Virulence genes and sanitizers resistance in Salmonella isolates from eggs in southern Brazil.

Salmonella spp. causes foodborne diseases related to the consumption of contaminated foods, especially poultry products. This study aimed to investigate the occurrence of Salmonella spp. serovars in raw eggs from supermarkets and street food markets in southern Brazil, to analyze virulence genes, resistance profiling to antimicrobials and sanitizers, and to determine the susceptibility of the isolates to Butia odorata extract. Among 160 samples analyzed, just two (1.25%) were positive for Salmonella spp.. One positive sample was from egg yolk (S. enterica serovar Gallinarum, isolate S28), and another one was from eggshell (S. enterica serovar Panama, isolate S37). Regarding the virulence genes, the isolate S37 harbored all the genes evaluated (hilA, invA, spvC, sefA, and pefA), while the isolate S28 did not harbor the pefA gene. The isolate S28 was resistant to tobramycin, azithromycin, and trimethoprim, while the isolate S37 showed resistance profile just to nalidixic acid. However, none of the resistance genes evaluated were identified. Both isolates showed resistance to benzalkonium chloride, chlorhexidine digluconate, sodium hypochlorite, and peracetic acid, presenting high MIC values for these sanitizers. In contrast, B. odorata extract showed antimicrobial activity against the isolates S28 and S37, however, more studies are needed to prove its potential as a natural antimicrobial compound.

Listeria monocytogenes in sliced cheese and ham from retail markets in southern Brazil.

The aims of this study were to evaluate the occurrence of Listeria monocytogenes and Salmonella spp. in sliced cheese and ham from retail markets in southern Brazil, as well as to perform molecular characterization and to assess the antimicrobial resistance profile of the isolates. Samples (n = 160) of sliced cheese and ham were collected at retail level from the city of Pelotas, Brazil. The isolation of L. monocytogenes and Salmonella spp. was performed and the isolates were confirmed by PCR, submitted to antimicrobial susceptibility testing and pulsed-field gel electrophoresis (PFGE). Listeria monocytogenes was found in 9.4% (15/160) of the samples. All L. monocytogenes isolates were positive for the prs, inlA, inlC and inlJ genes. Salmonella spp. was not isolated. Regarding the antimicrobial susceptibility, one (6.6%) L. monocytogenes isolate was resistant to streptomycin and four (26.6%) to clindamycin. Macrorestriction analysis with ApaI and AscI enzymes yielded two major PFGE groups I and II. All L. monocytogenes isolates showed virulence genes, and some of them were resistant to clinically used antimicrobials, representing a risk to public health. Moreover, PFGE patterns with high similarity were visualized in L. monocytogenes isolates at different times, demonstrating adaptability of the pathogen at retail level in the region.

Butia odorata Barb. Rodr. extract inhibiting the growth of Escherichia coli in sliced mozzarella cheese.

The aims of this study were to verify the occurrence of Escherichia coli in sliced mozzarella cheese marketed in Pelotas city, Brazil and perform the phenotypic and genotypic characterization of the isolates. Besides that, evaluate the susceptibility of E. coli to Butia odorata extract, characterize it chemically, and apply the extract in sliced mozzarella cheese contaminated experimentally with E. coli. Escherichia coli was isolated in 5% (4/80) of cheese samples, but no gene used as marker for E. coli O157:H7 or virulence genes were detected. The isolates were susceptible to B. odorata extract (MIC 15 mg mL-1 and MBC 29-58 mg mL-1), and the major compounds present in the extract were Z-10-Pentadecenol (80.1%) and Palmitic acid (19.4%). In cheese, after 72 h there was a significant difference between control (2.8 log CFU cm-2) and treated samples with MIC, 2 × MIC, 4 × MIC and 8 × MIC (1.3, 1.4, 1.6 and 0.5 log CFU cm-2, respectively). The isolation of E. coli in cheese indicates fecal contamination and poor hygienic practices. Butia odorata extract showed antimicrobial activity against E. coli both in vitro and in situ, indicating that it can be a good alternative for inhibiting the growth of this microorganism in sliced cheese.

