Sistema api campy para caracterização de amostras de campylobacter isoladas de descarga cecal, fezes, swabs cloacais e carcaças de frangos de corte

ABSTRACT The aim of the present study was to assess the API Campy system for characterization of Campylobacter strains. Forty-eight Campylobacter strains were isolated from 22 three-week-old broiler flocks: 15 from cecal droppings, 5 from feces, 3 from cloacal swabs, and 21 from carcasses. The strains were presumptively analyzed by phase contrast microscopy, Gram staining, catalase/oxidase activity, and latex agglutination test, and were then inoculated into the API Campy system, which consists of enzyme-linked and conventional assays under aerobic conditions and assimilation or inhibition tests under microaerophilic conditions, both incubated at 37º C for 2448 hours, the readings made with a computerized system. A total of 43 strains (89.58%) could be identified, whereas 5 (10.41%) yielded an unacceptable profile. The following species were identified: C. jejuni subsp. jejuni (68.8%), C. coli (8.3%), C. jejuni subsp doylei (6.3%), C. upsaliensis (4.2%) and C. fetus subsp. fetus (2.1%). An 81.8% prevalence was found for Campylobacter in broiler flocks, mainly Campylobacter jejuni subsp. jejuni, identified by the API Campy system, showing a larger number of species, subspecies and biotypes of Campylobacter among the strains isolated from carcasses than among the clinical strains obtained from poultry farms.

RESUMO O presente estudo verificou a aplicação do sistema API Campy para identificação de bactérias do gênero Campylobacter. Foram utilizadas 48 amostras, sendo 15 oriundas de descarga cecal, cinco de fezes, três de swabs cloacais e 21 de carcaças, isoladas de 22 lotes de frangos de corte com três semanas de idade. As amostras caracterizadas presuntivamente por microscopia em contraste de fase, coloração de Gram, catalase/oxidase e aglutinação em látex foram inoculadas no sistema API Campy, que consta de testes enzimáticos e convencionais em aerobiose e testes de assimilação ou inibição em microaerofilia, ambos incubados a 37º C por 24-48 horas, sendo a leitura realizada em sistema informatizado. Foi possível identificar 43 amostras (89,58%), enquanto cinco (10,41%) tiveram perfil inaceitável. Identificou-se as espécies C. jejuni subsp. jejuni (68,8%); C. coli (8,3%); C. jejuni subsp. doylei (6,3%); C. upsaliensis (4,2%) e C. fetus subsp. fetus (2,1%). A ocorrência de Campylobacter nos lotes de frango de corte estudados foi de 81,8% e as espécies identificadas principalmente como Campylobacter jejuni subesp. jejuni pelo sistema API Campy, apresentando um maior número de espécies, subespécies e biotipos deCampylobacter dentre as amostras isoladas de carcaças de frango do que dentre as amostras clínicas isoladas nas granjas.

OCURRENCE OF Campylobacter sp IN BROILER FLOCKSAND CORRESPONDING CARCASSES / OCORRÊNCIA DE Campylobacter sp EM LOTES DE FRANGOS DE CORTE E NAS CARCAÇAS CORRESPONDENTES

The aim of the present study was to assess the dissemination and levels of Campylobacter contamination in broiler flocks and related carcasses. Twenty-two flocks aged 3 weeks or older were assessed, and 110 cecal droppings and 96 carcasses (38 carcasses after defeathering and 58 after the last chilling operation) were enumerated. Bolton selective enrichment broth was used for enumeration of the organism. Additionally, the carcasses were submitted to pre-enrichment for the detection of the agent at low levels of contamination. Was noted that 18.18% of broiler flocks (4/22) was not colonized by Campylobacter; however, their carcasses were found to be contaminated after defeathering. The mean levels of colonization of positive flocks were 7.00 log10 cfu/g of cecal droppings, 5.15 log10 cfu per carcass after defeathering and 4.24 log cfu per carcass after the last chilling operation. The prevalence of Campylobacter in carcasses after defeathering and chilling were positively correlated, whereas a reduction of around 1 log10 was observed in the contamination initially found in the carcasses, showing that current measures, such as Hazard Analysis and Critical Control Points (HACCP) and Good Production Practices, are useful but insufficient for thoroughly eliminating Campylobacter from the end product. Key words: Chicken, contamination, slaughterhouse.

