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Salmonella Gallinarum is capable of causing high mortality in birds of the order Galliformes. This study aimed to relate the presence of clinical signs with the recovery of Salmonella Gallinarum from organs and c loacal swabs of Japanese quails (Coturnix coturnix) experimentally infected. A total of 70 female quails were housed in a pair per cage and divided in two groups (IG: quails inoculated with 1.5x106 CFU of Salmonella Gallinarum Nalr/mL and CG: control group). After the inoculation, birds were evaluated three times a day to verify the presence of clinical signs. Birds that presented ruffled feathers, eyes closed and remained quiet in the cage were removed for euthanasia, as well as the same number of birds from the inoculated groups that presented no clinical signs and from the control group. Cloacal swabbing was performed following euthanasia for the sampling of liver, spleen, caeca, ovarian follicles and lung for microbiological procedure. Quails with clinical signs and quails found dead presented positivity of 100%. While inoculated quails with no clinical signs presented a lower positivity (38.5%). Therefore, quails with septicemia caused by SG present clinical signs of the disease and the pathogen can be isolated and quantified in the organs. RESUMO Salmonella Gallinarum pode causar alta mortalidade em aves da ordem Galliformes. Objetivou-se neste estudo relacionar a presença de sinais clínicos com a recuperação de Salmonella Gallinarum de órgãos e swabs cloacais de codornas japonesas (Coturnix coturnix) experimentalmente infectadas. Um total de 70 codornas fêmeas foram alojadas em par por gaiola e divididas em dois grupos (IG: codornas inoculadas com 1,5x106 UFC de Salmonella Gallinarum Nalr / mL e CG: grupo controle). Após a inoculação, as aves foram avaliadas três vezes ao dia para se verificar a presença de sinais clínicos. As aves que se apresentaram com penas eriçadas, olhos fechados e permaneciam imóveis na gaiola foram removidas para a eutanásia, assim como o mesmo número de aves dos grupos inoculados que não apresentaram sinais clínicos e do grupo controle. O swab cloacal foi realizado após a eutanásia para a amostragem de fígado, baço, ceco, folículos ovarianos e pulmão para procedimento microbiológico. As codornas com sinais clínicos e as encontradas mortas apresentaram positividade de 100%, enquanto as codornas inoculadas sem sinais clínicos apresentaram menor positividade (38,5%). Portanto, codornas com septicemia causada por SG apresentam sinais clínicos da doença e o patógeno pode ser isolado e quantificado em diversos órgãos.

The psittacine Pyrrhura griseipectus is a threatened species currently only found in Ceará State, Brazil. A microbiological survey was conducted to determine the composition of the enteric microbiota of this species, as well as the phenotypic profiles of antimicrobial susceptibility presented by the isolates. Cloacal swabs were collected from individual birds and submitted to microbiological processing. Antimicrobial susceptibility profiles were evaluated using the disk diffusion method. Bacteria of the genus Escherichia, Proteus, Citrobacter, Pantoea, Klebsiella, Enterobacter, Morganella, Hafnia, Enterobacter, and Serratia were isolated. The most frequently isolated species were Escherichia coli, Proteus mirabilis and Proteus vulgaris, corresponding to 36.1%, 26.4%, and 8.3%, respectively. Isolates were more frequently resistant to azithromycin and tetracycline, while Escherichia coli was the main species presenting multidrug resistance. In conclusion, free-living grey-breasted parakeets may harbor enterobacteria with high antimicrobial resistance rates.

