Isolation of Salmonella spp. in cattle egrets (Bubulcus ibis) from Fernando de Noronha Archipelago, Brazil

Abstract The growth of the population of cattle egrets (Bubulcus ibis) in the archipelago of Fernando de Noronha constitutes a threat to public health and biological diversity because of their competition with and predation on native species and the possibility of transmission of pathogens to human beings, livestock and native wildlife. The aim here was to search for, isolate and identify serovars of Salmonella in clinically healthy local cattle egrets. Cloacal swabs were obtained from 456 clinically healthy cattle egrets of both sexes and a variety of ages. The swabs were divided into 51 pools. Six of these (11.7%) presented four serovars of Salmonella enterica subspecies enterica: Salmonella serovar Typhimurium; Salmonella serovar Newport; Salmonella serovar Duisburg; and Salmonella serovar Zega. One sample was identified as S. enterica subspecies enterica O16:y:-. Results in this study suggest that cattle egrets may be reservoirs of this agent on Fernando de Noronha and represent a risk to public health and biological diversity.

Isolation of Salmonella spp. in cattle egrets (Bubulcus ibis) from Fernando de Noronha Archipelago, Brazil.

The growth of the population of cattle egrets (Bubulcus ibis) in the archipelago of Fernando de Noronha constitutes a threat to public health and biological diversity because of their competition with and predation on native species and the possibility of transmission of pathogens to human beings, livestock and native wildlife. The aim here was to search for, isolate and identify serovars of Salmonella in clinically healthy local cattle egrets. Cloacal swabs were obtained from 456 clinically healthy cattle egrets of both sexes and a variety of ages. The swabs were divided into 51 pools. Six of these (11.7%) presented four serovars of Salmonella enterica subspecies enterica: Salmonella serovar Typhimurium; Salmonella serovar Newport; Salmonella serovar Duisburg; and Salmonella serovar Zega. One sample was identified as S. enterica subspecies enterica O16:y:-. Results in this study suggest that cattle egrets may be reservoirs of this agent on Fernando de Noronha and represent a risk to public health and biological diversity.

Antimicrobial susceptibility and serovars of salmonella circulating in commercial poultry carcasses and poultry products in Brazil.

Antimicrobial resistance was evaluated for 12,582 strains of Salmonella isolated by public and private laboratories from commercial poultry carcasses and poultry products from different regions in Brazil between 2007 and 2011. These isolates were submitted unsolicited to the National Reference Laboratory for Bacterial Enteroinfections, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil, for conclusive antigenic characterization. The prevalent Salmonella enterica serogroups were O:4 (B) (28.9%), O:9 (D1) (22.3%), O:21 (L) (15.9%), and O:7 (C1) (14.3%); the most prevalent serovars were Enteritidis, Minnesota, Typhimurium, Schwarzengrund, and Mbandaka. The antimicrobial susceptibility profiles of 1,234 strains indicated that 54.5% were resistant to up to eight antimicrobial drugs. Multidrug resistance (three or more classes) was observed in 16.4% of the strains, with 190 distinct patterns. The results showed increased resistance to ampicillin (12.4 to 18.9%), tetracycline (≈ 15.2 to ≈ 18.9%), and gentamicin (f7.0 to ≈ 9.6%) during the study period. Decreased resistance to nitrofurans (61.9 to 9.2%), quinolones (44.4 to 15.5%), and folate inhibitors (11.7 to 7.2%) was observed. Resistance to third-generation cephalosporins was detected in 5.5% of the strains throughout the study period; resistance to fluoroquinolones has been observed in 0.3% of the strains since 2009. These findings highlight the importance of surveillance along the food chain for detection of altered patterns among foodborne zoonotic bacteria that are important to public health.

