• Korean Journal of Veterinary Research
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• v.36 no.1
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• pp.119-129
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• 1996

The present study was conducted to rapidly detect Listeria monocytogenes in raw milk. Specificity and sensitivity of polymerase chain reaction(PCR) technique, and direct PCR were examinded in raw milk, also were compared the calssical culture methods with PCR technique. This method used a pair of primers based on a unique region in the 16S rRNA sequence of L nomocytogenes. In the PCR specificity tests, each of the 10 strains of L monocytogenes tested gave a single 70-bp band. But the other six Listera spp tested gave negative results. Results of the sensitivity tests showed that as few as 2 CFU of L monocytogenes in pure cultures could be detected with 16S rRNA-based primers, L-1 and L-2. In different PCR cycles, a PCR product was detected with $10^3$ cells of L monocytogenes from 25 cycles to 50 cycles and the concentration of PCR products was cycle-dependent. Raw milk samopes added L monocytogenes cells gave negative results. However, these samplers gave a single 70-bp band by pretreatment of pronase, and PCR products were detected with $10^1$ cells of L monocytogenes. To detemine the most sensitive culture protocol to use in conjunction with the PCR assay, raw milk samples were inoculated with L monocytogenes at concentrations ranging from 1 to $5.7{\times}10^4CFU/ml$. PCR assays from Listeria enrichment broth(LEB) containing raw milk samples added L monocytogene EGD could dtect 10 cells in pronase-pretreated samples without incubation, and 1 cell of L monocytogenes in both 12 hr and 24 hr incubation, respectively. Isolation raw of PCR assays was similar to that of classical culture methods, but required time for detection of L monocytogenes could remarkably be reduced compare to culture methods.

• Journal of Microbiology and Biotechnology
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• v.2 no.2
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• pp.73-77
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• 1992

The effect of a nisin-producing Lactococcus lactis spp. lactis (L. lactis) on the growth and attachment of Listeria monocytogenes Scott A and Brie 1 on stainless steel and their growth in Brain Heart Infusion broth was determined. Viable cells of Listeria decreased rapidly after 9~12 hr of incubation at $21^{\circ}C$ and after 6~9 hr of incubation at $32^{\circ}C$ in the presence of L. lactis. The number of L. monocytogenes Scott A attached to stainless steel in pure culture was $2.5{\times}10^3/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}2.3{\times}10^3/\textrm{cm}^2{\;}at{\;}32^{\circ}C$ after 48 hr of incubation, but was only $10/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}1.1{\times}10/\textrm{cm}^2{\;}at{\;}32^{\circ}C$ in the presence of L. lactis. Results from L. monocytogenes strain Brie 1 were similar to those from strain Scott A. The population of L. monocytogenes Scott A which attached to stainless steel with previously adherent L. lactis was $1.8{\times}10^2/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}8.2{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$, whereas the population attached to sterile stainless steel was $1.2{\times}10^3/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}2.1{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$. For L. monocytogenes Brie 1, the attached population of the control was $1.6{\times}10^4/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}3.2{\times}10^2/\textrm{cm}^2{\;}at{\;}32^{\circ}C$, and on stainless steel with adherent L. lactis, it was $1.1{\times}10/\textrm{cm}^2{\;}at{\;}21^{\circ}C{\;}and{\;}6.9{\times}10/\textrm{cm}^2{\;}at{\;}32^{\circ}C$. Surface adherent L. lactis was less inhibitory to attachment of L. monocytogenes on stainless steel than a liquid culture inoculum. Listeria attached to stainless steel survived dry storage for 20 days both in the presence and absence of adherent lactococci.

• Korean journal of food and cookery science
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• v.12 no.3
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• pp.312-319
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• 1996