This study aimed to isolate Escherichia coli and Salmonella enterica from captured feral pigeons in Fortaleza, Brazil, and, in addition to evaluate the antimicrobial susceptibility profiles and diagnose diarrheagenic E. coli strains. Pigeons were captured in four public locations in Fortaleza with three techniques. Individual cloacal swab samples were collected and submitted to bacterial isolation, biochemical identification and antimicrobial susceptibility test. Disk diffusion technique was used with twelve antibiotics. E. coli strains were submitted to DNA extraction followed by PCR to diagnose five diarrheagenic pathotypes. A total of 124 birds were captured. One bird was positive for Salmonella enterica (0.81%) and 121 (97.58%) were positive for E. coli. Among these, 110 isolates were submitted to antimicrobial susceptibility test and 28.18% (31/110) presented resistance to at least one antibiotic. Resistance to azithromycin was the most frequent (21.82%), followed by tetracycline (10.91%) and sulfamethoxazole with trimethoprim (8.9%). Multidrug resistance, calculated as a resistance to at least 3 antimicrobial classes, was identified in 3.64% (4/110) of strains. The maximum number of antimicrobial classes to which one strain was resistant was seven. Results demonstrated nine different resistance profiles and the most frequent was tetracycline and sulfamethoxazole with trimethoprim (4 strains), followed by chloramphenicol, azithromycin, tetracycline and sulfamethoxazole with trimethoprim (3 strains). Amoxicillin with clavulanic acid and tobramycin presented lowest levels of antimicrobial resistance, to which none of the tested strains were resistant. A single strain was positive for the eltB gene, which is a diagnostic tool to identify the Enterotoxigenic E. coli (ETEC) pathotype. None of the other investigated genes (stx1, stx2, estA, eaeA, ipaH, aatA and aaiC) were identified. The single isolate of S. enterica was a rough strain of Salmonella enterica subsp. enterica, but serotype identification was not possible. However, this isolate presented resistance to amoxicillin, amoxicillin with clavulanic acid, tetracycline and sulfamethoxazole with trimethoprim. Therefore, captured feral pigeons of Fortaleza presented a low prevalence of S. enterica and diarrheagenic E. coli. Considering the investigated pathogens, our results suggest a good health status and a low public health risk. However, important antimicrobial resistance profiles were identified.

Background: Rock pigeon (Columba livia) is an exotic and invasive species of the Columbidae family found in several cities around the world, including Brazilian cities, and close to humans. This species is found in large populations often in public locations, where people may have contact. Several studies have demonstrated the isolation of human pathogens from these birds. However, there are scarce studies describing infections by Gram-negative bacteria to which pigeons are susceptible. Therefore, this report aimed to describe a case of natural coinfection of Pseudomonas aeruginosa and Enteroaggregative Escherichia coli in a feral pigeon.Case: A sick feral pigeon was delivered at the Laboratory of Ornithological Studies, State University of Ceará, Fortaleza, Brazil. Due to poor prognosis, the individual was euthanized with ketamine via intravenous injection and submitted to necropsy, in which samples were collected for microbiological and histopathological procedure. The procedure was performed aseptically and samples were collected from intestine, liver, spleen, heart and lung. Bacterial isolation was performed with culture media selective for Gram negative bacteria and strains were identified biochemically. Histopathological examination was performed with conventional method and slides were stained with hematoxylin and eosin. DNA from E. coli isolates was extracted with simples boiling method and submitted to uniplex conventional polymerase chain reaction (PCR) to diagnose diarrheagenic pathotypes with specific primers for the following genes: aaiC, aatA, eaeA, stx1, stx2, eltB, estA and ipaH. Escherichia coli was isolated from the intestine, liver, spleen and lung, while Pseudomonas aeruginosa was present in liver, spleen and lung. E. coli strains from liver and spleen were positive for the aaiC gene, which is a diagnostic gene for Enteroaggregative Escherichia coli (EAEC) pathotype. Necropsy revealed the presence of several caseous lesions around the head of the bird, cellulitis in the abdominal region, hepatomegaly, splenomegaly, hemorrhagic intestine and pericarditis. Microscopical findings were most intense in liver and lung samples, presenting vascular and inflammatory alterations. Histopathological alterations indicated an acute presentation of the infections and several lesions in different organs, demonstrating the septicemic characteristic of the microorganisms involved in this case.Discussion: Both pathogens in this report were identified mostly in the same organs, which may indicate that there was an association in the pathogenesis. However, the entry routes of infection in this case may have been different, considering that Pseudomonas aeruginosa was not isolated from the intestine. The histopathological findings were not pathognomonic for either pathogen. However, some characteristic lesions were observed, such as cellulitis, which is commonly attributed to Escherichia coli. EAEC strains are human pathogens that cause acute and persistent diarrhea around the world. These microorganisms could be potentially transmitted to humans, as suggested by other studies with feral pigeons and enteric pathogens. However, the absence of reports of pigeon-transmitted diarrheagenic infections in humans may suggest that pigeons are only reservoirs of these pathogens. In addition, as this report demonstrates, these birds may also suffer from these infections. In conclusion, the free-living pigeon presented a natural occurring case of coinfection by Pseudomonas aeruginosa and Enteroaggregative Escherichia coli. Both are human opportunistic pathogens, and may have public health implications

