Sobrevivência de Campylobacter jejuni em amostras de leite pasteurizado e UHT artificialmente contaminados e mantidas sob refrigeração / Viability of Campylobacter jejuni in the refrigerated samples of artificially contaminated pasteurized and UHT milks

Campylobacter é o agente etiológico mais prevalente em gastroenterites de causa alimentar no mundo. Apesar de o leite cru ser fonte de infecção, pouco se conhece sobre as consequências da recontaminação do leite. A viabilidade de Campylobacter jejuni foi avaliada em leites pasteurizados e UHT mantidos sob refrigeração. Ambos os leites foram divididos em cinco porções de 100 mL, inoculados com 101 UFC.mL-1 de C. jejuni e mantidos de 4 ºC a 7 ºC por 48 horas. Repetiu-se o procedimento, utilizando-se inoculações de 102, 103 e 104 UFC.mL-1. As alíquotas foram analisadas imediatamente após inoculação e depois de 24 e 48 h quanto à viabilidade de C. jejuni. O micro-organismo manteve-se viável em todas as amostras, porém no leite pasteurizado houve redução de 1 ciclo log nas contagens após 24 h e baixas contagens após 48 h. Provavelmente,a redução ocorreu pela presença de microbiota neste leite, que competiu ou inibiu o crescimento de C. jejuni. O leite UHT ofereceu boas condições de sobrevivência em todos os períodos. O consumo de leite contaminado, mesmo em armazenamento refrigerado, pode ser fonte de infecção. O micro-organismo manteve-se mais viável no leite UHT quando comparado ao pasteurizado, provavelmente pela ausência de outros micro-organismos competidores.

Campylobacter is the most prevalent etiologic agent of foodborne gastroenteritis worldwide.Although raw milk is an important source of infection, little is known about the consequencesof milk recontamination. This study aimed at verifying the viability of Campylobacter jejuni inpasteurized and UHT milks stored under refrigeration. Both samples were divided into five portionsof 100 mL, inoculated with 101 CFU.mL-1 of C. jejuni and stored at 4 °C - 7 ºC for 48 h. The procedure was repeated using inoculations of 102, 103 and 104 CFU.mL-1. Aliquots were analyzed immediatelyafter inoculation and after 24 and 48 h to evaluate C. jejuni viability. The microorganism their viabilityin all of samples. In pasteurized milk a reduction of 1 log cycle occurred after 24 h and low countsafter 48 h. Probably, the reduction happened due to the occurrence of microbiota in this milk, whichcaused competition or inhibition of microorganism growth. UHT milk offered good conditions forbacteria survival in all of the periods. The consumption of contaminated milk, even stored under refrigeration might be a source of infection. Microorganism were more viable in UHT milk thanin pasteurized one, probably owing to the absence of competing microorganisms.

Campylobacter jejuni in commercial eggs

This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to assess the viability in the egg yolk. To determine whether the albumen and egg fertility interferes with the entry and survival of bacteria, we used varying concentrations of albumen and SPF and commercial eggs. C. jejuni was recovered in SPF eggs (fertile) after three hours in contact with contaminated wood shavings but not in infertile commercial eggs. The colonies isolated in the SPF eggs were identified by multiplex PCR and the similarity between strains verified by RAPD-PCR. The bacteria grew in different concentrations of albumen in commercial and SPF eggs. We did not find C. jejuni in commercial eggs inoculated directly into the air chamber, but the bacteria were viable during all periods tested in the wood shavings. This study shows that consumption of commercial eggs infected with C. jejuni does not represent a potential risk to human health.

Transmission of Campylobacter coli in chicken embryos

Campylobacter coli is an important species involved in human cases of enteritis, and chickens are carriers of the pathogen mainly in developing country. The current study aimed to evaluate the transmission of C. coli and its pathogenic effects in chicken embryos. Breeder hens were inoculated intra-esophageally with C. coli isolated from chickens, and their eggs and embryos were analyzed for the presence of bacteria using real-time PCR and plate culture. The viability of embryos was verified. In parallel, SPF eggs were inoculated with C. coli in the air sac; after incubation, the embryos were submitted to the same analysis as the embryos from breeder hens. In embryos and fertile eggs from breeder hens, the bacterium was only identified by molecular methods; in the SPF eggs, however, the bacterium was detected by both techniques. The results showed no relationship between embryo mortality and positivity for C. coli in the embryos from breeder hens. However, the presence of bacteria is a cause of precocious mortality for SPF embryos. This study revealed that although the vertical transmission is a possible event, the bacteria can not grow in embryonic field samples.

