Genomic characterization, antimicrobial resistance profiles, enterotoxin, and biofilm production of methicillin-resistant Staphylococcus aureus isolated from clinical and animal products origins in Eastern Turkey / Caracterização genômica perfis de resistência antimicrobiana, produção de enterotoxinas e biofilme Staphylococcus aureus resistente à meticilina isolado de origem clínica e de produtos de origem animal no leste da Turquia

Staphylococcus aureus is an opportunistic and ubiquitous pathogen found in the skin, nares, and mucosal membranes of mammals. Increasing resistance to antimicrobials including methicillin has become an important public concern. One hundred and eight (108) S. aureus strains isolated from a total of 572 clinical and animal products samples, were investigated for their biofilm capability, methicillin resistance, enterotoxin genes, and genetic diversity. Although only one strain isolated from raw retail was found as a strong biofilm producer, the percentage of antimicrobial resistance pattern was relatively higher. 17.59% of S. aureus strains tested in this study were resistant to cefoxitin and identified as methicillin-resistant S. aureus (MRSA) isolates. mecA and mecC harboring S. aureus strains were detected at a rate of 2.79% and 0.93%, respectively. In addition, staphylococcal enterotoxin genes including Sea, Seb, Sec, and Sed genes were found to be 18.5%, 32.4%, 6.5% and 3.7%, respectively. The phylogenetic relationship among the isolates showed relationship between joint calf and cow milk isolates. Multi locus sequence typing (MLST) revealed three different sequence types (STs) including ST84, ST829, and ST6238. These findings highlight the development and spread of MRSA strains with zoonotic potential in animals and the food chain throughout the world.

Staphylococcus aureus é um patógeno dúctil e ubíquo encontrado na pele, narinas e membranas mucosas de mamíferos. O aumento da resistência aos antimicrobianos, incluindo a meticilina, tornou-se uma importante preocupação pública. Cento e oito (108) cepas de S. aureus isoladas de um total de 572 amostras clínicas e de produtos animais foram investigadas por sua capacidade de biofilme, resistência à meticilina, genes de enterotoxinas e diversidade genética. Embora apenas uma cepa isolada do cru tenha sido encontrada como forte produtora de biofilme, a porcentagem do padrão de resistência antimicrobiana foi relativamente maior. Parte das cepas (17,59%) de S. aureus testadas neste estudo eram resistentes à cefoxitina e identificadas como isolados de MRSA. mecA e mecC abrigando cepas de S. aureus foram detectados a uma taxa de 2,79% e 0,93%, respectivamente. Além disso, verificou-se que os genes da enterotoxina estafilocócica, incluindo os genes Sea, Seb, Sec e Sed, eram 18,5%, 32,4%, 6,5% e 3,7%, respectivamente. A relação filogenética entre os isolados mostrou relação entre os isolados de bezerro e leite de vaca. A tipagem de sequência multiloco (MLST) revelou três tipos de sequência diferentes (STs), incluindo ST84, ST829 e ST6238. Essas descobertas destacam o desenvolvimento e a disseminação de cepas de MRSA com potencial zoonótico em animais e na cadeia alimentar em todo o mundo.

Perfil del riesgo de brotes alimentarios por intoxicación estafilocócica en Cuba / Risk profile of staphylococcal food poisoning outbreaks in Cuba

Introducción: Staphylococcus ocasiona con frecuencia intoxicaciones alimentarias; en Cuba es una de las principales causas de brotes. Objetivo: analizar el perfil del riesgo de brotes alimentarios por Staphylococcus en Cuba. Métodos: Se realizó un estudio observacional analítico sobre los factores higiénicos y epidemiológicos asociados al riesgo de la intoxicación alimentaria por Staphylococcus en Cuba, en el Instituto Nacional de Higiene Epidemiología y Microbiología, durante el periodo de enero a noviembre de 2018. La información se obtuvo mediante revisión bibliográfica sistémica, análisis de datos del Laboratorio de Referencia Nacional en Microbiología Sanitaria y informes de brotes de la Dirección Nacional de Salud Ambiental. Se estimó el riesgo microbiológico mediante el programa semicuantitativo Ross-Sumner. Resultados: Se identificó como principal peligro Stahylococcus aureus, productores de enterotoxina A y la determinación de aislamientos resistentes a los antimicrobianos, como Stahylococcus aureus meticilina resistentes. Los alimentos más implicados fueron los productos de repostería y la inocuidad se afectó en mayor frecuencia por la contaminación cruzada en el 31,4 por ciento brotes y la inadecuada conservación en 46,1 por ciento. La predicción de personas que enferman por año en la población de interés fue de 6 260. Conclusiones: El riesgo de la ocurrencia de brotes por Staphylococcus se estimó como alto; la vigilancia se recomienda en alimentos que requieren manipulación directa y no se aplica tratamiento térmico antes del consumo, con gestiones de riesgo dirigidas a productores y consumidores(AU)

Introduction: Staphylococcus is a frequent cause of food poisoning. In Cuba it is one of the main causes of outbreaks. Objective: Analyze the risk profile of staphylococcal food poisoning outbreaks in Cuba. Methods: An observational analytical study was conducted about hygienic and epidemiological factors associated to the risk for staphylococcal food poisoning in Cuba. The study was carried out at the National Institute of Hygiene, Epidemiology and Microbiology from January to November 2018. The information was obtained from a systematic bibliographic review, analysis of data from the Medical Microbiology National Reference Laboratory, and outbreak reports from the National Environmental Health Division. Microbiological risk was estimated with Ross-Sumner semi-quantitative software. Results: The main hazard identified was Staphylococcus aureus, enterotoxin A producers and determination of antimicrobial resistant isolates like methicillin-resistant Staphylococcus aureus. The food items most commonly involved were sweets, and safety was most frequently affected by cross-contamination in 31.4 percent of the outbreaks and inadequate preservation in 46.1 percent. Prediction of members of the population of interest becoming ill every year was 6 260. Conclusions: Risk for the occurrence of staphylococcal food poisoning outbreaks was considered to be high; surveillance is recommended of food items requiring direct manipulation and appropriate thermal treatment is not applied before consumption, with risk management actions aimed at manufacturers and consumers(AU)

