Cloacal enterobacteria isolated from captive roadside hawks (Rupornis magnirostris, GMELIN, 1788) and their antimicrobial susceptibility profile / Enterobactérias isoladas da cloaca de Gaviões-carijós (Rupornis magnirostris, GMELIN, 1788) cativos e seu perfil de susceptibilidade a antimicrobianos

Knowledge of the enterobacteria present in the roadside hawk can bring about an understanding of infectious diseases that can affect this bird, as well as other animals and/or humans, while also adding information of great ecological importance. Thus, the aim of the study was to determine the enterobacteria present in the cloaca of captive roadside hawks and antimicrobial susceptibility profile. Initially, cloacal samples from nine specimens were collected with the aid of swabs. Samples were placed in petri dishes with MacConkey agar, Hektoen agar, EMB agar and Salmonella-Shigella (SS) agar and incubated for 24 h at 35C. After incubation, the microorganisms were submitted to biochemical testing to confirm the presence of enterobacteria. Thereafter, the susceptibility profile of bacteria to antimicrobial agents was evaluated by a disk diffusion test according to the Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (77.8%), Klebsiella oxytoca (11.1%), Klebsiella pneumoniae (11.1%) and Salmonella spp. (55.6%) were isolated from the collected samples. Among the isolates, some bacteria showed resistance to up to three antimicrobial agents. This study has brought greater insight about the enterobacteria present in the roadside hawk (Rupornis magnirostris), exhibiting a significant percentage of enterobacteria important to public health; also, it showed the occurrence of strains with resistance profile to antimicrobial agents...

O conhecimento das enterobactérias presentes em Gavião-carijó pode trazer uma compreensão sobre as doenças infecciosas que podem acometer essa ave, como também outros animais e/ou humanos, além de trazer mais informações sobre essa espécie de grande importância ecológica. Desta forma, o objetivo do estudo foi determinar as enterobactérias presentes na cloaca de Gaviões-carijós cativos e seu perfil de susceptibilidade a antimicrobianos. Inicialmente, foram coletadas amostras cloacais de nove espécimes com o auxílio de swabs. As amostras foram plaqueadas em ágar MacConkey, ágar Hektoen, ágar EMB e ágar SS e incubadas por 24 h a 35C. Após incubação, as colônias foram submetidas às provas bioquímicas para confirmação da presença de enterobactérias. Posteriormente, o perfil de susceptibilidade das bactérias frente a agentes antimicrobianos foi avaliado através do teste da difusão em disco de acordo com o Clinical and Laboratory Standards Institute (CLSI). Escherichia coli (77,8%), Klebsiella oxytoca (11,1%), Klebsiella pneumoniae (11,1%) e Salmonella spp. (55,6%) foram isoladas das amostras coletadas. Dentre as cepas isoladas, algumas apresentaram resistência a até três antimicrobianos. O presente estudo trouxe um maior conhecimento sobre as enterobactérias presentes no Gavião-carijó (Rupornis magnirostris), mostrando um percentual significativo de enterobactérias de importância na saúde pública, evidenciando também a ocorrência de cepas com perfil de resistência a agentes antimicrobianos...

Identification and antimicrobial resistance of members from the Enterobacteriaceae family isolated from canaries (Serinus canaria) / Identificação e resistência antimicrobiana de membros da família Enterobacteriaceae isolados de canários (Serinus canaria)

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.

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.

Role of wild birds as carriers of multi-drug resistant Escherichia coli and Escherichia vulneris

Emergence and distribution of multi-drug resistant (MDR) bacteria in environments pose a risk to human and animal health. A total of 82 isolates of Escherichia spp. were recovered from cloacal swabs of migrating and non-migrating wild birds. All bacterial isolates were identified and characterized morphologically and biochemically. 72% and 50% of isolates recovered from non-migrating and migrating birds, respectively, showed positive congo red dye binding (a virulence factor). Also, hemolysin production (a virulence factor) was showed in 8% of isolates recovered from non-migrating birds and 75% of isolates recovered from migrating birds. All isolates recovered from non-migrating birds were found resistant to Oxacillin while all isolates recovered from migrating birds demonstrated resistance to Oxacillin, Chloramphenicol, Oxytetracycline and Lincomycin. Some bacterial isolates recovered from non-migrating birds and migrating birds exhibited MDR phenotype. The MDR isolates were further characterized by API 20E and 16S rRNA as E. coli and E. vulneris. MDR Escherichia isolates contain ~1-5 plasmids of high-molecular weights. Accordingly, wild birds could create a potential threat to human and animal health by transmitting MDR bacteria to water streams and other environmental sources through their faecal residues, and to remote regions by migration.