O presente trabalho teve por objetivo verificar a ocorrência de Campylobacter em lotes de frango de corte durante a criação e nas carcaças correspondentes após o abate. Foram avaliados 22 lotes a partir das três semanas de idade, com a análise de 110 conteúdos cecais e 96 carcaças (38 após a depenadeira e 58 após o último chiller). Para a enumeração do Campylobacter, utilizou-se o caldo Bolton, analisando-se as carcaças pré-enriquecimento. Quatro (18,18%) dos lotes analisados não estavam colonizados por Campylobacter, mas, após a depenadeira, carcaças correspondentes estavam contaminadas. Os níveis médios de colonização dos lotes positivos foram de 7 log10ufc/g de conteúdo cecal, 5,15 log10 ufc/carcaça após a depenadeira e 4,24 log10 ufc/carcaça após o último chiller. Os níveis de Campylobacter nas carcaças após a depenadeira e chiller tiveram correlação positiva. Observou-se redução de cerca de 1 log10 na contaminação inicial, demonstrando que medidas atuais, como análise de perigos e pontos críticos de controle e boas práticas de produção, são úteis, mas insuficientes para eliminar completamente Campylobacter do produto final. Palavras-chaveS: Abatedouro, aves, colonização bacteriana.

Antimicrobial resistance in Campylobacter spp isolated from broiler flocks

The aim of this study was to assess the antimicrobial susceptibility of 62 Campylobacter spp. strains obtained from broiler flocks using the agar diffusion method. The Campylobacter spp strains were isolated from 22 flocks aged between 3 and 5 weeks of life, isolated from cloacae swabs, stools and cecal droppings in the farm and from the carcass rinsing in the slaughterhouse. Campylobacter spp strains were tested on Mueller-Hilton (MH) agar (27 samples) and MH plus TTC agar (35 samples). The antimicrobial susceptibility test revealed a 62.5% resistance to at least one drug, especially to enrofloxacin (71%), neomycin (50%), lincomycin (50%), tetracycline (43%), penicillin (42%), ceftiofur (33%) amoxicillin (27%), spiramycin (20%), ampicillin (18%) and norfloxacin (14%), whereas a lower percentage of strains was resistant to erythromycin (10%) and doxycycline (10%). All strains were sensitive to gentamicin and lincomycin-spectinomycin and 80% of them to colistin. These results indicate that it is necessary to reduce the use of antimicrobials in veterinary and human medicine.

O objetivo deste estudo foi verificar a susceptibilidade antimicrobiana de 62 amostras de Campylobacter spp. em amostras isoladas de 22 lotes de frango de corte, pelo método de difusão em Agar. As amostras de Campylobacter spp foram isoladas de frangos com idade entre 3 e 5 semanas, isoladas a partir de swabs cloacais, fezes e descarga cecal obtidos nas granjas e de rinsagem de carcaças no abatedouro. Das 62 amostras avaliadas, 27 foram testadas em ágar MH e 35 no ágar MH com TTC. O perfil de susceptibilidade antimicrobiana apresentou 62,5% de resistência para, no mínimo, uma droga, sobretudo para enrofloxacina (71%), neomicina (50%), lincomicina (50%), tetraciclina (43%), penicilina (42%), ceftiofur (33%) amoxicilina (27%), espiramicina (20%), ampicilina (18%) e norfloxacina (14%), enquanto uma percentagem menor foi observada frente eritromicina (10%) e doxiciclina (10%). Todas as amostras foram sensíveis a gentamicina e linco-espectinomicina e 80% à colistina. Pelo exposto, faz-se necessária a redução do uso dos antimicrobianos na medicina veterinária e em humanos.