Research of Salmonella spp. and evaluation of pathogenicity, cytotoxicity of Escherichia coli isolates proceeding from sparrows (Passer domesticus) / Pesquisa de Salmonella spp. e avaliação da patogenicidade, citotoxidade de isolados de Escherichia coli procedentes de pardais (Passer domesticus)

The aim of this study was to research the occurrence of Salmonella spp. and Escherichia coli in feces samples of sparrows, as well as to identify the pathogenicity, cytotoxicity and sensitivity profile of the isolates to antimicrobial use. Two hundred and twenty eight sparrows were captured in eight farms. The in vitro pathogenicity test was performed by the isolates culture on congo red-magnesium oxalate Agar, whilst the in vivo pathogenicity test was performed in one day-old chicks. In order to study the cytotoxic effects of indicators, samples were inoculated into Vero cells. The results obtained for Escherichia coli isolation confirmed the presence of this microorganism in 30 (13.2%) of the evaluated samples. Out of those isolates, 10 (33.3%) presented the capacity of absorbing ongo red. As for in vivo pathogenicity a 68.0% of mortality rate of the evaluated samples was observed. Out of 20 isolates tested for cytotoxin production, none of them presented cytotoxic effect in the Vero cells. The Salmonella spp was isolated only in one sample (0.04%), and it was identified as Salmonella enterica subspecies houtenae. Results obtained through this research indicate the need for new studies to identify other virulence factors of E. coli samples and to delineate the phylogenetic profile of the isolates in order to establish a relation with colibacillosis outbreaks in chickens and broilers in the studied region, as well as to analyze the critical points in the aviculture productive chain to identify the source of Salmonella enterica subspecies houtenae.

Objetivou-se com este estudo pesquisar a ocorrência de Salmonella spp. e Escherichia coli em amostras de fezes de pardais, além de avaliar a patogenicidade, citotoxicidade e perfil de sensibilidade dos isolados frente a antimicrobianos. Foram capturados 228 pardais em oito granjas. O teste de patogenicidade in vitro foi realizado por meio do cultivo dos isolados em ágar oxalato de magnésio acrescido de vermelho de congo, enquanto o teste de patogenicidade in vivo foi realizado em pintos de um dia. Para o estudo dos indicadores dos efeitos citotóxicos, as amostras foram inoculadas em células Vero. Os resultados obtidos quanto ao isolamento de Escherichia coli confirmaram a presença deste microorganismo em 30 (13,2%) amostras analisadas. Destes isolados, dez (33,3%) apresentaram capacidade de absorção do vermelho congo. Quanto à patogenicidade in vivo observou-se uma taxa de mortalidade de 68,0% das amostras analisadas. Dos 20 isolados testados quanto à produção de citotoxina, nenhum apresentou efeito citotóxico nas células Vero. Obteve-se o isolamento de Salmonella spp. em apenas uma amostra (0,04%), sendo tipificada em Salmonella enterica subespécie houtenae. Os resultados obtidos nesta pesquisa indicam a necessidade da realização de novos estudos para identificar outros fatores de virulência das amostras de E. coli e traçar o perfil filogenético dos isolados para estabelecer uma relação com surtos de colibacilose em galinhas e frango de corte na região estudada, além de analisar os pontos críticos na cadeia produtiva da avicultura para identificar a origem da Salmonella enterica subespécie houtenae.

Effects of gamma irradiation on the viability and phenotypic characteristics of Salmonella Enteritidis inoculated into specific-pathogen-free eggs.

