Serogroup identification, phenotypic detection of hemolysis and extended spectrum beta-lactamases of Escherichia coli isolated from psittacine of illegal wildlife trade in Fortaleza, Brazil / Identificação de sorogrupo e detecção fenotípica de betalactamase de espectro estendido e produção de hemolisina em Escherichia coli isoladas de psitacídeos do tráfico de animais silvestres em Fortaleza-CE

This study aimed to identify serogroups of Escherichia coli important for human health in isolates from psittacine of illegal wildlife trade in Ceará State. In addition, hemolysis and production of Extended Spectrum Beta-Lactamases (ESBL) was assessed in the isolates. A total of 78 E. coli strains isolated from different Psittaciformes species from a wildlife rehabilitation center in Fortaleza, Brazil. The isolates used in this study were previously identified and stored. Serogroup identification was performed using polyvalent sera for EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) and EHEC (O157). ESBL detection was performed with double disk synergy method. For hemolysis detection, isolates were inoculated in blood agar base enriched with ovine blood. Only 31 (39.7%) isolates were seropositive and the most frequent were O127, O114, O128 and O111. There was no agglutination for serogroups O55, O124, O136 or O157. Considering both seropositive and seronegative isolates, 9 (11.5%) and 35 (44.9%) presented hemolysis and ESBL production, respectively. In conclusion, the investigated psittacine from illegal wildlife trade hosted ESBL-producing E. coli strains and some belong to important serogroups often linked to severe human infections.(AU)

Este trabalho teve como objetivo identificar sorogrupos de E. coli importantes para a saúde humana, oriundos de psitacídeos provenientes do tráfico no estado do Ceará, assim como detectar atividade hemolítica e produção de betalactamase de espectro estendido (ESBL). Foram testadas 78 cepas de Escherichia coli provenientes de psitaciformes do Centro de Triagem de Animais Silvestres, Fortaleza, CE. Para a identificação dos sorogrupos, utilizaram-se soros polivalentes EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) e EHEC (O157). Para detecção de ESBL, as cepas foram submetidas ao método de aproximação de disco e, para a detecção de hemolisina, foram plaqueadas em ágar sangue base enriquecido com sangue de carneiro. No geral, 31 (39,7%) das amostras foram soropositivas. Os sorogrupos mais frequentemente detectados foram O127, O114, O128 e O111. Não houve positividade para os sorogrupos O55, O124, O136 e O157. Considerando-se as amostras sororreagentes e não sororreagentes, observou-se que nove (11,5%) e 35 (44,9%) cepas de E. coli apresentaram produção de hemolisinas e de ESBL, respectivamente. Em conclusão, constatou-se que psitacídeos provenientes do tráfico de animais silvestres albergam cepas de E. coli produtoras de ESBL e providas de importantes sorogrupos implicados em graves infecções humanas.(AU)

Isolation and antimicrobial resistance profiles of enterobacteria from nestling grey-breasted parakeets (Pyrrhura griseipectus)

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.(AU)

Serogroup identification, phenotypic detection of hemolysis and extended spectrum beta-lactamases of Escherichia coli isolated from psittacine of illegal wildlife trade in Fortaleza, Brazil / Identificação de sorogrupo e detecção fenotípica de betalactamase de espectro estendido e produção de hemolisina em Escherichia coli isoladas de psitacídeos do tráfico de animais silvestres em Fortaleza-CE

This study aimed to identify serogroups of Escherichia coli important for human health in isolates from psittacine of illegal wildlife trade in Ceará State. In addition, hemolysis and production of Extended Spectrum Beta-Lactamases (ESBL) was assessed in the isolates. A total of 78 E. coli strains isolated from different Psittaciformes species from a wildlife rehabilitation center in Fortaleza, Brazil. The isolates used in this study were previously identified and stored. Serogroup identification was performed using polyvalent sera for EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) and EHEC (O157). ESBL detection was performed with double disk synergy method. For hemolysis detection, isolates were inoculated in blood agar base enriched with ovine blood. Only 31 (39.7%) isolates were seropositive and the most frequent were O127, O114, O128 and O111. There was no agglutination for serogroups O55, O124, O136 or O157. Considering both seropositive and seronegative isolates, 9 (11.5%) and 35 (44.9%) presented hemolysis and ESBL production, respectively. In conclusion, the investigated psittacine from illegal wildlife trade hosted ESBL-producing E. coli strains and some belong to important serogroups often linked to severe human infections.(AU)

