ABSTRACT During the last years, Brazilian government control programs have detected an increase of Salmonella Heidelberg in poultry slaughterhouses a condition that poses a threat to human health However, the reasons remain unclear. Differences in genetic virulence profiles may be a possible justification. In addition, effective control of Salmonella is related to an efficient epidemiological surveillance system through genotyping techniques. In this context, the aim of this study was the detection of 24 virulence-associated genes in 126 S. Heidelberg isolates. We classified the isolates into 56 different genetic profiles. None of the isolates presented all the virulence genes. The prevalence of these genes was high in all tested samples as the lowest number of genes detected in one isolate was 10/24. The lpfA and csgA (fimbriae), invA and sivH (TTSS), and msgA and tolC (intracellular survival) genes were present in 100% of the isolates analyzed. Genes encoding effector proteins were detected in the majority of SH isolates. No single isolate had the sefA gene. The pefA gene was found in only four isolates. We have also performed a screening of genes associated with iron metabolism: 88.9% of isolates had the iroN geneand 79.4% the sitC gene . Although all the isolates belong to the same serotype, several genotypic profiles were observed. These findings suggest that there is a diversity of S. Heidelberg isolates in poultry products. The fact that a single predominant profile was not found in this study indicates the presence of variable sources of contamination caused by SH. The detection of genetic profiles of Salmonella strains can be used to determine the virulence patterns of SH isolates.

RESUMO: Extratos de taninos (com e sem enxague) foram comparados com o uso de hipoclorito de sódio e água em alface crespa minimamente processada, sob atmosfera modificada. Foram avaliadas diferença total de cor e análises microbiológicas após a sanitização e durante o armazenamento dos produtos (9 dias à 3°C). O extrato de tanino SM(r) com enxague apresentou resultados semelhantes aos da solução de cloro para todos os parâmetros analisados e, além disso, apresentou uma elevada redução da contagem inicial microbiológica de alface minimamente processada. No entanto, durante o armazenamento, SM(r) não apresentou melhores resultados do que o uso de água. Portanto, SM(r) poderia ser utilizado na desinfecção da água de lavagem das alfaces, reduzindo a microbiota inicial e, assim, impedir a contaminação cruzada durante o processamento mínimo de vegetais.

ABSTRACT: In the present study, tannin extracts (rinsed or not) were compared to the use of sodium hypochlorite and tap water on fresh cut crisp leaf lettuce stored under modified atmosphere packaging. Effects of these sanitizers on total color difference and microbial levels of the product after sanitization and storage for 9 days at 3°C were evaluated. Performance of rinsed SM(r) tannin extract was comparable to the results of chlorine solution for all the analyzed parameters and; furthermore, that extract presented a high reduction in the initial bacterial count of minimally processed lettuce. However, storage of tannin extracts, did not impart better outcomes than the use of tap water. Therefore, the tannin extract storage SM(r) could be used in washing water to reduce the initial microbiological load, avoiding cross contamination in vegetables minimally processed.

A total of 120 beef carcasses were analyzed during processing at a slaughterhouse in southern Brazil. The carcasses were sampled by swab at three different steps of the slaughter line and then they were tested for Salmonella and E. coli. The Salmonella isolates were also examined for antimicrobial susceptibility. Salmonella prevalence distribution was modeled and the probability of contamination was simulated using @Risk program and 10,000 interactions. Results demonstrated that 4 beef carcasses (3.3%) were positive for Salmonella only in the first point. The six isolates of Salmonella were classified: S. Newport (n = 3), S. Saintpaul (n = 2) and S. Anatum (n = 1). No Salmonella strains exhibited resistance to any of the antimicrobials tested. As expected, the most contaminated point with E. coli was the first point (hide), presenting counts from 0.31 to 5.07 log cfu/100 cm². Much smaller E. coli counts were observed in the other points. Results indicated low levels of Salmonella and E. coli on the beef carcasses analyzed and also low probability of contamination of the carcasses by Salmonella, suggesting adequate microbiological quality.

