Serotyping and Antimicrobial Susceptibility Profiling of Glaesserella parasuis Isolated from Diseased Swine in Brazil.

Glaesserella parasuis is one of the major pathogens in swine intensive production systems. To date, 15 serovars have been described, and the prevalence of these serotypes in different geographical regions has been identified by several methods. G. parasuis outbreaks could be controlled with vaccination if it were not for serovar diversity and limited cross-serovar protection; consequently, antibiotic therapy continues to be necessary for infection control. Here, we present the isolation, identification, serotyping, and antibiotic susceptibility profiling of G. parasuis from diseased swine in Brazil. A total of 105 G. parasuis strains, originating from nine different Brazilian states, were evaluated, and serotypes 4 and 5 were found to be the most prevalent (27.6% and 24.8% respectively). Aminoglycosides, florfenicol, tiamulin, and ß-lactams were tested, and they presented lower resistant rates against G. parasuis strains. The highest resistance rates were observed against tylosin (97.1%), sulfadimethoxine (89.5%), danofloxacin (80%), trimethoprim/sulfamethoxazole (62.5%), enrofloxacin (54.3%), and clindamycin (50.5%). Multidrug resistance was detected in 89.5% of tested strains, and a total of sixty resistance profiles were identified. The cluster analysis of resistance patterns showed no correlation with the isolation year or G. parasuis serotype.

Vagococcus sp. a porcine pathogen: molecular and phenotypic characterization of strains isolated from diseased pigs in Brazil.

INTRODUCTION: Vagococcus spp. is known for its importance as a systemic and zoonotic bacterial pathogen even though it is not often reported in pigs. This is related to the pathogen misidentification due to the lack of usage of more discriminatory diagnostic techniques. Here we present the first report of Vagococcus lutrae in swine and the characterization of Vagococcus fluvialis and Vagococcus lutrae isolated from diseased animals. METHODOLOGY: Between 2012 and 2017, 11 strains with morphological characteristics similar to Streptococcus spp. were isolated from pigs presenting different clinical signs. Bacterial identification was performed by matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry and confirmed by 16S rRNA sequencing and biochemical profile. Strains were further genotyped by single-enzyme amplified fragment length polymorphism (SE-AFLP). Broth microdilution was used to determine the minimal inhibitory concentration of the antimicrobials of veterinary interest. RESULTS: Ten strains were identified as V. fluvialis and one was identified as V. lutrae. The SE-AFLP analysis enabled the species differentiation with specific clustering of all V. fluvialis separately from the V. lutrae strain. Most strains presented growth in the maximum antibiotic concentration values tested for eight of the 10 analyzed antimicrobial classes. CONCLUSIONS: The observed resistance pattern can represent a problem for veterinary and producers in the treatment of diseases associated Vagococcus spp. in swine production. Vagococcus species may also be a risk for pig industry workers. The data described here will be of great value in further understanding the behavior of this pathogen in animal production.

Identificação de Klebsiella spp. em fezes de psitacídeos cativos

No Brasil são encontradas mais de 80 espécies de aves da família Psittacidae, incluindo papagaios, araras e periquitos. A manutenção destas aves em zoológicos contribui para a conservação das espécies, mas o ambiente de cativeiro pode favorecer a colonização intestinal por enterobactérias. O objetivo deste trabalho foi identificar a presença de Klebsiella spp. nas fezes de psitacídeos cativos. Foram analisadas 34 amostras de fezes de aves alojadas em um zoológico do estado de São Paulo em maio de 2017. As amostras foram coletadas com auxílio de suabe estéril, no mês de maio de 2017 e submetidas ao cultivo bacteriológico, utilizando-se ágar MacConkey com incubação a 37oC por 24 horas. Foram identificadas 11 (32,35%) amostras positivas para Klebsiella spp., incluindo as espécies: K. pneumoniae e K. oxytoca. Os resultados demonstraram a presença de aves colonizadas por Klebsiella spp. Novos estudos são necessários para determinação da patogenicidade destas estirpes, perfil de resistência antimicrobiana e riscos de saúde animal, uma vez que as enterobactérias não são componentes da microbiota de psitacídeos saudáveis.(AU)

More than 80 species of birds belonged to the family Psittacidae in Brazil, including parrots, macaws and parakeets. The maintenance of these birds in Zoos may help the conservation of the species, but the captivity can favor intestinal colonization by enterobacteria. The aim of this study was to identify the presence of Klebsiella spp. in the fecal samples of captive parrots. Thirty -four fecal samples of psittacine birds from a Zoo of São Paulo state were analyzed. The samples were collected with a sterile swab during May 2017, and subjected to bacteriological culture on MacConkey agar, with incubation at 37oC for 24 hours. The results showed that 11 (32.35%) were positive for Klebsiella spp., with identification of the species: K. pneumoniae and K. oxytoca. This study showed the presence of birds colonized by Klebsiella spp. Further studies are required to determine the pathogenicity, the antimicrobial resistance profile and animal health risks, since enterobacteria do not belong to microbiota of psittacine birds.(AU)