Caracterización de cepas de Escherichia coli enteropatogénico (EPEC) aisladas durante el proceso de faena de pollos / Characterization of enteropathogenic Escherichiia coli (EPEC) strains isolated during the chicken slaughtering process

En Argentina, Escherichia coli enteropatogénico (EPEC) es uno de los agentes más prevalentes aislados de niños con diarrea. Debido a que la contaminación con este patotipo en productos de pollo podría ocurrir durante el proceso de faena, nos planteamos como objetivo aislar y caracterizar EPEC de muestras de animales vivos (cloacas), carcasas evisceradas sin lavar, carcasas lavadas y agua del tanque de enfriamiento. Se caracterizaron 29 aislamientos de EPEC que presentaron una amplia variedad de serotipos, algunos de los cuales (O2:H40, O8:H19 y O108:H9) han sido informados en otras especies animales. También se encontró el serotipo O45:H8, aislado con anterioridad de niños con diarrea. Se detectaron aislamientos de los serotipos O2:H40, O108:H9 y O123:H32 en distintas etapas del proceso de faena, lo que sugiere que el procesamiento no se realiza en forma adecuada. Se torna necesario reforzar las medidas de control e higiene en las distintas etapas del proceso para disminuir la contaminación microbiana

In Argentina, EPEC is one of the most prevalent agents isolated from children with diarrhea. Because contamination with this pathotype could occur during slaughter, the aim of this study was to isolate and characterize EPEC strains obtained from live animals (cloacae), eviscerated carcasses, washed carcasses and water from chillers. Twenty nine isolates of atypical EPEC were characterized. These isolates presented a wide variety of serotypes, some of which (O2:H40, O8:H19 and O108:H9) had been reported in other animal species. Serotype O45:H8, previously isolated from children with diarrhea was also found. Isolates of serotypes O2:H40, O108:H9 and O123:H32 were detected at different stages of the slaughtering process, suggesting that the process is not adequately performed. This latter fact highlights the importance of reinforcing control and hygienic measures at different stages of the chicken slaughtering process in order to reduce microbial contamination

Detection of vanC 1 gene transcription in vancomycin-susceptible Enterococcus faecalis

Here we report the presence and expression levels of the vanC 1 and vanC 2/3 genes in vancomycin-susceptible strains of Enterococcus faecalis. The vanC 1 and vanC 2/3 genes were located in the plasmid DNA and on the chromosome, respectively. Specific mRNA of the vanC 1 gene was detected in one of these strains. Additionally, analysis of the vanC gene sequences showed that these genes are related to the vanC genes of Enterococcus gallinarum and Enterococcus casseliflavus. The presence of vanC genes is useful for the identification of E. gallinarum and E. casseliflavus. Moreover, this is the first report of vanC mRNA in E. faecalis.

Molecular detection of enteropathogenic Escherichia coli in asymptomatic captive psittacines / Detecção molecular de Escherichia coli enteropatogênica em psitacídeos assintomáticos em cativeiro

Psittaciformes are one of the most endangered groups of birds, and several Brazilian species are classified between vulnerable and critically endangered. It is thus necessary to identify agents that cause infections in captive wild animals and to assess the risks posed thereof and to design interventions to minimize the possibility of disease outbreaks, leading to the conservation of endangered species. The purpose of this study was to identify enteropathogenic Escherichia coli (EPEC) cloacal isolates from asymptomatic psittacines in captivity and evaluate the distribution of the EPEC pathotype. Cloacal swabs were obtained from 46 asymptomatic birds, and resulting isolates were tested by polymerase chain reaction (PCR) for the presence of the attaching and effacing gene (eae) and bundle-forming pilus structural gene (bfpA) of EPEC. Samples from several species were tested, and three samples were found to be positive for the eae and bfpA genes and characterized as typical EPEC. This is the first report of this pathotype in asymptomatic psittacines. Although certain E. coli strains are more pathogenic than others, various factors should be considered when determining the potential of E. coli isolates to cause disease in captive psittacines. Birds that are positive for the EPEC (typical) strain could be zoonotic sources of infection, and may have acquired these strains through contact with humans or domestic animals. These findings may also be valuable for the long-term management of endangered species ex situ as one EPEC sample was isolated from a Red-tailed Amazon (Amazona brasiliensis).