Resistência a oxacilina em Staphylococcus spp isolado de leite mastítico / Oxacillin-resistance Staphylococcus spp isolated from mastitic milk

A mastite ocupa lugar de destaque entre as doenças que acometem o rebanho leiteiro, em virtude de problemas econômicos e de saúde pública. Staphylococcus spp são os agentes infecciosos mais envolvidos na etiologia da doença. Staphylococcus spp resistentes à oxacilina, isolados de amostras de leite provenientes de animais com histórico de mastite recorrente, foram analisadas utilizando-se os testes de difusão em gel, ágar de triagem, concentração mínima inibitória (CMI) e pesquisa do gene mecA. De 134 amostras de leite analisadas, Staphylococcus spp foi isolado em 55,22% das amostras (74/134). O teste de difusão em disco demonstrou que a maioria dos isolados apresentaram multiresistência, sendo 51,35% (38/74) resistentes à oxacilina. Esse perfil foi confirmado em paralelo pela CMI e pelo ágar de triagem. A análise molecular demonstrou que 33,78% dos isolados (25/74) possuíam o gene mecA, sendo mais frequentemente isolado em Staphylococcus aureus e Staphylococcus coagulase negativa, com índices de 48% (12/25) e 32% (8/25),respectivamente, seguido de S. intermedius com 16% (4/25) e de S. hyicus com 4% (1/25). O presente estudo denota o grave problema associado à Staphylococcus aureus resistente à oxacilina, bem como no meio rural, o que justifica a dificuldade de tratamento e a recorrência da infecção.

Mastitis is of a major concern among the diseases that affect the dairy herd due to the economic problemsand public health concerns. Staphylococcus spp has been the infectious bacterium and the etiologic agentof this disease. The oxacillin resistance of Staphylococcus spp isolated from milk samples of animals witha history of recurrent mastitis was investigated. This study was performed by means of gel diffusion test,agar screening, minimum inhibitory concentration (MIC), and the mecA gene searching. Of 134 milksamples, Staphylococcus spp was isolated from 55.22% samples (74/134). By means of disk diffusion test,the majority of the isolates showed multiresistance, and 51.35% (38/74) were resistant to oxacillin. Thisprofile was confirmed in parallel by CMI and the agar screening. By molecular analysis 33.78% of isolates(25/74) revealed the mecA gene, being frequently isolated from Staphylococcus aureus and coagulase negativeStaphylococcus at rates of 48% (12/25) and 32% (8/25 ), respectively, and followed by S. intermedius in 16%(4 / 25) and S. hyicus in 4% (1/25). The serious problem associated with oxacillin-resistant Staphylococcusaureus was found this study, including in rural environment and these findings justifie the treatmentdifficulty and the recurrence of infection.

Fatores de Risco Relacionadas à Contaminação Microbiana de massa de quibe / Risk factors related to microbial masses of kibe

A massa de quibe passa por apreciável manuseio, ficando exposta a uma série de perigos ou oportunidades de contaminações microbianas que podem levar à deterioração do produto ou oferecer risco à saúde do consumidor. As inadequadas condições de higiene e armazenamento podem acentuar esse problema. Este estudo objetivou averiguar a contaminação por coliformes termotolerantes, Staphylococcus coagulase positiva e Salmonella spp. em massa de quibe comercializadas nos cinco setores geográficos da cidade de Uberlândia-MG e sua adequação em comparação com as especificações da ANVISA, e também o risco de procedimentos relacionados ao seu preparo e armazenamento para a contaminação deste alimento. Das amostras analisadas, 42% (21/50) foram consideradas insatisfatórias. Destas, 38% (19/50) apresentavam coliformes termotolerantes acima dos permitidos pela legislação e 4% (2/50) a presença de Salmonella spp. Nenhuma amostra mostrou inadequação quanto aos números de Staphylococcus coagulase positiva. Os fatores de risco relacionados positivamente à contaminação por coliformes termotolerantes foram: distribuição e organização dos alimentos no balcão frigorífico (P=0,002; OR=3,21), a higiene e conservação de utensílios (P=0,26; OR=2,02) e a climatização (P=0,09; OR=2,72). Nenhum dos riscos analisados foi diretamente associado à presença de Salmonella spp.

