Monitoring of Pig Slaughter Stages and Correlation in the Prevalence of Pathogens and Levels of Microorganisms That Indicate Microbiological Quality and Hygiene Using a Predictive Model.

Pig production is relevant to the Brazilian economy. Different stages of the raising and slaughtering process influence the microbiological quality of pig products and by-products. Microbiological analysis and hazard analysis and critical control points (HACCPs) are tools for monitoring microbiological quality indicator microorganisms. The construction of predictive models can assist the process of monitoring the microbiological quality of pig products. This study aimed to map the slaughter stages and develop a model to predict the absence or presence of Salmonella based on the process variables (distance from the farm to the slaughterhouse and aerobic mesophilic) and analyze their influence on contamination indicator microorganisms. A total of 810 samples were collected at nine stages of the slaughter process (bleeding, scalding, dehairing, singeing, washing, evisceration, inspection, final washing, and chilling). The binary class predictive model was used as a microbiological quality predictor at the slaughter stages. Salmonella was identified at all process stages, with lower contamination levels at the scalding and chilling stages, whereas the highest levels were found at the dehairing and bleeding stages. The predictive model revealed an accuracy of about 85% for Salmonella being a tool to monitor the microbiological quality of pig slaughter.

Characteristics of virulence, resistance and genetic diversity of strains of Salmonella Infantis isolated from broiler chicken in Brazil / Características de virulência, resistência e diversidade genética de estirpes de Salmonella Infantis isoladas de frangos de corte no Brasil

Salmonella Infantis is frequently associated with human infections worldwide and is transmitted by consumption of contaminated foods, particularly those of animal origin, especially the chicken meat. We aimed to evaluate virulence characteristics, antimicrobial resistance and the genetic similarity of 51 strains of S. Infantis isolated from samples of poultry origin. The strains were isolated from 2009 to 2010 in a company with full cycle of broiler's production in the state of São Paulo, Brazil. The antimicrobial susceptibility test was performed and, by PCR, we evaluated the presence of the genes lpfA (hem-adhesion), agfA (hem-biofilm) and sefA (hem-adhesion) and resistance genes to beta-lactams (blaTEM, blaSHV, bla CTX-M and blaAmpC ). The phylogenetic relationship was determined by RAPD-PCR method. Among the drugs tested, the highest percentages of resistance were to amoxicillin (35.3%) and to sulfonamide (15.7%). Eleven antimicrobial resistance patterns were identified (A1 to A11), none of them presented a multiresistance profile (> 3 antimicrobials classes). There was 100% of positivity for the agfA gene, 92.2% for the lpfA gene, and no strain presented the sefA gene. Most of the isolates showed similarities in virulence potential, since they were simultaneously positive for two studied genes, agfA and lpfA (92.2%, 47/51). Of the 18 (35.3%) strains resistant to antimicrobials of the β-lactam class, 10 (55.5%) were positive to blaAmpC gene, five (27.8%) for blaCTX-M , two (11.1%) to blaSHV and no strain presented the blaTEM gene. The phylogenetic evaluation has shown the presence of five clusters (A, B, C, D and E) with similarity greatSalmonella Infantis is frequently associated with human infections worldwide and is transmitted by consumption of contaminated foods, particularly those of animal origin, especially the chicken meat. We aimed to evaluate virulence characteristics, antimicrobial resistance and the genetic similarity of 51 strains of S. Infantis isolated from samples of poultry origin. The strains were isolated from 2009 to 2010 in a company with full cycle of broiler's production in the state of São Paulo, Brazil. The antimicrobial susceptibility test was performed and, by PCR, we evaluated the presence of the genes lpfA (hem-adhesion), agfA (hem-biofilm) and sefA (hem-adhesion) and resistance genes to beta-lactams (blaTEM, blaSHV, bla CTX-M and blaAmpC ). The phylogenetic relationship was determined by RAPD-PCR method. Among the drugs tested, the highest percentages of resistance were to amoxicillin (35.3%) and to sulfonamide (15.7%). Eleven antimicrobial resistance patterns were identified (A1 to A11), none of them presented a multiresistance profile (> 3 antimicrobials classes). There was 100% of positivity for the agfA gene, 92.2% for the lpfA gene, and no strain presented the sefA gene. Most of the isolates showed similarities in virulence potential, since they were simultaneously positive for two studied genes, agfA and lpfA (92.2%, 47/51). Of the 18 (35.3%) strains resistant to antimicrobials of the ß-lactam class, 10 (55.5%) were positive to blaAmpC gene, five (27.8%) for blaCTX-M , two (11.1%) to blaSHV and no strain presented the blaTEM gene. The phylogenetic evaluation has shown the presence of five clusters (A, B, C, D and E) with similarity greater than 80%, and three distinct strains which were not grouped in any cluster. Cluster B grouped 33 strains, all positive for lpfA and agfA genes, from both, the broiler farming facility and the slaughterhouse, persistent throughout all the study period. This cluster also grouped 18 strains clones with genetic similarity greater than 99%, all isolated in the slaughterhouse. The presence of virulence genes associated with persistent strains clones for a long period, warns to the possibility of S. Infantis to form biofilm, and should be constantly monitored in broilers' production chain, in order to know the profile of the strains that may contaminate the final product and evaluate the hazards that represents to public health.er than 80%, and three distinct strains which were not grouped in any cluster. Cluster B grouped 33 strains, all positive for lpfA and agfA genes, from both, the broiler farming facility and the slaughterhouse, persistent throughout all the study period. This cluster also grouped 18 strains clones with genetic similarity greater than 99%, all isolated in the slaughterhouse. The presence of virulence genes associated with persistent strains clones for a long period, warns to the possibility of S. Infantis to form biofilm, and should be constantly monitored in broilers' production chain, in order to know the profile of the strains that may contaminate the final product and evaluate the hazards that represents to public health.(AU)