Identificação de Klebsiella spp. em fezes de psitacídeos cativos

No Brasil são encontradas mais de 80 espécies de aves da família Psittacidae, incluindo papagaios, araras e periquitos. A manutenção destas aves em zoológicos contribui para a conservação das espécies, mas o ambiente de cativeiro pode favorecer a colonização intestinal por enterobactérias. O objetivo deste trabalho foi identificar a presença de Klebsiella spp. nas fezes de psitacídeos cativos. Foram analisadas 34 amostras de fezes de aves alojadas em um zoológico do estado de São Paulo em maio de 2017. As amostras foram coletadas com auxílio de suabe estéril, no mês de maio de 2017 e submetidas ao cultivo bacteriológico, utilizando-se ágar MacConkey com incubação a 37oC por 24 horas. Foram identificadas 11 (32,35%) amostras positivas para Klebsiella spp., incluindo as espécies: K. pneumoniae e K. oxytoca. Os resultados demonstraram a presença de aves colonizadas por Klebsiella spp. Novos estudos são necessários para determinação da patogenicidade destas estirpes, perfil de resistência antimicrobiana e riscos de saúde animal, uma vez que as enterobactérias não são componentes da microbiota de psitacídeos saudáveis.

More than 80 species of birds belonged to the family Psittacidae in Brazil, including parrots, macaws and parakeets. The maintenance of these birds in Zoos may help the conservation of the species, but the captivity can favor intestinal colonization by enterobacteria. The aim of this study was to identify the presence of Klebsiella spp. in the fecal samples of captive parrots. Thirty -four fecal samples of psittacine birds from a Zoo of São Paulo state were analyzed. The samples were collected with a sterile swab during May 2017, and subjected to bacteriological culture on MacConkey agar, with incubation at 37oC for 24 hours. The results showed that 11 (32.35%) were positive for Klebsiella spp., with identification of the species: K. pneumoniae and K. oxytoca. This study showed the presence of birds colonized by Klebsiella spp. Further studies are required to determine the pathogenicity, the antimicrobial resistance profile and animal health risks, since enterobacteria do not belong to microbiota of psittacine birds.

Arcobacter spp. in fecal samples from Brazilian farmed caimans (Caiman yacare, Daudin 1802).

The aim of this study was to perform the identification and molecular characterization of Arcobacter cryaerophilus and Arcobacter butzleri isolated from caiman (Caiman yacare), kept at a production farm, in Brazil. Forty fecal samples were analyzed. After isolation and identification, 21/40 strains of A. butzleri and 19/40 strains of A. cryaerophilus were subjected to PCR for potential virulence gene detection. The results of the PCR showed 38/40 strains positive for the cadF, cj1349, ciaB, and tlyA genes, 39/40 strains positive for the pldA gene, and 40/40 strains positive for the mviN gene. None of the strains presented the irgA gene. Hemagglutinin (hecA gene) and hemolysin (hecB) genes were detected in 21/40 and 16/40 strains, respectively. The SE-AFLP showed a great genetic diversity, but some clonally groups were disseminated in various tanks. These data reveal that the strains presented the same virulence traits described from Arcobacter isolated from food-borne disease in humans.

Pesquisa de bactérias gram negativas em papagaio do congo (Psittacus erithacus) mantidos em criatório comercial / Research for gram-negative bacteria in congo's parrot (Psittacus erithacus) held in commercial breeding

A microbiota intestinal de psitacídeos é constituida principalmente por bactérias Gram positivas e tem menor prevalência de Gram negativas. Embora existam poucos estudos descrevendo a microbiota destas aves, uma série de agentes bacterianos com potencial zoonótico pode ser transmitida das aves para os humanos. O objetivo deste trabalho foi pesquisar a presença de bactérias Gram negativas em um criadouro de papagaios do congo (Psittacus erithacus). Foram coletadas e analisadas amostras de fezes e de coana de 24 animais. Os resultados obtidos demonstraram a presença de bactérias Gram negativas nas fezes (45,8%) e na coana (62,5%). Foi isolado um total de 26 colônias bacterianas, com predomínio de agentes da família Enterobacteriaceae (21/26). As espécies encontradas foram Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis e Aeromonas caviae. Os dados deste trabalho revelaram uma elevada frequência de bactérias Gram negativas na microbiota destas aves. Estudos futuros serão realizados para avaliar a virulência e a resistência antimicrobiana, determinando o impacto potencial da presença destes agentes para a saúde dos animais e os riscos zoonóticos.(AU)