Salmonella has been identified as the main aetiological agent responsible for foodborne diseases in several countries worldwide, including Brazil. In the State of Rio Grande do Sul (RS), southern Brazil, previews studies analysed official foodborne illnesses data, identifying Salmonella as the main bacterial agent of foodborne diseases during the period of 1997 to 2001. The present study aimed to analyse the official epidemiological data on salmonelloses occurred in the State of RS, during the period of 2002 to 2004. Even though data on recent salmonelloses were available, only data concerning the period comprising in 2002 to 2004 were analysed because the official worksheet records presented more consistent information about the salmonellosis outbreaks. Results indicated that, among the 624 foodborne outbreaks officially investigated, 202 (32.37%) were confirmed as salmonellosis. Among them 23,725 people were involved, 4,148 became sick, 1,878 were hospitalized and one person died. The season with the highest incidence of salmonelloses was spring, and the most affected age group was composed of people aged between 20 to 49 years old (56.66%). Animal origin foods -especially eggs and meat products -were very often involved with the outbreaks, however homemade mayonnaise was identified as the main food vehicle for salmonelloses (53.51%). The majority of the cases occurred inside private homes (55.81%) and food services (12.1%), and the main factors contributing to the occurrence of the outbreaks were the consumption of products without sanitary inspection (26.7%) and exposure of food at room temperature for more than two hours (18.58%). Similarly to what was previously reported for the period of 1997 to 2001, Salmonella spp. was the most prevalent foodborne disease agent in the State of RS during the years of 2002 to 2004.

Staphylococcal food poisoning is one of the most frequent foodborne illnesses worldwide and it is caused by the ingestion of food contaminated with enterotoxins produced by some strains of Staphylococcus (S.) aureus. In the State of Rio Grande do Sul (RS), Southern Brazil, S. aureus has been identified as the second most frequent agent of foodborne illnesses in the last two decades. The aim of the present study was to assess and analyse the epidemiological data of S. aureus food poisoning occurred in the State of RS during the years of 2000 to 2002. The official records of epidemiological investigations carried out by the Sanitary Surveillance Services of the State of RS were analysed. Among foodborne outbreaks for which aetiology was determined, S. aureus was identified as the responsible agent of 57 foodborne outbreaks, being 42 (74%) confirmed by microbiological analyses and 15 (26%) confirmed by clinical symptoms and/or epidemiological data. Staphylococcal outbreaks were responsible for the exposition of 5,991 persons, of which 1,940 (32%) were interviewed by the Sanitary Surveillance officers. The most affected age group corresponded to people with 20 to 49 years old (48%), where men (48%) and women (52%) were affected similarly. The main involved food vehicles were meats servings (35%), followed by pastries (25%), cheese (23%), pasta (11%) and potato salad with homemade mayonnaise (11%). The majority of the outbreaks occurred inside private homes (33%) followed by commercial food establishments (28%). Inadequate control of temperature and failures in general hygiene practices were identified as the main factors responsible for the outbreaks. In conclusion, S. aureus was an important food poisoning etiological agent in the State of RS during 2000 to 2002 and its prevention depends on control measures involving different parts of the food chain.

In the State of Rio Grande do Sul (RS), Southern Brazil, a good manufacturing practices regulation was published recommending two disinfection methods for cleaning cloths used in food services. The aim of the present study was to evaluate the efficacy of those methods. Cleaning cloths were sampled without prior notice at food services, on common working days. For the analyses, the cloths were divided in two sub-samples, being one of them microbiologically analyzed. The second sub-sample was further divided in two pieces and submitted to hand washing for two minutes. After that, one piece was boiled in water for 15 min and the other one was soaked in a 200 ppm sodium hypochlorite solution for 15 min. Both pieces of cloth were submitted to microbiological analyses. Cleaning cloths presented total aerobic mean counts of 6.9 ± 6.7 log/cm². All cleaning cloths presented coliform contamination, and 40% demonstrated mean counts of 6.2 ± 5.6 log/cm². Presumptive S. aureus mean counts of 5.5 ± 4.9 log/cm² were found. No statistic correlation was observed among the number of meals served daily in the food services and the microbiological contamination levels. After washing and disinfection, microbiological counts were significantly (p < 0.05) reduced by both methods, achieving an approximately 5 log reduction. The reductions achieved by the sodium hypochlorite soaking method and the boiling method were not significantly different. Thus, it was possible to conclude that both recommended methods were suitable to disinfect cleaning cloths used in food services.