Detection of virulence factors in coagulase-negative Staphylococcus spp. strains isolated from Emmental cheese / Detecção de fatores de virulência em cepas de Staphylococcus spp. coagulase negativos isoladas de queijo Emmental

Food prepared with products derived from animals are involved in most cases of staphylococcal poisoning; therefore, the research of Staphylococcus spp. in Emmental cheese is more applicable. The objective of this study was to identify coagulase-negative Staphylococcus spp. (CNS) in cheese using biochemical and molecular techniques to detect the presence of nine genes responsible for the production of enterotoxins. From 180 samples analyzed, 204 CNS strains were obtained and identified as being 46 (22.6%) S. saprophyticus strains, 27 (13.2%) S. hominis spp. hominis strains, 22 (10.8%) S. sciuri strains, 21 (10.3%) S. xylosus strains, 19 (9.3%) S. epidermidis strains, 19 (9.3%) S. haemolyticus strains, 17 (8.3%) S. lentus strains, 17 (8.3%) S. warneri strains, 11 (5.4%) S. equorum strains and 5 (2.5%) S. cohnni . Using the PCRm protocol, 14 (6.9%) strains with the presence of the genes on the enterotoxin E (SEE)11 (78.6%), J (SEJ) 1 (7%), C (SEC) 1 (7%) and I (SEI) 1 (7%) were detected. Based on the results, the type of package is not interfered of growth and isolated that Staphylococcus spp. in cheese. It was observed that bacteria capacity to produce coagulase cannot be understood as an indicative of enterotoxigenicity; therefore, the CNS should be considered as a target of importance in the epidemiology of staphylococcal intoxications. It can be concluded that CNS need to be included in bacterial foodborne disease research, since the genes responsible for the production of toxins were detected and none of the studied samples presented Staphylococcus spp. counting above the limits allowed by legislation.(AU)

Os alimentos preparados com produtos de origem animal são os mais envolvidos em casos de intoxicação alimentar estafilocócica; portanto a pesquisa do Staphylococcus spp. em queijos tipo Emmental é relevante. O objetivo foi isolar e identificar espécies de Staphylococcus coagulase negativas (CNS)de queijo Emmental acondicionado em vários tipos de embalagem, por meio de técnicas bacteriológicas e bioquímicas e detectar, por PCR, a presença de nove genes responsáveis pela produção de enterotoxinas. Das 180 amostras, foram isoladas 204 cepas de CNS, que foram identificadas por provas bioquímicas como: 46 (22,6%) S. saprophyticus, 27 (13,2%) S. hominis spp. hominis, 22 (10,8%) S. sciuri, 21 (10,3%) S. xylosus, 19 (9,3%) S. epidermidis , 19 (9,3%) S. haemolyticus , 17 (8,3%) S. lentus , 17 (8,3%) S. warneri , 11(5,4%) S. equorum e 5 (2,5%) S. cohnii . Na PCR multiplex, em 14 (6,9%) isolados foi detectada a presença dos genes para enterotoxina E (SEE), em 11 (78,6%) J (SEJ), em 1 (7%) C (SEC) e em 1 (7%) I (SEI). Com base nos resultados, o tipo de embalagem não interferiu na multiplicação dos Staphylococcus spp. isolados dos queijos. Neste estudo, verificou-se que a capacidade para a produção de coagulase pela bactéria não pode ser concebida como indicativa de enterotoxigenicidade, portanto devem-se considerar os CNS como objeto de importância na epidemiologia das intoxicações estafilocócicas, fazendo-se necessária a atenção com relação à pesquisa dos CNS nos alimentos, uma vez que foram detectados genes responsáveis pela produção de toxinas, e nenhuma das amostras apresentou contagem para Staphylococcus spp. acima do limite permitido pela legislação.(AU)

La implementación de procedimientos estandarizados en la prevención de enfermedades transmitidas por los alimentos. Conteo microbiológico del Staphylococcus aureus en quesos frescos / Implementation of standardized procedures to prevent diseases transmitted by foodstuffs. Microbiological count of Staphylococcus aureus in freshcheese

Introducción: las enfermedades transmitidas por alimentos son una problemática mundial a la que la Organización Mundial de la Salud y los países brindan una atención especial. Dentro de los grupos de alto riesgo se encuentran los productos de origen animal, como la leche y sus derivados. Staphylococcus aureus es un microorganismo causante de brotes de las enfermedades transmitidas por alimentos, lo que genera la necesidad de estudiar la carga microbiana y establecer los controles adecuados. Objetivo: evaluar la calidad microbiológica en el proceso manufacturero en el área de producción de la Cooperativa de Producción Agropecuaria Chone LTDA, con el fin de asegurar la ausencia del Staphylococcus aureus en el queso fresco 100% chonero. Materiales y métodos: Se realizó un estudio de evaluación de la calidad en cuatro fases. Se realizaron exámenes microbiológicos iniciales, se identificaron las causas esenciales de contaminación identificadas por observación directa, revisión de documentos y entrevistas, como base para la elaboración e implementación de POE (Procedimientos Operativos Estandarizados) y POES (Procedimientos Operativos Estandarizados de Saneamiento). Se cumplieron los requerimientos éticos y se realizaron los análisis estadísticos utilizando porcentaje e índices como medida resumen y con el uso de IBM SPSS, versión libre, realizándose regresión logística binaria para determinar la probabilidad de éxito al aplicar dichos procedimientos. Resultados: los análisis microbiológicos iniciales (antes de implementación de POE y POES), dieron como resultado: 98, 116, 79, 120 y 13 UFC/g para las muestras: DiQ1, DiQ2, DiQ3, DiQ4 y DiQ5, respectivamente. Las causas esenciales de contaminación fueron falta de control microbiológico, falta de control del aspecto del personal y falta de control del agua) Una vez implementados los POE y POES, el análisis mostró ausencia en todas las muestras analizadas. Conclusiones: se concluye que la aplicación de POE y POES tiene un efecto significativo sobre la ausencia del Staphylococcus aureus, con probabilidad de éxito del 100% (AU).