The intestinal microbiota of parrots consists mainly by Gram positive bacteria and has a lower prevalence of Gram negative. Although there are few studies describing the microbiota of these birds, many of these bacteria have a zoonotic potential and can be transmitted from birds to humans. The aim of this study was to investigate the presence of Gram negative in congo parrots (Psittacus erithacus). Fecal and coana samples collected from 24 birds were analyzed. The results showed the presence of Gram negative bacteria in feces (45.8%) and coana (62.5%). A total of 26 strains were isolated, with predominance of members of the Enterobacteriaceae family (21/26). The species identified were Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis and Aeromonas caviae. The data of this study revealed a high frequency of Gram negative bacteria in the microbiota of these birds. Future studies will be conducted to assess virulence and antimicrobial resistance profile, looking for the potential impact of these agents over animal health and zoonotic risks.(AU)

Isolamento e identificação de Pasteurella multocida e Bordetella spp. em coelhos (Oryctolagus cuniculus) / Isolation and identification of Pasteurella multocida and Bordetella spp. in rabbits (Oryctolagus cuniculus)

As doenças respiratórias representam uma das principais causas de morbidade e mortalidade em coelhos. A pasteurelose é a doença bacteriana de maior impacto na cunicultura brasileira. Infecções mistas por Pasteurella multocida e Bordetella spp. resultam em um quadro clínico grave, com descarte de matrizes e elevado prejuízo econômico. O objetivo desse trabalho foi pesquisar a presença de Pasteurella multocida e Bordetella spp. em uma granja de coelhos comerciais com histórico de doença respiratória crônica. Foram avaliados 34 animais doentes. Na cultura bacteriológica foram identificados 6 animais positivos para Pasteurella multocida. e 3 positivos para Bordetella spp. Estes achados confirmam a presença destes agentes na granja e apontam a necessidade de adoção de medidas sanitárias para controle da infecção.(AU)

Respiratory diseases represent a major cause of morbidity and mortality in rabbits. Pasteurellosis is the most important bacterial disease on Brazilian cuniculture flocks. The infection associated by P. multocida and Bordetella spp. results in a severe clinical picture, and becomes responsible for waste matrices and high economic damage. The aim of this study was to search the presence of Pasteurella multocida and Bordetella spp in a commercial farm of rabbits, with a history of chronic respiratory disease. Thirty-four sick animals were investigated. The results of bacteriological tests showed six animals positive for Pasteurella multocida and 3 positive for Bordetella spp. These results confirm the presence of both pathogens and highlight the need of sanitary management for infection control.(AU)

Isolamento e identificação de Pasteurella multocida e Bordetella spp. em coelhos (Oryctolagus cuniculus) / Isolation and identification of Pasteurella multocida and Bordetella spp. in rabbits (Oryctolagus cuniculus)

As doenças respiratórias representam uma das principais causas de morbidade e mortalidade em coelhos. A pasteurelose é a doença bacteriana de maior impacto na cunicultura brasileira. Infecções mistas por Pasteurella multocida e Bordetella spp. resultam em um quadro clínico grave, com descarte de matrizes e elevado prejuízo econômico. O objetivo desse trabalho foi pesquisar a presença de Pasteurella multocida e Bordetella spp. em uma granja de coelhos comerciais com histórico de doença respiratória crônica. Foram avaliados 34 animais doentes. Na cultura bacteriológica foram identificados 6 animais positivos para Pasteurella multocida. e 3 positivos para Bordetella spp. Estes achados confirmam a presença destes agentes na granja e apontam a necessidade de adoção de medidas sanitárias para controle da infecção.

Respiratory diseases represent a major cause of morbidity and mortality in rabbits. Pasteurellosis is the most important bacterial disease on Brazilian cuniculture flocks. The infection associated by P. multocida and Bordetella spp. results in a severe clinical picture, and becomes responsible for waste matrices and high economic damage. The aim of this study was to search the presence of Pasteurella multocida and Bordetella spp in a commercial farm of rabbits, with a history of chronic respiratory disease. Thirty-four sick animals were investigated. The results of bacteriological tests showed six animals positive for Pasteurella multocida and 3 positive for Bordetella spp. These results confirm the presence of both pathogens and highlight the need of sanitary management for infection control.

