Multi-virulence of Campylobacter jejuni carried by chicken meat in Brazil.

Campylobacter jejuni is the most frequent cause of bacterial gastroenteritis; therefore, the characteristics of its epidemiology must be continuously investigated to support possible mitigating measures. This is particularly important when evaluating representative strains from the world's leading chicken meat exporter, Brazil. We evaluated a panel of 14 virulence genes in 359 strains of C. jejuni isolated from chilled broiler carcasses in Brazil. The genes were classified into five virulence categories (B: biofilm/motility; SS: secretion/cytotoxicity system; CI: invasion/colonization; GB: Guillain-Barré; and AE: adaptation to stress). The percentage of strains with stress adaptation genes (86.07%) indicates the ability to survive in unfavorable environments; in addition, the strains showed a risk of causing infections in humans due to the frequency of the hcp gene (97.77%). Genes related to Guillain-Barre syndrome (GBS) in 77.44% of strains are an additional concern, which must be monitored. The gene panel showed the presence of 124 virulence profiles. Individual analyses by carcass, slaughter establishment, and municipalities in which they were located showed high index variabilities (I.Var.) of 0.82, 0.87, and 0.78, respectively. Georeferencing indicated the state of Paraná as a hotspot for virulent strains. Higher levels of isolation and multi-virulence were identified in the summer, which is hot and humid in Brazil. Together, our results showed that the studied strains are a potential danger to public health and that there is an urgent need for their surveillance and the adoption of control measures, especially in the state of Paraná.

Alternative use of phage display: phage M13 can remain viable in the intestines of poultry without causing damage.

Phage display (PD) is a tool for developing new molecules to control pathogens. Peptides selected by PD are commonly synthesised and tested, but the use of phage M13 displaying the selected peptides as a direct biding in the intestinal tract has not yet been tested. This study evaluated whether phage M13 can remain viable in the chicken gastrointestinal tract and whether it causes injury or humoral immune response. We inoculated phage M13 or E. coli ER2738 (ECR) infected with M13 into birds at different ages. We found the virus in faeces at 5 or 13 days after inoculation, just when it infected the ECR. The presence of phage M13 or ECR did not result in gut injuries and had no impacts on weight gain and bird health. Furthermore, the levels of IgY were similar in all treatments, which indicates that the virus can be used in chicken until 42 days without being recognised by the immune system. This work provides a scientific basis for the use of PD as a tool in numerous applications to control different pathogens.

First Report of Genetic Variability of Erysipelothrix sp. Strain 2 in Turkeys Associated to Vero Cells Morphometric Alteration.

Erysipelas is a disease caused by the Erysipelothrix genus, whose main species is the E. rhusiopathiae, the causative agent of animal erysipelas and human erysipeloid. We isolated Erysipelothrix sp. strain 2 (ES2) from turkey's organs during an outbreak in Brazilian commercial and breeder flocks with sepsis and high mortality levels. We studied 18 flocks, accounting for 182 samples, being eight flocks (84 samples) as ES2 positive with individuals demonstrating clinical symptoms and high mortality. We obtained the genetic variability of 19 samples with PFGE and found two clones, both from the same flock but different samples, and two clusters. Interestingly, we found 15 strains with high genetic variability among and within flocks. We have found a positive association between the proximity of ES2 positive turkey flocks and commercial swine sites through epidemiological analysis. We infected Vero cells with two different isolates and three distinct concentrations of ES2. After performing the morphometry, we recorded enlargement of the nucleus and nucleolus. Moreover, we performed fluorescence assays that resulted in apoptotic and necrotic cells. We demonstrated that ES2 could multiply in the extracellular medium and invade and survive inside Vero cells. For the first time, our finds show that ES2 may have similar behavior as E. rhusiopathiae as a facultative intracellular microorganism, which may represent a hazard for humans.