Introduction: Diseases transmitted by foodstuffs (DTF) are a worldwide problem that receives special attention by WHO and different countries. High risk groups include foodstuff of animal origin, like milk and its by-products. Staphylococcus aureus is a microorganism responsible for DTF outbreaks which generates the necessity of studying its microbial burden and establishing the adequate controls. Objective: to evaluate the microbiological quality of the manufacturing process in the production area of the Farming and Stockbreeding Cooperative Chone LTDA, to assure the absence of Staphylococcus aureus in the manufacturing process of fresh cheese. Materials and methods: A study to assess the quality in four stages was carried out: initial microbiological tests, identification of the essential contamination causes by direct observation, documental review and inquires. They were used to elaborate and implement the Standardized Operational Procedures (SOP) and the Standardized Operational Procedures of Sanitation (SOPS). The ethical requirements were fulfilled and the statistical tests were performed using percentages and indexes as summary measures through IBM SPSS, free version; binary logistic regression was made to determine the success when applying those procedures. Results: initial microbiological controls (before implementing SOP and SOPS) gave these results 98, 116, 79, 120, and 13 UFC/g for samples DiQ1, DiQ2, DiQ3, DiQ4 and DiQ5 respectively. The main contamination causes were lack of microbiological control, lack of control on the staff's cleanliness and lack of control on the water. After implementing SOP and SOPS microbiological control found particularly clean samples. Conclusions: SOP and SOPS have a significant effect on the amount of Staphylococcus aureus, with 100% success probability (AU).

Portación de Staphylococcus aureus enterotoxigénico tipo A, en frotis nasofaríngeos en manipuladores de alimentos / Carriage of Staphylococcus aureus among food service workers

Background Staphylococcus aureus produces 11 serotypes of endotoxins that may cause food poisoning. Aim To determine the prevalence of type A enterotoxigenic Staphylococcus aureus carriage among food service workers in Chillan, Chile. Material and Methods Pharyngeal swabs were obtained from 100 food service workers and were cultured in Agar plates. After identifying the presence of Staphylococcus aureus, DNA was extracted to identify type A toxin by conventional PCR. Results Thirty eight percent of samples were colonized with Staphylococcus aureus. Among these, 26% were toxin A producers. Conclusions Half of the sampled workers carried Staphylococcus aureus and a quarter of these produced type A enterotoxin.

Habituation of enterotoxigenic Staphylococcus aureus to Origanum vulgare L. essential oil does not induce direct-tolerance and cross-tolerance to salts and organic acids

Enterotoxigenic Staphylococcus aureus strains that were isolated from foods were investigated for their ability to develop direct-tolerance and cross-tolerance to sodium chloride (NaCl), potassium chloride (KCl), lactic acid (LA) and acetic acid (AA) after habituation in sublethal amounts (1/2 of the minimum inhibitory concentration - 1/2 MIC and 1/4 of the minimum inhibitory concentration - 1/4 MIC) of Origanum vulgare L. essential oil (OVEO). The habituation of S. aureus to 1/2 MIC and 1/4 MIC of OVEO did not induce direct-tolerance or cross-tolerance in the tested strains, as assessed by modulation of MIC values. Otherwise, exposing the strains to OVEO at sublethal concentrations maintained or increased the sensitivity of the cells to the tested stressing agents because the MIC values of OVEO, NaCl, KCl, LA and AA against the cells that were previously habituated to OVEO remained the same or decreased when compared with non-habituated cells. These data indicate that OVEO does not have an inductive effect on the acquisition of direct-tolerance or cross-tolerance in the tested enterotoxigenic strains of S. aureus to antimicrobial agents that are typically used in food preservation.

Occurrence of enterotoxin-encoding genes in Staphylococcus aureus causing mastitis in lactating goats / Ocorrência de genes codificadores de enterotoxinas em Staphylococcus aureus associados a mastite em cabras em lactação

Staphylococcal enterotoxins are the leading cause of human food poisoning worldwide. Staphylococcus spp. are the main mastitis-causing agents in goats and frequently found in high counts in goat milk. This study aimed to investigate the occurrence of enterotoxin-encoding genes in Staphylococcus aureus associated with mastitis in lactating goats in Paraiba State, Brazil. Milk samples (n=2024) were collected from 393 farms. Staphylococcus aureus was isolated in 55 milk samples. Classical (sea, seb, sec, sed, see) and novel (seg, seh, sei) enterotoxin-encoding genes were investigated by means of polymerase chain reaction (PCR). From thirty-six tested isolates, enterotoxin-encoding genes were detected in 7 (19.5 percent) S. aureus. The gene encoding enterotoxin C (seC) was identified in six isolates, while seiwas observed in only one isolate. The genes sea, seb, sed, see, seg and seh were not observed amongst the S. aureus investigated in this study. In summary, S. aureus causing mastitis in goats can harbor enterotoxin-encoding genes and seC was the most frequent gene observed amongst the investigated isolates. This finding is important for surveillance purposes, since enterotoxin C should be investigated in human staphylococcal food poisoning outbreaks caused by consumption of goat milk and dairy products.

As enterotoxinas estafilocócicas são as principais causas de intoxicação alimentar em humanos em todo o mundo. O principal agente causador da mastite caprina são os Staphylococcus spp., frequentemente encontrado em altas contagens no leite caprino. Este estudo objetivou investigar a ocorrência de genes codificadores de enterotoxinas em Staphylococus aureus associados com mastite em cabras em lactação no estado da Paraíba, Brasil. As amostras de leite (n=2024) foram coletadas em 393 propriedades. Foram isolados 55 S. aureus em amostras de leite. Os genes codificadores de enterotoxinas clássicas (sea, seb, sec, sed, see) e as novas (seg, seh, sei) foram investigadas por meio de reação em cadeia de polimerase (PCR). Foram testados trinta e seis isolados, foram detectados 7 (19,5 por cento) genes codificadores de enterotoxinas de S. aureus. O gene codificador da enterotoxina C (sec) foi identificado em seis isolados, enquanto sei foi observado em apenas um isolado. Os genes sea, seb, sed, see, seg e seh não foram observados entre os S. aureus investigados neste estudo. Em síntese, a mastite caprina causada por S. aureus pode abrigar genes codificadores de enterotoxinas e sec foi o gene mais frequentemente observado entre os isolados investigados. Essa descoberta é importante para fins de vigilância e a enterotoxina C deve ser investigada em surtos de intoxicações alimentares estafilocócicas em humanos causadas pelo consumo de leite caprino e seus derivados.

