Survey on pathogenic Escherichia coli and Salmonella spp. in captive cockatiels (Nymphicus hollandicus)

Abstract We surveyed healthy captive cockatiels (Nymphicus hollandicus) for Escherichia coli and Salmonella spp. Cloacal swabs were collected from 94 cockatiels kept in commercial breeders, private residencies and pet shops in the cities of São Paulo/SP and Niterói/RJ (Brazil). Three strains of E. coli from each individual were tested for the presence of ExPEC-, APEC- and DEC-related genes. We evaluated the blaTEM, blaSHV, blaOXA, blaCMY, blaCTX-M, tetA, tetB, aadA, aphA, strAB, sul1, sul2, sul3, qnrA, qnrD, qnrB, qnrS, oqxAB, aac (6)′-Ib-cr, qepA resistance genes and markers for plasmid incompatibility groups. Salmonella spp. was not detected. E. coli was isolated in 10% of the animals (9/94). Four APEC genes (ironN, ompT, iss and hlyF) were detected in two strains (2/27-7%), and iss (1/27-4%) in one isolate. The highest resistance rates were observed with amoxicillin (22/27-82%), ampicillin (21/27-79%), streptomycin (18/27-67%), tetracycline (11/27-41%). Multiresistance was verified in 59% (16/27) of the isolates. We detected strAB, bla TEM, tetA, tetB, aadA, aphaA, sul1, sul2, sul3 resistance genes and plasmid Inc groups in 20 (74%) of the strains. E. coli isolated from these cockatiels are of epidemiological importance, since these pets could transmit pathogenic and multiresistant microorganisms to humans and other animals.

Survey on pathogenic Escherichia coli and Salmonella spp. in captive cockatiels (Nymphicus hollandicus)

Abstract We surveyed healthy captive cockatiels (Nymphicus hollandicus) for Escherichia coli and Salmonella spp. Cloacal swabs were collected from 94 cockatiels kept in commercial breeders, private residencies and pet shops in the cities of São Paulo/SP and Niterói/RJ (Brazil). Three strains of E. coli from each individual were tested for the presence of ExPEC-, APEC- and DEC-related genes. We evaluated the blaTEM, blaSHV, blaOXA, blaCMY, blaCTX-M, tetA, tetB, aadA, aphA, strAB, sul1, sul2, sul3, qnrA, qnrD, qnrB, qnrS, oqxAB, aac (6)-Ib-cr, qepA resistance genes and markers for plasmid incompatibility groups. Salmonella spp. was not detected. E. coli was isolated in 10% of the animals (9/94). Four APEC genes (ironN, ompT, iss and hlyF) were detected in two strains (2/277%), and iss (1/274%) in one isolate. The highest resistance rates were observed with amoxicillin (22/2782%), ampicillin (21/2779%), streptomycin (18/2767%), tetracycline (11/2741%). Multiresistance was verified in 59% (16/27) of the isolates. We detected strAB, bla TEM, tetA, tetB, aadA, aphaA, sul1, sul2, sul3 resistance genes and plasmid Inc groups in 20 (74%) of the strains. E. coli isolated from these cockatiels are of epidemiological importance, since these pets could transmit pathogenic and multiresistant microorganisms to humans and other animals.

Captive wild birds as reservoirs of enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC)