Consumption of minas frescal cheese may be a source of human infection by Campylobacter jejuni / Consumo de queijo minas frescal pode ser fonte de infecção humana por Campylobacter jejuni

Campylobacter spp. is an emerging pathogen that causes gastroenteritis in humans and the consumption of dairy food can characterize sources of infection. We aimed to verify the viability and a presence of transcripts associated with characteristics of virulence and adaptation of C. jejuni isolated from Minas Frescal cheeses, produced with contaminated milk and stored under refrigeration for up to ten days. The samples were analyzed for bioindicators, Campylobacter spp., pH, acidity, moisture and sodium chloride. Campylobacter spp. recovered were evaluated for the production of transcripts of: ciaB, dnaJ, p19 and sodB. The results were correlated with the viability of C. jejuni and changes in their transcriptome. Storage at lowtemperatures reduced C. jejuni from the first to the fourth day. The variations in humidity, pH and acidity influenced the decreasing of C. jejuni. There was a reduction in transcripts' production of the four genes, more pronounced on the fourth day, indicating the inability of the microorganism to perform its metabolic activities, due to the conditions of injury. Despite the presence of mechanisms of virulence and adaptation, C. jejuni could not remain viable four days after production. However, consumption of fresh cheese contaminated with Campylobacter jejuni can be a source of infection when consumed up to four days after production.

Campylobacter spp. é um patógeno emergente que causa gastroenterite em seres humanos e o consumo de produtos lácteos pode caracterizar fontes de infecção. O objetivo deste estudo foi verificar a viabilidade e a presença de transcritos associadas a características de virulência e adaptação de C. jejuniisoladas de queijos frescos, produzidos com leite contaminado e mantidos refrigeradas por dez dias. Foram analisados bioindicadores, Campylobacter spp., pH, acidez, umidade e cloreto de sódio. Campylobacter spp. recuperados foram avaliados quanto à produção dos transcritos: ciaB, dnaJ, p19 e sodB. Os resultados foram correlacionados com a viabilidade de C. jejuni e alterações no transcriptoma. O armazenamento em baixas temperaturas reduziu C. jejuni do primeiro ao quarto dia. As variações na umidade, pH e acidez influenciaram a queda de C. jejuni. Houve uma redução na produção de transcritos dos quatro genes, mais pronunciada no quarto dia, indicando a incapacidade do micro-organismo em realizar suas atividades metabólicas, devido às condições de injúria. Apesar da presença de mecanismos de virulência e adaptação, C. jejuni não permaneceu viável quatro dias após a produção. Porém, o consumo de queijo fresco contaminado com Campylobacter jejunipode ser uma fonte de infecção quando consumido até quatro dias após a produção.

Viability and gene transcription of Campylobacter jejuni in infant milk formulas / Viabilidade e transcrição genética de Campylobacter jejuni em fórmulas lácteas infantis

This study simulated the contamination of two varieties of infant milk formulas (homemade and commercial) with 103 and 104 CFU/mLof Campylobacter jejuni, that were kept under refrigeration (4-7ºC) for up to 48 hours. The aim of this study was to verify the maintenance of the viability and ability of Campylobacter jejuni to produce transcripts of virulence and resistance to stress conditions during periods of 0 (after preparation), 24 and 48 hours. C. jejuni remained viable during all analyzed stages and thepresence of coliforms was not detected. In general, the counts reduced 1 log cycle after 48 hours for all samples, except the 104 CFU/mL inoculum of commercial formula, which reduced 2 log cycles, indicating greater injury of C. jejuni in this food matrix. C. jejuni showed to be more adapted to homemade matrix, due to high transcription of the gene related to cell invasion, ciaB, and more susceptible in the commercial matrix, due to the high transcription of genes related to conditions of stress tolerance (dnaJ, p19, sodB). The low infective dose of C. jejuni coupled with greater vulnerability of children less than five years indicate the need for care in the preparation and maintenance of infant formulas, to prevent the use of contaminated raw material and cross-contamination, especially in homemade formulations