The kibe mass pass to an intensive handling, being exposed to several hazards on microbial contamination opportunities. These contaminations can conduct to product deterioration or offer risks to the costumers' health. The inadequate hygiene conditions and storage emphasize this problematic system. The study aimed to determine the kibe mass contamination by bacterial from thermotolerants coliforms, Staphylococcus positive coagulase and Salmonella spp., of the kibe masses commercialized in five geographic sectors of the Uberlândia-MG; ascertain the suitability of the product's specifications to ANVISA, and the main risks related to contamination by thermotolerant. 42% of the analyzed samples (21/50) were found as unsatisfactory due termotolerant coliforms (38%) above permissible legislation values and Salmonella spp. presence (4%). No samples showed inadequate counting of Staphylococcus positive coagulase. Risk factors positively related to contamination by fecal coliform were: distribution and organization of food at the counter refrigerator (P = 0.002, OR = 3.21), hygiene and maintenance of vessels (P = 0.26, OR = 2, 02) and air (P = 0.09, OR = 2.72). None of the risk analysis was directly associated with the presence of Salmonella spp...

Viability of Campylobacter jejuni in commercial eggs / Viabilidade de Campylobacter jejuni em ovos comerciais

The aim of this study was to verify the penetration and permanence of C. jejuni inside commercial eggs. Thirty eggs were submerged in peptone water artificially contaminated with 10 5 UFC.mL-1 of C. jejuni. An equal number of eggs were submerged in an identical solution, however, without addition of the microorganism. Parallel to this, 60 eggs were inoculated by means of a small incision in the larger pole, without lesion to the internal membrane, inserted into the air chamber with a sterile insulin needle containing 10 5 CFU.mL-1 of C. jejuni. Thirty eggs were inoculated with a sterile solution of 0.85% sodium chloride and kept as negative control. After storage at 25ºC, and after 1, 6 and 24 hours, 10 eggs from each of the two treatments were separated into the albumen and yolk contents, and each portion was individually analyzed by the traditional culture method for the presence/absence of Campylobacter. In another nine eggs, obtained from the same lot of animals, the albumens were separated to verify their inhibitory activity against the microorganism. The albumen in different concentrations was incorporated into Bolton broth and inoculated with 10 6 CFU.mL-1 of C. jejuni, and after incubation at 37ºC for 24 hours, striated in selective agar. In none of the analyzed samples (treatments and controls) was the growth of Campylobacter observed. The results demonstrated that under the experimental conditions used, Campylobacter jejuni is incapable of surviving, colonizing and multiplying inside commercial eggs. The white of the eggs, as an isolated factor, even at the concentration of 84.9% was incapable of completely inhibiting the growth of the microorganism in Bolton broth at 37ºC, in a microaerophilic atmosphere. The consumption of commercial eggs is not a risk factor for the acquisition of Campylobacteriosis by humans.

O objetivo desse estudo foi verificar a passagem e a permanência de C. jejuni no interior de ovos comerciais. Trinta ovos foram submersos em água peptona artificialmente contaminada com 105 CFU.mL-1 de C. jejuni e igual número de ovos foi submerso em solução idêntica, porém, sem adição do microrganismo. Em paralelo, 60 ovos foram inoculados por meio de uma pequena incisão no pólo maior, sem lesionar a membrana interna, inseridos na câmara de ar com uma agulha de insulina estéril contendo 10 5 CFU.mL-1 de C. jejuni. Trinta ovos foram inoculados com solução estéril de cloreto de sódio 0,85% e mantidos como controle negativo. Após armazenamento a 25ºC, e após 1, 6 e 24 horas, 10 ovos de cada um dos dois tratamentos foram separados nos conteúdos de albume e gema, e cada porção individualmente analisada pelo método tradicional de cultivo para presença/ausência de Campylobacter. Em outros nove ovos, obtidos do mesmo lote de animais, foram separados albume, para verificar sua atividade inibitória frente ao microrganismo. O albume em diferentes concentrações foi incorporada em caldo Bolton e inoculada com 10 6 CFU.mL-1 de C. jejuni, e após incubação a 37ºC por 24 horas, estriadas em ágar seletivo. Em nenhuma das amostras analisadas (tratamentos e controle) foi observado o crescimento de Campylobacter. Os resultados demonstraram que nas condições experimentais utilizadas, Campylobacter jejuni não é capaz de sobreviver, colonizar e se multiplicar no interior de ovos comerciais. O albume, como fator isolado, mesmo na concentração de 84,9% não foi capaz de inibir completamente o crescimento do microrganismo em caldo Bolton a 37ºC em microaerofilia. O consumo de ovos não é um fator de risco para a aquisição de campilobacteriose por humanos.