Este trabalho teve como objetivo identificar sorogrupos de E. coli importantes para a saúde humana, oriundos de psitacídeos provenientes do tráfico no estado do Ceará, assim como detectar atividade hemolítica e produção de betalactamase de espectro estendido (ESBL). Foram testadas 78 cepas de Escherichia coli provenientes de psitaciformes do Centro de Triagem de Animais Silvestres, Fortaleza, CE. Para a identificação dos sorogrupos, utilizaram-se soros polivalentes EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) e EHEC (O157). Para detecção de ESBL, as cepas foram submetidas ao método de aproximação de disco e, para a detecção de hemolisina, foram plaqueadas em ágar sangue base enriquecido com sangue de carneiro. No geral, 31 (39,7%) das amostras foram soropositivas. Os sorogrupos mais frequentemente detectados foram O127, O114, O128 e O111. Não houve positividade para os sorogrupos O55, O124, O136 e O157. Considerando-se as amostras sororreagentes e não sororreagentes, observou-se que nove (11,5%) e 35 (44,9%) cepas de E. coli apresentaram produção de hemolisinas e de ESBL, respectivamente. Em conclusão, constatou-se que psitacídeos provenientes do tráfico de animais silvestres albergam cepas de E. coli produtoras de ESBL e providas de importantes sorogrupos implicados em graves infecções humanas.(AU)

Isolation and antimicrobial resistance profiles of enterobacteria from nestling grey-breasted parakeets (Pyrrhura griseipectus)

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.

Detecção e avaliação do perfil de sensibilidade antimicrobiana de enterobactérias isoladas de periquitos cara-suja (Pyrrhura griseipectus) em cativeiro / Detection and evaluation of the antimicrobial sensibility profile of enterobacteria isolated from captive Grey-breasted parakeet (Pyrrhura griseipectus)

This study was conducted to detect enterobacteria in captive Grey-breasted parakeet from a commercial breeding located in the state of Ceara. 25 individual cloacal swabs were collected and this material was subjected to microbiological processing to obtain the bacterial isolation, identification, and evaluation of the sensitivity profile to antimicrobials. The bacterial isolation in ten samples was determined, belonging to the genera Enterobacter, Hafnia, Serratia, Escherichia, Pantoea and Klebsiella, in which these last three corresponded to the bacteria most isolated, with respective percentages of 28.6%, 21.4%, and 21.4%. As for the sensitivity profile, antibiotics to which the bacteria showed more resistance were trimethoprim-sulfamethoxazole (57%), tetracycline (21%), and azithromycin (43%). Thus, it was shown that the parakeets were carriers of Enterobacteriaceae with resistance characteristics.(AU)

Detecção e avaliação do perfil de sensibilidade antimicrobiana de enterobactérias isoladas de periquitos cara-suja (Pyrrhura griseipectus) em cativeiro / Detection and evaluation of the antimicrobial sensibility profile of enterobacteria isolated from captive Grey-breasted parakeet (Pyrrhura griseipectus)

This study was conducted to detect enterobacteria in captive Grey-breasted parakeet from a commercial breeding located in the state of Ceara. 25 individual cloacal swabs were collected and this material was subjected to microbiological processing to obtain the bacterial isolation, identification, and evaluation of the sensitivity profile to antimicrobials. The bacterial isolation in ten samples was determined, belonging to the genera Enterobacter, Hafnia, Serratia, Escherichia, Pantoea and Klebsiella, in which these last three corresponded to the bacteria most isolated, with respective percentages of 28.6%, 21.4%, and 21.4%. As for the sensitivity profile, antibiotics to which the bacteria showed more resistance were trimethoprim-sulfamethoxazole (57%), tetracycline (21%), and azithromycin (43%). Thus, it was shown that the parakeets were carriers of Enterobacteriaceae with resistance characteristics.(AU)

Detecção e avaliação do perfil de sensibilidade antimicrobiana de enterobactérias isoladas de periquitos cara-suja (Pyrrhura griseipectus) em cativeiro

ABSTRACT This study was conducted to detect enterobacteria in captive Grey-breasted parakeet from a commercial breeding located in the state of Ceara. 25 individual cloacal swabs were collected and this material was subjected to microbiological processing to obtain the bacterial isolation, identification, and evaluation of the sensitivity profile to antimicrobials. The bacterial isolation in ten samples was determined, belonging to the genera Enterobacter, Hafnia, Serratia, Escherichia, Pantoea and Klebsiella, in which these last three corresponded to the bacteria most isolated, with respective percentages of 28.6%, 21.4%, and 21.4%. As for the sensitivity profile, antibiotics to which the bacteria showed more resistance were trimethoprim-sulfamethoxazole (57%), tetracycline (21%), and azithromycin (43%). Thus, it was shown that the parakeets were carriers of Enterobacteriaceae with resistance characteristics.

