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Abstract: The aim of this study was to evaluate the occurrence of Campylobacter in meat samples (120 chicken and 100 beef) and determine the antimicrobial susceptibility of the isolates. A total of 220 samples from retail outlets were purchased in the city of São Paulo, Brazil. Campylobacter detection was performed according to the International Organization for Standardization (ISO) method ISO‐10272‐1:2006. A PCR assay based on nucleotide sequence differences in the lpxA gene was used to distinguish between C. jejuni and C. coli. Antimicrobial resistance was determined by agar disc diffusion method. Campylobacter was isolated from 17 (7.7%) of 220 samples. Breast fillets exhibited the highest contamination rate (25%; 5/20), followed by wings (15%; 6/40), whole leg (15%; 3/20), drumstick (10%; 2/20), and drumette (5%; 1/20). All beef samples were negative for Campylobacter. The most prevalent species found was C. coli, followed by C. jejuni. The isolates were commonly resistant to nalidixic acid and ciprofloxacin. Data obtained confirm the need of monitoring and control of Campylobacter in poultry production chain. Practical applications: Campylobacter spp. cause foodborne illness in humans commonly through the consumption of contaminated poultry meat. Although Brazil is the world's largest poultry meat exporter, data regarding this pathogen are limited in our country. In the present study, chicken cuts purchased from retail stores in São Paulo may occasionally be contaminated with Campylobacter. This underline the importance of surveillance of foodborne pathogens in retail meats, and the data can contribute to risk analyses or control measures in the meat production chain. [ABSTRACT FROM AUTHOR]

This study evaluated the effects of irradiation on the reduction of Shiga toxin-producing Escherichia coli (STEC), Salmonella strains, and Listeria monocytogenes, as well as on the sensory characteristics of minimally processed spinach. Spinach samples were inoculated with a cocktail of three strains each of STEC, Salmonella strains, and L. monocytogenes, separately, and were exposed to gamma radiation doses of 0, 0.2, 0.4, 0.6, 0.8, and 1.0 kGy. Samples that were exposed to 0.0, 1.0, and 1.5 kGy and kept under refrigeration (4°C) for 12 days were submitted to sensory analysis. D10-values ranged from 0.19 to 0.20 kGy for Salmonella and from 0.20 to 0.21 for L. monocytogenes; for STEC, the value was 0.17 kGy. Spinach showed good acceptability, even after exposure to 1.5 kGy. Because gamma radiation reduced the selected pathogens without causing significant changes in the quality of spinach leaves, it may be a useful method to improve safety in the fresh produce industry. [ABSTRACT FROM AUTHOR]

This study was aimed at determining the effects of different storage scenarios on the growth potential of Salmonella strains and Listeria monocytogenes in ready-to-eat (RTE) mixes of iceberg and crisp lettuces (Lactuca sativa) and collard greens (Brassica oleracea). Vegetables were submitted to minimal processing, experimentally contaminated to achieve 10¹ and 10² CFU/g, packed under modified atmosphere and in perforated film, and submitted to the following storage scenarios: I = 100% of the shelf life (6 days) at 7°C; II = 70% of shelf life at 7°C and 30% at 15°C; III = 30% at 7°C and 70% at 15°C; IV = 100% at 15°C. Higher populations of Salmonella were observed in lettuce mixes than in collard greens; the opposite occurred with L. monocytogenes. Keeping the RTE vegetables at 15°C during the whole shelf life (scenario IV) or part of it (scenarios II and III) markedly influenced the growth of both pathogens in most of the scenarios studied (P < 0.05). Growth potentials of strains of Salmonella and L. monocytogenes were significantly different depending on the scenarios in samples packed with perforated film in comparison to those stored under modified atmosphere (P < 0.05). The findings indicate that even contamination as low as 10¹ CFU/g can lead to high populations if there is temperature abuse during storage (15°C). This study of the behavior of Salmonella and L. monocytogenes in RTE vegetables provides insights that may be useful in the development of strategies to control pathogen growth in these products. [ABSTRACT FROM AUTHOR]