The goal of this study was to determine the effects of various levels of gamma irradiation on the phenotypic characteristics of 20 strains of Salmonella Enteritidis inoculated separately into specific-pathogen-free shell eggs. Bacterial strains were inoculated into egg yolks and exposed to (60)Co radiation at doses of 0.49 to 5.0 kGy. The eggs were maintained at 25°C and analyzed for the presence of Salmonella on days 1, 2, 4, and 7, and the recovered Salmonella isolates were characterized biochemically. All strains were resistant to doses of 0.49, 0.54, 0.59, 0.8, and 1 kGy; colony counts were ≥10(5) CFU/ml of egg yolk except for one strain, which was detected at 96 h and at 7 days after irradiation at 1 kGy, with a population reduction of 2 log CFU/ml. For the other evaluated doses, 12 strains (60.0%) were resistant at 1.5 kGy and 7 strains (35.0%) were resistant at 3.0 kGy. Among all analyzed strains, 5.0 kGy was more effective for reducing and/or eliminating the inoculated bacteria; only two (10%) strains were resistant to this level of irradiation. Salmonella colony counts were significantly reduced (P < 0.01) with increasing doses from the day 1 to 7 of observation, when microbial growth peaked. Loss of mobility, lactose fermentation, citrate utilization, and hydrogen sulfide production occurred in some strains after irradiation independent of dose and postirradiation storage time. Increases in antibiotic susceptibility also occurred: seven strains became sensitive to ß-lactams, two strains became sensitive to antifolates, and one strain each became sensitive to fluoroquinolone, phenicol, nitrofurans, tetracyclines, and aminoglycosides. The results indicate that up to 5.0 kGy of radiation applied to shell eggs inoculated with Salmonella Enteritidis at 4 log CFU per egg is not sufficient for complete elimination of this pathogen from this food matrix.

Enteropatógenos bacterianos em peixes criados em uma estação de reciclagem de nutrientes e no ecossistema relacionado / Bacterial enteropathogens from fishes of a nutrient recycle system and its ecosystem

Avaliou-se a presença de enteropatógenos bacterianos em 72 amostras obtidas a partir de peixes criados em sistema de reciclagem de nutrientes, em estação experimental, no município de Petrópolis, RJ. Paralelamente, foram obtidas amostras do lodo utilizado como adubo orgânico e da cama de aves localizada na área interna dos tanques criatórios. A metodologia empregada incluiu o pré-enriquecimento em Caldo Lactosado e Agua Peptonada Tamponada, seguido de enriquecimento em Agua Peptonada Alcalina (pH 8,4-8,6) e subseqüente semeadura em Agar GSP para o isolamento de Aeromonas spp. e Plesiomonas shigelloides. Para os demais microrganismos, alíquotas de 1ml foram inoculadas nos meios de enriquecimento Caldo Rappaport-Vassiliadis e Caldo Tetrationato de Kauffmann com posterior semeadura em Agar Entérico Hektoen e Agar Salmonella-Shigella. Com a finalidade de monitorar o índice de coliformes fecais, visando conhecer a qualidade da água para este sistema, paralelamente à coleta de peixes foram avaliadas amostras de água dos tanques criatórios e de macrófitas. No cômputo geral foram isoladas 116 cepas de enteropatógenos bacterianos, destacando-se Aeromonas spp (67,2 por cento) com 9 espécies (A. veronii biogrupo sobria, A. hydrophila, A. sobria, A. trota, A.eucrenophila, A. veronii biog. veronii, A. media, A. caviae e A jandaei) e Aeromonas spp., seguido de Edwardsiella tarda (16,4 por cento), Plesiomonas shigelloides (12,9 por cento) e Salmonella (3,4 por cento). A análise da qualidade da água empregada no sistema revelou, de um modo geral, índices mais elevados de coliformes fecais nos tanques dos peixes (>1800/100 ml).

The presence of bacterial enteropathogens from fishes of a nutrient recycle system from a Experimental Station in Petropolis, RJ, was evaluated in 72 samples from april 2000 to july 2001 Simultaneously was collected the mud used as organic manure and poultry beds localized next to the tanks. The isolation procedures included preenrichment in Peptone Water followed by enrichment with alcaline Peptone Water (pH 8.4-8.6), and streaked onto GSP Agar for the isolation of Aeromonas spp. and Plesiomonas shigelloides. For enteropathogenic bacteria of the Enterobacteriaceae family, 1ml samples were transferred for enrichment in Rappaport- Vassiladis broth and Kauffmann Tetrathionate Broth followed by streak onto Hektoen Enteric Agar and Salmonella-Shigella Agar. Simultaneously at each visit samples of water from fish and macrophyte tanks were collected for monitoring faecal coliforms (MPN) using A1 medium. Among the 116 isolates, Aeromonas spp. were the most frequent (67.2 percent) with 9 species (A. veronii, biogroup sobria, A. hidrophila, A. sobria, A. trota, A. eucrenophila, A. veronii biog. veronii, A. media, A. cavia and A. jandaei), followed by Edwardsiella tarda (16.4 percent), Plesiomonas shigelloides (12.9 percent) and Salmonella spp. (3.4 percent). The NMP of fecal coliforms showed higher values in the fish tanks (>1800/100ml).

