An Evaluation Of Immunomagnetic Separation-Real-Time Pcr (Ims-Rtipcr) Combined Assay For Rapid And Specific Detection Of Escherichia Coli O157:H7 In Raw Milk And Ground Beef

The aim of this study is to optimize a rapid and specific method for detection of E. coli O157:H7 (EHEC) from food samples with high background flora using an immunomagnetic separation-real-time polymerase chain reaction assay (IMS-RTiPCR). For this, EHEC cells were recovered from raw milk and ground meat samples that were artificially contaminated to the final concentrations of 10(1), 10(3) and 10(5) cfu EHEC/mL or g, after a non-selective pre-enrichment at 35 degrees C for 8 h following either with or without capturing by micro-sized beads. Then, EHEC cells were identified by RTiPCR. The study was also carried out without any enrichment to evaluate the enrichment efficiency of the assay. By comparing the assay with and without pre-enrichment, we showed that even the usage of specific micro-sized beads did not improve EHEC detection in raw milk and ground beef samples and so gave false negative results unless an 8-h pre-enrichment was applied. Besides, the assay can be completed in less than 9 h with a minimum detection limit of 10(3) cfu EHEC/mL or g in foods and hence was found to be simple routine-based method that exhibits great potential for implementing as a rapid screening of EHEC in microbial tests as compared to existing techniques.WoSScopu

Characterization Of Antibiotic Resistance In Salmonella Enterica Isolates Determined From Ready-To-Eat (Rte) Salad Vegetables

In the last decade, ready-to-eat (RTE) salad vegetables are gaining increasing importance in human diet. However, since they are consumed fresh, inadequate washing during processing can bring on some foodborne illnesses, like salmonellosis, since these food items have natural contamination from soil and water. During 2009–2010, a total of 81 samples were purchased arbitrarily from local markets in Ankara, and were examined for Salmonella contamination. Salmonella screening was performed by using anti-Salmonella magnetic beads system and polymerase chain reaction (PCR) identification of the suspected colonies. Then, the antibiotic resistance profiles of four Salmonella strains identified (strains RTE-1, RTE-2, RTE-3, and RTE-4) were also investigated, since the mechanism by which Salmonella spp. have accumulated antibiotic resistance genes is of interest. All strains showed resistance against sulfonamides (MIC > 128 mg/L). Further results suggested that associated sulfonamide resistance genes were encoded by the 55.0 kb plasmid of strain RTE-1 that involves no integrons. As a result of using two primers (P1254 and P1283) in randomly amplified polymorphic DNA-PCR (RAPD-PCR) analysis, two common amplicons (364 bp and 1065 bp) were determined. The findings of this study provide support to the adoption of guidelines for the prudent use of antibiotics in order to reduce the number of pathogens present on vegetable and fruit farms. Besides, since it is shown that these bacteria started to gain resistance to antibiotics, it is necessary to further investigate the prevalence of them in foods.PubMedWoSScopu

Application Of Bacteriocin-Like Inhibitory Substances (Blis)-Producing Probiotic Strain Of Lactobacillus Plantarum In Control Of Staphylococcus Aureus In White-Brined Cheese Production

The aim of this study was to investigate the antimicrobial activity of an autochthonous probiotic strain of bacteriocin-like inhibitory substances (BLIS)-producing Lactobacillus plantarum, previously isolated from a Tulum cheese and satisfied technological criteria as adjunct culture in cheese production, in reducing Staphylococcus aureus during production and ripening of white-brined cheeses. Cheeses were manufactured in two trials from pasteurized milk artificially contaminated with S. aureus to the mean level of 6.243 log MPN mL(-1). Lb. plantarum BG33 was added at 1% as adjunct to the starter culture. The study was also carried out with control group cheeses produced without the adjunct culture. S. aureus counts were monitored for up to 90 days by RAM's 5-tube MPN method and each positive tube of MPN (most probable number) method was confirmed by PCR amplification of a 400 bp fragment of the nuc gene. which encodes the thennostable nuclease of S. aureus. The capacity of lb. plantarunt BG33 to reduce S. aureus count was found as 0.9 log unit on the 18th day of ripening. After 39 and 59 days of ripening, Lb. plantarum BG33 lowered S. aureus count by 1.9 and 2.0 log units, respectively, when compared to control group cheeses in which it was lowered by 0.5 and 1.0 log units, respectively. As a result, the BLIS activity of Lb. plantarum BG33 throughout ripening of white-brined cheese could make it useful as bioprotective adjunct culture in white-brined cheese production to prevent S. aureus growth which is an important foodborne pathogen in respect of safe cheese production.WoSScopu