Salmonella Infantis é frequentemente associada a infecções humanas no mundo todo sendo transmitida pelo consumo de alimentos contaminados, principalmente aqueles de origem animal, com destaque para a carne de frango. Objetivou-se avaliar características de virulência, resistência antimicrobiana e a similaridade genética de 51 estirpes de S. Infantis isoladas em amostras de origem avícola. As estirpes foram isoladas no período de 2009 a 2010 em uma empresa com ciclo completo de produção de frango de corte, localizada no estado de São Paulo, Brasil. Foi realizado o teste de susceptibilidade antimicrobiana e pela técnica de PCR, foi avaliada a presença dos genes lpfA (fímbria-adesão), agfA (fímbria-biofilme) e sefA (fímbria-adesão) e os genes de resistência aos beta-lactâmicos (bla TEM, blaSHV, blaCTX-M e blaAmpC ). A relação filogenética foi determinada pelo método de RAPD-PCR. Dentre as drogas testadas, os maiores percentuais de resistência foram para amoxacilina com 35,3% e sulfonamida com 15,7%. Onze perfis de resistência aos antimicrobianos foram identificados (A1 a A11), sendo que nenhum deles apresentou perfil de multirresistência (>3 classes de antimicrobianos). Houve 100% de positividade para o gene agfA, 92,2% para o gene lpfA e nenhuma estirpe apresentou o gene sefA. A maioria dos isolados apresentaram semelhanças no potencial de virulência, pois foram positivos simultaneamente para dois genes estudados, agfA e lpfA (92,2% - 47/51). Das 18 (35,3%) estirpes resistentes aos antimicrobianos da classe dos ß-lactâmicos, 10 (55,5%) foram positivas para o gene blaAmpC , cinco (27,8%) para blaCTX-M , duas (11,1%) para blaSHV e nenhuma estirpe apresentou o gene bla TEM . A avaliação filogenética demonstrou a presença de cinco clusters (A, B, C, D e E) com similaridade superior a 80%, e três estirpes distintas que não foram agrupadas em nenhum dos clusters. O cluster B agrupou 33 estirpes, todas positivas para os genes lpfA e agfA, provenientes tanto do aviário quanto do matadouro frigorífico, persistentes durante todo o período do estudo. Este cluster ainda agrupou 18 estirpes clones com similaridade genética superior a 99%, todas isoladas no matadouro frigorífico. A presença dos genes de virulência, associada à persistência das estirpes clones durante um longo período do estudo, alertam para a possibilidade de S. Infantis em formar biofilme, devendo ser constantemente monitorada na cadeia de produção avícola, especialmente no ambiente de abate, de forma a conhecer o perfil das estirpes que podem contaminar o produto final e assim avaliar os perigos que representam para a saúde pública.(AU)

