The microbial inhibition of foodborne pathogens was determined in brain heart infusion broth with 10% (wt/vol) water-soluble extracts of green, jasmine, black, dungglre, and oolong tea against Escherichia coli O157:H7, Salmonella enterica serovar Enteritidis, Listeria monocytogenes, and Staphylococcus aureus. The mixed culture (approximately 6.0 log CFU/ml), which was composed of the four pathogens, was inoculated into brain heart infusion broth with and without tea extracts. After incubation at 35 degrees C for 0, 1, 3, and 5 days, proper dilution of each sample was spiral plated on each selective agar. Viable cell counts were performed after incubation at 35 degrees C for 24 to 36 h. Green, jasmine, and black tea exhibited an approximately 5.0 log suppression of S. aureus compared with the control from days 1 to 5. Green and jasmine tea also suppressed the growth of L. monocytogenes by approximately 3.0 log CFU/ml on day 5. In contrast, no tea extracts inactivated E. coli O157:H7 and Salmonella Enteritidis. Based on the result in liquid medium, green and jasmine teas of 0.1% (vol/wt) were individually evaluated for their antimicrobial activity against L. monocytogenes and S. aureus in a food model (ground beef) stored at 7 degrees C for 0, 1, 3, 5, and 7 days. Viable cell counts of total bacteria, L. monocytogenes, and S. aureus in ground beef were not significantly different among green and jasmine tea and the control.
A colony polymerase chain reaction (PCR) technique was applied with an established Fung's double tube (FDT) method for rapid detection and confirmation of Clostridium perfringens. Published sequences of PCR primers for C. perfringens alpha toxin gene were used and PCR conditions were optimized. From the detection of C. perfringens by FDT tube to the confirmation by a colony PCR assay took as short as 16–18 h. The method was applied to 147 isolates of anaerobic sulfite reducing bacteria isolated from foods, sewages and animal clinical specimens. The results were compared with standard methods for the confirmation of C. perfringens. Of those 147 suspected isolates, 97 and 99 were confirmed as C. perfringens by standard methods and the colony PCR technique, respectively. We found the developed method simple, rapid, cost‐effective, and most importantly, very reliable for the detection and confirmation of C. perfringens. PRACTICAL APPLICATIONS With Fung's double tube method and a colony polymerase chain reaction technique, a completed determination of Clostridium perfringens contamination can be accomplished within 16–18 h. This saves at least 2–3 days when compared with standard methods. Moreover, it minimizes the cost and labor needed since an anaerobic chamber as well as steps of Gram staining and biochemical testing can be avoided. The developed method is a powerful tool and an alternative for the enumeration of C. perfringens. It can be applied to samples from various sources and highly reliable results are expected. Most microbiological laboratories have a thermocycler and reagents as parts of their basic instruments. Therefore, the developed method can be easily applied without massive investment.
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