Pesquisa de bactérias gram negativas em papagaio do congo (Psittacus erithacus) mantidos em criatório comercial / Research for gram-negative bacteria in congo's parrot (Psittacus erithacus) held in commercial breeding

A microbiota intestinal de psitacídeos é constituida principalmente por bactérias Gram positivas e tem menor prevalência de Gram negativas. Embora existam poucos estudos descrevendo a microbiota destas aves, uma série de agentes bacterianos com potencial zoonótico pode ser transmitida das aves para os humanos. O objetivo deste trabalho foi pesquisar a presença de bactérias Gram negativas em um criadouro de papagaios do congo (Psittacus erithacus). Foram coletadas e analisadas amostras de fezes e de coana de 24 animais. Os resultados obtidos demonstraram a presença de bactérias Gram negativas nas fezes (45,8%) e na coana (62,5%). Foi isolado um total de 26 colônias bacterianas, com predomínio de agentes da família Enterobacteriaceae (21/26). As espécies encontradas foram Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis e Aeromonas caviae. Os dados deste trabalho revelaram uma elevada frequência de bactérias Gram negativas na microbiota destas aves. Estudos futuros serão realizados para avaliar a virulência e a resistência antimicrobiana, determinando o impacto potencial da presença destes agentes para a saúde dos animais e os riscos zoonóticos.

The intestinal microbiota of parrots consists mainly by Gram positive bacteria and has a lower prevalence of Gram negative. Although there are few studies describing the microbiota of these birds, many of these bacteria have a zoonotic potential and can be transmitted from birds to humans. The aim of this study was to investigate the presence of Gram negative in congo parrots (Psittacus erithacus). Fecal and coana samples collected from 24 birds were analyzed. The results showed the presence of Gram negative bacteria in feces (45.8%) and coana (62.5%). A total of 26 strains were isolated, with predominance of members of the Enterobacteriaceae family (21/26). The species identified were Escherichia coli, Enterobacter asburiae, Klebsiella oxytoca, Citrobacter freundii, Citrobacter amolonaticus, Enterobacter cloacae, Providencia alcalifaciens, Providencia rettgeri, Citobacter koseri, Citrobacter braakii, Pseudomonas mendocina, Alcaligenes faecalis and Aeromonas caviae. The data of this study revealed a high frequency of Gram negative bacteria in the microbiota of these birds. Future studies will be conducted to assess virulence and antimicrobial resistance profile, looking for the potential impact of these agents over animal health and zoonotic risks.

Draft Genome Sequence of Non-O1 and Non-O139 Vibrio cholerae Strain VCC19.

Vibrio cholerae O1 is the causative agent of cholera and is ubiquitous in the aquatic environment, while V. cholerae strains non-O1 and non-O139 are recognized as causative agents of sporadic and localized outbreaks of diarrhea. Here, we report the complete sequence of a non-O1 and non-O139 V. cholerae strain (VCC19), which was isolated from the environment in Brazil. The sequence includes the integrative conjugative element (ICE). This paper is the first report of the presence of such an element in a V. cholerae strain isolated in Brazil.

Avaliação da performance de metodologias de detecção de Cryptosporidium spp. e Giardia spp. em água destinada ao consumo humano, para o atendimento às demandas da Vigilância em Saúde Ambiental no Brasil / Performance evaluation of different methodologies for detection of Cryptosporidium spp. and Giardia spp. in water for human consumption to meet the demands of the Environmental Health Surveillance in Brazil

Objetivo: avaliar a performance de três protocolos de concentração de cistos e oocistos em amostras de água bruta de rios brasileiros. Métodos: os protocolos estudados foram precipitação química, filtração em membranas de 47mm de diâmetro e 3μm de porosidade nominal e filtração com o sistema Filta-Max®. Amostras de água bruta coletadas de rios nos estados de São Paulo e Minas Gerais foram analisadas após contaminação artificial. Os resultados de precisão inicial e recuperação de organismos marcados (Color-Seed®) foram comparados com os critérios da Agencia de Proteção Ambiental dos Estados Unidos da Améri¬ca (USEPA). Resultados: Nos ensaios de precisão inicial para cistos de Giardia, a filtração em membranas e com Filta-Max® atingiram os critérios da USEPA para recuperação de cistos. Para Cryptosporidium, somente o procedimento com Filta-Max® alcançou os critérios. Conclusão: o sistema Filta-Max® foi o único método que atingiu todos os critérios para identificação de Cryptosporidium e Giardia em água.

Objective: this study aimed to evaluate three different concentration protocols of cysts and oocysts in raw water samples in Brazilian rivers. Methods: the protocols studied were chemical precipitation, filtration in membranes of 47mm of diameter versus nominal porosity of 3µm and filtration using Filta-Max® system. The raw water samples collectedfrom rivers in the states of São Paulo andMinas Gerais were analyzed after being seeded with Color-Seed®. The results of initial precision using filtration in membranes and recovery of Color-Seed® organisms were compared to the acceptance criteria established by the United States Environmental Protection Agency (USEPA). Results: The study of initial precision to Giardia using filtration in membranes and recovery of Color-Seed® met the USEPA criteria; for Cryptosporidium only the Filta-Max® achieved the established criteria. Conclusion: Filta-Max® system showed to be the only method that achieved all the performance criteria for identification of Giardia and Cryptosporidium in water.