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.

High pressure spray with water shows similar efficiency to trimming in controlling microorganisms on poultry carcasses.

A study was conducted to evaluate a high pressure spray (HPS) with water as an alternative to trimming to remove gastrointestinal contamination on poultry carcasses and improve microbiological quality. The study was conducted under commercial conditions in 5 slaughter plants with one plant presenting approximately 5% of carcasses with visible gastrointestinal contamination (VGC), and the others showing approximately 12% of VGC. In all 5 plants, carcasses were sampled from the slaughter line and separated into 6 groups corresponding to 3 different treatments: A) carcasses with VGC before and after trimming; B) carcasses with VGC before and after HPS; and C) carcasses with no VGC before and after HPS. At the end of Trial A and prior to Trials B and C, an HPS equipment was installed before the end of the slaughter line. The HPS equipment was operated with 10 kgf/cm² of pressure and 1.5 L of potable water per carcass. Carcasses were analyzed using a rinsing procedure, and the following microbiological parameters were evaluated: the prevalence of Salmonella and Campylobacter, the abundance of Escherichia coli (EC), Enterobacteriaceae (EB), and the Total Viable Count (TVC). Salmonella was found in all plants at a prevalence ranging from 0.8% (plant 1) to 17.3% (plant 5), and the difference between plants was significant (P < 0.05%). The prevalence of Campylobacter ranged from 2.1 (plant 1) to 18.6% (plant 4) (P < 0.05%). The prevalence of Campylobacter was similar in plants 2, 3, and 5, and a significant difference (P < 0.05%) was observed compared to plants 1 and 4. In all plants, the EC, EB, and TVC counts did not show a significant difference (P > 0.05%) in any treatments. These results demonstrate that HPS with water is an alternative method for removing VGC and improving or maintaining the microbiological quality and safety of broiler carcasses.

Prevalence of Salmonella and Campylobacter on broiler chickens from farm to slaughter and efficiency of methods to remove visible fecal contamination.

A study was conducted to investigate the prevalence of Salmonella and Campylobacter from farm to slaughter. The efficiency of trimming and water spray (490 to 588 kPa pressure) on the removal of visible fecal contamination from broiler carcasses before chilling was also investigated. Drag swabs were used to sample litter from the farm houses. Samples of ceca and carcasses without and with visible fecal contamination before and after trimming or spray washing of fecal contamination were taken during slaughter of the flocks previously visited at the farms. There was a low prevalence of Salmonella on the litter from the farms (5%) and cecum and carcasses (0%). However, Campylobacter jejuni and Campylobacter coli were present in farms' litter (100 and 58.8%, respectively), cecum samples (100 and 70.6%, respectively), and carcasses with (58.8 and 11.6%, respectively) and without (17.6 and 9.8%, respectively) visible fecal contamination. There was high prevalence of C. jejuni but at low counts and low prevalence and high counts of C. coli. Campylobacter lari was not detected in any sample. Trimming the visible fecal contamination decreased the prevalence of C. jejuni but increased occurrence of C. coli. Trimming did not reduce the counts of Campylobacter and of hygiene indicator microorganisms on the carcasses. Water shower reduced the counts of hygiene indicator microorganisms by 20%. Therefore, control measures for preventing introduction of Campylobacter and the use of good hygienic conditions are needed to warrant the microbiological quality and safety of broiler carcasses.