Pesquisa de estafilococos e Escherichia coli em mãos de colaboradores, em unidades de alimentação e nutrição na cidade do Rio de Janeiro, RJ / Staphylococcus and Escherichia coli research in the hands of employees at food and nutrition units in the city of Rio de Janeiro, RJ

As Doenças Veiculadas por Alimentos ­ DVA são motivos constantes de preocupação no mundo todo, tendo em vista o aumento do consumo de alimentos fora dos domicílios. As DVA podem ser causadas pela ingestão de alimentos contaminados por bactérias patogênicas, cuja contaminação geralmente decorre do modo inapropriado de manusear, preparar ou estocar alimentos em Unidades de Alimentação e Nutrição ­ UAN como restaurantes e hotéis. O presente trabalho teve como objetivo avaliar a contaminação por estafilococos e Escherichia coli nas mãos de 107 colaboradores de alimentos (global e separado por sexo) em seis diferentes UAN da cidade do Rio de Janeiro. Os materiais foram coletados através de impressão digital em meios de cultura específicos e encaminhados ao laboratório. Os resultados avaliados de acordo com os laudos fornecidos demonstraram que 66% dos colaboradores foram suspeitos de portar estafilococos sendo 65% do sexo masculino e 55% do sexo feminino. A Escherichia coli não foi detectada nas mãos dos colaboradores na presente pesquisa. Sugere-se treinamento visando as boas práticas de manipulação e adequação dos processos operacionais padrão de higienização de mãos, visando minimizar a ocorrência de estafilococos com mais ênfase para os colaboradores do sexo masculino.

The Foodborne Diseases - FD are a constant concern throughout the world in order to increase the consumption of food outside of homes. The FD can be caused by eating food contaminated with pathogenic bacteria, which contamination usually arises from the inappropriate way to handle, prepare or store food in Food and Nutrition Units ­ FNU like restaurants and hotels. This study aimed to evaluate the contamination by staphylococci and Escherichia coli in the hands of 107 employees of food (overall and separated by sex) in six different FNU in city of Rio de Janeiro. The materials were collected through fingerprint specific culture media and sent to the laboratory. The results evaluated according to the reports provided showed that 66% of employees were suspected of carrying staphylococci and 65% of men and 55% of women. Escherichia coli were not detected in the hands of the collaborators in this research. It is suggested that training aimed at the good handling practices and the adequacy of standard operating procedures for hand hygiene to minimize the occurrence of staphylococci with more emphasis on male employees.

Contaminação microbiana de sanduíches em lanchonetes: estudo transversal realizado em Brasília / Bacterial contamination of sandwiches sold in snack bars: cross-sectional study carried out in Brasilia, Brazil

OBJETIVO: investigar as condições higiênico-sanitárias de sanduíches disponíveis para consumo humano comercializados na região de Brasília, Distrito Federal (DF). MÉTODOS: realizou-se um inquérito transversal de amostragem representativa para as lanchonetes na região de Brasília em 2006. A amostra foi constituída de 167 lanchonetes, sorteadas aleatoriamente de uma lista de estabelecimentos cadastrados na Secretaria de Saúde do Distrito Federal. Em cada lanchonete, um sanduíche foi preparado e analisado imediatamente para a contagem de coliformes, estafilococos, Bacillus cereus e detecção de Salmonella. RESULTADOS: dentre as 167 amostras, 55 (33 por cento; intervalo de confiança de 95 por cento: 26 por cento-40 por cento) apresentaram contaminação acima dos limites estabelecidos pela legislação vigente para as seguintes bactérias: coliformes, estafilococos, Bacillus cereus e Salmonella. Ao considerar qualquer nível de contaminação, esse valor sobe para 92 por cento. CONCLUSÃO: devido ao grau de contaminação encontrado sugere-se realização de atividades educativas e de vigilância com intuito de melhorar a qualidade higiênico-sanitária dos sanduíches avaliados.

OBJECTIVE: to investigate the hygienic conditions of sandwiches available for human consumption sold in the region of Brasilia, Federal District. METHOD: a cross sectional survey of a representative sample of snack bars in the Brasilia region was conducted in 2006. A sample of 167 snack bars was randomly selected from a list of existing establishments registered with the Federal District's Health Department. In each snack bar a sandwich was prepared and analyzed immediately for coliforms, staphylococcus, Bacillus cereus counts and Salmonella detection. RESULTS: of the total of 167 samples, 55 (33 per cent; 95 per cent confidence interval: 26 per cent-40 per cent) had contamination levels above the limits established by prevailing law for the following bacteria: coliforms, staphylococcus, Bacillus cereus and Salmonella. When considering any kind of contamination, this percentage increases to 92 per cent. CONCLUSION: given the level of contamination found, educational and health surveillance activities are needed to improve hygiene procedures in the establishments that prepare sandwiches.

Prevalence of enterotoxin-encoding genes and antimicrobial resistance in coagulase-negative and coagulase-positive Staphylococcus isolates from black pudding / Prevalência de genes codificadores de enterotoxinas e resistência antimicrobiana em estafilococos coagulase-negativo e coagulase-positivo isolados de morcilhas no sul do Brasil

INTRODUCTION: Staphylococcal species are pathogens that are responsible for outbreaks of foodborne diseases. The aim of this study was to investigate the prevalence of enterotoxin-genes and the antimicrobial resistance profile in staphylococcus coagulase-negative (CoNS) and coagulasepositive (CoPS) isolates from black pudding in southern Brazil. METHODS: Two hundred typical and atypical colonies from Baird-Parker agar were inoculated on mannitol salt agar. Eighty-two mannitol-positive staphylococci were submitted to conventional biochemical tests and antimicrobial susceptibility profiling. The presence of coagulase (coa) and enterotoxin (se) genes was investigated by polymerase chain reaction. RESULTS: The isolates were divided into 2 groups: 75.6% (62/82) were CoNS and 24.4% (20/82) were CoPS. The biochemical tests identified 9 species, of which Staphylococcus saprophyticus (37.8%) and Staphylococcus carnosus (15.9%) were the most prevalent. Antimicrobial susceptibility tests showed resistance phenotypes to antibiotics widely administered in humans, such as gentamicin, tetracycline, chloramphenicol, and erythromycin. The coa gene was detected in 19.5% (16/82) of the strains and 4 polymorphic DNA fragments were observed. Five CoNS isolates carrying the coa gene were submitted for 16S rRNA sequencing and 3 showed similarity with CoNS. Forty strains were positive for at least 1 enterotoxin-encoding gene, the genes most frequently detected were sea (28.6%) and seb (27.5%). CONCLUSIONS: The presence of antimicrobial resistant and enterotoxin-encoding genes in staphylococci isolates from black pudding indicated that this fermented food may represent a potential health risk, since staphylococci present in food could cause foodborne diseases or be a possible route for the transfer of antimicrobial resistance to humans.