ABSTRACT Psittacine birds have been identified as reservoirs of diarrheagenic Escherichia coli, a subset of pathogens associated with mortality of children in tropical countries. The role of other orders of birds as source of infection is unclear. The aim of this study was to perform the molecular diagnosis of infection with diarrheagenic E. coli in 10 different orders of captive wild birds in the state of São Paulo, Brazil. Fecal samples were analyzed from 516 birds belonging to 10 orders: Accipitriformes, Anseriformes, Columbiformes, Falconiformes, Galliformes, Passeriformes, Pelecaniformes, Piciformes, Psittaciformes and Strigiformes. After isolation, 401 E. coli strains were subjected to multiplex PCR system with amplification of genes eae and bfp (EPEC), stx1 and stx2 for STEC. The results of these tests revealed 23/401 (5.74%) positive strains for eae gene, 16/401 positive strains for the bfp gene (3.99%) and 3/401 positive for stx2 gene (0.75%) distributed among the orders of Psittaciformes, Strigiformes and Columbiformes. None of strains were positive for stx1 gene. These data reveal the infection by STEC, typical and atypical EPEC in captive birds. The frequency of these pathotypes is low and restricted to few orders, but the data suggest the potential public health risk that these birds represent as reservoirs of diarrheagenic E. coli.

Gram-negative bacterium isolated from cardinals (Paroaria coronata and P. dominicana) seized from illegal wildlife trade / Bactérias gram-negativas em cardeais (Paroaria coronata e Paroaria dominicana) apreendidos do tráfico de animais silvestres

Illegal wildlife trade removes millions of birds from nature every year. Among the most trafficked species of birds are redcrested cardinals (Paroaria coronata) and red-cowled cardinals (Paroaria dominicana). The cloacal microbiota of free-living passerines consists mainly of gram-positive bacilli and cocci, and gram-negative bacilli predominate in captive birds. Under stress and low immunity, gram-negative species may cause opportunistic infections. This study identified bacteria from cloacal microbiota of 49 specimens of P. coronata and P. dominicana seized from illegal wildlife trade in São Paulo (SP). In this study, 13 species of gram-negative bacteria, including Salmonella spp. and Pseudomonas aeruginosa were isolated. An increased occurrence of Escherichia coli was identified in 42/49 (85.7 %) of fecal samples. Among the E. coli strains, 21/42 belonging to the phylogenetic groups B2 and D, were related to extraintestinal pathogenic strains causing disease in humans. Klebsiella pneumoniae were isolated in 28/49 (57.1 %) samples. These results reinforce the fact that stressful conditions of illegal trade can favor the colonization of cloacal microbiota of these birds by pathogens, which represents a risk for their reintroductioninto the wild, including the transmission of diseases to humans and other animals...

Anualmente o tráfico de animais silvestres retira milhões de aves da natureza. Os cardeais (Paroaria coronata) e cardeais-donordeste (Paroaria dominicana) estão incluídos entre as espécies de aves mais traficadas. A microbiota cloacal de passeriformes de vida livre é composta principalmente por bacilos e cocos gram-positivos, já os bacilos gram-negativos predominam em aves de cativeiro. Em situações de estresse e baixa de imunidade as bactérias gram-negativas podem causar infecções oportunistas. O presente trabalho identificou bactérias da microbiota da cloaca de 49 espécimes de P. coronata e P. dominicana apreendidas do tráfico de animais silvestres em São Paulo (SP). Foram isoladas treze espécies de bactérias gram-negativas, incluindo Salmonella spp. e Pseudomonas aeruginosa. A maior frequência de ocorrência foi de Escherichia coli, identificada em 42/49 (85,7%) das amostras fecais. Dentre os isolados de E. coli, 21/42 pertenciam aos grupos filogenéticos B2 e D, relacionados a estirpes patogênicas que causam doença extraintestinal em humanos. Klebsiella pneumoniae foi isolada em 28/49 (57,1%) das amostras. Esses resultados reforçam que as condições estressantes a que esses animais são submetidos em situações de tráfico, incluindo o contato com humanos, podem favorecer a colonização da microbiota cloacal das aves por patógenos, o que representa um risco para a sua reintrodução na natureza considerando-se o possível contato com humanos e outros animais...