Os psitacídeos são um dos grupos de aves mais ameaçadas no mundo e diversas espécies brasileiras são classificadas desde vulneráveis à criticamente ameaçadas de extinção. Torna-se, portanto, necessário identificar os agentes que causam infecções em animais selvagens em cativeiro e determinar os riscos relacionados de modo a intervir sobre os fatores envolvidos para diminuir a possibilidade de surtos de doenças e promover a conservação de espécies ameaçadas. O objetivo deste estudo foi identificar Escherichia coli Enteropatogência (EPEC) de isolados cloacais de psitacídeos assintomáticos em cativeiro e avaliar a distribuição do patotipo EPEC. Suabes cloacais foram coletados de 46 psitacídeos assintomáticos e os isolados foram testados pela reação em cadeia pela polimerase (PCR) para a presença do gene attaching and effacing (eae) e bundle forming pilus (bfpA) de EPEC. Amostras oriundas de diversas espécies foram testadas e três amostras resultaram positivas para os genes eae e bfp e caracterizadas como EPEC típicas. Esse é o primeiro relato em psitacídeos assintomáticos para esse patotipo. Apesar de que algumas cepas de E.coli serem mais patogênicas do que outras, diversos fatores devem ser considerados para determinar o potencial de isolados de E.coli de causar doença em psitacídeos em cativeiro. Aves positivas para cepas de EPEC (típicas) poderiam ser fontes de infecção zoonóticas e adquirir essas cepas através do contato com humanos e animais domésticos. Esses achados também podem ser valiosos para o manejo a longo prazo de espécies ameaçadas ex situ já que uma amostra de EPEC foi isolada de um Papagaio-de-cara-roxa (Amazona brasiliensis).

Detection of the ompA gene of Chlamydophila pecorum in captive birds in Argentina / Detección del gen ompA de Chlamydophila pecorum en aves en cautiverio, en la Argentina

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, Including humans, other mammals and birds. However, very little is known about chlamydial infections in birds in our region. In the present study, we examined 28 clinically normal birds In illegal captivity that were confiscated in the province of Córdoba, Argentina. The objective was to detect Chlamydophila spp. in cloacal swabs by genetic analysis of the ompA gene. Nested-PCR of the ompA gene identified five samples as Chlamydophila pecorum and the sequence analysis demonstrated the presence of the ompA gene of C. pecorum In these birds. On the other hand, Chlamydophila psittaci was not detected. These birds could be either asymptomatic reservoirs or subclinical carriers of C. pecorum. This is the first report of the detection of C. pecorum in Argentina.

Las bacterias que pertenecen a la familia Chlamydiaceae causan un extenso espectro de enfermedades en una amplia gama de huéspedes, incluidos los seres humanos, otros mamíferos y aves. Sin embargo, se sabe muy poco acerca de las infecciones por clamidias en aves de nuestra reglón. Esta Investigación examinó 28 aves clínicamente normales mantenidas en cautiverio ¡legal, que fueron confiscadas en Córdoba, Argentina. El objetivo fue detectar Chlamydophila spp. en hisopados cloacales por análisis del gen ompA. La PCR anidada del gen ompA reveló la presencia de Chlamydophila pecorum en cinco muestras. El análisis de secuencias demostró la presencia del gen ompA de C. pecorum en estas aves. Por el contrario, Chlamydophila psittaci no se detectó. Estas aves pueden ser reservónos asintomáticos o portadores subclínlcos de C. pecorum. Este es el primer informe de la detección de C. pecorum en la Argentina.

Comparison of wildlife and captivity rattlesnakes (Crotalus durissus terrificus) microbiota / Comparação da microbiota de cascavéis (Crotalus durissus terrificus) de vida-livre e cativeiro

The study evaluated and compared the aerobic microbiota from the oral cavity, cloaca and venom of Crotalus durissus terrificus snakes, recently caught from the wild and kept under quarantine (WQ), individual captivity (IC) and collective captivity (CC). Antimicrobial drug effectiveness on isolated agents also was assayed. From group I, II and III were isolated, respectively, 29 (63.04 percent), 38 (90.48 percent) and 21 (42.86 percent) microorganisms from the cloaca; 15 (32.61 percent), 3 (7.14 percent) and 25 (51.02 percent) microorganisms from the oral cavity; and, 2 (4.35 percent), 1 (2.38 percent) and 3 (6.12 percent) microorganisms from venom. The most frequent bacteria were Pseudomonas aeruginosa, Proteus vulgaris and Morganella morganii, with sensitivity to amikacin, gentamicin, norfloxacin, sulfazotrin and tobramycin. Snakes kept in semi-open captivity exhibited the fewest microorganisms in oral cavities, perhaps due to the environment in captivity, with different temperature gradients, running water, absence of daily handling, circulating air, possibility of moving around, daily cleaning, and sunlight access.

Este estudo avaliou e comparou a microflora aeróbica da cavidade oral, cloaca e veneno de serpentes Crotalus durissus terrificus recém-capturadas da natureza e mantidas sob quarentena (WQ), mantidas em cativeiro coletivo (CC) e em cativeiro individual (IC). A eficácia de drogas antimicrobianas de agentes isolados foi também avaliada. Foram isolados microorganismos dos grupos I, II e III respectivamente: 29 (63.04 por cento), 38 (90.48 por cento) e 21 (42.86 por cento) da cloaca; 15 (32.61 por cento), 3 (7.14 por cento) e 25 (51.02 por cento) da cavidade oral, e finalmente 2 (4.35 por cento), 1 (2.38 por cento) e 3 (6.12 por cento) do veneno. As bactérias mais frequentes foram Pseudomonas aeruginosa, Proteus vulgaris e Morganella morganii, com sensibilidade para amikacina, gentamicina, norfloxacina, sulfazotrina e tobramicina. Serpentes mantidas no cativeiro semi-aberto mostraram menor número de agentes infecciosos em cavidade oral, talvez devido ao ambiente de cativeiro com diferentes gradientes de temperatura, água corrente, ausência de manejo diário, ampla circulação de ar, possibilidade de movimentação pelos animais, limpeza diária e acesso ao Sol.

