Enterobacterias en tortugas silvestres y cautivas del Amazonas, Podocnemis expansa (Testudines: Podocnemididae)

a tortuga amazónica Podocnemis expansa Schweigger, 1812 es un recurso muy importante para las poblaciones ribereñas de fauna de la región amazónica, además de ser una de las principales especies enumeradas para la producción en cautiverio. El consumo de esta especie como alimento en la región ha generado una demanda de estudios sobre salud animal y sus posibles impactos en la salud pública. El objetivo principal de este estudio fue evaluar la microbiota gastrointestinal de las tortugas amazónicas en cautiverio y vida libre, y la influencia del hábitat en la composición de la flora. Un total de 116 tortugas adultas, de ambos sexos fueron elegidas, y 51 fueron capturados en la isla de São Miguel, Santarém (PA), 50 animales en cautiverio para comercio y 15 provenían de un criadero de reproducción para la conservación, que se encuentra en la región metropolitana de Belém. De cada animal se recogió material biológico de la cloaca y se envió al laboratorio de la Universidad Federal de Pará, Brazil. De 116 muestras se obtuvieron 245 aislamientos bacterianos en el que 83 (33.87 %) eran de animales de vida libre, y 162 (65.72 %) de cautiverio. Especies de Klebsiella pneumoniae fueron los aislamientos más frecuentes de las 52 muestras, 21.22 % del crecimiento total de bacterias, seguido de Enterobacter cloacae 29 %, (35/14), Serratia marcescens 84 % (11/29) y Salmonella spp. 80 % (24/9). En las tortugas de vida libre, los microorganismos aislados corresponden a los géneros: Enterobacter, Klebsiella, Citrobacter y Aeromonas. Klebsiella pneumoniae, S. marcescens, E. cloacae y Salmonella spp. presentaron frecuencias altas en animales de cautiverio. Este resultado muestra una mayor diversidad de microorganismos en animales de vida libre y muestras con alta contaminación en animales de cautiverio. Las especies de Salmonella spp., E. coli y Acinetobacter spp., pueden ser sugeridas como indicadores de la calidad sanitaria de las poblaciones de la tortuga amazónica. Sin embargo el estudio reveló que el hábitat influyó en la composición de la flora gastrointestinal de las tortugas. El conocimiento de la flora gastrointestinal de los animales es de suma importancia en la identificación de los agentes patógenos presentes en la fauna nativa de la región amazónica.

The turtle Podocnemis expansa is an important wildlife species from the Amazon rainforest of Brazil. This also represents an important resource for coastal communities, as it has been historically consumed as food. Nevertheless, besides the sustainability issues, recent concerns have been raised over the health of the animals and possible impacts on public health. The aim of this study was to compare the occurrence of Enterobacteriaceae in the intestinal tract of captive and free living Amazon turtles. We examined a total of 116 adult turtles, including 51 free individuals from the island of Sao Miguel, in Santarém (Pará-PA) town, 50 captive business, and 15 from a conservation breeding area, located in the metropolitan area of Belém (PA). In total we obtained 245 bacterial growths in which 83 (33.8 %) were from the free ranging turtles, and 162 (65.7 %) isolates from captive animals. The species Klebsiella pneumoniae was the most frequent, with 52 isolates, totaling 21.2 % of bacterial growth, followed by Enterobacter cloacae 29 % (35/14), Serratia marcescens 84 % (29/11), and Salmonella spp. 80 % (24/9). In free ranging turtles the most commonly isolated microorganisms were Enterobacter spp., Klebsiella spp., and Citrobacter spp.; while Aeromonas spp., Klebsiela pneumoniae, S. marcescens, E. cloacae and Salmonella spp. were the most frequently identified microorganisms in captive animals. Results showed a greater diversity of microorganisms among the wild animals, and a high contamination per sample on captive animals. The species of Salmonella spp., E. coli and Acinetobacter spp. can be used as indicators of the sanitary quality of Amazon turtle populations. The habitat influenced the composition of the gastrointestinal flora of turtles. Knowledge of the gastrointestinal flora of animals is important for the identification of pathogens present in the native fauna of the Amazon region. Rev. Biol. Trop. 63 (4): 1083-1089. Epub 2015 December 01.

Tissue apoptosis in mice infected with Leptospira interrogans serovar Icterohaemorrhagiae.

BACKGROUND: This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi). METHODS: The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed. RESULTS: Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2, 4 and 7 (p = 0.025; p < 0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2, 4, 7 and 10 days (p = 0.0069; p < 0.05). CONCLUSIONS: Our results suggest that infection of L. interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.

Tissue apoptosis in mice infected with Leptospira interrogans serovar Icterohaemorrhagiae

Background: This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi). Methods: The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed. Results: Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2,4 and 7 (p = 0.025; p <0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2,4, 7 and 10 days (p = 0.0069; p<0.05). Conclusions: Our results suggest that infection of L interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.