Detecção e avaliação do perfil de sensibilidade antimicrobiana de enterobactérias isoladas de periquitos cara-suja (Pyrrhura griseipectus) em cativeiro

ABSTRACT This study was conducted to detect enterobacteria in captive Grey-breasted parakeet from a commercial breeding located in the state of Ceara. 25 individual cloacal swabs were collected and this material was subjected to microbiological processing to obtain the bacterial isolation, identification, and evaluation of the sensitivity profile to antimicrobials. The bacterial isolation in ten samples was determined, belonging to the genera Enterobacter, Hafnia, Serratia, Escherichia, Pantoea and Klebsiella, in which these last three corresponded to the bacteria most isolated, with respective percentages of 28.6%, 21.4%, and 21.4%. As for the sensitivity profile, antibiotics to which the bacteria showed more resistance were trimethoprim-sulfamethoxazole (57%), tetracycline (21%), and azithromycin (43%). Thus, it was shown that the parakeets were carriers of Enterobacteriaceae with resistance characteristics.

Experimental infection of Salmonella Enteritidis in quails submitted to forced molting by feed fasting

This study aimed at evaluating bacterial shedding, as detected in swabs, feces, and eggs of quails submitted to forced molting by feed fasting and experimentally infected with a Salmonella Enteritidis (SE) strain. In the experiment, 84 40-week-old Italian female quails were distributed in the following groups: FI (quails induced to molt by fasting and inoculated with Salmonella Enteritidis - SE); CI (quails fed with a laying diet and inoculated with SE); FNI (quails induced to molt by fasting and not inoculated with SE); and CNI (quails fed with a laying feed and not inoculated with SE). Feces, cloacal swabs, and eggs were collected on day 1, 3, 7 and 14 post-inoculation (dpi) and submitted to bacteriological analyses. All samples obtained from cloacal swabs were negative for SE. None of the quails of the non-inoculated groups (FNI and CNI) were positive for SE in the fecal samples. Among the inoculated quails, the FI group presented significantly higher (p 0.05) SE shedding in the feces on 1 dpi than the CI group. On 4 dpi, no significant difference was observed (p 0.05) in SE shedding between the inoculated quail groups. On 7 dpi, only the FI group shed SE in the feces, whereas on 14 dpi, none of the groups shed SE. According to the results, we concluded that quails submitted to molting by fasting have higher possibility of shedding SE in the feces.(AU)

Recovery of Salmonella Gallinarum in the Organs of Experimentally-Inoculated Japanese Quails (Coturnix coturnix)

Salmonellosis is an infection caused by specific or non specific serotypes of theSalmonella genus, responsible for losses in the poultry industry. Fowl typhoid, caused by S. Gallinarum (SG) is important because it causes elevated mortality in adult birds, leading to economic losses in the poultry industry. This study aimed at quantifying the number of viable SG cells in the liver, spleen, lung, cecum, and reproductive tract (ovary and testicles) of experimentally inoculated Japanese quails (Coturnix coturnix), as well as SG shedding in their feces. One hundred and two Japanese quails, with four months of age at the beginning of the experiment, were used. The birds were inoculated with three bacterial cultures containing different concentrations (6x104CFU/0.1mL, 2x105 CFU/0.4mL, or 5x106CFU/0.2mL) of SG resistant to nalidixic acid. On days 1, 4, 7, and 14 after the inoculation (dpi) individual cloacal swabs were collected from six birds per group, which were subsequently sacrificed for organ sampling. The swab samples were streaked directly on plates containing brilliant green agar and nalidixic acid (VBNal). Samples that were negative after 24h, were streaked again. The collected organs were individually macerated and transferred to buffered peptone water at 0.1%. The solutions were immediately diluted serially for CFU counting in VBNal. SG was successfully recovered from one quail, which was inoculated with 2x105 CFU/0.4mL, and from five quails of the group inoculated with 5x106CFU/0.2mL inoculum. All of the analyzed cloacal swab samples were negative. Therefore, this study demonstrated it was difficult to isolate SG from the analyzed organs and that it was not possible to recover thepathogen in the cloacal swabs collected from inoculated quails. These results may be explained by the absence of flagella in SG, inducing weak intestinal immune response in the beginning of the infection and preventing its isolation in cloacal swab samples. The low positivity rate of the analyzed organs may be due to the immune status of the euthanized birds, since the SG dissemination in the animal organism occurs mostly close to death, which was observed in the birds found dead during the experiment.(AU)