RESUMO: O tráfico de animais selvagens consiste numa atividade ilegal exercida em diversas partes do mundo, e o Brasil, em função de sua grande extensão territorial e vasta biodiversidade, é um país bastante atingido por essa prática. O grupo de animais em perigo que mais se destaca é o das aves e, entre elas, as da ordem Psittaciformes. Essas são as mais traficadas por serem consideradas inteligentes e possuidoras de grandes habilidades de comunicação. Por isso, são retiradas indiscriminadamente da natureza. No entanto, as condições em que essas aves são capturadas e mantidas em cativeiro favorecem o aparecimento de diversas enfermidades, entre elas as causadas pelas enterobactérias. As bactérias que mais causam mortes e problemas em psitacídeos são as do gênero Salmonella e Escherichia coli , responsáveis pela salmonelose e pela colibacilose, respectivamente. Este trabalho apresenta uma revisão da literatura sobre essas enterobactérias em Psittaciformes, ressaltando o potencial zoonótico desses agentes. ABSTRACT: The illegal wildlife trade is widespread among several countries, and Brazil is one of the most frequently involved due to its great territorial extension and large diversity of animal species. Birds are the main target, and among them the Psittaciformes order, which are removed indiscriminately from nature, is one of the most frequently traded due to their intelligence and great communicating skills. However, the conditions in which these birds are captured and maintained in captivity favor several diseases, among them the ones caused by some Enterobacteriaceae. In this family, the most frequently associated bacteria to illnesses and deaths in psittacine are Salmonella and Escherichia coli , which cause salmonellosis and colibacillosis, respectively. Therefore, this study presents a review of the literature concerning these enterobacteria in Psittaciformes, emphasizing the zoonotic potential of these bacteria.

The Enterobacteriaceae family contains potentially zoonotic bacteria, and their presence in canaries is often reported, though the current status of these in bird flocks is unknown. Therefore, this study aimed to identify the most common genera of enterobacteria from canaries (Serinus canaria) and their antimicrobial resistance profiles. From February to June of 2013, a total of 387 cloacal swab samples from eight domiciliary breeding locations of Fortaleza city, Brazil, were collected and 58 necropsies were performed in canaries, which belonged to the Laboratory of Ornithological Studies. The samples were submitted to microbiological procedure using buffered peptone water and MacConkey agar. Colonies were selected according to their morphological characteristics on selective agar and submitted for biochemical identification and antimicrobial susceptibility. A total of 61 isolates were obtained, of which 42 were from cloacal swabs and 19 from necropsies. The most isolated bacteria was Escherichia coli with twenty five strains, followed by fourteen Klebsiellaspp., twelve Enterobacterspp., seven Pantoea agglomerans, two Serratiaspp. and one Proteus mirabilis. The antimicrobial to which the strains presented most resistance was sulfonamides with 55.7%, followed by ampicillin with 54.1% and tetracycline with 39.3%. The total of multidrug-resistant bacteria (MDR) was 34 (55.7%). In conclusion, canaries harbor members of the Enterobacteriaceae family and common strains present a high antimicrobial resistance rate, with a high frequency of MDR bacteria. Resumo: A família Enterobacteriaceae possui bactérias com potencial zoonótico e a presença destas bactérias em canários é relatada na literatura, porém a realidade dos plantéis de criadores de canários é desconhecida. Portanto, este trabalho teve como objetivo isolar enterobactérias de canários belga (Serinus canarius) com o intuito de conhecer os gêneros mais comuns nestas aves e suas respectivas resistências a antimicrobianos. De fevereiro a junho de 2013 foram coletadas 387 amostras de swabs cloacais de canários de oito propriedades da cidade de Fortaleza, Brasil e de 58 necropsias de aves do acervo próprio do Laboratório de Estudos Ornitológicos. As amostras foram submetidas a isolamento microbiológico utilizando-se água peptonada e ágar MacConkey. As colônias foram selecionadas de acordo com suas características morfológicas nas placas, submetidas à tipificação bioquímica para identificação e ao teste de sensibilidade a antimicrobianos. Foram isoladas 61 cepas, sendo 42 de suabes cloacais e 19 de necropsias. A bactéria mais isolada foi Escherichia coli com vinte e cinco cepas, seguida por catorze Klebsiella spp., doze Enterobacter spp., sete Pantoea agglomerans, duas Serratiaspp. e uma cepa de Proteus mirabilis. As cepas apresentaram maior resistência a sulfonamidas com 55,7%, seguidas por ampicilina com 54,1% e tetraciclina com 39,3%. Além disso, o total de cepas resistentes a múltiplas drogas (RMD) foi 34 (55,7%). Portanto, conclui-se que os canários albergam enterobactérias e que as cepas apresentam alto índice de resistência a antimicrobianos, com alta frequência de cepas RMD.