This study was the first conducted in Brazil to evaluate the presence of Enterobacter sakazakii in milk-based powdered infant formula manufactured for infants 0 to 6 months of age and to examine the conditions of formula preparation and service in three hospitals in São Paulo State Brazil. Samples of dried and rehydrated infant formula, environments of milk kitchens, water, bottles and nipples, utensils, and hands of personnel were analyzed, and E. sakazakii and Enterobacteriaceae populations were determined. All samples of powdered infant formula purchased at retail contained E. sakazakii at <0.03 most probable number (MPN)/100 g. In hospital samples, E. sakazakii was found in one unopened formula can (0.3 MPN/100 g) and in the residue from one nursing bottle from hospital A. All other cans of formula from the same lot bought at a retail store contained E. sakazakii at <0.03 MPN/100 g. The pathogen also was found in one cleaning sponge from hospital B. Enterobacteriaceae populations ranged from 10¹ to 105 CFU/g in cleaning aids and <5 CFU/g in all formula types (dry or rehydrated), except for the sample that contained E. sakazakii, which also was contaminated with Enterobacteriaceae at 5 CFU/g. E. sakazakii isolates were not genetically related. In an experiment in which rehydrated formula was used as the growth medium, the temperature was that of the neonatal intensive care unit (25°C), and the incubation time was the average time that formula is left at room temperature while feeding the babies (up to 4 h), a 2-log increase in levels of E. sakazakii was found in the formula. Visual inspection of the facilities revealed that the hygienic conditions in the milk kitchens needed improvement. The length of time that formula is left at room temperature in the different hospitals while the babies in the neonatal intensive care unit are being fed (up to 4 h) may allow for the multiplication of E. sakazakii and thus may lead to an increased health risk for infants. [ABSTRACT FROM AUTHOR]

In Brazil, the incidence of Bacillus cereus outbreaks is unknown, and there is little information about B. cereus occurrence in food. In addition, data on toxin production and genetic characterization of the B. cereus isolates cannot be found. This pathogen causes two distinct types of toxin-mediated foodborne illnesses known as diarrheal and emetic syndromes. Diarrheal syndrome has been linked to three different enterotoxins: two protein complexes, hemolysin BL (HBL) and nonhemolytic enterotoxin (NHE); and an enterotoxic protein, cytotoxin K (cytK). Emetic syndrome is related to cereulide, a toxin encoded by the ces gene. In this study, NHE and HBL production capacities of 155 strains of B. cereus isolated from Brazilian food products were evaluated with an immunoassay. Strains were also tested for the presence of the genes of the HBL and NHE complexes, cytK, cytK-1, cytK-2, and ces, using PCR. HBL was detected in 105 (67.7%) strains and NHE in 154 (99.4%) strains. All the strains harbored at least one gene of the NHE complex, while 96.1% of them were positive for at least one of those of the HBL complex. Genes cytK1 and ces were not detected. All strains showed toxigenic capacity and could represent a risk for consumers if good practices are not followed. This is the first report on toxigenic and genetic profiles of B. cereus strains isolated in Brazil. [ABSTRACT FROM AUTHOR]

The increasing demand for fresh fruits and vegetables and for convenience foods is causing an expansion of the market share for minimally processed vegetables. Among the more common pathogenic microorganisms that can be transmitted to humans by these products are Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella. The aim of this study was to evaluate the microbial quality of a selection of minimally processed vegetables. A total of 181 samples of minimally processed leafy salads were collected from retailers in the city of Sao Paulo, Brazil. Counts of total coliforms, fecal coliforms, Enterobacteriaceae, psychrotrophic microorganisms, and Salmonella were conducted for 133 samples. L. monocytogenes was assessed in 181 samples using the BAX System and by plating the enrichment broth onto Palcam and Oxford agars. Suspected Listeria colonies were submitted to classical biochemical tests. Populations of psychrotrophic microorganisms > 106 CFU/g were found in 51% of the 133 samples, and Enterobacteriaceae populations between 105 and 106 CFU/g were found in 42% of the samples. Fecal coliform concentrations higher than 10² CFU/g (Brazilian standard) were found in 97 (73%) of the samples, and Salmonella was detected in 4 (3%) of the samples. Two of the Salmonella-positive samples had < 10² CFU/g concentrations of fecal coliforms. L. monocytogenes was detected in only 1 (0.6%) of the 181 samples examined. This positive sample was simultaneously detected by both methods. The other Listeria species identified by plating were L. welshimeri (one sample of curly lettuce) and L. innocua (2 samples of watercress). The results indicate that minimally processed vegetables had poor microbiological quality, and these products could be a vehicle for pathogens such as Salmonella and L. monocytogenes. [ABSTRACT FROM AUTHOR]