A survey on gram-negative bacteria in saffron finches (Sicalis flaveola) from illegal wildlife tradein Brazil / Pesquisa de bactérias gram-negativas em canários-da-terra (Sicalis flaveola) resgatados do tráfico ilegal de aves no Brasil

Passerines such as canaries or finches are the most unlawfully captured species that are sent to wildlife centers in São Paulo, Brazil. Captured birds may have infection by opportunistic bacteria in stressful situations. This fact becomes relevant when seized passerine are reintroduced. The aim of this study was to evaluate the health state of finches from illegal wildlife trade using microbiological approaches. Microbiological samples were collected by cloacal and tracheal swabs of 100 birds, captured during 2012 and 2013. The results indicate high frequency of gram-negative bacteria in feces and oropharynx, especially from the Enterobacteriaceae family (97.5%). The most frequent genera were Escherichia coli (46.5%) and Klebsiella pneumoniae (10.4%). Enterobacter cloacae, Serratia liquefaciens, Serratia spp. Klebsiella oxytoca and Citrobacter freundii were isolated with lower frequency from asymptomatic birds. The presence of enteropathogenic Escherichia coli (EPEC) and Shiga toxinproducing strain (STEC) confirm the zoonotic risks and public health concern.(AU)

No Estado de São Paulo, Brasil, os pássaros como os canários-da-terra têm sido uma das espécies mais frequentemente resgatadas do tráfico ilegal e enviadas aos centros de vida selvagem. Em situações de estresse estas aves podem ser acometidas por infecções causadas por bactérias oportunistas. Este fato é de grande importância quando é planejada da reintrodução das aves na natureza. O presente trabalho foi delineado para avaliar o estado de saúde de canários-da-terra resgatados do tráfico ilegal. Foram colhidas soabes da traqueia e da cloaca de 100 aves resgatadas durante os anos de 2012 e 2013. Os resultados obtidos revelaram alta frequência de bactérias gram-negativas nas fezes e no orofaringe dos animais, com maior frequência para os membros da família Enterobacteriaceae (97,5%). Os gêneros mais frequentes foram Escherichia coli (46,55) e Klebsiella pneumoniae (10,4%). Outros microorganismos incluindo Enterobacter cloacae, Serratia liquefaciens, Serratia spp, Klebsiella oxytoca e Citrobacter freundii também foram isolados em menor frequencia de aves assintomáticas. A presença de estirpes de Escherichia coli enteropagênicas (EPEC) e as produtoras da toxina de Shiga confirmam o risco de zoonose e a importância para saúde pública deste tipo de ave.(AU)

Salmonella Agona em Perus (Meleagris gallopavo) de criações comerciais no Brasil / Salmonella Agona in Turkeys (Meleagris gallopavo) in commercial farms from Brazil

Salmonella spp. é um dos principais agentes envolvidos em casos de doenças de origem alimentar em humanos, responsável por perdas significativas na avicultura. O presente trabalho investigou a presença de Salmonella spp. em fezes de perus comerciais no Brasil. Foram colhidos suabes fecais de 14 lotes de perus comerciais (pool de seis aves/lote). Os suabes foram submetidos aos procedimentos de isolamento bacteriológico convencionais e a detecção de DNA do agente foi realizada com a técnica de PCR. Salmonella spp. foi detectada em um total de nove lotes dos 14 avaliados. As amostras foram negativas na identificação molecular dos sorovares Enteritidis e Typhimurium. Os isolados foram encaminhados ao laboratório de referência para sorotipagem e identificados como S. Agona; um patógeno considerado emergente em vários países.

Abstract Salmonella spp. is one of the major players involved in cases of foodborne diseases in humans and is responsible for significant losses in the poultry industry. The aim of this study was to investigate the presence of Salmonella spp. in feces of commercial turkeys from Brazil. Fecal swabs from 14 turkey farms (pool of six poults/flocks) were collected. The swabs were subject to the conventional bacteriological isolation procedures and to DNA detection of the agent trough PCR. Salmonella spp. was present in a total of nine from 14 turkey farms evaluated. The samples were negative on molecular identification for serovars Enteritidis and Typhimurium. Isolated strains submitted to the reference laboratory for serotyping were identified as S. Agona that has been described as emergent pathogen in several countries.