The objective of the present study was to assess the growth and the recovery of Salmonella (S.) Enteritidis SE86 in different diluents, culture media and using different plating methods after the exposure to 200 mg/kg sodium dichloroisocyanurate (NaDCC). Before and after NaDCC exposure, SE86 was cultured at 30 °C and 7 °C in the following diluents: Peptone water (P), Saline solution (SaS), Peptone water+Saline solution (P+SaS), Peptone water+Tween 80+Lecithin+Sodium thiosulfate (P+N) and Saline solution+Tween 80+Lecithin+Sodium thiosulfate (SaS+N). The SaS diluent was chosen because it was able to maintain cells viable without growth and was further used for plating SE86 on non selective medium (Tryptic Soy Agar-TSA) and on selective media (Mannitol Lysine Crystal Violet Brilliant Green Agar-MLCB; Brilliant Green Agar-BGA; Salmonella Shigella Agar-SS and Xylose Lysine Dextrose-XLD). The Thin Agar Layer method (TAL) i.e., selective media overlayed with non selective TSA was also evaluated. Results indicated that SE86 not exposed to NaDCC was able to grow in P, P+N, SaS+N and P+SaS, but not in SaS, that was able to maintain cells viable. SE86 exposed to NaDCC demonstrated similar counts after dilution in SaS and the plating on non selective TSA, selective media MLCB, BGA, SS and XLD and on TAL media. SE86, S. Typhimurium and S. Bredeney, exposed or not exposed to NaDCC, showed no significant differences in counts on TSA, XLD and XLD overlayed with TSA, suggesting that all those media may be used to quantify NaDCC-exposed Salmonella by plating method.

Este estudo tem por objetivo avaliar a presença de microrganismos indicadores higiênico-sanitários em amostras de alimentos servidos em escolas públicas de Porto Alegre. Foram analisados todos os alimentos servidos na refeição do turno da visita, quanto à presença de Escherichia coli, Staphylococcus coagulase positiva, Salmonella sp. e Shigella sp. No total de 196 alimentos analisados de 120 escolas, 4 apresentavam contagem de Escherichia coli acima do permitido e dois tinham a presença de Staphylococcus coagulase positiva. Os gêneros Shigella e Salmonella não foram encontrados. Foi observado que a maioria das escolas estudadas servia alimentos dentro de padrões higiênico-sanitários adequados. Foi evidenciado que somente escolas municipais contavam com a orientação de responsável técnico pela alimentação escolar. Das escolas estaduais 60% nunca haviam recebido visita de nutricionista nas quais foram encontrados procedimentos em desacordo com as exigências da legislação. Na maioria das escolas, os alimentos servidos estavam dentro de padrões adequados, porém os problemas detectados demonstram a necessidade da implantação das Boas Práticas no ambiente escolar.

The objective of this study was to evaluate the presence of hygienic and sanitary indicator microorganisms in samples of food served in public schools in Porto Alegre. All the food served in the meal of the session visited was analyzed for Escherichia coli, coagulase-positive Staphylococcus, Salmonella sp. and Shigella sp. Of the total of 196 food products analyzed in 120 schools, 4 contained and Escherichia coli score above the permitted level, and 2 contained coagulase-positive Staphylococcus. Neither Shigella nor Salmonella genus were detected. In the majority of schools studied, it was found that food was of an adequate hygienic-sanitary standard. However, only municipal schools had the supervision of a technician responsible for school food. In the state schools, 60% had never been visited by a nutritionist and in these schools several procedures failed to comply with legal requirements. In most of the schools studied, the food served to students was within adequate standards, though the problems detected revealed the need for the implementation of Best Practices in the school environment.

Este estudo tem por objetivo avaliar a presença de microrganismos indicadores higiênico-sanitários em amostras de alimentos servidos em escolas públicas de Porto Alegre. Foram analisados todos os alimentos servidos na refeição do turno da visita, quanto à presença de Escherichia coli, Staphylococcus coagulase positiva, Salmonella sp. e Shigella sp. No total de 196 alimentos analisados de 120 escolas, 4 apresentavam contagem de Escherichia coli acima do permitido e dois tinham a presença de Staphylococcus coagulase positiva. Os gêneros Shigella e Salmonella não foram encontrados. Foi observado que a maioria das escolas estudadas servia alimentos dentro de padrões higiênico-sanitários adequados. Foi evidenciado que somente escolas municipais contavam com a orientação de responsável técnico pela alimentação escolar. Das escolas estaduais 60% nunca haviam recebido visita de nutricionista nas quais foram encontrados procedimentos em desacordo com as exigências da legislação. Na maioria das escolas, os alimentos servidos estavam dentro de padrões adequados, porém os problemas detectados demonstram a necessidade da implantação das Boas Práticas no ambiente escolar.