INTRODUÇÃO: Estafilococos são patógenos responsáveis por surtos de doenças transmitidas por alimentos. O estudo investigou a prevalência de genes de enterotoxinas e o perfil de resistência aos antimicrobianos em estafilococos coagulase-negativo (CoNS) e estafilococos coagulase-positivo (CoPS) isolados de morcilhas no sul do Brasil. MÉTODOS: Duzentas colônias típicas e atípicas do ágar Baird-Parker foram inoculadas em ágar sal-manitol. Oitenta e dois estafilococos manitol-positivos foram submetidos a testes bioquímicos e perfil de susceptibilidade antimicrobiana. A presença dos genes da coagulase (coa) e enterotoxinas (se) foi investigada por reação em cadeia da polimerase (PCR). RESULTADOS: Os isolados foram divididos em dois grupos: 75,6% (62/82) CoNS e 24,4% (20/82) CoPS. Através dos testes bioquímicos, 9 espécies foram determinadas, Staphylococcus saprophyticus (37,8%) e Staphylococcus carnosus (15,9%) foram as mais prevalentes. Testes de susceptibilidade demostraram fenótipos de resistência aos antibióticos administrados em humanos, como gentamicina, tetraciclina, cloranfenicol e eritromicina. O gene coa foi detectado em 19,5% (16/82) das cepas e quatro fragmentos de DNA polimórficos foram observados. Cinco CoNS contendo o gene coa foram submetidos ao sequenciamento do 16S rRNA e três mostraram similaridade com CoNS. Quarenta amostras foram positivas para pelo menos um gene se, os mais frequentes foram sea (28,6%) e seb (27,5%). CONCLUSÕES: A presença de resistência aos antimicrobianos e de genes se nos isolados de morcilha indicou que este alimento pode representar um risco potencial à saúde, já que a presença nos alimentos pode causar doenças de origem alimentar ou ser uma possível rota de transferência de estafilococos resistentes aos humanos.

Brotes por Salmonella spp., Staphylococcus aureus y Listeria monocytogenes asociados al consumo de pollo / Outbreaks of Salmonella spp., Staphylococcus aureus and Listeria monocytogenes associated with poultry consumption. Systematic review.

Introducción. Las enfermedades transmitidas por alimentos son un serio problema de salud pública y, el pollo, uno de los alimentos asociados con ellas. Objetivo. Determinar la distribución y frecuencia de brotes alimentarios asociados al consumo de pollo contaminado por Salmonella spp., Listeria monocytogenes y Staphylococus aureus, mediante una revisión sistemática de la literatura científica. Materiales y métodos. Se buscaron los estudios de brotes asociados a Salmonella spp., S. aureus y L. monocytogenes, en las bases de datos Medline, Pubmed, Science Direct,SciELO,Librería Cochrane (CCRT),Biblioteca Virtual en Salud (BVS), Highwire,HINARI y MedicLatina. Se obtuvieron los datos para el cálculo de odds ratios (OR) mediante la elaboración de tablas de contingencia en el programa RevMan5™. Resultados. Siete artículos cumplieron con los criterios de inclusión y no se encontraron reportes de L. monocytogenes. El OR global fue de 3,01 (IC95% 2,37-3,81), lo que se interpreta como una asociación significativa entre el consumo de pollo contaminado y la infección alimentaria. Se presentó heterogeneidad en los estudios incluidos (p=0,03), por lo que fue necesario un análisis por subgrupos de microorganismos; para el caso de Salmonella spp., el OR fue de 2,67 (IC95% 2,09-3,41). No se hizo análisis para S. aureus por reportarse un solo artículo. Conclusiones. Se encontró un OR de 2,61, lo que indica que hay una fuerte asociación entre el consumo de pollo y la adquisición de salmonelosis. El principal factor de riesgo para adquirir salmonelosis es el consumo de pollo de asadero en los restaurantes.

Introduction. Food borne diseases are a serious public health problem. Poultry are often associated with these outbreaks. Objective. A systematic review of the literature is provided concerning the distribution and frequency of food borne outbreaks associated with consumption of chicken contaminated with Salmonella spp., Listeria monocytogenes and Staphylococcus aureus. Materials and methods. The search for studies of outbreaks associated with Salmonella, S. aureus and L. monocytogenes was conducted in Medline, Pubmed, Science Direct, Scielo, Cochrane Library (CCRT), Virtual Health Library (VHL), Highwire, HINARI and MedicLatina. Data were obtained for the calculation of odds ratio (OR) by preparing contingency tables using the RevMan5 program. Results. Seven articles met the inclusion criteria; however, no reports of L. monocytogenes were obtained. The overall OR was 3.01 (95% CI: 2.37, 3.81); this was interpreted as a significant association between the consumption of contaminated chicken and food poisoning. In the included studies heterogeneity (p= 0.03) was presented, so it took a subgroup analysis of microorganisms, in the case of Salmonella OR was 2.67 (95% CI: 2.09 -3.41). No analysis was made for S. aureus reported a single article. Conclusions. The OR indicated a strong association between chicken consumption and acquisition of salmonellosis. The main risk factor for acquiring salmonellosis is the consumption of chicken from grill restaurants.