An outbreak of staphylococcal food poisoning involving approximately 180 people occurred in Brodowski, Sao Paulo State, Brazil, in April 1998. Strains of Staphylococcus aureus isolated from foods and food handlers, implicated as the etiologic agent, were characterized with phenotypic (phage typing, antibiotic susceptibility test, and enterotoxin production), and genotypic (random amplified polymorphic DNA) characterization. Strains isolated from vegetable salad with mayonnaise sauce, broiled chicken, pasta in tomato sauce, and from the oropharyngeal secretions of five food handlers--A, B, C, H, and I--showed the same phage profile and antibiotic resistance. Random amplified polymorphic DNA generated 17 combined profiles with primers OPE-20 and OPA-7. The similarity of strains was analyzed by generating a dendrogram that classified the 59 strains of S. Aureus into four major clusters (I, II, III, and IV). Strains from four food handlers (A, B, H, and I) and from vegetable salad with mayonnaise, broiled chicken, and pasta in tomato sauce showing the same phage type profile and resistance to antibiotics belonged to the same cluster and produced staphylococcal enterotoxin A. Therefore, these foods and food handlers were incriminated in the outbreak. [ABSTRACT FROM AUTHOR]

Data on the prevalence and growth of Listeria monocytogenes in lightly preserved fish products from subtropical and tropical regions are very scarce. Our research describes L. monocytogenes that was detected in 5% of the packages of cold-smoked surubim, a native Brazilian freshwater fish that we analyzed, and shows that the strains isolated were of the same random amplified polymorphic DNA subtype as the strains that were isolated from the same factory 4 years earlier. A bacteriocinogenic strain of Carnobacterium piscicola (strain C2), isolated from vacuum-packed cold-smoked surubim, and two C. piscicola strains, isolated from vacuum-packed, cold-smoked salmon, were capable of limiting or completely inhibiting the growth of an L. monocytogenes (strain V2) isolated from surubim in fish peptone model systems incubated at 10°C. Monocultures of L. monocytogenes reached 108 CFU/ml (g), whereas the growth of L. monocytogenes was completely inhibited by C. piscicola C2. The bacteriocinogenic C. piscicola A9b+ and its nonbacteriocinogenic mutant A9b- reduced maximum Listeria levels by 2 to 3 log units. Both bacteriocinogenic C. piscicola strains prevented listerial growth in cold-smoked fish juices (surubim and salmon). Although the carnobacteria grew poorly on cold-smoked surubim at 10°C, the strains were able to reduce maximum Listeria counts by 1 to 3 log units in an artificially inoculated product (surubim). We conclude that Brazilian smoked fish products harbor L monocytogenes and should be stabilized against the growth of the organism. C. piscicola C2 has the potential for use as a bioprotective culture in surubim and other lightly preserved fish, but further studies are required to optimize its effect. [ABSTRACT FROM AUTHOR]

Control of Salmonella spp. in food production chains is very important to ensure safe foods and minimize the risks of foodborne disease occurrence. This study aimed to identify the prevalence and main contamination sources of Salmonella spp. in a pig production chain in southern Brazil. Six lots of piglets produced at different farms were tracked until their slaughter, and samples were subjected to Salmonella spp. detection. The obtained isolates were serotyped, subjected to antimicrobial resistance testing, and pulsed field gel electrophoresis (PFGE). Salmonella spp. was detected in 160 (10.2%) samples, and not detected in pig carcasses after final washing or chilling. Among the 210 Salmonella spp. isolates, S. Typhimurium was the most prevalent (n = 101) and resistant to at least one antimicrobial. High resistance rates were detected against tetracycline (83.8%), chloramphenicol (54.3%), and trimethoprim-sulfamethoxazole (33.3%). The isolates that were non-susceptible to three or more classes of antimicrobials (n = 60) were considered multidrug-resistant (MDR), and isolates resistant to up to six of the tested antimicrobials were found. PFGE allowed the identification of genetic diversity and demonstrated that farm environment and feed supply may be sources for the dissemination of Salmonella spp. along the production chain. The results revealed the sources of Salmonella contamination in the pig production chain and highlighted the risks of antimicrobial resistance spread. [ABSTRACT FROM AUTHOR]