Risk factors and likelihood of Campylobacter colonization in broiler flocks

Campylobacter was investigated in cecal droppings, feces, and cloacal swabs of 22 flocks of 3 to 5 week-old broilers. Risk factors and the likelihood of the presence of this agent in these flocks were determined. Management practices, such as cleaning and disinfection, feeding, drinkers, and litter treatments, were assessed. Results were evaluated using Odds Ratio (OR) test, and their significance was tested by Fisher's test (p 0.05). A Campylobacter prevalence of 81.8% was found in the broiler flocks (18/22), and within positive flocks, it varied between 85 and 100%. Campylobacter incidence among sample types was homogenous, being 81.8% in cecal droppings, 80.9% in feces, and 80.4% in cloacal swabs (230). Flocks fed by automatic feeding systems presented higher incidence of Campylobacter as compared to those fed by tube feeders. Litter was reused in 63.6% of the farm, and, despite the lack of statistical significance, there was higher likelihood of Campylobacter incidence when litter was reused. Foot bath was not used in 45.5% of the flocks, whereas the use of foot bath associated to deficient lime management increased the number of positive flocks, although with no statiscal significance. The evaluated parameters were not significantly associated with Campylobacter colonization in the assessed broiler flocks.

Risk factors and likelihood of Campylobacter colonization in broiler flocks

Campylobacter was investigated in cecal droppings, feces, and cloacal swabs of 22 flocks of 3 to 5 week-old broilers. Risk factors and the likelihood of the presence of this agent in these flocks were determined. Management practices, such as cleaning and disinfection, feeding, drinkers, and litter treatments, were assessed. Results were evaluated using Odds Ratio (OR) test, and their significance was tested by Fisher's test (p 0.05). A Campylobacter prevalence of 81.8% was found in the broiler flocks (18/22), and within positive flocks, it varied between 85 and 100%. Campylobacter incidence among sample types was homogenous, being 81.8% in cecal droppings, 80.9% in feces, and 80.4% in cloacal swabs (230). Flocks fed by automatic feeding systems presented higher incidence of Campylobacter as compared to those fed by tube feeders. Litter was reused in 63.6% of the farm, and, despite the lack of statistical significance, there was higher likelihood of Campylobacter incidence when litter was reused. Foot bath was not used in 45.5% of the flocks, whereas the use of foot bath associated to deficient lime management increased the number of positive flocks, although with no statiscal significance. The evaluated parameters were not significantly associated with Campylobacter colonization in the assessed broiler flocks.

Pesquisa de Salmonella sp na cadeia produtiva de avestruzes no Sudeste do Brasil

As criações de avestruzes vêm se desenvolvendo no Brasil e em outros países. De acordo com a literatura, Salmonella sp está envolvida na mortalidade de filhotes de avestruzes, sendo também importantes causadoras de toxinfecção alimentar em seres humanos. Apesar disso, existem poucos estudos envolvendo a pesquisa de Salmonella sp nesta espécie. Desse modo, elaborou-se o presente trabalho com o objetivo de pesquisar Salmonella sp em avestruzes de uma propriedade da região sudeste do Brasil. Para isso, foram realizados exames bacteriológicos, em 80 amostras de fezes de avestruzes (colhidas nas varias fases da criação), 90 ovos não eclodidos, 30 amostras de ração e em 30 amostras de fezes de roedores presentes na propriedade. Em seguida, investigou-se Salmonella sp em fezes, conteúdo cecal, fígado, baço e carcaça de 90 avestruzes abatidos. Aliado a isso, colheu-se o soro sanguíneo destes animais para a realização do ?teste da Pulorose?. Os exames microbiológicos demonstraram ausência de Salmonella sp em amostras de fezes de roedores e em fezes, ovos, conteúdo cecal, fígado, baço e carcaça dos avestruzes analisados. Por outro lado, Salmonella enterica subespécie enterica 4, 12: i:- foi isolada de uma amostra de ração e Salmonella Javiana de outras duas. O teste sorológico foi negativo nas 90 amostras. O sistema de criação, o bom manejo sanitário adotado na propriedade e a adoção dos programas de Boas Práticas de Fabricação (BPF) e Análise de Perigos e Pontos Críticos de Controle (APPCC) no abatedouro contribuíram para a ausência de Salmonella sp nas amostras examinadas

