• The Plant Pathology Journal
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• v.16 no.5
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• pp.286-289
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• 2000

Mechanol extracts of three cold-tolerant eucalyptus trees-Eucalyptus darlympleana, E. gunnii and E. unigera were screened for antimicrobial activity against twenty two phyto-pathogenic fungi and six food-borne bacterial pathogens. E. unigera showed the antagonistic activity against all the tested pathogens. Among the tested fungal pathogens, Pythium species were highly sensitive to the leaf extracts. Especially, P. vanterpoolii, a causal agent of leaf blight in creeping bentgrass (Agrostis palustris), was completely inhibited by the extracts. The eucalyptus extracts were also effective in inhibiting the fungal growth of Botrytis cinerea and Phomopsis sp. isolated from the lesions of kiwifruit soft rot during post-harvest storage. Escherichia coli O-157 was less sensitive to the inhibition than the other bacterial pathogens tested. It was likely that Gram positive bacteria-Bacillus subtilis and Streptococcus mutans were more sensitive to the eucalyptus extracts than Gram negative bacteria-Escherichia coli, Salmonella enteritidis and Pseudomonas aeruginosa. Our findings suggest that the cold-tolerant eucalyptus species have antimicrobial properties that can serve the development of novel fungitoxic agents or food preservatives.

• Food Science and Biotechnology
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• v.15 no.1
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• pp.138-142
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• 2006

Various levels of antibacterial activity have been identified for water and ethanol extracts of corn silk, particularly against Salmonella typhimurium KCTC 2515. In general, the water extract was more effective than the ethanol extract. The minimum inhibitory concentration (MIC) for the water extract was 7.5 mg/disc for S. typhimurium KCTC 2515 and B. cereus KCTC 1092, as well as for the ethanol extract against S. typhimurium KCTC 2515 and S. typhimurium KCTC 1925. However, the MICs for the water extract were lower than those for the ethanol extract against all bacteria tested, except S. typhimurium KCTC 1925 and B. cereus KCTC 1014. The growth of the tested organisms in the synthesized broth medium was inhibited with the addition of 5-fold levels of MIC. Using sterilized milk as the model food system, we found that the lag phase for these microorganisms was extended up to 3 days at $20^{\circ}C$, but was not affected at $4^{\circ}C$. These results indicate that bacterial growth was strongly inhibited by corn silk extract at $20^{\circ}C$.

• Korean Journal of Food Science and Technology
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• v.37 no.4
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• pp.611-617
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• 2005

Dispersed repetitive DNA elements in genomes of microorganisms differ among and within species. Because distances between repetitive sequences vary depending on bacterial strains, genomic fingerprinting with interspersed repetitive sequence-based probes can be used to distinguish unrelated organisms. Among well-known bacterial repetitive sequences, Repetitive Extragenic Palindromic (REP) sequence has been used to identify environmental bacterial species and strains. We applied REP-PCR to detect and differentiate four major Gram-negative food-borne bacterial pathogens, E. coli, Salmonella, Shigella, and Vibrio. Target DNA fragments of these pathogens were amplified by REP-PCR method. PCR-generated DNA fragments were separated on 1.5% agarose gel. Dendrograms for PCR products of each strain were constructed using photo-documentation system. REP-PCR reactions with primer pairs REP1R-I and REP2-I revealed distinct REP-PCR-derived genomic fingerprinting patterns from E. coli, Salmonella, Shigella, and Vibrio. REP-PCR method provided clear distinctions among different bacterial species containing REP-repetitive elements and can be widely used for typing food-borne Gram-negative strains. Results showed established REP-PCR reaction conditions and generated dendrograms could be used with other supplementary genotyping or phenotyping methods to identify isolates from outbreak and to estimate relative degrees of genetic similarities among isolates from different outbreaks to determine whether they are clonally related.