The objective of this study was to evaluate the presence of hygienic and sanitary indicator microorganisms in samples of food served in public schools in Porto Alegre. All the food served in the meal of the session visited was analyzed for Escherichia coli, coagulase-positive Staphylococcus, Salmonella sp. and Shigella sp. Of the total of 196 food products analyzed in 120 schools, 4 contained and Escherichia coli score above the permitted level, and 2 contained coagulase-positive Staphylococcus. Neither Shigella nor Salmonella genus were detected. In the majority of schools studied, it was found that food was of an adequate hygienic-sanitary standard. However, only municipal schools had the supervision of a technician responsible for school food. In the state schools, 60% had never been visited by a nutritionist and in these schools several procedures failed to comply with legal requirements. In most of the schools studied, the food served to students was within adequate standards, though the problems detected revealed the need for the implementation of Best Practices in the school environment.

Salmonella enterica serovar Enteritidis is one of the main pathogens responsible for foodborne illness in Brazil. Probiotic bacteria can play a role in defense and recovery from enteropathogenic -infections. In this study, the ability of Lactobacillus acidophilus LA10 to colonise and exert anta-gonistic effects in the gastrointestinal tract was tested before and during experimental infection in conventional mice contaminated with S. Enteritidis (SE86). A dose of 0.1 mL containing 10(8) viable cells of SE86 and L. acidophilus LA10 was orally administered by gavage to mice. The experiment was divided into groups. As a negative control, Group 1 was administered only sterile saline solution. As a positive control, Group 2 was administered only SE86. Group 3 was first administered SE86, and after 10 days, treated with L. acidophilus LA10. Group 4 was first administered L. acidophilus LA10,and after 10 days, challenged with SE86.The results demonstrated that a significant number of SE86 cells were able to colonize the gastrointestinal tract of mice, specifically in the colon and ileum. L. acidophilus LA10 demonstrated an antagonistic effect against SE86, with better results observed for Group 3 over Group 4. Thus, L. acidophilus LA10 shows potential antagonistic effects against S. Enteritidis SE86, especially if administered after infection.

Little information about Shigella responsible for foodborne shigellosis is available in Brazil. The present study aimed to investigate the antimicrobial resistance and PCR-ribotyping patterns of Shigella isolates responsible for foodborne outbreaks occurred in Rio Grande do Sul State (RS), Southern Brazil in the period between 2003 and 2007. Shigella strains (n=152) were isolated from foods and fecal samples of victims of shigellosis outbreaks investigated by the Surveillance Service. Identification of the strains at specie level indicated that 71.1% of them were S. flexneri, 21.5% S. sonnei, and 0.7% S. dysenteriae. Ten strains (6.7%) were identified only as Shigella spp. An increasing occurrence of S. sonnei was observed after 2004. Most of the strains were resistant to streptomycin (88.6%), followed by ampicillin (84.6%), and sulfamethoxazole/trimethoprim (80.5 %). Resistant strains belonged to 73 patterns, and pattern A (resistance to ampicillin, sulfamethoxazole/trimethoprim, tetracycline, streptomycin, chloramphenicol, and intermediate resistance to kanamycin) grouped the largest number of isolates (n=36). PCR-ribotyping identified three banding patterns (SH1, SH2, and SH3). SH1 grouped all S. flexneri and SH2 grouped all S. sonnei. The S. dysenteriae strain belonged to group SH3. According to the results, several Shigella isolates shared the same PCR-rybotyping banding pattern and the same resistance profile, suggesting that closely related strains were responsible for the outbreaks. However, other molecular typing methods need to be applied to confirm the clonal relationship of these isolates.