Evaluation of three sampling methods for the microbiological analysis of broiler carcasses after immersion chilling.

Countries have different official programs and implement different sampling methods for the detection of Salmonella on poultry carcasses. In Brazil, a 25-g sample of skin and muscle excision (SME) from the wings, neck, and pericloacal parts is used; in the European Union (EU), a 25-g sample of neck skin (NSE) is used; and, in the United States, the whole carcass is rinsed with 400 ml of diluent (WCR). In the present study, these methods were evaluated to compare Salmonella occurrence and counts of hygiene indicator microorganisms (Escherichia coli, Enterobacteriaceae, and total viable count of aerobic mesophilic bacteria) using different carcasses from the same flock and also using different analytical units taken from the same carcass. Eighty flocks, with four broiler carcasses from each, were included in this study; three broilers were sampled according to protocols from Brazil, the EU, and the United States, and the last one by all three methods. SME, NSE, and WCR provided equivalent results (P > 0.05) for Salmonella detection on broiler carcasses when using different carcasses from the same flock and when using the same carcass. The predominant serovar was Salmonella Enteritidis. For the enumeration of hygiene indicator microorganisms, WRC provided higher counts than SME or NSE (P < 0.05), when using both the same or different carcasses. Therefore, it is possible to directly compare Salmonella results in poultry carcasses when using the methods recommended by the legislative bodies of Brazil, the United States, and the EU. However, WCR provides the best results for hygiene indicator microorganisms.

Ocorrência de bactérias aeróbias mesófilas e de Bacillus cereus em pimenta-do-reino / Presence of mesophilic aerobes and Bacillus cereus in black pepper

Foram analisadas 37 amostras de pimenta-do-reino moída, coletadas no comércio de Viçosa (MG) e Säo Paulo (SP) e, em 85 por cento destas, constatou-se a presença de Bacillus cereus em números de até 4,6 x 104 UFC/g. A populaçäo de bactérias aeróbias mesófilas variou de 2,8 x 105 a 1,9 x 108 UFC/g.

Avaliação do método tradicional para detecção de Salmonella spp. em carnes in natura / Evaluation of the traditional method for detection of Salmonella spp. in raw meat products

Avaliou-se o efeito da temperatura de pré-enriquecimento e o desempenho de três caldos de enriquecimento seletivo e dois ágars de isolamento, utilizados para detecção de Salmonella em carnes in natura. As amostras foram pré-enriquecidas em água tamponada e peptonada 1 por cento e incubadas a 35ºC e a 42ºC por 24 horas. Alíquotas de cada amostra pré-enriquecida foram inoculadas em caldos Tetrationato, Rappaport-Vassiliadis e Selenito Cistina. Os ágars Rambach e Verde Brilhante foram usados para o isolamento de Salmonella. Os resultados mostraram que o maior índice de resultados positivos foi obtido com pré enriquecimento a 42ºC. Entre os meios de enriquecimento seletivo, o caldo Tetrationato com pré-enriquecimento a 42ºC proporcionou o maior índice de isolamento de Salmonella spp.

Avaliaçäo do método tradicional para detecçäo de Salmonella spp. em carnes in natura / Evaluation of the traditional method for detection of Salmonella spp. in raw meat products

Avaliou o efeito da temperatura de pré-enriquecimento e o desempenho de três caldos de enriquecimento seletivo e dois agares de isolamento, utilizados para detecçäo de Salmonella em carnes in natura. Amostras de carne suíça, naturalmente contaminadas com Salmonella, foram pré-enriquecidas em água peptonada tamponada a 1 por cento e incubadas a 35§C e 42§C por 24 horas. Alíquotas de cada amostra pré-enriquecida foram inoculadas em caldos Tetrationato, Rappaport-Vassiliadis e Selenito Cistina. Os agares Rambach e Verde Brilhante foram usados para o isolamento de Salmonella. Os resultados revelaram que o caldo Tetrationato com pré-enriquecimento a 42§C proporcionou o maior índice de isolamentos de Salmonella spp.