Antimicrobial resistance and R-plasmid in Salmonella spp from swine and abattoir environments

Sorovares de Salmonella isolados de suinos são de particular interesse não só pelo potencial patogênico para esta espécie animal, como também pela sua relevância em Saúde Pùblica. Com base no perfil de resistência aos antimi-crobianos foram selecionadas 13 amostras de Salmonella pertencentes aos sorovares Muenster (7 amostras), Derby (4), Typhimurium (1) e Braenderup (1), isoladas de suinos sadios e do ambiente de abatedouro no Estado do Rio de Janeiro. As amostras foram submetidas a conjugação bacteriana, utilizando como receptora E.coli K12 55 Nal r Lac+ F -, com a finalidade de verificar a capacidade da transferência de marcos de resistência. O fenômeno de transferência gênica foi detectado em 7 amostras e, com exceção de Salmonella Typhimurium que transconjugou para Sm, Tc e Su, as demais se caracterizaram por transferir somente o marco Su. Na análise plasmidial das amostras doadoras e suas respectivas transconjugantes foi revelado um plasmídio de 63 Kb, provavelmente relacionado com a multirresistência de S. Typhimurium.

Padrão de susceptibilidade a antimicrobianos e perfil plasmidial em Salmonella Muenster isoladas de suínos e do ambiente de abatedouros / Antimicrobial susceptibility pattern and plasmid profile in Salmonella Muenster isolated from swine and abattoir environment, Brazil

Foram analisadas para a presença de plasmídios 38 amostras de Salmonella Muenster isoladas de suínos e do ambiente de abatedouros localizados no Estado do Rio de Janeiro, no período de março de 1991 a fevereiro de 1992. A seleção das amostras teve como orientação o perfil apresentado frente aos antimicrobianos estreptomicina, tetraciclina, sulfonamida e sulfametoxazol-trimetoprim, sendo analisadas 13 cepas resistentes a um ou mais antimicrobianos, 18 com grau intermediário e sete sensíveis. Plasmidios variando em tamanho de 1,2 Kb a 42 Kb foram detectados em 37 (97,36 por cento) das 38 amostras, correspondendo a 11 perfis distintos (P1 a P11), variando em número de 1 a 6 plasmidios por modelo. O número e diversidade de plasmidios foi maior que os marcos de resistência por cepa. O plasmidio de 2,85 Kb foi o mais freqüente, estando presente em 83,78 por cento das 37 cepas; somente o de 7,95 Kb foi detectado nos dois abatedouros. Não houve paralelismo entre padrão de resistência e perfil plasmidial, onde um mesmo antibiotipo foi encontrado em vários perfis plasmidiais. Os resultados na presente investigação permitiram concluir que a caracterização de plasmidios constituiu-se uma ferramenta útil e simples no rastreamento epidemiológico deste sorovar.

Conventional and molecular typing of Salmonella typhi strains from Brazil

Phenotypic and genotypic characteristics of Salmonella Typhi were studied in 30 strains, isolated in different years, from some areas in Brazil. Conventional typing methods were performed by biochemical tests, Vi phage-typing scheme, and antimicrobial susceptibility test. Molecular typing methods were performed by analysis of plasmid DNA and by random amplified polymorphic DNA (RAPD-PCR). For the latter, an optimization step was performed to ensure the reproducibility of the process in genetic characterization of S. Typhi. The predominance of 76.7 percent of biotype I (xylose +, arabinose -) was noticed in all studied areas. Three phage types were recognized, with prominence for the phage types A (73.3 percent) and I+IV (23.3 percent). All the strains were susceptible to the drugs used. However, 36.7 percent of the strains contained plasmids, with predominance of the 105 Kb plasmid. RAPD was capable of grouping the strains in 8 genotypic patterns using primer 784, in 6, using primer 787 and in 7, using primer 797. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented nonsignificant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and performance, being considered as a promising alternative typing method for S. Typhi

Conventional and molecular typing of Salmonella typhi strains from Brazil.