Este estudo simulou a contaminação de duas variedades de fórmulas de leite infantil com Campylobacter jejuni (caseiras e comerciais), que foram mantidas sob refrigeração (4-7ºC) por até 48 horas. O objetivo do estudo foi verificar a manutenção da viabilidade e capacidade de produzir transcritos de virulência e resistência a condições de estresse durante os períodos de 0 (após preparação), 24 e 48 horas. C. jejunipermaneceu viável durante todas as etapas analisadas e a presença de coliformes não foi detectada. Em geral, as contagens reduziram 1 ciclo log após 48 horas para todas as amostras, exceto o inóculo de 104 CFU/mL na fórmula comercial, que reduziu 2 ciclos logarítmicos, indicando maior lesão de C. jejuni nesta matriz alimentar. C. jejuni mostrou-se mais adaptado à matriz popular, devido à alta transcrição do gene relacionado à invasão celular, ciaB, e mais suscetível na matriz comercial, devido à alta transcrição de genes relacionados a tolerância a condições de estresse (dnaJ, p19, sodB). A baixa dose infectante de C. jejuni, juntamente com maior vulnerabilidade de crianças menores de cinco anos, indicam a necessidade de cuidados na preparação e manutenção de fórmulas infantis, para prevenir o uso de matéria-prima contaminada e contaminação cruzada, especialmente em formulações caseiras.

Ação de diferentes sanitizantes em biofilmes de Salmonella Minnesota / Action of different sanitizers in biofilmes of Salmonella Minnesota

Objetivou-se avaliar a eficiência do contato por 15 minutos do hipoclorito de sódio 1%, e ácido peracético 0,8% na inibição de biofilmes formad os por três cepas distintas de Salmonella Minnesota em aço, poliuretano e polipropileno e determinar a recuperação das células remanescentes. Qualitativamente houve influência na classificação dos biofilmes de acordo com a superfície, que foram diferentes dependendo do tipo de cepa e de material, porém a quantidade de células sésseis permanece constante, pois independente da cepa e da superfície, os biofilmes mantiveram uma contagem de 5,18 Log UFC, indicando que as diferenças referem-se à matriz polimérica. O uso do hipoclorito foi capaz de destruir as células, que não foram recuperadas após reincubação. Já o ácido peracético não promoveu redução significativa. O uso do hipoclorito de sódio 1% é eficiente na sua remoção.

Características de virulência de Salmonella Heidelberg isoladas na cadeia avícola brasileira / Virulence characteristics of Salmonella Heidelberg isolated in the Brazilian poultry chain

S. Heidelberg é frequentemente isolada em produtos avícolas e associada a infecções alimentares humanas. Objetivou-se determinar a presença de genes de virulência em 62 cepas de S. Heidelberg isoladas na cadeia de produção avícola brasileira e inferir o seu potencial em causar infecções em humanos. Foi avaliado, por PCR, a presença dos genes avrA, invA, lpfA, agfA, sefA, luxS e sodC. Houve 100% de positividade para os genes avrA e invA, 98,4% para agfA, 96,8% para sodC, 87,1% para lpfA, 77,4% para luxS e ausência d o gene sefA. Os genes de virulência encontrados nas amostras alertam para a possível gravidade das infecções causadas por este sorovar. Estes resultados são importantes para maior compreensão dos perfis de patogenicidade de cepas circulantes de S. Heidelberg nos sistemas de produção avícola brasileiros.

Prevalência e virulência de Campylobacter spp. em diferentes matrizes cárneas / Prevalence and virulence of Campylobacter spp. in different types of meats

Objetivou-se determinar a prevalência de Campylobacter em fígado de frango e bovino, pernil suíno e carne moída e suas características de virulência. Nos isolados, determinou-se a espécie e a presença de genes de virulência por PCR. Foram analisadas 660 amostras representativas das comercializadas no Brasil, compostas por: pernil suíno resfriado (138), fígado de frango (138), patinho bovino moído (138) e fígado bovino (246), provenientes de 53 marcas e sob diferentes serviços de inspeção durante o período de março de 2014 a maio de 2016. A identificação da espécie e a presença dos genes flaA, pldA, ciaB, cadF e cdtABC, luxS, danJ e sodB foram realizadas por PCR. Das amostras, 35/660 (5,3%) foram positivas para o gênero Campylobacter, sendo 9/35 (25,71%) identificadas como C. jejuni e 6/35 (17,14%) como C. coli. Os resultados indicam a necessidade de monitoramento das diferentes matrizes cárneas quanto a presença de Campylobacter.