Abstract: Listeria monocytogenes is a pathogen capable of adhering to many surfaces and forming biofilms, which may explain its persistence in food processing environments. This study aimed to genetically characterise L. monocytogenes isolates obtained from bovine carcasses and beef processing facilities and to evaluate their adhesion abilities. DNA from 29 L. monocytogenes isolates was subjected to enzymatic restriction digestion (AscI and ApaI), and two clusters were identified for serotypes 4b and 1/2a, with similarities of 48% and 68%, respectively. The adhesion ability of the isolates was tested considering: inoculum concentration, culture media, carbohydrate source, NaCl concentration, incubation temperature, and pH. Each isolate was tested at 108 CFUmL−1 and classified according to its adhesion ability as weak (8 isolates), moderate (17) or strong (4). The isolates showed higher adhesion capability in non-diluted culture media, media at pH 7.0, incubation at 25°C and 37°C, and media with NaCl at 5% and 7%. No relevant differences were observed for adhesion ability with respect to the carbohydrate source. The results indicated a wide diversity of PFGE profiles of persistent L. monocytogenes isolates, without relation to their adhesion characteristics. Also, it was observed that stressing conditions did not enhance the adhesion profile of the isolates. [Copyright &y& Elsevier]

Several alternative analytical methods are currently available for the rapid microbiological testing of food. Due to their many advantages, particularly their convenience of use, the popularity of ready-to-use systems for the enumeration of hygiene indicator microorganisms is increasing. However, the ability of these systems to enumerate stressed microorganisms, such as those that may be found growing in acidic foods, is unknown. Therefore, the aim of this study was to evaluate the performance of Petrifilm™ and SimPlate™ plates for the enumeration of total aerobes and fungi (yeasts and molds) in acidic fruit juices, using non-pasteurized orange juice as an experimental model. The samples were analyzed before and after neutralization of pH, and the results were compared with those obtained using conventional procedures, i.e. pour-plates containing Standard Methods Agar, acidified potato dextrose agar, or dichloran-glycerol agar. The results obtained with Petrifilm and SimPlate for counts of mesophilic aerobes as well as for yeast and mold correlated well with those obtained using conventional procedures. Although no statistically significant differences were observed between counts of non-neutralized and neutralized samples (α ≤ 0.05), better correlation indexes were observed in the neutralized samples. Both Petrifilm and SimPlate proved to be good alternative methods for testing the microbiological quality of acidic fruit juices. [Int Microbiol 2005; 8(1):49-53] [ABSTRACT FROM AUTHOR]

Lactobacillus sake 2a is a bacteriocinogenic strain isolated from “lingu¨ic¸a frescal”, a Brazilian sausage. The combined effect of modified-atmosphere (MA) packaging (100% CO2 and 50% CO2/50% N2) and addition of L. sake 2a on inhibition of growth of Listeria monocytogenes was evaluated in “lingu¨ic¸a” stored at 6 °C. By the end of the first week, the inhibition of L. monocytogenes due to MA was significant (P&les;0.05) while the presence of L. sake 2a did not influence significantly the growth of the pathogen. After 14 days, a reduction of 1.3–1.4 log in counts of L. monocytogenes was observed in samples containing L. sake 2a only or MA packaged only, while a reduction of 3.5 log was detected in those submitted to both treatments. Results indicate that inhibition of L. monocytogenes in “lingu¨ic¸a frescal” by the bacteriocinogenic L. sake 2a is enhanced by the packaging of the product in MA. [Copyright &y& Elsevier]