Incidência de Salmonella e Escherichia coli em carne suína in natura e salame colonial, consumidos pela populaçäo de Chapecó - SC / Presence of Salmonella and Escherichia coli in pork in natura and 'colonial' sausage, which the Chapecó - SC population consumed

Bactérias como a Salmonella e a Escherichia coli, há muito tempo, säo reconhecidas como importantes patógenos causadores de intoxicaçöes alimentares, sendo que a contaminaçäo de alimentos com microrganismos de origem fecal deve ser sempre ser assumida como um sério problema. Investigou a presença de Salmonella e E. coli em carne suína in natura e salame colonial consumidos pela populaçäo de Chapecó-SC. Foi analisado um total de 50 amostras de carne suína e 50 amostras de salame colonial. Os resultados deste estudo revelaram a presença de Salmonella em 6,0 por cento das amostras de carne suína in natura e em 6,0 por cento das amostras de salame colonial, sendo que os sorovares isolados em ambos os produtos foram: typhimurium, agona e schazengrund. Entretanto, 82,0 por cento das amostras de carne suína e 84,0 por cento das amostras de salame colonial apresentaram contaminaçäo por E. coli sendo que 44,0 por cento e 72,0 por cento, respectivamente, estavam acima do número máximo permitido para occonsumo humano. Estes resultados indicam que tais produtos säo inadequados para o consumo humano e os riscos de intoxicaçöes alimentares por estes microrganismos säo elevados.

Método tradicional clássico para detecçäo de Salmonella em alimentos: um problema técnico bastante complexo / Classic traditonal method for detection of Salmonella in foods

A Salmonella é uma bactéria entérica responsável por graves surtos de intoxicaçöes alimentares sendo que os produtos de origem animal säo os principais veículos transmissores deste patógeno. A busca de uma metodologia adequada e eficiente para detectar Salmonella em alimentos tem sido uma busca constante de todos os profissionais que trabalham na pesquisa desta bactéria. Apesar de nos últimos anos muitos métodos com tecnologia avançadas terem surgido para diagnosticar Salmonella em alimentos, o método cultural clássico, apesar de sua morosidade e labor, continua sendo amplamente utilizado em laboratórios de monitoramento de Salmonella nas indústrias de alimentos e sua eficiência representa confiabilidade nos resultados obtidos e a segurança nos programas de controle desta bactéria. No entanto, muitos estudos têm revelado diferenças significativas entre resultados obtidos com este método, principalmente no que se refere aos meios de cultura e as temperaturas de incubaçäo aplicadas nas etapas de seu procedimento. Desta forma, verifica as principais diferenças encontradas na literatura científica referente aos meios de cultura e suas temperaturas de incubaçöes, visando buscar informaçöes que auxiliem todos os profissionais que utilizam esta metodologia.

Qualidade microbiológica da água proveniente de poços e fontes de dois municípios da regiäo do Alto Uruguai Catarinense / Microbiology quality of water destinated to human consume, from two towns of Alto Uruguai Catarinense region

Entre julho de 1994 e julho de 1996, foram realizadas colheitas de amostras de água dos poços e fontes em dois municípios da regiäo do Alto Uruguai Catarinense, Oeste de Santa Catarina. As mesmas foram colhidas assepticamente em frascos estéreis, acondicionadas em caixas isotérmicas e transportadas ao laboratório para análise imediata. Foram analisadas 183 amotras, sendo 84 do município 01 e 99 do município 02.