Resistência aos antimicrobianos de Salmonella heidelberg isoladas na cadeia de produção avícola / Antimicrobial resistance of Salmonella heidelberg isolated in the poultry chain production

S. Heidelberg é responsável por causar doença grave em humanos, e por apresentar alta resistência aos antimicrobianos, sendo considerada um problema emergente na saúde pública mundial. Objetivou-se determinar a resistência aos antimicrobianos em 67 cepas de S. Heidelberg isoladas na cadeia de produção avícola no ano de 2016. Utilizou-se a técnica de disco difusão frente ao ácido nalidíxico, amoxacilina/ácido clavulânico, ceftazidima, ceftiofur, ciprofloxacina, enrofloxacina, gentamicina, imipenem, sulfametoxazol/trimetoprim, sulfato de colistina, tetraciclina. Os maiores índices de resistência foram para o ácido nalidíxico e tetraciclina (100%), seguido do sulfato de colistina (98,5%).Todas as cepas foram classificadas como multirresistentes, alarmando para o perigo que representam para a saúde pública.

Spread of the serotypes and antimicrobial resistance in strains of Salmonella spp. isolated from broiler.

Serotypes and antimicrobial susceptibility of Salmonella's isolates from broiler production chain were determined. A total of 239 isolated strains from chicken, carcasses, breeding environments, and slaughter was analyzed by disk diffusion test, in the period of 2009 to 2010. For antibiotics with a high number of resistant strains, the minimal inhibitory concentration (MIC) was performed. We identified 24 serotypes, being the most frequent, Minnesota (31.4%) and Infantis (22.6%). The highest percentages of resistance were obtained for sulfonamide (42.7%), followed by tetracycline with 37.6% and amoxicillin with 27.6%. From the total, 32 resistance profiles were identified, being 60.7% of the strains were resistant to at least one antibiotic. Of these, 31.7% of the isolates showed multidrug resistance profiles belonging to serovar Minnesota, Saintpaul, and S. enterica. The highest resistance was found in isolates from slaughterhouse (66.9%) and aviary (58.7%). A large number of strains showed MIC above the maximum tested concentration for the antibiotics amoxicillin and sulfamethoxazole. The high number of Salmonella's resistant strains indicates the need for prudent use of these drugs in poultry production in order to reduce the occurrence and spread of antibiotic resistance profiles, and the risk that multiresistant strains isolated from broilers may pose a risk to human health.

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.

Campylobacteriosis: an emerging zoonosis, underdiagnosed and underreported by public health agencies in Brazil / Campilobacteriose: uma zoonose emergente, subdiagnosticada e subnotificada pelos órgãos de saúde pública no Brasil

The infection by Campylobacter spp. constitutes a zoonosis of worldwide distribution, with serious repercussions for public health and a significant socio-economic impact. The Campylobacter species that most often are implicated in outbreaks of gastrointestinal disease are C. jejuni and C. coli. Both are recognized as the most important and with greater pathogenic potential from the viewpoint of food safety. This review aims to clarify and discuss, based on literature, the epidemiology of Campylobacter spp. and the main challenges in their control, to assist in better understanding of the risk posed to human health. Due to the difficulty of isolating and cultivating of Campylobacter spp., the high cost and not mandatory, laboratory diagnosis by traditional cultivation techniques is not widely used in Brazil, resulting in lack of accurate data on the occurrence of the disease in the country. This factor has been identified as responsible for underdiagnosing and underreporting of the disease, which means that data on contamination of animal products or outbreaks are not statements of actual national situation. The epidemiology of Campylobacter spp. is considered complex presenting multiple determinants in the spread of this microorganism. Chickens are important reservoirs of Campylobacter sp. and the main target of control measures, however, other vehicles of transmission, such as raw cow milk, drinking water and other foods of animal origin must also be considered in the set of measures to be taken to control disease. The infection by Campylobacter spp. cause gastroenteritis in humans and occurred by oral contact with infected animals. In fact, are pets or production and food consumption, the most important pathways. The clinical manifestations of the disease can be from mild to severe symptoms, including watery diarrhea, sometimes with blood, abdominal pain, fever, headache and nausea. Sometimes the infection can get complications after the diarrhea stops and starts a progressive paralysis that can result in respiratory failure and severe neurological dysfunction and may even lead to death. The occurrence of campylobacteriosi's outbreaks should be mandatory to the authorities of surveillance, to provide an investigation of common sources and transmission control through preventive measures , thereby minimizing the potential risks to human health.