Experimental infection of Salmonella Enteritidis in quails submitted to forced molting by feed fasting

This study aimed at evaluating bacterial shedding, as detected in swabs, feces, and eggs of quails submitted to forced molting by feed fasting and experimentally infected with a Salmonella Enteritidis (SE) strain. In the experiment, 84 40-week-old Italian female quails were distributed in the following groups: FI (quails induced to molt by fasting and inoculated with Salmonella Enteritidis - SE); CI (quails fed with a laying diet and inoculated with SE); FNI (quails induced to molt by fasting and not inoculated with SE); and CNI (quails fed with a laying feed and not inoculated with SE). Feces, cloacal swabs, and eggs were collected on day 1, 3, 7 and 14 post-inoculation (dpi) and submitted to bacteriological analyses. All samples obtained from cloacal swabs were negative for SE. None of the quails of the non-inoculated groups (FNI and CNI) were positive for SE in the fecal samples. Among the inoculated quails, the FI group presented significantly higher (p 0.05) SE shedding in the feces on 1 dpi than the CI group. On 4 dpi, no significant difference was observed (p 0.05) in SE shedding between the inoculated quail groups. On 7 dpi, only the FI group shed SE in the feces, whereas on 14 dpi, none of the groups shed SE. According to the results, we concluded that quails submitted to molting by fasting have higher possibility of shedding SE in the feces.

Recovery of Salmonella Gallinarum in the Organs of Experimentally-Inoculated Japanese Quails (Coturnix coturnix)

Salmonellosis is an infection caused by specific or non specific serotypes of theSalmonella genus, responsible for losses in the poultry industry. Fowl typhoid, caused by S. Gallinarum (SG) is important because it causes elevated mortality in adult birds, leading to economic losses in the poultry industry. This study aimed at quantifying the number of viable SG cells in the liver, spleen, lung, cecum, and reproductive tract (ovary and testicles) of experimentally inoculated Japanese quails (Coturnix coturnix), as well as SG shedding in their feces. One hundred and two Japanese quails, with four months of age at the beginning of the experiment, were used. The birds were inoculated with three bacterial cultures containing different concentrations (6x104CFU/0.1mL, 2x105 CFU/0.4mL, or 5x106CFU/0.2mL) of SG resistant to nalidixic acid. On days 1, 4, 7, and 14 after the inoculation (dpi) individual cloacal swabs were collected from six birds per group, which were subsequently sacrificed for organ sampling. The swab samples were streaked directly on plates containing brilliant green agar and nalidixic acid (VBNal). Samples that were negative after 24h, were streaked again. The collected organs were individually macerated and transferred to buffered peptone water at 0.1%. The solutions were immediately diluted serially for CFU counting in VBNal. SG was successfully recovered from one quail, which was inoculated with 2x105 CFU/0.4mL, and from five quails of the group inoculated with 5x106CFU/0.2mL inoculum. All of the analyzed cloacal swab samples were negative. Therefore, this study demonstrated it was difficult to isolate SG from the analyzed organs and that it was not possible to recover thepathogen in the cloacal swabs collected from inoculated quails. These results may be explained by the absence of flagella in SG, inducing weak intestinal immune response in the beginning of the infection and preventing its isolation in cloacal swab samples. The low positivity rate of the analyzed organs may be due to the immune status of the euthanized birds, since the SG dissemination in the animal organism occurs mostly close to death, which was observed in the birds found dead during the experiment.

Isolation of Salmonella enterica subsp. enterica (O: 4, 5: i) and Salmonella enterica subsp. Typhimurium from free-living domestic pigeons (Columba livia) / Detecção de Salmonella enterica subsp enterica (O: 4, 5: i) e Salmonella enterica subsp. Typhimurium em pombo doméstico de vida livre (Columba livia)

The present study reports the isolation of Salmonella enterica in organs of free-living domestic pigeons. In the clinic examination, the presence of feces in the peri-cloacal and abdominal regions were observed, as well as symptoms such as cachexy, incoordination and opisthotonos. Before any therapeutic protocol was applied the bird died and a necropsy was then performed for the removal of spleen, liver, kidney and intestine for bacteriological examination and antibiotic sensitivity test. Salmonella enterica subsp.enterica (O:4,5:i-) and Salmonella enterica subsp. enterica serovar Typhimurium were isolated from the liver and intestine and the sensitivity test demonstrated that these strains are sensitive to several antibiotics.(AU)