Abstract: Minimally processed vegetables (MPV) may be important vehicles of Salmonella spp. and cause disease. This study aimed at detecting and enumerating Salmonella spp. in MPV marketed in the city of São Paulo, Brazil. A total of 512 samples of MPV packages collected in retail stores were tested for Salmonella spp. and total coliforms and Escherichia coli as indication of the hygienic status. Salmonella spp. was detected in four samples, two using the detection method and two using the counting method, where the results were 8.8 × 102 CFU/g and 2.4 × 102 CFU/g. The serovars were Salmonella Typhimurium (three samples) and Salmonella enterica subsp. enterica O:47:z4,z23:- (one sample). Fourteen samples (2.7%) presented counts of E. coli above the maximum limit established by the Brazilian regulation for MPV (102 CFU/g). Therefore, tightened surveillance and effective intervention strategies are necessary in order to address consumers and governments concerns on safety of MPV. [Copyright &y& Elsevier]

Abstract: The bacteriocin-producing strain Enterococcus faecium ST5Ha was isolated from smoked salmon and identified by biomolecular techniques. Ent. faecium ST5Ha produces a pediocin-like bacteriocin with activity against several lactic acid bacteria, Listeria spp. and some other human and food pathogens, and remarkably against HSV-1 virus. Bacteriocin ST5Ha was produced at high levels in MRS broth at 30 °C and 37 °C, reaching a maximum production of 1.0 × 109 AU/ml, checked against Listeria ivanovii ATCC19119 as target strain and surrogate of pathogenic strain Listeria monocytogenes. The molecular weight of bacteriocin ST5Ha was estimated to be 4.5 kDa according to tricine-SDS-PAGE data. Ent. faecium ST5Ha harbors a 1.044 kb chromosomal DNA fragment fitting in size to that of pediocin PA-1/AcH. In addition, the sequencing of bacteriocin ST5Ha gene indicated 99% of DNA homology to pediocin PA-1/AcH. The combined application of low levels (below MIC) of ciprofloxacin and bacteriocin ST5Ha resulted in a synergetic effect in the inhibition of target strain L. ivanovii ATCC19119. Bacteriocin ST5Ha displayed antiviral activity against HSV-1, an important human pathogen, with a selectivity index of 173. To the best of our knowledge, this is the first report on Ent. faecium as a potential producer of pediocin-like bacteriocin with antiviral activity. [ABSTRACT FROM AUTHOR]

Abstract: Linguiça is a highly popular and appreciated pork product in Brazil, frequently consumed undercooked. Aiming at collection of data for a future risk assessment, this study evaluated the prevalence and counts of Listeria monocytogenes in linguiça samples collected at retail level in Sao Paulo, SP, Brazil. ISO methods were used for detection and enumeration of the pathogen (11290-1 and 11290-2, respectively). Isolates were submitted to Simplex-PCR for hlyA gene and those with biochemical features of L. monocytogenes and hlyA positive were serotyped using a Multiplex PCR. Ninety percent of the samples were positive for Listeria spp., and L. monocytogenes was detected in 42% of the samples, with counts below 102 CFU/g in all samples. A prevalence of uncommon serotypes 4a and 4c was observed. [Copyright &y& Elsevier]

The routine methods for detecting Listeria sp. in foods are time consuming and involve using selective enrichments and plating on agars. In this study, the presence of Listeria sp. in 120 meat and meat product samples was investigated by two rapid immunoassays (TECRA Listeria Visual Immunoassay [VIA] and BioControl Visual Immunoprecipitate Assay [VIP] for Listeria ) and a cultural procedure. The cultural method of detecting Listeria sp. followed Canada's Health Protection Branch Method, and the rapid tests followed the manufacturers' instructions. The agreement between the cultural and the rapid tests was established at a confidence limit of 95%. Seventy-nine samples (65.8%) were Listeria sp. positive in at least one of the three tests. There was no statistically significant difference between the cultural procedure and any of the rapid immunoassays. The agreement rates between the VIA and the cultural method and between the VIP and the cultural method were 87 and 84%, respectively. Both tests – the VIA and VIP – proved to be rapid, efficient and easy to perform. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a bacterium capable to adhere to the surfaces of equipment and utensils and subsequently form biofilms. It can to persist in the food processing environmental for extended periods of time being able to contaminate the final product. The aim of this study was to trace the contamination route of L. monocytogenes on a fresh mixed sausage processing line, from raw material to the final product. The isolates obtained were characterized by serotyping and molecular typing by pulsed-field gel electrophoresis (PFGE) using the restriction enzymes ApaI and AscI. L. monocytogenes was detected in 25% of the samples. The samples of raw material were not contaminated, however, the microorganism was detected in 21% of the environmental samples (food contact and non-food contact), 20.8% of the equipments, 20% of the food worker's hands, 40% of the mass ready to packaging and in all the final products samples, demonstrating that the contamination of final product occurred during the processing and the importance of cross contamination. PFGE yielded 22 pulsotypes wich formed 7 clusters, and serotyping yielded 3 serotypes and 1 serogroup, however, the presence of serotypes 4b and 1/2b in the final product is of great concern for public health. The tracing of contamination showed that some strains are adapted and persisted in the processing environment in this industry.