Occurrence and characterization of Listeria monocytogenes from beef jerky processing line.

Beef jerky is a ready-to-eat product that does not require refrigeration at the point of sale. Here, we evaluated the occurrence of Listeria monocytogenes in the production process of beef jerky, the presence of virulence genes and the genomic relatedness of the isolates, to assess the safety of the final product. The raw material, surfaces with and without contact with the product and the final product were evaluated along the beef jerky processing line. The samples were evaluated by VIDAS immunoassay system, and the L. monocytogenes isolates were confirmed and evaluated for the presence of several virulence genes by PCR. Listeria monocytogenes was identified in six of the 84 samples (7.14%), and no genetic relationship was observed among isolates. Samples of raw material (2/7), food contact surface (1/56), and work surfaces without contact with food (3/14) presented contamination by L. monocytogenes. The final product was not contaminated, demonstrating that barriers to multiplication of pathogens used during the production process were effective for its control.

Tolerance to benzalkonium chloride and antimicrobial activity of Butia odorata Barb. Rodr. extract in Salmonella spp. isolates from food and food environments.

Salmonellosis, caused by the consumption of contaminated foods, is a major health problem worldwide. The aims of this study were to assess the susceptibility of Salmonella spp. isolates to benzalkonium chloride (BC) disinfectant and the antimicrobial activity of Butia odorata Barb. Rodr. extract against the same isolates from food and food environments. Moreover, phenotypic and genotypic resistance profiles, the presence of virulence genes and biofilm forming ability were determined. The minimum inhibitory concentration (MIC) of B. odorata extract against Salmonella spp. ranged from 10 to >19 mg.mL-1. Resistance to ampicillin, streptomycin, nalidixic acid, sulfonamide, trimethoprim/sulfamethoxazole, trimethoprim, tetracycline, and chloramphenicol was observed. In addition, multidrug resistance was observed in seven isolates (26.92%). The MIC of BC ranged from 32 to 64 mg.L-1, higher concentrations in comparison with wild-type MICs, and therefore were considered tolerant. Several resistance genes were detected, of which the most common were aadA, qacEΔ1, blaTEM, int1, sul1, and tetA. All isolates carried at least one virulence gene and produced biofilms on stainless steel surfaces at 10 and 22 °C. On the other hand, the B. odorata extract showed activity against Salmonella spp., and it has the potential to be used as a natural antimicrobial to control this important foodborne pathogen, despite its virulence potential and antimicrobial resistance profile.

Antibacterial activity of Timsen® (n-alkyl dimethyl benzyl ammonium chloride-40%) in scalding and precooling water in poultry slaughterhouses.

The objective of this study was to evaluate the efficacy of a product based on n-alkyl dimethyl benzyl ammonium chloride-40%, marketed as Timsen®, during scalding and precooling of poultry carcasses in slaughterhouses. To this end, three poultry slaughterhouses (A, B and C) were evaluated. The product was added (200 ppm) to the scalding (58 °C) and precooling water (4 °C), and microbiological analyses were performed of the water and the poultry carcasses before and after Timsen® addition. The product controlled the multiplication of aerobic mesophilic microorganisms, both in the scalding as in the precooling water. In a comparison of carcasses soaked in Timsen®-treated scalding and precooling water with carcasses soaked in untreated water, the count of aerobic mesophilic microorganisms in the later was higher and thermotolerant coliform was not detected in samples of carcasses soaked in Timsen®-treated water. When the scalding and precooling water was not treated with the product, Listeria spp. was isolated from poultry carcasses of two slaughterhouses (A and C), while these microorganisms were not detected when Timsen® was applied. The use of Timsen® in the scalding and precooling water enhanced the safety and control microbial contamination of poultry carcasses.