Portación y caracterización de Staphylococcus aureus en manipuladores de alimentos / Carriage and characterization of Staphylococcus aureus in food handlers

Staphylococcus aureus es una causa de intoxicaciones alimentarias por su capacidad de producir enterotoxinas. Los manipuladores de alimentos que portan S. aureus productores de enterotoxinas pueden provocar intoxicaciones alimentarias. Se estudiaron muestras tomadas de fosas nasales de 88 manipuladores de alimentos en la provincia de Misiones. El 37,5 % de los individuos analizados eran portadores de S. aureus. Mediante técnicas de amplificación (PCR), se detectaron genes que codifican la producción de enterotoxinas en 13 de los 33 aislamientos obtenidos (39,4 %) y en el 14,7 % de los manipuladores. De estos aislamientos, 10 portaban el gen sea y 3 el gen sec. El estudio de sensibilidad a los antibióticos mostró un 100 % de sensibilidad a teicoplanina, gentamiclna y rifampicina; 2 aislamientos fueron resistentes a clindamicina y a eritromicina y 4 resultaron resistentes a la meticilina. Estos resultados son un alerta e indicarían la necesidad de desarrollar medidas racionales para reducir el riesgo potencial de intoxicaciones alimentarias.

Staphylococcus aureus causes food poisoning due to its ability to produce enterotoxins. Food handlers carrying enterotoxin-producing S. aureus can contaminate food, thus leading to food poisoning. Samples were obtained from 88 food handlers in the Province of Misiones, Argentina. S. aureus was isolated from nasal swaps and PCR amplification was performed for genes encoding staphylococcal enterotoxins. A total of 37.5 % food handlers were positive for S. aureus. Expression of enterotoxin genes was found in 13 of the 33 (39.4 %) S. aureus isolates studied, accounting for 14.7 % of food handlers. Gene sea was detected in 10 isolates followed by gene sec in 3 isolates. All isolates were susceptible to teicoplanin, gentamicin and rifampicin. Four isolates were resistant to methicillin whereas 2 isolates were resistant to clindamycin and erythromycin. These results constitute a critical alert and indicate the need for developing rational measures to reduce the potential risk of food poisoning.

Portación nasal de Staphylococcus aureus en manipuladores de alimentos del Mercado Nº 4 de Asunción, Paraguay / Nasal carriage of Staphylococcus aureus in food handlers in Market Number 4 in Asuncion, Paraguay

INTRODUCCIÓN: La correcta manipulación de los alimentos es indispensable para proteger nuestra salud; sin embargo, las enfermedades transmitidas por alimentos son bastante frecuentes, siendo el Staphylococcus aureus uno de los agentes causales más comunes. OBJETIVO: Determinar la prevalencia de portación de nasal de Staphylococcus aureus en manipuladores de alimentos del Mercado Nº4 de Asunción - Paraguay, y evaluar su susceptibilidad al antibiótico oxacilina. MATERIAL Y MÉTODO: Estudio observacional, descriptivo y de corte transversal, con componentes analíticos. Se realizaron hisopados nasales en una muestra compuesta por 105 manipuladores de alimentos en el Mercado Nº4 de Asunción - Paraguay con el fin de detectar la portación nasal de Staphylococcus aureus y sus sensibilidad a la oxacilina. También se determinó la asociación entre el sexo y la portación nasal de Staphylococcus aureus mediante la prueba de X2 (Chi cuadrado) con un nivel de significación de 0,05. RESULTADOS: Staphylococcus aureus fue aislado de las muestras nasales provenientes de 33,3 por ciento de los manipuladores de alimentos estudiados. El estudio de susceptibilidad a antibióticos mostró que 88,6 por ciento fueron sensibles a oxacilina, con 5,7 por ciento de sensibilidad intermedia y 5,7 por ciento de resistencia. No se observó asociación entre sexo y portación nasal de Staphylococcus aureus: Prevalencia Relativa =1,23 (Intervalo de Confianza del 95 por ciento: 0,45 – 1,99; valor p=0,9). DISCUSIÓN: Los resultados de nuestro estudio revelaron una portación nasal de Staphylococcus aureus similar a la comunicada por otros trabajos científicos. Estos resultados confirman la necesidad de educación y programas de capacitación sobre seguridad alimentaria dirigida a manipuladores de alimentos.

INTRODUCTION: Proper food handling is essential to protect our health. However food-borne diseases are common, Staphylococcus aureus being one of the most common causative agents. OBJECTIVE: Determine the prevalence of nasal carriage of Staphylococcus aureus in food handlers of Market No. 4 in Asunción - Paraguay, and assess their susceptibility to oxacillin. MATERIAL AND METHOD: Observational, cross-sectional, descriptive study with analytical components. Nasal swabs were performed on a sample of 105 food handlers in the No. 4 market in order to detect nasal carriage of Staphylococcus aureus and their susceptibility to oxacillin. We also determined the association between sex and nasal carriage of Staphylococcus aureus by the X2 test (Chi square) with a significance level of 0.05. RESULTS: Staphylococcus aureus was isolated from nasal swabs from 33.3 percent samples of food handlers studied. The antibiotic susceptibility study showed that 88.6 percent were susceptible to oxacillin, with 5.7 percent sensitivity and 5.7 percent intermediate resistance. There was no association between gender and nasal carriage of Staphylococcus aureus: Relative Prevalence = 1.23 (95 percent of Confidence Interval: 0.45 to 1.99, p value = 0.9). DISCUSSION: The results of our study revealed a Staphylococcus aureus nasal carriage similar to that reported by other scientific studies. These results confirm the need for education and training programs focused on food safety for food handlers.

Estafilococos em alimentos / Staphilococci in food

Este artigo de revisão tem como objetivo descrever as principais características do Staphylocococus aureus e a relação deste micro-organismo com as doenças transmitidas por alimentos. S. aureus é a principal espécie do gênero e o homem e os animais são os reservatórios. Este micro-organismo se multiplica com facilidade em vários alimentos e produz enterotoxinas (EE) termorresistentes. Leite cru e derivados, creme, tortas recheadas com creme, salada de batata, atum, carne de frango, presunto, carnes e produtos a base de ovos já foram incriminados nos surtos alimentares envolvendo os estafilococos. Os principais sintomas são náuseas, vômito, cãibras abdominais e diarréia. O período de incubação varia de 2 a 4 horas. São conhecidos 20 tipos diferentes de EE e sua produção é influenciada pela temperatura, pH, atividade de água, tamanho do inóculo, fonte de carbono e nitrogênio, concentração de sal e condições atmosféricas do substrato. A maioria das intoxicações é produzida por EEA e EED. As EE são detectáveis nos alimentos que apresentam populações de S. aureus acima de 105 UFC/mL de alimento. A dose mínima de EE ingerida para causar intoxicação é 100 ng. Para detecção de EE, existem métodos como Sensibilidade Ótima em Placas (OSP- Optimun Sensitivity Place), microslide, Aglutinação Reversa Passiva em Látex (RPLA- Reversed Passive Latex Agglutination) e ELISA. A Reação em Cadeia pela Polimerase (PCR- Polimerase Chain Reaction), apesar de não diferenciar células viáveis de não viáveis, pode ser aplicada para detecção de diversos tipos de estafilococos enterotoxigênicos.