In recent years, the ostrich farming has expanded considerably in Brazil and all over the world. According to literature, Salmonella sp causes mortality in ostrich chicks, being also responsible for human food borne diseases. Even though, little research has focused on Salmonella in ostriches. This study was elaborated with the purpose to search for Salmonella sp in ostriches of a farm located on Brazilian southwest region. By conventional microbiological method, Salmonella sp was researched in 80 samples of ostrich?s droppings, 90 eggs, 30 samples of feed and 30 samples of droppings from resident rodent population. In sequence, this bacterium was searched in droppings, caecal content, spleen, liver and carcass from 90 slaughtered ostriches. Also, blood serum of those animals were harvested and submitted to test for Pullorum Disease (Rapid Serum Blood Agglutination Test). No Salmonella sp was detected in eggs, caecal content, liver, spleen, carcass and droppings from ostriches, nor in droppings from rodents. On the other hand, Salmonella Javiana was isolated from two samples of feed and Salmonella enterica enterica 4, 12: i:- from another one. The serologic test was negative for all samples. The management of ostrich farming, the good sanitary management adopted on farm and the application of Hazard Analysis and Critical Control Point (HACCP) principles and Good Manufacturing Practices (GMP) during the slaughter process contributed for absence of Salmonella sp on samples assayed

OCURRENCE OF Campylobacter sp IN BROILER FLOCKSAND CORRESPONDING CARCASSES / OCORRÊNCIA DE Campylobacter sp EM LOTES DE FRANGOS DE CORTE E NAS CARCAÇAS CORRESPONDENTES

The aim of the present study was to assess the dissemination and levels of Campylobacter contamination in broiler flocks and related carcasses. Twenty-two flocks aged 3 weeks or older were assessed, and 110 cecal droppings and 96 carcasses (38 carcasses after defeathering and 58 after the last chilling operation) were enumerated. Bolton selective enrichment broth was used for enumeration of the organism. Additionally, the carcasses were submitted to pre-enrichment for the detection of the agent at low levels of contamination. Was noted that 18.18% of broiler flocks (4/22) was not colonized by Campylobacter; however, their carcasses were found to be contaminated after defeathering. The mean levels of colonization of positive flocks were 7.00 log10 cfu/g of cecal droppings, 5.15 log10 cfu per carcass after defeathering and 4.24 log cfu per carcass after the last chilling operation. The prevalence of Campylobacter in carcasses after defeathering and chilling were positively correlated, whereas a reduction of around 1 log10 was observed in the contamination initially found in the carcasses, showing that current measures, such as Hazard Analysis and Critical Control Points (HACCP) and Good Production Practices, are useful but insufficient for thoroughly eliminating Campylobacter from the end product. Key words: Chicken, contamination, slaughterhouse.

O presente trabalho teve por objetivo verificar a ocorrência de Campylobacter em lotes de frango de corte durante a criação e nas carcaças correspondentes após o abate. Foram avaliados 22 lotes a partir das três semanas de idade, com a análise de 110 conteúdos cecais e 96 carcaças (38 após a depenadeira e 58 após o último chiller). Para a enumeração do Campylobacter, utilizou-se o caldo Bolton, analisando-se as carcaças pré-enriquecimento. Quatro (18,18%) dos lotes analisados não estavam colonizados por Campylobacter, mas, após a depenadeira, carcaças correspondentes estavam contaminadas. Os níveis médios de colonização dos lotes positivos foram de 7 log10ufc/g de conteúdo cecal, 5,15 log10 ufc/carcaça após a depenadeira e 4,24 log10 ufc/carcaça após o último chiller. Os níveis de Campylobacter nas carcaças após a depenadeira e chiller tiveram correlação positiva. Observou-se redução de cerca de 1 log10 na contaminação inicial, demonstrando que medidas atuais, como análise de perigos e pontos críticos de controle e boas práticas de produção, são úteis, mas insuficientes para eliminar completamente Campylobacter do produto final. Palavras-chaveS: Abatedouro, aves, colonização bacteriana.