• Food Science of Animal Resources
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• v.30 no.4
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• pp.590-596
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• 2010

Meat and meat products are a potential source of food-borne pathogens, including Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7, and Bacillus cereus. A sensitive and specific PCR assay for the detection of these pathogens in meat and meat products was developed in this study, as part of a broader effort to reduce the potential health hazards posed by these pathogens. Initially, PCR conditions were standardized with purified DNA. Under standard conditions, the detection level for PCR was as low as 10 pg of purified bacterial DNA. After overnight growth of bacteria in a broth medium, as few as $10^2$ CFU of bacteria were detected by PCR assay. The primers employed in the PCR assay were found to be highly specific for individual organisms, and evidenced no cross-reactivity with heterologous organisms. Additionally, the multiplex PCR assays also amplified some target genes from the four pathogens, and multiplex amplification was obtained from as little as 10 pg of DNA, thus illustrating the excellent specificity and high sensitivity of the assay. In conclusion, this PCR-based technique provides a sensitive and specific method for the detection of S. aureus, Salmonella spp., E. coli O157:H7, and B. cereus in meat and meat products.

• Korean Journal of Food Science and Technology
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• v.37 no.6
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• pp.1005-1011
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• 2005

Widespread distributions of repetitive DNA elements in bacteria genomes are useful for analysis of genomes and should be exploited to differentiate food-borne pathogenic bacteria among and within species. Enterobacterial repetitive intergenic consensus (ERIC) sequence has been used for ERIC-PCR genomic fingerprinting to identify and differentiate bacterial strains from various environmental sources. ERIC-PCH genomic fingerprinting was applied to detect and differentiate four major Gram-negative food-borne bacterial pathogens, Esherichia coli, Salmonella, Shigella, and Vibrio. Target DNA fragments of pathogens were amplified by ERIC-PCR reactions. Dendrograms of subsequent PCR fingerprinting patterns for each strain were constructed, through which relative similarity coefficients or genetic distances between different strains were obtained numerically. Numerical comparisons revealed ERIC-PCR genotyping is effective for differentiation of strains among and within species of food-borne bacterial pathogens, showing ERIC-PCR fingerprinting methods can be utilized to differentiate isolates from outbreak and to determine their clonal relationships among outbreaks.

• Journal of Applied Biological Chemistry
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• v.51 no.6
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• pp.285-291
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• 2008

A potential probiotics bacterial strain, CS90, was isolated from Korean soybean paste (doenjang). The strain CS90 showed antimicrobial activity against food-borne pathogenic bacteria including Salmonella enterica, Salmonella enteritids, Salmonella typhymurium, Bacillus cereus, Listeria ivanovii, Listeria. monocytogenes, Sthaphylococcus aureus, and Sthaphylococcus epidermidis and showed a significant survival rate of 35.7 to 57.8% under the artificial gastric acidic condition (pH 2 to 3). The strain CS90 was classified as Bacillus subtilis based on morphological, physiological, chemotaxonomic features and phylogenetic analysis based on 16S rDNA sequence and designated as B. subtilis CS90. B. subtilis CS90 can be used as a potential probiotics.

• Journal of Environmental Health Sciences
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• v.37 no.2
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• pp.133-140
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• 2011

Antimicrobial activity and bactericidal activity of Caesalpinia sappan L. extracts were investigated against five food-borne pathogens, E. coli, S. aureus, S. typhimurium, B. cereus and L. monocytogenes. Methanol extract of Caesalpinia sappan L. revealed antimicrobial activities against five pathogens. In particular, by paper disc diffusion the highest activity was shown against L. monocytogenes. Antimicrobial activities of methanol extracts showed the most potent activities, but hexane fraction had no activity. Fractions of ethyl acetate and butanol turned out to have higher antimicrobial activities against Gram(+) bacteria than Gram(-) bacteria. The minimum inhibitory concentration against five food-borne pathogens was 1.563 mg/ml on Gram(+) bacteria and 3.125 mg/ml on Gram(-) bacteria. The result of antimicrobial activity in a shaking flask method showed that bacterial growth rate fell by more than 99.999% at 3.125 mg/ml of methanol extract. The highest rate of viable reduction (99.998%) was shown at 0.781 mg/ml of methanol extract against L. monocytogenes. After five minutes of reaction between test strains and methanol extracts, the growth rates of five kinds of bacteria were reduced by more than 99.999% at a concentration of 100 mg/ml. Therefore, it is suggested that methanol extracts of Caesalpinia sappan L. can be developed as a natural sanitizer or disinfectant.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.675-690
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• 2007