This review article aims to describe the main characteristics of Staphylocococus aureus and the relation of this microorganism with food-borne disease. S. aureus is the main species of this genus, and man and animals are the main reservoirs. These microorganisms can easily grow in food and produce thermo-resistant enterotoxins. Whole milk and dairy products, cream, cream-filled pies, potato salad, tuna, chicken, ham, meat and egg-based products have been incriminated in food-borne outbreaks involving Staphylococcus. The main symptoms are nausea, vomit, abdominal cramps and diarrhea. The incubation period varies from 2 to 4 hours. Twenty different enterotoxins are known and their production is influenced by temperature, pH, water activity, inoculum size, carbon and nitrogen availability, salt concentration and atmospheric conditions. The majority of food poisoning is caused by enterotoxins A and E. The enterotoxins are detectable in food containing S. aureus populations above 105 CFU/mL. The minimum dose of enterotoxin necessary to cause staphylococcal poisoning is 100 ng. To detect enterotoxins, methods like optimum sensitivity plate (OSP), microslide, reversed passive latex agglutination (RPLA) and ELISA can be applied. Although polimerase chain reaction (PCR) cannot differentiate viable and nonviable cells, it can be used to detect different kinds of enterotoxigenic staphylococci.

Occurrence of staphylococcus aureus in raw milk produced in dairy farms in São Paulo state, Brazil

The objective of the present study was to evaluate the occurrence of Staphylococcus aureus in milk produced in 37 farms located in the regions of Ribeirão Preto and São Carlos, state of São Paulo, Brazil. Two-hundred and eight samples of milk from individual cows showing subclinical mastitis, and 37 samples of bulk tank milk were analyzed. S. aureus strains were detected in 18 (7.3 percent) milk samples: 14 (6.7 percent) from samples of individual cows, and 4 (10.8 percent) from bulk tank milk. Two individual milk samples (14.3 percent) and two bulk milk samples contained enterotoxigenic S. aureus. PFGE analysis revealed the genetic heterogeneity of the strains isolated from raw milk, which presented to 13 S. aureus patterns. Results confirmed the potential transmission of staphylococcal food poisoning to consumers via milk of cows affected by subclinical mastitis, mainly when raw milk is ingested.

[Molecular and serological detection of enterotoxigenic staphylococcus aureus from traditionally dairy products]

Staphylococcus aureus is one of the most causes of food poisoning [FP] in dairy products. The main etiologic agent of FP is staphylococcal enterotoxins [SE]. There are different types of SE, but type A [SEA] and type B [SEB] are the most important types. Because traditional dairy products are still produced and sold without a permit from the Ministry of Health, this study was conducted to evaluate molecular and serological detection of enterotoxigenic Staphylococcus aureus SEA and SEB from traditionally dairy products. In the current study, 100 samples of dairy products, which were produced by traditional methods, were transported to the laboratory under sterile conditions and were assessed. Samples were cultured and identified by routine bacteriological methods. The isolated bacteria were evaluated by PCR tests for diagnosis of the gene encoding of SEA and SEB. Subsequently, the ability of above mentioned strains to produce enterotoxin were examined by Sac's culture method and were confirmed by SRID. Data were analyzed using chi-square test. The results indicated that 32% of dairy products were contaminated by Staphylococcus aureus [18% cream, 10% cheese, 4% milk]. The PCR results showed that 15.6% of Staphylococcus aureus isolates possessed the SEA gene, 9.3% had the SEB gene and 6.2% possessed both genes. The ability of enterotoxin production indicated that 80% of SEA and 33% of SEB genes were expressed. Enterotoxins SEA and SEB are heat stable; therefore heating has no effect on dairy products contaminated by entertoxins and gastritis may occur in a short period of time. As PCR is a rapid, sensitive, specific and inexpensive methods, we suggest that it can be replaced to traditionally assays for detecting SE

Factores relacionados con enfermedades transmitidas por alimentos en restaurantes de cinco ciudades de Colombia, 2007 / Factors related to foodborne diseases in restaurants from 5 Colombian cities, 2007

Antecedentes. En Colombia existe subregistro de la notificación de casos de enfermedades transmitas por alimentos. La Organización Mundial de la Salud plantea que la mayoría de las contaminaciones de alimentos que generan enfermedades transmitidas por alimentos ocurren por problemas sanitarios y de manipulación, dentro del hogar y en sitios en donde se elaboran para la venta. Objetivo. Identificar los factores relacionados con la transmisión de enfermedades transmitidas por alimentos en expendios de alimentos. Materiales y métodos. En cinco ciudades de Colombia se encuestaron 300 establecimientos y 1.522 manipuladores de alimentos a quienes se les hizo control microbiológico de manos y, a 1.286, examen coprológico y coprocultivo. Resultados. Veinticinco establecimientos (8,3 por ciento) no tenían una ubicación adecuada, 113 (37,7 por ciento) no contaban con planes de saneamiento y sólo 26 (8,7 por ciento) realizaban prácticas apropiadas de almacenamiento. En los manipuladores se halló que 765 (50,3 por ciento) ingresaron con examen médico y 924 (60,7 por ciento) realizaron curso de manipulación de alimentos. En sus prácticas de trabajo se evidenció manejo simultáneo de dinero y alimentos (17 por ciento), uso de joyas (15,2 por ciento), uñas largas y con esmalte (8,9 por ciento), y 15,2 por ciento refirieron no lavarse las manos cuando manipulaban dinero y en los no capacitados se halló 1,3 veces más frecuente este hábito (RR=1,36 IC 95 por ciento=1,10 – 1,69). Se encontraron parásitos intestinales en 26,9 por ciento; 49 (3,8 por ciento) fueron positivos para parásitos patógenos, 6 (0,46 por ciento) para enterobacterias patógenas y 8 (0,52 por ciento) cultivos de manos, para Staphylococcus aureus.