A infecção por Campylobacter spp. constitui em uma zoonose de distribuição mundial, com graves repercussões em saúde pública e com um impacto sócio-econômico significativo. Das espécies de Campylobacter sp., as que estão mais frequentemente implicadas nos surtos de doença gastrointestinal são C. jejuni e C. coli, sendo reconhecidas como as mais importantes e com maior potencial patogênico do ponto de vista da segurança alimentar. Esta revisão visa esclarecer e discutir, com base nos dados da literatura, a epidemiologia de Campylobacter sp. e os principais desafios no seu controle, para auxiliar no melhor entendimento do risco que representa para a saúde humana. Devido à dificuldade de isolamento e cultivo de Campylobacter sp., ao alto custo da análise e da mesma não ser obrigatória, o diagnóstico laboratorial por meio de técnicas tradicionais de cultivo não é muito utilizado no Brasil, acarretando falta de dados precisos sobre a ocorrência da doença no país. Este fator tem sido identificado como responsável por subdiagnóstico e subnotificação da enfermidade, o que significa que dados sobre contaminação de produtos de origem animal ou surtos não são demonstrativos da real situação nacional. A epidemiologia das infecções por Campylobacter spp. é considerada complexa, apresentando múltiplos fatores determinantes na disseminação desse microrganismo. As aves são importantes reservatórios de Campylobacter sp. e o principal alvo de medidas de controle, porém, outros veículos de transmissão, como o leite cru, a água potável e outros alimentos de origem animal também precisam ser considerados no conjunto de medidas a serem adotadas para o controle da doença. A infecção por Campylobacter sp. causadoras de gastroenterite no homem acontece principalmente por via oral, mas pode ocorrer também pelo contato com animais de estimação ou de produção infectados. O quadro clínico da doença varia desde sintomas leves a graves, incluindo diarreia aquosa, algumas vezes com sangue, dor abdominal, febre, dor de cabeça e náuseas. Porém, pode haver complicações posteriores devidoresposta auto-imune levando a paralisia progressiva que pode resultar em insuficiência respiratória e disfunção neurológica grave, podendo até mesmo levar a morte. A ocorrência de casos ou surtos de campilobacteriose devem ser notificados às autoridades de vigilância epidemiológica, para que se desencadeie a investigação das fontes comuns e o controle da transmissão por meio de medidas preventivas, minimizando assim os potenciais riscos para a saúde humana.

Dinâmica química, microbiológica e física da silagem de farelo úmido de glúten de milho / Physical, chemical and microbiological dynamics of wet corn gluten feed silage

Objetivou-se avaliar aspectos físico-químicos e microbiológicos do farelo úmido de glúten de milho (FUGM) em função do tempo de ensilagem. O FUGM foi avaliado em silos experimentais e, nos dias um, três, sete, 14, 21, 28 e 42, foram realizadas as avaliações físicas, químicas e microbiológicas da silagem. O delineamento experimental foi inteiramente casualizado com quatro repetições e obtidas equações de regressão em função do tempo. O tempo de ensilagem não afetou (P>0,05) o percentual de fibra em detergente neutro, fibra em detergente ácido, proteína bruta, matéria mineral, extrativos não-nitrogenados, nutrientes digestíveis totais e contagem de clostrídios. Os teores de matéria seca, extrato etéreo, perdas por efluentes, perdas por gases e população de bactérias láticas variaram entre 46,54 e 49,05%; 0,96 e 1,71%; 17,97 e 35,14g kg-1; 1,90 e 5,68% e 5,0 e 6,9log UFCg-1 de silagem, respectivamente, todos com equações com r2≤0,6.O pH (Y=4,12926-0,00894dias+0,00024126dias2,r2=0,75) e a temperatura (Y=19,82657+0,11734dias-0,00502dias2,r2=0,73) tiveram seus menores e maiores valores, respectivamente, em 18,5 e 11,7 dias. A contagem de bolores e leveduras sofreu redução em função do tempo de ensilagem (Y=6,8983-0,0363dias,r2=0,84). A ensilagem permite eficiente conservação dos aspectos químicos, microbiológicos e físicos do FUGM até 42 dias de ensilagem.