O presente trabalho relata o isolamento de Salmonella enterica em órgãos de um pombo doméstico de vida livre. No exame clínico foi observada a presença de fezes pericloacal na região ventral, caquexia, incoordenação motora e opistótono. Antes de iniciar um protocolo terapêutico, a ave foi a óbito, e, em seguida, foi realizada uma necropsia para remoção do baço, fígado, rim e intestino para exame bacteriológico e teste de sensibilidade a antibióticos. Foi isolado Salmonella enterica subsp. enterica (O:4,5:i-) e Salmonella enterica subsp. enterica sorovar Typhimurium do fígado e intestino, e o teste de sensibilidade demonstrou que essas cepas são sensíveis a vários antimicrobianos.(AU)

Isolation of Salmonella spp. in captive Psittaciformes from zoos and a commercial establishment of Fortaleza, Brazil / Isolamento de Salmonella spp. em Psittaciformes de zoológicos e estabelecimento comercial de Fortaleza, Brasil

This study showed a low prevalence of Salmonella spp. in captive psittacines from zoos and a commercial establishment of Fortaleza. None of the isolated serotypes (S. Lexington, S. Saintpaul and S. Newport) have yet been reported in Amazona aestiva, Ara chloroptera or Melopsittacus undulatus. However, the fact that most birds presented negative for Salmonella spp. may not imply the absence of this pathogen in these birds, since the intermittent excretion is a well-known characteristic of this microorganism.(AU)

A manutenção de aves em cativeiro reúne condições que favorecem a disseminação de doenças infecciosas, sendo a Salmonella uma dessas doenças infecciosas que acomete os psitacídeos. Portanto o objetivo do presente estudo foi isolar e identificar Salmonella spp. em Psittaciformes mantidos em criatórios comerciais e conservacionistas da Região Metropolitana de Fortaleza. Para o estudo, foram coletados swabs cloacais de 182 psitacídeos clinicamente sadios. Os resultados mostraram que três psitacídeos avaliados (1,65%) foram positivos: Amazona aestiva (Salmonella Lexington), Ara chloroptera (Salmonella Saintpaul) e Melopsittacus undulatus (Salmonella Newport). De acordo com a literatura científica, não há registro desses sorotipos em psitacídeos. Esta pesquisa evidenciou uma baixa prevalência de Salmonella spp. em Psittaciformes mantidos em criatórios comerciais e conservacionistas da Região Metropolitana de Fortaleza.(AU)

Dissemination of Salmonella enteritidis by experimentally-infected pigeons

Two groups of domestic pigeons (Columba livia) were experimentally infected orally with doses of 9.5 x10(7) and 9.5 x10(9) CFU/mL (group A and B, respectively) of a Salmonella Enteritidis (SE) strain isolated from chickens. None of the used doses caused mortality of the inoculated birds; however, the pathogen was successfully recovered from the liver and spleen of group B birds on day 7 post-inoculation (dpi). Pathogen shedding, as evaluated through cloacal swabs, occurred in both groups until the 14th day of observation (p 0.05). Among all fecal samples collected from group B (n=4), three different birds shed the pathogen in their feces, out of which two were positive on 3 dpi and one on 7 dpi. The same number of fecal samples was evaluated in group A and only one bird shed the pathogen, on 7 and 14 dpi. The concentration of the microorganism in the feces was lower in group A than any sample from Group B. Salmonella Enteritidis isolated from chickens, when inoculated in pigeons, may be recovered from feces, cloacal swabs and organs, and these birds may contaminate poultry causing economic losses as well as posing a risk to the public health.(AU)

Dissemination of Salmonella enteritidis by experimentally-infected pigeons

Two groups of domestic pigeons (Columba livia) were experimentally infected orally with doses of 9.5 x10(7) and 9.5 x10(9) CFU/mL (group A and B, respectively) of a Salmonella Enteritidis (SE) strain isolated from chickens. None of the used doses caused mortality of the inoculated birds; however, the pathogen was successfully recovered from the liver and spleen of group B birds on day 7 post-inoculation (dpi). Pathogen shedding, as evaluated through cloacal swabs, occurred in both groups until the 14th day of observation (p 0.05). Among all fecal samples collected from group B (n=4), three different birds shed the pathogen in their feces, out of which two were positive on 3 dpi and one on 7 dpi. The same number of fecal samples was evaluated in group A and only one bird shed the pathogen, on 7 and 14 dpi. The concentration of the microorganism in the feces was lower in group A than any sample from Group B. Salmonella Enteritidis isolated from chickens, when inoculated in pigeons, may be recovered from feces, cloacal swabs and organs, and these birds may contaminate poultry causing economic losses as well as posing a risk to the public health.