Flora bacteriana cloacal y nasal de Lepidochelys olivacea (Testudines: Cheloniidae) en el pacífico norte de Costa Rica / Cloacal and nasal bacterial flora of Lepidochelys olivacea (Testudines: Cheloniidae) from the North Pacific coast of Costa Rica

The aerobic cloacal and nasal bacterial flora of 45 apparently healthy female olive ridley sea turtles (Lepidochelys olivacea )was studied at Nancite nesting beach,in Santa Rosa National Park (Costa Rican North Pacific)during July and August 2002.Bacterial samples were obtained by inserting sterile swabs directly into the cloaca and the nasal cavities of the turtles.Ninety-nine aerobic bacterial isolates, including 10 Gram-negative and 5 Gram-positive bacteria, were recovered.The most common bacteria cultured were Aeromonas spp. (13/45) and Citrobacter freundi (6/45)from cloacal samples and Bacillus spp.(32/45), Staphylococcus aureus (6/45)and Corynebacterium spp.(5/45)from nasal ducts.The results of the present study showed that the aerobic bacterial flora of nesting female olive ridleys was composed of several potential human and animal microbe pathogens.

Con el objetivo de determinar la flora normal aerobia, cloacal y nasal de la tortuga lora (Lepidochelys olivacea ), entre los meses de julio y agosto del 2002,se colectaron muestras bacteriológicas de 45 quelonios aparentemente sanos,durante el desove en Playa Nancite,Parque Nacional Santa Rosa, Costa Rica, a través del uso de hisopos estériles que se introdujeron en la cloaca y en uno de los conductos nasales. De las muestras recolectadas se obtuvieron e identificaron un total de 99 aislamientos, incluyendo 10 grupos de Gram-negativos y 5 de Gram-positivos. De cada tortuga se obtuvo un promedio de 0.7 bacterias de la cloaca y 1.4 de las cavidades nasales. Las bacterias más frecuente halladas fueron Aeromonas spp.(13/45) y Citrobacter freundi (6/45) en la cloaca, y Bacillus spp. (32/45),Staphylococcus aureus (6/45)y Corynebacterium spp.(5/45)en las cavidades nasales. En este investigación, la flora microbiana de las tortugas lora resultó constituida por microorganismos potencialmente patógenos para el ser humano y las tortugas.

Patogenicidade dos virus de influenza equina para pintos / Pathogenicity of equine influenza virus for chickens

Considerando a importância das aves na epidemiologia dos vírus de Influenza tipo "A",procurou-se investigar a patogenicidade para pintos dos vírus de Influenza Equina. Cepas isoladas no Brasil, representativas dos subtipos A/equi 1 (H7N7) e A/equi 2(H3N8), foram usadas. Oito experimentos foram realizados com amostra A/equi 2/RJ/85, utilizando 89 pintos de 04 a 18 dias de idade. Seiscentos e trinta e três amostras de material cloacal, coletadas entre 01 e 14 dias pós-infecçäo (p.i) foram inoculadas em ovos embrionados, ensejando a recuperaçäo de 12 amostras de vírus. Provas de inibiçäo de hemoaglutinaçäo com soros conhecidos mostraram que os vírus recuperados eram inibidos pelos soros imunes homólogos (A/equi 2). O exame dos soros sangüíneos dos pintos usados nestes experimentos, obtidos antes da inoculaçäo de vírus, mostrou ausência de anticorpos para A/equi 1 e A/equi 2. Dos 89 pintos infectados com vírus A/equi 2, o exame dos soros de sangrias realizadas entre 18 e 21 dias p.i. revelou presença de anticorpos para o vírus inoculado em 71 casos (79,8 por cento). Setenta pintos foram inoculados com a cepa A/equi 1; 543 amostras de material cloacal inoculadas em ovos embrionados deram resultados negativos. Entretanto, todos os pintos desenvolveram anticorpos para o vírus inoculado. Näo foi registrado qualquer sinal clínico nos pintos inoculados

Isolation of DNA methylase modification enzyme

Type II restriction endonuclease was isolated from locally isolated strain Enterobacter cloaca of recognition sequence CGATCG. Its cognate methylase was isolated and its 5 methyl Cytosine was confirmed by methylation and DNA hydrolysis