Phenotypic and genotypic characteristics of Salmonella Typhi were studied in 30 strains, isolated in different years, from some areas in Brazil. Conventional typing methods were performed by biochemical tests, Vi phage-typing scheme, and antimicrobial susceptibility test. Molecular typing methods were performed by analysis of plasmid DNA and by random amplified polymorphic DNA (RAPD-PCR). For the latter, an optimization step was performed to ensure the reproducibility of the process in genetic characterization of S. Typhi. The predominance of 76.7% of biotype I (xylose +, arabinose -) was noticed in all studied areas. Three phage types were recognized, with prominence for the phage types A (73.3%) and I+IV (23.3%). All the strains were susceptible to the drugs used. However, 36.7% of the strains contained plasmids, with predominance of the 105 Kb plasmid. RAPD was capable of grouping the strains in 8 genotypic patterns using primer 784, in 6, using primer 787 and in 7, using primer 797. Conventional phenotypic typing methods, as well as the DNA plasmid analysis, presented nonsignificant discriminatory power; however, RAPD-PCR analysis showed discriminatory power, reproducibility, easy interpretation and performance, being considered as a promising alternative typing method for S. Typhi.

Microbial contamination of sand from major beaches in Fortaleza, Ceará State, Brazil

No presente trabalho, avaliou-se a presença de contaminação fecal e de microrganismos patogênicos em areia seca e areia úmida de três praias de Fortaleza, Ceará: Praia do Mucuripe, Praia do Futuro e Praia do Caça e Pesca. A praia que apresentou maior índice de contaminação com E.coli em areia seca foi a do Caça e Pesca (56 per center), seguido das praias do Mucuripe (28 per center) e Futuro (16 per center). Em areia úmida a ordem decrescente de contaminação por E. coli foi Praia da Caça e Pesca, Futuro e Mucuripe com 48 per center, 28 per center e 24 per center de contaminação, respectivamente. Salmonella foi isolada de duas amostras da Praia do Futuro, uma de areia seca e outra de areia úmida. V. parahaemolyticus foi isolado em 2 amostras de areia seca e 2 de úmida da Praia da Caça e Pesca. Esse patógeno foi isolado de uma amostra de areia úmida da Praia do Futuro e de três de areia úmida e quatro de areia seca da Praia do Mucuripe. As leveduras encontradas em todas as amostras pertenciam ao gênero Candida. O índice de contaminação por leveduras foi maior em amostras de areia seca que de areia úmida. A praia que apresentou maior contaminação por leveduras foi a Praia do Futuro (41 per center), seguida da Praia do Caça e Pesca e (33 per center) e da Praia do Mucuripe (26per center).

Helicobacter pylori in dyspeptic children and adults: endoscopic, bacteriologic and histologic correlations

Using different bacteriological (urease test, Gram staining and culture) and histological (Steiner staining and modified Giemsa staining) techniques, we searched for the presence of Helicobacter pylori in the gastric antrum of 200 dyspeptic Brazilian patients (106 females and 94 males aged 19 days to 81 years). The presence of bacteria was then correlated with the endoscopic and histological findings. H. pylori was present in 59.5 of the population studied. In Brazil, colonization occurs early, involving 37 of the dyspeptic population by 20 years of age. The presence of H. pylori in the gastric antrum was strongly associated with duodenal ulcer (P < 0.001) and a normal endoscopic examination did not exclude the possibility of colonization of the gastric antrum by H. pylori. The most sensitive test was the preformed urease test (89). We conclude that more than one diagnostic method should preferably be used for the detection of H. pylori and that the presence of H. pylori is closely correlated with active chronic gastritis (P < 0.001).