Campylobacter sp em mecônio de pintainhos e em cloaca de reprodutoras de corte / Campylobacter sp in meconium of one day old chicks and swabcloacal from breeder hens

Campylobacter sp é um importante patógeno veiculado pelos alimentos e os produtos de origem aviária são os mais implicados na transmissão desses microrganismos aos humanos. O conhecimento das formas de transmissão de bactérias do gênero Campylobacter entre as aves irá permitir o estabelecimento de formas de controle eficientes nas granjas para impedir a disseminação desse agente. A principal via de transmissão nos aviários é a horizontal, porém a transmissão vertical deve ser objeto de estudo por ser uma via ainda não comprovada. O objetivo desse estudo foi verificar em um mesmo lote de aves a presença da Campylobacter sp em galinhas matrizes e em mecônio de pintainhos de corte de um dia. O diagnóstico, foi realizado por PCR automatizado, o BAX Systen da DuPont. Foram coletados e analisados suabes cloacais de 33 aves (pool de três aves), totalizando 11 amostras e mecônio de pintainhos recémeclodidos (pool de três), totalizando 10 amostras. A positividade foi de 80% (8/10) para as amostras de mecônio e 54,55% (6/11) das amostras obtidas de galinhas. Esses resultados representam indícios da transmissão vertical, mas outras pesquisas devem ser conduzidas utilizando técnicas moleculares para detecção de Campylobacter sp em amostras de fezes, assim como técnicas de genotipificação dos espécimes isolados para comprovação da transmissão vertical.

Campylobacter sp is an important agent that causes food infection and the consume of avian products was the principal way of transmission of this organism to human been. The knowledge of the forms of transmission of the Campylobacter between the birds will allow the establishment of efficient forms of control in the farms to hinder the dissemination of this agent. The main transmission routes on chicken farms is horizontal however, the vertical transmission must be object of study still, it’s not proven the objective of this research, that was to verify the presence of Campylobacter sp in breeder hens and meconium. The used diagnostic method, was the automatized system PCR, the BAX Systen of the DuPont. The microbiological analyses were performed, using cloacal swabs from 33 breeder hens (pool of 3 hens each sample) and meconium samples from 30 one day old chicks (pool of 3 broilers each sample). Analysis of the meconium showed 80% (8/10) positive and breeder hens by the cloacal swab method 54,55% (6/11). These results represent indications of the vertical transmission, but other research must be lead using molecular techniques for detention of Campylobacter sp in excrement samples, as well as genetics techniques of isolated specimens to evidence the vertical transmission.

Campylobacter sp in eggs from cloacal swab positive breeder hens

Campylobacter sp is a microaerophilic, thermotolerating Gram negative bacterium, known to be one of the main causes of food-borne human infections. Among the foods that carry these microorganisms, the chicken is outstanding. In Brazil, a large chicken exporting country, few researches are conducted about their prevalence in breeder hens and the transmission through eggs. The aim of this research was to verify the presence of Campylobacter sp in the shells and within the eggs from positive cloacal swab breeder hens. Microbiological analyses were made on cloacal swabs of 140 weighed breeder hens. The positive breeder hens were set aside and in a total of 244 of their eggs, Campylobacter sp was present in macerated shells and yolk contents during 7 weeks. Out of the 140 researched breeder hens, 25 (17.8 percent) were positive from cloacal swabs, however the eggs were not positive. The physiological characteristics of the birds, their eggs and Campylobacter sp favor the bacterium entering and surviving in the eggs, but in this study, no positive result was found in macerated shells or in the yolks, indicating that vertical transmission is probably an unusual event.

Campylobacter sp é reconhecida como uma das principais causas de gastrenterite humana de origem alimentar. Dentre os alimentos veiculadores desses microrganismos, a carne de frango tem sido a mais implicada. Os estudos existentes sobre a transmissão vertical da Campylobacter são escassos e não conclusivos. O objetivo desse estudo foi verificar a presença de Campylobacter sp na casca e interior de ovos de matrizes positivas em swabs cloacais e a possibilidade de transmissão vertical. Foram analisados swabs cloacais de 140 matrizes pesadas e seus ovos colhidos para análise durante 7 semanas consecutivas. Dos 244 ovos colhidos, 129 foram fumigados e 115 analisados sem tratamento. Foram analisados o macerado da casca e a gema. Das 140 matrizes pesquisadas, 25 (17,8 por cento) foram positivas em swabs cloacais. Não houve positividade em nenhuma das amostras de ovos, independente da fumigação ou período de coleta. Apesar das características fisiológicas das matrizes, dos ovos e da Campylobacter sp serem favoráveis à entrada e sobrevivência da bactéria nos ovos, nesse estudo, nenhuma positividade foi encontrada, tanto no macerado da casca quanto na gema dos ovos, indicando que a transmissão vertical é possivelmente um evento raro.