Brote de enfermedad alimentaria en la localidad de El Huecú, provincia de Neuquén / Foodborne disease outbreak in El Huecú community, province of Neuquén

En febrero de 2006 ocurrió un brote epidémico de gastroenteritis aguda de origen alimentario, en ocasión de un festejo popular en una pequeña localidad de la provincia de Neuquén, Argentina. Aproximadamente 800 personas participaron de un almuerzo en las instalaciones del Gimnasio Municipal, y unas tres horas después de finalizado, cerca de 150 asistentes consultaron al hospital local, afectados por síndrome gastroentérico agudo. Se realizó una investigación epidemiológica caso-control a través de un muestreo representativo no probabilístico. Los resultados epidemiológicos establecieron un brote de ETA a fuente común, con una relación caso-control de 1:1,8. Los principales síntomas fueron cólicos abdominales (88%), vómitos (73,5%) y diarrea (60%). La torta que se sirvió en ese evento fue identificada como el alimento causal (OR 9,79; IC 95%; 2,66-36,00; valor p = 0,0001), sujeto a condiciones higiénico-sanitarias insatisfactorias en los diferentes procesos de elaboración, conservación y manipulación. De una porción de la torta se aisló una cepa de Staphylococcus aureus subespecie aureus, coagulasa positiva, enterotoxigénica, con un recuento de 2,4x10(6) UFC/g, y también se aisló este microorganismo de tres muestras de manos y narinas de personas involucradas en la preparación y el servicio. Las cepas aisladas de un operador y de la torta portaron el gen sea y presentaron el mismo patrón de SmaI-PFGE. Se atribuyó el brote de ETA a la contaminación durante el proceso de preparación de la torta consumida durante ese almuerzo popular, lo que podría estar relacionado con deficiencias en aspectos higiénicos y con la falta de refrigeración y de mantenimiento de la cadena de frío.

In the summer of 2006, an epidemic outbreak of acute gastrointestinal illness related to food consumption occurred in a small town in the province of Neuquén, Argentina. During a popular feast, approximately 800 local residents attended lunch held in the facilities of the Municipal Gymnasium. About three hours later, nearly 150 attendees sought medical assistance at the local hospital due to acute gastroenteritis. A case-control epidemiological investigation was conducted using representative non-probability sampling. The epidemiological investigation showed a common-source foodborne disease outbreak with a case-control ratio of 1:1.8. The main symptoms were abdominal cramps (88%), vomiting (73.5%) and diarrhea (60%). The cake was identified as the source of infection (OR 9.79; IC 95%, 2.66-36.00; p = 0.0001), and unsatisfactory hygienic conditions in food production, conservation and handling steps were identified. Coagulase positive, enterotoxigenic Staphylococcus aureus, subspecies aureus was detected in a piece of cake, with a count of 2.4x10(6) CFU/g, and in samples from the hands and nostrils of three people involved in food preparation and service. The strains isolated from both the cake and one of the food handlers carried the sea gene, and presented the same SmaI-PFGE pattern. The foodborne disease outbreak was considered to be due to contamination in the preparation process of the cake consumed at the feast, which was related to inadequate hygienic conditions, lack of refrigeration and cold chain disruption.

Avaliação das boas práticas, relacionadas ao tempo e temperatura, na distribuição de pescado em restaurantes / Evaluation of best practices related to time and temperature, the distribution of fish in restaurants

Atualmente, o pescado constitui a quinta parte da proteína de origem animal consumida em nível mundial; contudo, a explosão demográfica (1950-1993) levou que a produção do pescado aumentasse de 20 para 1000 milhões de toneladas por ano. Em países desenvolvidos, os registros demonstram que 60 por cento dos surtos de toxinfecção alimentar são decorrentes do consumo de alimentos contaminados servidos em restaurantes. Sendo assim, a padronização do processamento e a manutenção das temperaturas têm sido citadas como parâmetros importantes para se evitar cada vez mais as freqüentes toxinfecções alimentares. Com isso, nosso objetivo foi avaliar as Boas Práticas nas etapas de distribuição do pescado, conceituando, investigando e identificando sua importância para garantia da inocuidade do produto a ser consumido. Esta pesquisa abrangeu três diferentes Unidades de Alimentação, sendo um restaurante comercial e dois restaurantes terceirizados do tipo self-service. Entre julho e dezembro de 2003 foram coletados dados referentes a tempo e temperatura de exposição das preparações a base de pescados, assim como Boas Práticas referentes a esta etapa. Nossos resultados sugerem que, na unidade A (comercial), o produto servido pode estar oferecendo riscos à saúde do consumidor, visto que seu tempo e temperatura de exposição foram inadequados, segundo a Legislação vigente. O mesmo ocorreu com as demais unidades, porém, com tempos de exposição bem mais curtos (alta rotatividade), não oferecendo tantos riscos ao seu consumo.

Molecular subtyping of Staphylococcus aureus isolated from a severe food-poisoning / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the molecular types of Staphylococcus aureus isolated from a severe food-poisoning and to trace the possible strains.</p><p><b>METHODS</b>Real-time PCR was applied to detect nuc gene as a specific marker for S. aureus, mecA gene encoding methicillin resistance and 5 other genes encoding staphylococcal enterotoxins (sea, seb, see, sed, see). Isolates were also performed with 16S rRNA oligonucleotide sequence analyzing by DNAStar MegAlign 5.0 software and pulse-field gel electrophoresis (PFGE) by BioNumerics Version 4.0 software.</p><p><b>RESULTS</b>The nuc gene was detected from the 10 isolated strains, sea and seb genes were detected from 7 strains. There were 4 16 S rRNA types and 5 PFGE types found from all the strains.</p><p><b>CONCLUSIONS</b>Three relative S. aureus strains were involved in the severe food-poisoning at least. Molecular subtyping might give a molecular epidemiological evidence and support the source tracing of an outbreak.</p>