Tissue apoptosis in mice infected with Leptospira interrogans serovar Icterohaemorrhagiae

Background:This investigation aimed to evaluate the occurrence of some apoptotic features induced by Leptospira interrogans serovar Icterohaemorrhagiae infection in young BALB/c mice during 2, 4, 7, 10, 14 and 21 days post-infection (dpi).Methods:The animals were euthanized and lung, liver and kidneys were harvested to histopathology analysis and immunohistochemistry to caspase-3 antigen detection was performed.Results:Chromatin condensation in kidney and liver tissues, but not in lung tissue, was observed. Caspase-3 reactive cells, mainly characterized as renal epithelial cells, were detected in the days 14 and 21 at high levels when compared to days 2,4 and 7 (p = 0.025; p <0.05). Lung sections revealed caspase-3 labeled alveolar cells in 10 and 14 days post-infection was higher than observed at 7 days (p = 0.0497; p < 0.05). Liver sections demonstrated reactive cells at a highest level at 14 and 21 days post-infection when comparison to 2,4, 7 and 10 days (p = 0.0069; p<0.05).Conclusions:Our results suggest that infection of L interrogans induce in kidney, liver and lung an activation of apoptosis mediated by caspase-3 dependent pathway in later phases of infectious process.(AU)

Isolation and molecular characterization of Brazilian turkey reovirus from immunosuppressed young poults.

In this study, we investigated turkey reovirus (TReoV) in tissue samples from young birds, aged 15 days. RT-PCR for TReoV detected 3.3 % positive samples and TReoV was successfully isolated in Vero cells. Histological analysis of positive bursa of Fabricius (BF) revealed atrophied follicles and lymphocyte depletion. The number of CD8+, CD4+ and IgM+ cells was lower in infected BF. Phylogenetic analysis based on S3 gene showed that the Brazilian TReoV isolates clustered in a single group with 98-100 % similarity to TReoV strains circulating in the United States. This is the first indication that TReoV infection may be a contributing factor to immunosuppression in young birds.

Validation of a reference control for an SYBR-Green fluorescence assay-based real-time PCR for detection of bovine herpesvirus 5 in experimentally exposed bovine embryos.

The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced in vitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced in vitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves (p < 0.05). On the other hand, no differences were found in the development of bovine embryos in vitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs.

Estudo da mcrobiota fúngica gastrintestinal de morcegos (Mammalia, Chiroptera) da região noroeste do estado de São Paulo: potencial zoonótico / Study of gastrointestinal fungal flora of bats (Mammalia, Chiroptera) of the northwest region of São Paulo state: zoonotic potential

Os morcegos são hospedeiros de uma rica diversidade de microrganismos. Muitos trabalhos apontam uma estreitaligação entre quirópteros e fungos com potencial patogênico, principalmente por habitarem ambientes como cavernas, grutas e ocos de árvores, favoráveis à manutenção e propagação dos fungos. O objetivo do trabalho foi estudar a microbiota fúngica gastrintestinal de morcegos. Das 98 amostras pertencentes a 11 espécies de morcegos procedentes de 15 cidades estudadas, 20% são da espécie Carollia perspicillata, 19% Artibeus lituratus, 17% Molossus rufus, 13% Glossophaga soricina, 9% Nyctinomops macrotis, 8% Molossus molossus, 7% Desmodus rotundus, 2% Lasiurus ega, e 1%Eptesicus furinalis, Myotis nigricans e Tadarida brasiliensis. O gênero Aspergillus sp. foi isolado de 29% das amostras, seguidos por 6% Microsporum sp. e Penicillium sp., 4% Tricophyton sp. e zigomicetos e 2% Fusarium sp. Das espécies deleveduras, 14% foram de Rhodotorula sp., 10% Candida sp. e 2% Cryptococcus sp., 22% dos isolados permaneceram sem identificação. Todos os 82 cultivos de vísceras foram negativos para Histoplasma capsulatum. Houve associação estatísticasignificativa entre os resultados do cultivo microbiológico e as espécies de morcegos (p < 0,05). Concluímos que osmorcegos podem atuar como agentes veiculadores de fungos com potencial patogênico, entretanto outros trabalhosdevem ser realizados a fim de estabelecer estratégias que permitam identificar os principais fatores correlacionados com o crescimento e a disseminação dos microrganismos na natureza e qual a implicação dos quirópteros no cicloepidemiológico.

Bats are hosts of a rich diversity of microorganisms. Many studies indicate a close link between bats and fungi with pathogenic potential, especially for living in environments such as caves, caverns and hollow trees, favorable to the maintenance and spread of fungi. The objective was to study the gastrointestinal mycoflora of bats. Of the 98 samples belonging to 11 species of bats coming from 15 studied cities, 20% of the species were Carollia perspicillata, 19% Artibeus lituratus, 17% Molossus rufus, 13% Glossophaga soricina, 9% Nyctinomops macrotis, 8% Molossus molossus, 7% Desmodus rotundus, 2% Lasiurus ega and 1% Eptesicus furinalis, Myotis nigricans and Tadarida brasiliensis. The genus Aspergillus sp. was isolated from 29% of the samples, followed by 6% Microsporum sp. and Penicillium sp. 4% Trichophyton sp. and zygomycetes and 2% Fusarium sp. Of yeast species, 14% were from Rhodotorula sp., 10% Candida sp. and 2% Cryptococcus sp., 22% of isolates remained unidentified. All 82 cultures of organs were negative for Histoplasma capsulatum. There was a statistically significant association between the results of microbiological culture and bat species (p < 0.05). We conclude that the bats can act as disperser agents of fungi with pathogenic potential, although other studies should be performed to establish strategies to identify the main factors correlated with the growth and spread of microorganisms in nature and implication of bats in the epidemiological cycle.