This study aimed to evaluate physicochemical and microbiological aspects of wet corn gluten feed (WCGF) according to time of ensilage. The WCGF was evaluated in experimental silos and after one, three, seven, 14, 21, 28 and 42 days, physical, chemical and microbiological evaluation of the silage was carried out. The results were analyzed as a completely randomized design with four replications and regression equations as a function of time. The number of days of ensiling did not affect neutral detergent fiber, acid detergent fiber, crude protein, mineral matter, non-nitrogenous extract and total digestive nutrients percentages, as well as clostridia count. Dry matter, ether extract, effluents losses and percentages of gas losses and lactic acid bacteria count with variations of 46.54 and 49.05%; 0.96 and 1.71%; 17.97 and 35.14g kg-1; 1.90 and 5.68% and 5.0 and 6.9log CFUg-1 of silage, respectively, all had low coefficient of determination (r2≤0.6).The pH (Y=4.12926-0.00894days+0.00024126days2,r2=0.75) and temperature (19.82657+0.11734days-0.00502days2,r2=0.73) had their minor and major values, respectively, in 18.5 and 11.7 days. Moulds and yeasts count decrease with time of ensilage (Y=6.8983-0.0363days,r2=0.84). The ensiling allows efficient conservation of the chemical, microbiological and physical aspects of WCGF until 42 days of ensilage.

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.

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.

Virulência de Salmonella spp. de origem avícola e resistência a antimicrobianos / Virulence of Salmonella spp. of poultry products origin and antimicrobial resistance

A habilidade do gênero Salmonella em causar doenças depende de vários fatores genéticos determinantes, dentre eles destacam-se os genes que codificam alguma característica de virulência nessas bactérias. Objetivou-se com esse trabalho pesquisar genes associados à virulência em isolados de Salmonella spp. de origem avícola, em sua fase de aderência e invasão celular, e avaliar a resistência desses isolados a 11 antimicrobianos. Foram analisadas 18 amostras de Salmonella spp. isoladas durante o ano de 2009 a partir de suabes de arrasto provenientes de aviários de frango de corte do estado de São Paulo. Todas as amostras foram testadas através da técnica de PCR para os genes associados à virulência: invA, lpfA e agfA. A maioria das amostras apresentou alta positividade para os genes de virulência, e 88,9% das amostras apresentaram os três genes estudados. Verificou-se que 17 amostras (94,4%) apresentaram fragmentos específicos para os genes invA e lpfA e o gene agfA foi positivo em todas as amostras avaliadas (100%). Todas as cepas apresentaram resistência pelo menos a um antimicrobiano testado. Os dados indicaram que das amostras analisadas a maior resistência foi à amoxacilina com 27,7%. Os resultados demonstram a patogenicidade dos sorovares de Salmonella, que representam potenciais desafios sanitários na avicultura, pelo risco da disseminação de espécimes virulentos e multirresistentes entre os animais e que, consequentemente, podem acometer o homem.

The ability of Salmonella to cause disease depends on several genetic determinant factors, including genes that encodes some virulence trait of these bacteria. The aim of this work was to research genes associated with virulence in Salmonella isolates of poultry products in its phase of adhesion and cell invasion, and to evaluate the resistance of these isolates to 11 antimicrobials. Were analyzed 18 samples of Salmonella spp. isolated during the 2009 year using drag swabs from poultry broiler in the state of São Paulo. All samples were tested using PCR technique for genes that were associated to virulence: invA, lpfA and agfA. Most samples had high positivity to virulence genes, and 88.9% of the samples had the three genes. It was verified that 17 samples (94.4%) had specific fragments to the invA and lpfA genes and the gene agfA was positive in all samples (100%). All samples were resistant to at least one antibiotic tested, the data indicated that the samples analyzed had greatest resistance to amoxacilin with 27.7%. The results demonstrate the pathogenicity of the Salmonella serovars which represent potential health challenges in poultry, the risk of the spread of multidrug resistant and virulent specimens among the animals and therefore can affect humans.

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.

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.