This study evaluated the growth of naturally occurring L. monocytogenes in sliced, vacuum-packed mortadella samples during storage at 5°C until the expiration date. Tukey's test indicated that counts of L. monocytogenes on 0, 10, 20, 30 and 40 days of storage were significantly different (p<0.05), indicating growth during shelf life. In three trials, the mean increase was 1.72 log cycles. Vacuum packing and storage under refrigeration were not effective in controlling the growth of L. monocytogenes in sliced mortadella, indicating that good manufacturing practices and implemented HACCP programs are essential to assure safety of this product.

This work studied the radiation resistance of Listeria monocytogenes and Salmonella species and the effect of irradiation on leaf flavonoid content and sensory acceptability of minimally processed arugula. Immersion in ozone-treated water reduced the analyzed microorganisms by 1 log. L. monocytogenes and Salmonella were not isolated from samples. Samples of this vegetable were inoculated with a cocktail of Salmonella spp. and L. monocytogenes and exposed to gamma irradiation. D10 values for Salmonella ranged from 0.16 to 0.19 kGy and for L. monocytogenes from 0.37 to 0.48 kGy. Kaempferol glycoside levels were 4 and ca. 3 times higher in samples exposed to 1 and 2 kGy, respectively, than in control samples. An increase in quercetin glycoside was also observed mainly in samples exposed to 1 kGy. In sensory evaluation, arugula had good acceptability, even after exposure to 2 and 4 kGy. These results indicate that irradiation has potential as a practical processing step to improve the safety of arugula.

Several alternative analytical methods are currently available for the rapid microbiological testing of food. Due to their many advantages, particularly their convenience of use, the popularity of ready-to-use systems for the enumeration of hygiene indicator microorganisms is increasing. However, the ability of these systems to enumerate stressed microorganisms, such as those that may be found growing in acidic foods, is unknown. Therefore, the aim of this study was to evaluate the performance of Petrifilm(tm) and SimPlate(tm) plates for the enumeration of total aerobes and fungi (yeasts and molds) in acidic fruit juices, using non-pasteurized orange juice as an experimental model. The samples were analyzed before and after neutralization of pH, and the results were compared with those obtained using conventional procedures, i.e. pour-plates containing Standard Methods Agar, acidified potato dextrose agar, or dichloran-glycerol agar. The results obtained with Petrifilm and SimPlate for counts of mesophilic aerobes as well as for yeast and mold correlated well with those obtained using conventional procedures. Although no statistically significant differences were observed between counts of non-neutralized and neutralized samples (alpha >/== 0.05), better correlation indexes were observed in the neutralized samples. Both Petrifilm and SimPlate proved to be good alternative methods for testing the microbiological quality of acidic fruit juices.

The presence of Salmonella in 200 raw food samples of animal origin was investigated by means of the rapid immunoassays Tecra Salmonella VIA and Tecra Salmonella UNIQUE (Tecra Diagnostics, Rosewille, New South Wales, Australia) and a cultural procedure. Forty-five samples (22.5%) were Salmonella positive by at least one of the three methods. The number of positive samples according to the analytical method was 34 (75.6%) for the cultural procedure, 29 (64.4%) for Tecra Salmonella VIA, and 27 (60.0%) for Tecra Salmonella UNIQUE. Tecra Salmonella UNIQUE detected three positive samples that were not detected by the two other methods. The cultural method also detected three positive samples that both rapid methods were unable to detect. McNemar's chi-square tests indicated that the differences between results given by the rapid immunoassays when compared with those of the cultural method were not significant (P > 0.05).