Genetic similarity between APEC and Escherichia coli strains isolated from Guaruba guarouba in a survey on healthy captive psittacine birds / Similaridade genética entre APEC e cepas de Escherichia coli isoladas de Guaruba guarouba em um estudo com psitacídeos hígidos de cativeiro

The role of psittacine birds as a reservoir of avian pathogenic Escherichia coli (APEC) is not known but would be helpful in understanding the human – animal interface, since the enteric microbiota of these birds consists of Gram positive bacteria. The aim of this study was to identify the presence of APEC in feces of clinically healthy Guaruba guarouba. To do this, we isolated and analyzed E. coli from cloacal fecal samples taken from 87 psittacine birds from six zoologic parks, three commercial breeders and one conservation breeder. Of the 87 birds examined, 46 (52.87%) presented E. coli in feces. The presence of the following eight virulence genes was determined by the polymerase chain reaction (PCR): irp2, iucD, iss, vat, cvi/cva, tsh, astA, and papC, and 29 (63.04%) of 46 E. coli isolates tested were positive at least one of the eight genes studied. The frequency of virulence genes observed in isolates of E. coli were 32.6% (15/46) irp2, 26% (12/46) iucD, 19.5% iss (9/46), 17.4% vat (8/46), 17.4% cvi/cva (8/46), 8.7% tsh (4/46), 4.4% astA (2/46) and 0% papC (0/46). The isolates were grouped in 13 genotypic profiles according to virulence gene combinations, but only 2 isolates were classified as APEC, with the pattern iuc, iss, cvi/cva, irp + and iuc, iss, cvi/cva, irp, tsh, vat +. This study reveals the presence of APEC in clinically healthy captive G. guarouba, suggesting that these psittacine birds may act as reservoir for pathogenic microorganisms. Epidemiological studies are needed to determine the relevance of this species as a reservoir and the implications for conservation of endangered species G. guarouba.

O papel dos psitacídeos como reservatório de Escherichia coli patogênicas para aves (APEC) não é conhecido, mas será útil para a compreensão da interface humano-animal, uma vez que a microbiota entérica destas aves é composta por bactérias Gram-positivas. O objetivo deste estudo foi identificar a presença de APEC em fezes de Guaruba guarouba clinicamente saudáveis. Para isso, foram isoladas e analisadas E. coli presentes em fezes cloacais coletadas de 87 psitacídeos, alojados em seis zoológicos, três criatórios comerciais e um criatório conservacionista. Das 87 aves examinadas, 46 (52,87%) apresentaram E. coli nas fezes. A presença de oito genes de virulência foi determinada pela reação em cadeia pela polimerase (PCR): irp2, iucD, iss, vat, cvi/cva, tsh, astA, e papC, e 29 (63,04%) dos 46 isolados foram positivos para pelo menos um dos oito genes estudados. A frequência dos genes de virulência observada nos isolados de E. coli foi 32.6% (15/46) irp2, 26% (12/46) iucD, 19.5% iss (9/46), 17.4% vat (8/46), 17.4% cvi/cva (8/46), 8.7% tsh (4/46), 4.4% astA (2/46) e 0% papC (0/46). Os isolados foram agrupados em 13 perfis genotípicos de acordo com combinações de genes de virulência, mas apenas duas amostras foram classificadas como APEC, com o perfil iuc, iss, cvi/cva, irp + e iuc, iss, cvi/cva, irp, tsh, iuc, iss, +. Este estudo revela a presença de APEC em aves de cativeiro (G. guarouba) clinicamente saudáveis, sugerindo que estes psitacídeos possam atuar como reservatórios de micro-organismos patogênicos. Estudos epidemiológicos são necessários para determinar a relevância desta espécie como reservatório e as implicações para a conservação da espécie ameaçada G. guarouba.