para pesquisa de anticorpos anti-B. abortus foi utilizada a técnica de Soroaglutinação Rápida com antígeno acidificado tamponado (AAT). Para pesquisa de anticorpos contra-Leptospira spp. utilizou-se a Técnica de Aglutinação Microscópica (MAT). Nenhum animal reagiu para Brucella abortus e um animal foi reagente para B. canis na IDGA, porém foi negativo na IDGA-2ME. Dezessete por cento dos cães (47/274) apresentaram anticorpos contra-Leptospira spp., com predominância do sorovar Canicola. Os cães de Belém e Castanhal não são fontes de infecção para B. abortus e B. canis, no entanto são reservatórios de diferentes sorovares de Leptospira spp.