Elisângela de Souza Lopes

Isolation of Salmonella spp. in captive Psittaciformes from zoos and a commercial establishment of Fortaleza, Brazil

A manutencao de aves em cativeiro reune condicoes que favorecem a disseminacao de doencas infecciosas, sendo a Salmonella uma dessas doencas infecciosas que acomete os psitacideos. Portanto o objetivo do presente estudo foi isolar e identificar Salmonella spp. em Psittaciformes mantidos em criatorios comerciais e conservacionistas da Regiao Metropolitana de Fortaleza. Para o estudo, foram coletados swabs cloacais de 182 psitacideos clinicamente sadios. Os resultados mostraram que tres psitacideos avaliados (1,65%) foram positivos: Amazona aestiva (Salmonella Lexington), Ara chloroptera (Salmonella Saintpaul) e Melopsittacus undulatus (Salmonella Newport). De acordo com a literatura cientifica, nao ha registro desses sorotipos em psitacideos. Esta pesquisa evidenciou uma baixa prevalencia de Salmonella spp. em Psittaciformes mantidos em criatorios comerciais e conservacionistas da Regiao Metropolitana de Fortaleza.

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 14 th 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.

Bacteriological investigation of microorganisms (Salmonella sp. and other Enterobacteriaceae) in common quails (Coturnix coturnix) submitted to different forced-molting procedures

The aim of this study was to investigate the presence of Salmonella in common quails submitted to forced molting. A total of 240 quails were divided at 40 weeks of age into four groups: CG (control, quails not submitted to molting); FM (fasting method); WM (fed wheat midds ad libitum); and ZM (zinc oxide method). From each group, 10 cloacal swabs, 10 fecal samples, and 20 egg samples were collected before molting (two weeks) and after molting (two weeks). The microbiological procedures for Salmonella spp. identification were performed in four steps. The agglutination test, using somatic and flagellar antigens, was used to confirm Salmonella-suspected colonies. According to the methodology applied, none of the samples was positive for Salmonella spp. The results showed that 20.0% of the egg samples from birds submitted to forced molting were contaminated with enterobacteria. It was concluded that, under the conditions of the present experiment, the stress caused by forced molting did not induce infection by Salmonella spp. or increased Enterobacteriaceae contamination levels in the eggs.

Salmonella Gallinarum virulence in experimentally-infected Japanese quails (Coturnix japonica)

Salmonella Gallinarum is the agent of fowl typhoid in poultry and infects mainly adult galliforms, causing significant economic losses in poultry production. Because quails are susceptible to this disease and quail production is becoming increasingly important in Brazil, this study was carried out to evaluate the virulence of Salmonella Gallinarum strain to quails. The inoculum was prepared from S. Gallinarum strain resistant to nalidixic acid. Forty eight 16-week-old Japanese quails were randomly distributed in three groups. Before the experiment, cloacal swabs were collected from all birds in order to confirm they were free from Salmonella spp. Cloacal swabs and fecal samples were collected on days 03, 06, 09, 12, and 15 post-inoculation. Birds that died during the experiment were submitted to post-mortem examination, and had their organs aseptically collected for bacteriological examination. All eggs produced during the experiment were also examined. The mortality rate recorded during the experiment was 43.75% (21/48). S. Gallinarum was recovered from the organs of the birds that naturally died during the experiment, but the agent was not isolated from the organs of sacrificed birds. No egg sample was positive for Salmonella Gallinarum. It was concluded that S. Gallinarum may be recovered from the organs of experimentally-infected Japanese quails.

Isolation and antimicrobial resistance of Escherichia coli and Salmonella enterica subsp. enterica (O:6,8) in broiler chickens

Background: The third largest poultry flock in Northeast Brazil is located in Ceara 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 Ceara 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.

Identification and antimicrobial resistance of members from the Enterobacteriaceae family isolated from canaries (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.

Isolation of Salmonella enterica subsp. enterica (O:4,5:i) and Salmonella enterica subsp. Typhimurium from free-living domestic pigeons (Columba livia)

O presente trabalho relata o isolamento de Salmonella enterica em orgaos de um pombo domestico de vida livre. No exame clinico foi observada a presenca de fezes pericloacal na regiao ventral, caquexia, incoordenacao motora e opistotono. Antes de iniciar um protocolo terapeutico, a ave foi a obito, e, em seguida, foi realizada uma necropsia para remocao do baco, figado, rim e intestino para exame bacteriologico e teste de sensibilidade a antibioticos. Foi isolado Salmonella enterica subsp. enterica (O:4,5:i-) e Salmonella enterica subsp. enterica sorovar Typhimurium do figado e intestino, e o teste de sensibilidade demonstrou que essas cepas sao sensiveis a varios antimicrobianos.

Isolamento de Salmonella spp. e Escherichia coli de psittaciformes: relevância em saúde pública

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.

Pesquisa de enterobactérias em patos domésticos (Cairina moschata) de propriedades localizadas em quatro municípios do Ceará, Brasil

This study aimed to isolate and identify members of the Enterobacteriaceae that were present in domestic ducks (Cairina moschata) from properties located in four cities in the State of Ceara, Brazil. Therefore, 65 stool samples and 47 cloacal swabs were collected from farms located in the following cities: Fortaleza, Boa Agua, Eusebio and Cascavel. The samples were submitted to bacteriological processing. In pre-enrichment, all of the stool and swab samples were cultured in buffered peptone water 0.1%. For selective enrichment, aliquots from the tubes of the prior step after incubation were transferred to tubes containing Rappaport-Vassiliadis and Selenite Cystine broths. Plates with MacConkey and Brilliant Green agars were streaked with the content from the enrichment tubes after incubation. Colonies were chosen based on their morphological characteristics for the biochemical tests (TSI: Triple-Sugar-Iron; LIA: Lysine-Iron-Agar; and SIM: Sulfide-indole-motility). The bacteria were identified based on their biochemical characteristics. The mostly isolated bacteria were Citrobactersp., Proteussp., and Enterobactersp. In a lower frequency, isolated enterobacteria were Klebsiellasp., Hafniasp., Escherichia coli, Edwardsiellasp., Providenciasp. and Serratiasp. With the methodology applied, no Salmonellawas isolated from the evaluated ducks, which is a genus commonly associated with this avian species; however the microbiota were composed by the main enterobacteria that are common to other species of birds, some of which are potentially pathogenic both to animals and humans.

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 greybreasted parakeets may harbor enterobacteria with high antimicrobial resistance rates.