The adhesion of Salmonella (S.) strains to stainless steel and polyethylene and their inactivation by biocides used in food industry was investigated. Coupons of stainless steel and polyethylene were immersed in bacterial suspensions of S. Enteritidis, S. Typhimurium, and S. Bredeney during 15, 30, and 60 minutes, and submitted to different concentrations of peracetic acid (PAA), sodium hypochlorite (NaOCl), and quaternary ammonium (Quat) sanitizers. Hydrophobicity of the surfaces was evaluated by contact angle measurements using the sessile drop method and bacterial adhesion was accompanied through bacterial counts and scanning electron microscopy (SEM). Results indicated that the three serovars of Salmonella presented similar adhesion to both materials (5.0 to 6.5 log cfu cm-2). The time of exposure did not influence the counts of adhered cells on both surfaces, however SEM revealed larger clusters of S. Enteritidis on both materials, not found for the other serovars. S. Enteritidis presented lower sessile drop angle on polyethylene, indicating hydrophilic properties of this material. The biocides were not able to inactivate all the microorganisms adhered on both surfaces. At least 1 log cfu cm-2 of all serovars tested remained viable after the exposure to different biocide concentrations. In general, higher counts of survivors were observed on polyethylene disinfected with different concentrations of biocides. S. Bredeney e S. Typhimurium were more resistant than S. Enteritidis to PAA, whilst S. Enteritidis presented smaller reduction rates to NaOCl. This last biocide was able to reduce Salmonella counts in approximately 3.0 to 4.0 log cm-2. When adhered to polyethylene, the serovars S. Typhimurium and S. Enteritidis were more resistant to Quat than S. Bredeney in all concentrations tested, and the numbers of S. Enteritidis remained almost unaltered. On stainless steel disinfected by Quat, S. Bredeney presented higher numbers of survivors.

No período de 1999 a 2002, uma linhagem geneticamente caracterizada de Salmonella Enteritidis esteve envolvida em mais de 90% das salmoneloses ocorridas no RS. Este trabalho teve por objetivo avaliar a cinética de crescimento dessa linhagem, comparando-a com o crescimento de uma linhagem de S. Typhimurium e de S. Bredeney não envolvida em surtos alimentares. Para tanto, cada linhagem foi semeada separadamente em caldo nutriente (CN) e em salada de batata com maionese caseira (SMC), os quais foram mantidos a 30 °C e 9,5 °C. Os resultados obtidos em laboratório foram comparados com os resultados modelados pelo Pathogen Modelling Program, USDA. Em CN, a cinética de crescimento a 30 °C foi semelhante para todas as linhagens, as quais atingiram aproximadamente 8 log UFC.mL-1. Em SMC, a 30 °C, a linhagem de S. Enteritidis apresentou maior quantidade de células nas primeiras 6 horas de crescimento, sendo que somente depois de 12 horas todos os microrganismos testados atingiram quantidades semelhantes de células, ou seja, aproximadamente 6 log UFC g-1. Em CN e em SMC, na temperatura de 9,5 °C, não foi detectado crescimento de nenhuma das linhagens testadas de Salmonella durante as primeiras 24 horas, sugerindo que essa temperatura foi suficiente para controlar a multiplicação desses microrganismos. A modelagem bacteriana confirmou a maioria dos resultados obtidos.

From 1999 to 2002, a genetically characterized strain of Salmonella Enteritidis was involved in more than 90% of foodborne salmonellosis in the state of Rio Grande do Sul, southern Brazil. This work aimed to evaluate the growth kinetics of this strain in comparison with that of other Salmonella serovar strains (S. Typhimurium and S. Bredeney) which were not involved in salmonellosis outbreaks. Each strain was inoculated separately in nutrient broth (NB) and in potato salad prepared with homemade mayonnaise (MS), and incubated at 30 and 9.5 °C. The experimental results were compared with results obtained by the Pathogen Modeling Program, USDA. All the strains showed similar growth characteristics in NB at 30 °C, reaching approximately 8 log CFU.mL-1. In MS, the S. Enteritidis strain showed higher counts in the first 6 hours when cultivated at 30 °C. Only after 12 hours of incubation did all the microorganisms reach similar counts (approximately 6 log CFU g-1). No growth of any strain was detected in the first 24 hours in NB and MS at 9.5 °C, suggesting that this temperature was adequate to control the multiplication of the tested Salmonella strains. The bacterial modeling confirmed most of the experimental results.