Background: The third largest poultry flock in Northeast Brazil is located in Ceará State. Some pathogens are commonly disseminated in broiler chicken flocks, such as the bacteria from the Enterobacteriaceae family. Among these, some strains of Escherichia coli are frequently associated with different pathological manifestations in domestic animals, while bacteria from the genus Salmonella are considered the most frequent enteric pathogens reported causing foodborne infections in humans. Therefore, this study aimed to evaluate the prevalence and antimicrobial resistance of Salmonella sp. and Escherichia coli strains isolated from broiler chickens in the Metropolitan Region of Fortaleza city, Brazil.Materials, Methods & Results: Samples were collected from July-2014 to March-2015 in ten broiler chicken farms located in the Metropolitan Region of Fortaleza city, Brazil, with birds in pre-slaughter age. From each farm, 100 individual cloacal swabs were randomly collected from broilers independent of clinical status. Distinct methodologies were used in order to provide optimal isolation conditions for both the bacterial species. For Escherichia coli, the methodology consisted in enrichment with BHI broth, plating in EMB agar and biochemical identification, after which some isolates were maintained in nutrient agar for antimicrobial resistance evaluation. For the isolation of Salmonella sp., a standard method was used with pre-enrichment, selective enrichment, selective plating and biochemical identification steps. Antimicrobial susceptibility test (AST) was performed with disk diffusion technique and the following antibiotics were tested: ampicillin, ceftiofur, ciprofloxacin, trimethoprim-sulfamethoxazole, polymyxin B, gentamycin, cloranfenicol, tetracycline, azithromycin and fosfomycin. According to the methodology used, 95.9% of samples were positive for Escherichia coli and the most frequent resistance was to trimethoprim-sulfamethoxazole. Salmonella sp. was isolated from 0.2% of the samples, which were identified as the serotype Salmonella enterica subsp. enterica O:6,8. Both isolates presented the same antimicrobial resistance profile, which were resistant to six, out of ten tested antibiotics (trimethoprim-sulfamethoxazole, tetracycline, ciprofloxacin, azithromycin, chloramphenicol and ceftiofur).Discussion: The low prevalence of Salmonella observed in this study have also been reported by other studies performed in poultry farms in Ceará State, which suggests a good status for this pathogen in the local industry, however further efforts in order to eradicate this pathogen must be applied. The salmonella serotype detected in this study is rarely reported in the literature, especially from the poultry industry. In Brazil, the use of tetracyclines, quinolones and penicillins as feed additives or growth promoters is prohibited; however, a high resistance to drugs from these groups was detected. In addition, multidrug resistant E. coli isolates presented more elevated rates than other studies reported in the literature with antibiotics commonly used in the poultry industry and this may indicate an excessive use of these drugs in the production routine. These results should serve as a warning for surveillance programs to evaluate the incidence of these microorganisms as well as their antimicrobial resistance rates, which may be an important tool for control and prevention in meat poultry production.