Bacteriocins in Gram-positive bacteria have attracted much attention because many have a strong antimicrobial activity also against bacteria outside the genera of the producers. Lantibiotics and the pediocin-like bactericins have attracted most attention since they kill a broad spectrum of Gram-positive bacteria including important pathogens. But many other promising Gram-positive bacteriocins have been thoroughly characterized. Recent studies have shown that bacteriocins may playa role in the intestinal flora to protect us against the food-borne pathogens. Bacterial genome sequencing has demonstrated that there may be an arsenal of such compounds and we are only seeing the top of the iceberg. The present review gives a short outlook of the field of bacteriocins with focus on lactic acid bacteria and includes recent findings.

• Korean Journal of Veterinary Research
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• v.49 no.4
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• pp.319-328
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• 2009

This study investigated the epidemiology of Listeria (L.) monocytogenes, a food-borne pathogen. The epidemiology of food-borne pathogens is of great importance for clarifying bacterial origin and preventing bacterial contamination and infection. This work examined 68 L. monocytogenes strains, including 11 reference strains and 57 isolates from imported US beef, domestic meats (beef, pork, chicken meat), raw milk, and milk plants. The random amplified polymorphic DNA (RAPD) techniques were optimized to develop a standard molecular epidemiological analysis of L. monocytogenes. There was great genetic variability among the isolates, which produced 24 and 34 RAPD patterns with primer HLWL85 and HLWL74, respectively. The discriminatory power of the RAPD methods with HLWL85 and HLWL74 primer were very high (DI = 0.957; S ${\geq}$ 80%, S ${\geq}$ 95%). Some RAPD types were specific to origin. A few RAPD types were specific for L. monocytogenes strains belonging to a particular serotype. Using the HLWL85 primer, the strains isolated from milk plants could be distinguished from the other strains. And using the HLWL74 primer, the strains isolated from imported beef (US) could be distinguished completely from the other strains.

• Journal of Food Hygiene and Safety
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• v.26 no.2
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• pp.114-119
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• 2011

In this study, two duplex real-time PCR approach with melting curve analysis is presented for the detection of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus, which are important food-borne bacterial pathogens usually present in fresh and/or minimally processed vegetables. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the ${\beta}$-glucuronidase (uidA, E. coli), thermonuclease (nuc, S. aureus), hemolycin (hly, L. monocytogenes) and tetrathionate reductase (ttr, Salmonella spp.) genes. Melting curve analysis using a SYBR Green I real-time PCR approach showed characteristic $T_m$ values demonstrating the specific and efficient amplification of the four pathogens; $80.6{\pm}0.9^{\circ}C$, $86.9{\pm}0.5^{\circ}C$, $80.4{\pm}0.6^{\circ}C$ and $88.1{\pm}0.11^{\circ}C$ for S. aureus, E. coli O157:H7, L. monocytogenes and Salmonella spp., respectively. For all the pathogens, the two duplex, real-time PCR was equally sensitive to uniplex real-time PCR, using same amounts of purified DNA, and allowed detection of 10 genome equivalents. When our established duplex real-time PCR assay was applied to artificially inoculated fresh lettuce, the detection limit was $10^3$ CFU/g for each of these pathogens without enrichment. The results from this study showed that the developed duplex real-time PCR with melting curve analysis is promising as a rapid and cost-effective test method for improving food safety.