Neste trabalho, um ELISA indireto baseado em um anticorpo monoclonal (MAb) especifico para proteína de membrane externa de Salmonellaenterica serovar Enteritidis foi usado para detecção de Salmonella em 154 amostras de carne de frango. Sua eficiência foi determinada através de comparação com os resultados obtidos pela metodologia convencional. A prevalência de amostras contaminadas com Salmonella foi de 23% pelo método de cultivo convencional, e 26% pelo ELISA. De 35 amostras positivas para Salmonella pela metodologia convencional, 32 também foram positivas no ELISA. Outras sete amostras foram positivas somente no ELISA. Comparando os resultados obtidos nos dois métodos, o ELISA demonstrou sensibilidade e especificidade de 94%, e valor preditivo positivo e negativo de 82% e 98% respectivamente. A sorotipagem dos isolados revelou 31 Salmonella enterica serovar Enteritidis, 2 Salmonella enterica serovar Heidelberg, 1 Salmonella enterica serovar Choleraesuis e 1 Salmonella enterica sorovar 6,7:-:-.

In this work, an indirect ELISA based on a monoclonal antibody (MAb) specific for an outer membrane protein of Salmonellaenterica serovar Enteritidis was used for detection of Salmonella in 154 samples of chicken meat. Its efficiency was determined through comparison with the results obtained from the conventional method. The prevalence of samples contaminated with Salmonella was 23% with the conventional culture method, and 26% with the ELISA. From thirty-five samples positive for Salmonella by the conventional method, 33 were also positive by ELISA. Seven other samples were only positive in the ELISA. Comparison of the results obtained in the two methods showed an ELISA sensitivity and specificity of 94%, and positive and negative predictive values of 82% and 98% respectively. The serotyping of the isolates revealed 31 Salmonella enterica serovar Enteritidis, 2 Salmonella enterica serovar Heidelberg, 1 Salmonella enterica serovar Choleraesuis and 1 Salmonella enterica sorovar 6,7:-:-.

Salmonella spp. foi a principal causa de doenças transmitidas por alimentos nos últimos anos no estado do Rio Grande do Sul, sul do Brasil. Foram avaliados dados epidemiológicos de salmoneloses ocorridas durante o período de 1997 a 1999 fornecidos pela Divisão de Vigilância Sanitária do Rio Grande do Sul. Os seguintes fatores foram investigados: número total de surtos confirmados, número de pessoas envolvidas, incidência dos surtos de acordo com a estação do ano, idade e sexo das pessoas envolvidas, principais alimentos envolvidos, local de armazenagem dos alimentos envolvidos e local de ocorrência dos surtos e suas causas prováveis. Os resultados demonstraram 8217 pessoas envolvidas, das quais 1557 foram hospitalizadas. O maior número de surtos ocorreu durante a primavera, enquanto as pessoas na faixa etária de 16 a 50 anos foram mais frequentemente afetadas. O alimento mais comumente relacionado aos surtos foi a maionese caseira (42,45%), enquanto as principais causas das salmoneloses foram a utilização de matéria-prima sem inspeção (22,92%), na grande maioria ovos, e alimentos mantidos à temperatura ambiente por mais de 2 horas (20,55%). A maioria dos surtos ocorreu dentro de residências (43,70%) e estabelecimentos comerciais (25,21%).

Salmonella spp. was the major cause of reported foodborne diseases in the last years in the State of Rio Grande do Sul, South of Brazil. Epidemiological data on salmonellosis, occurred in the period of 1997 to 1999 and supplied by the Division of Sanitary Surveillance (DVS/RS), were analysed according to the following factors: total number of confirmed outbreaks, number of people involved, outbreaks incidence according to the period of the year, age and sex of involved people, food vehicle, food storage conditions, local where the disease occurred, and possible causes of the outbreaks. The results indicated that 8217 people were involved, and 1557 had to be hospitalized. The highest number of outbreaks occurred during springtime and the principal age group affected was between 16 and 50 years. The most common food vehicle was salad prepared with homemade mayonnaise (42.45%). The principal causes of salmonellosis were raw-materials not submitted to regulatory inspection (22.92%), mainly eggs, and foods maintained at room temperature for more than 2 hours (20.55%). The majority of the outbreaks occurred in private homes (43.70%) and commercial food establishments (25.21%).