This study reports a co-infection of Escherichia coli and Salmonella in a free-living ruddy ground dove (Columbina talpacoti) received at the Laboratory of Ornithological Studies of the State University of Ceará, Brazil. The bird presented diarrhea, leg paralysis and anorexia, and died shortly after. Necropsy was then performed and samples from lung, kidney, liver and intestine were collected for microbiological and histopathological analyses. Escherichia coli was isolated from cloacal swab, lung and kidney samples. Salmonella ser. Saintpaul was identified in liver and spleen samples. Escherichia coli isolates were tested for the presence of eight diagnostic genes for diarrheagenic pathotypes (STEC, ETEC, EPEC, EIEC, EAEC) with conventional polymerase chain reaction (PCR). EAEC was detected in the lung and kidney, and STEC in the intestine. In conclusion, Columbina talpacoti is susceptible to enteroaggregative Escherichia coli and Salmonella ser. Saintpaul infection, which may have public health implications. Este estudo relata um caso de coinfecção por Escherichia coli e Salmonella ser. Saintpaul em uma rolinha-roxa (Columbina talpacoti) recebida pelo Laboratório de Estudos Ornitológicos da Universidade Estadual do Ceará, Brasil. A ave apresentava diarreia, paralisia nas pernas e anorexia, indo a óbito rapidamente. A necropsia foi realizada e amostras de pulmão, rim, fígado e intestino foram coletados para isolamento microbiológico e análise histopatológica. Escherichia coli foi identificada em amostras de suabe cloacal, pulmão e rim. Salmonella ser. Saintpaul foi identificada no fígado e baço. Isolados de E. coli foram testados para a presença de oito genes de diagnóstico para patotipos diarreiogênicos (STEC, ETEC, EPEC, EIEC, EAEC) através de reação em cadeia de polimerase (PCR) convencional. EAEC foi detectada no pulmão e rim, e STEC foi identificada no intestino. Em conclusão, Columbina talpacoti é suscetível a infecção por Escherichia coli enteroagregativa e Salmonella ser. Saintpaul, o que pode implicar em risco para a saúde pública.

This study aimed to verify the presence of members from the Enterobacteriaceae family and determine antimicrobial susceptibility profiles of the isolates in canaries bred in northeastern Brazil; in addition, the presence of diarrheagenic Escherichia coli (DEC) and avian pathogenic Escherichia coli (APEC) was also verified in these birds. Samples were collected during an exhibition organized by the Brazilian Ornithological Federation in July 2015 in Fortaleza, Brazil. A total of 88 fecal samples were collected and submitted to pre-enrichment step using buffered peptone water, followed by enrichment with the following broths: brain-heart infusion, Rappaport-Vassiliadis, and Selenite-Cystine. Subsequently, aliquots were streaked on MacConkey, brilliant green and salmonella-shigella agar plates. Colonies were selected according to morphological characteristics and submitted to biochemical identification and antimicrobial susceptibility tests with disk-diffusion technique. E. coli strains were evaluated for the presence of eight DEC genes and five APEC genes through conventional polymerase chain reaction (PCR) screening. The most frequent species observed were Pantoea agglomerans (25%), Serratia liquefaciens (12.5%), and Enterobacter aerogenes (9.1%). A single rough strain of Salmonella enterica subsp. enterica was identified in one sample (1.1%). High resistance rates to amoxicillin (78.7%) and ampicillin (75.4%) were identified. Polymyxin B (9.8%), gentamycin (6.6%), and enrofloxacin (6.6%) were the most efficient antibiotics. The total number of multidrug-resistant strains (isolates resistant to more than three antimicrobial classes) was 23 (37.7%). Four E. coli strains were tested for the virulence genes, and two were positive for APEC virulence genes: one strain was positive for iutA and the other for hlyF. In conclusion, canaries in northeastern Brazil participating in exhibitions may present Salmonella spp., Escherichia coli and other enterobacteria in the intestinal microbiota with antimicrobial resistance. These results indicate that, although the E. coli strains recovered from canaries in this study have some virulence genes, they still do not fulfill all the requirements to be considered APEC.