Oyster (Crassostrea virginica) and Weakfish (Cynoscion regalis) were stored at 6, 0, -4 and -20$^{\circ}C$ for up to 45 days and examined for changes in microflora. Aerobic plate counts (incubated at 21$^{\circ}C$) were performed at selected times during storage and 495 isolates (255 isolates from oyster and 240 isolates from Weakfish) were randomly selected from the plates during the storage. Before the storage of the fishes, viable counts of oyster were 4.9${\times}$10$\^$5/ CFU/g of meat and those of Weakfish were 1.5${\times}$10$^4$ CFU/cm$^2$of skin. Microflora of oyster before storage, the major isolates identified as Pseudomonas spp. (67%) and Vibrio spp. (20%). Pseudomonas ll1/1V-H and Flavobacterium/Cytophaga were predominant genus in the microflora of oyster during cold storage at 6, 0, -4 and -20$^{\circ}C$. The composition of the microflora of Weakfish before storage, Acinetobacter (40%) and Moraxella (33%) were the major species, with Pseudomonas and Vibrio constituting a small percentage of the total isolates. The microflora shifted to predominantly Pseudomonas spp. during storage at 6. 0 and -4$^{\circ}C$, making up from 60 to 100％ of isolated strains. During frozen storage, the percentage of isolates identified as Mnraxella increased to 40-60% of the total isolates. During cold storage, halophilic bacteria (Pseudomonas lII/IV-H and Vibrio) were predominant in oyster while nonhalophilic bacteria (Pseudomonas III/IV-NH and Moraxella) were predominant in Weakfish. Vibrio spp. were higher in oyster than in Weak fish. Listeria spp. were not isolated but unidentified ${\beta}$-hemolytic bacteria were islolated from both of the fishes during cold storage.

• Food Science of Animal Resources
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• v.27 no.1
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• pp.127-131
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• 2007

In order to investigate the presence of pathogenic bacteria in fresh pig loin and the growth changes of microorganism in raw ham during storage at 10 and $25^{\circ}C$. These hams were manufactured according to a short-ripening procedure being completed in 4 weeks with dry-curing followed by wet-curing and ripening. The result regarding the contamination level of microorganism in the fresh raw pig loin showed that the count of total aerobes was $3.11\;log\;CFU/cm^2$, and the population of lactic acid bacteria, Pseudomonas spp., Clostridium spp., and yeast and mould had not risen over $2\;log\;CFU/cm^2$ on the storage time. However, the average count ofEnterobacteriaceae in pork loin was $3.11\;log\;CFU/cm^2$, which represented the predominant species. The pathogenic bacteria including Salmonella spp, Staphylococcus aureus, Vibrio parahaemolyticus, Clostridium perfringene, Listeria monocytogenes, and Escherichia coli O157:H7 were not detected either in fresh pork loin or in raw ham products stored at 10 and $25^{\circ}C$. The initial count of total aerobes in raw ham samples was 3.06 log CFU/g, and increased slightly after 90 days at 10 and $25^{\circ}C$ to 4.6 and 4.69 log CFU/g, respectively. The predominant species in raw ham products during storage time were lactic acid bacteria and Staphylococcus spp.

• Korean Journal of Food Science and Technology
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• v.35 no.1
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• pp.97-102
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• 2003

Raw and washed spinaches were tested to evaluate the incidences of Aeromonas hydrophila, Escherichia coli O157:H7, Plesiomonas shigelloides, Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Yersinia enterocolitica, Bacillus cereus, Campylobacter jejuni, Clostridium perfringens, Listeria monocytogenes, and Staphylococcus aureus. Four pathogenic bacteria were isolated from spinach samples, and identified by morphological and biochemical methods, including API and ATB identification systems. Isolates from MacConkey, Cereus Selective, Clostridium Perfringens, and Baird-Parker agar media were in 99.9, 99.8, 99.9, and 97.8% agreements with A. hydrophila, B. cereus, C. perfringens, and S. aureus at the species level, respectively. SET-RPLA revealed, among the five strains of S. aureus isolates, two produced type A enterotoxin. All five strains of B. cereus isolates produced enterotoxin as revealed with CRET-RPLA.

• Food Science and Biotechnology
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• v.17 no.6
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• pp.1368-1371
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• 2008

In this study, 50 rump samples of raw beef obtained from local Korean supermarkets were analyzed to survey microbial distributions. As results, mesophilic microorganisms ranged from $(1.4{\pm}0.01){\times}10^2$ to $(1.6{\pm}0.05){\times}10^5\;CFU/g$, and total coliforms ranged from 0 to $(1.3{\pm}0.04){\times}10^4\;CFU/g$. Major foodborne pathogens, including Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella spp., were not found among the samples. However, Staphylococcus aureus was isolated with 4% frequency. Other isolated microorganisms included Enterobacter amnigenus (4%), Enterobacter cloacae (24%), E. coli (24%), Listeria innocua (8%), Staphylococcus saprophyticus (56%), Staphylococcus xylosus (10%), and Staphylococcus warneri (8%).

• Journal of Food Hygiene and Safety
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• v.30 no.4
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• pp.303-308
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• 2015

The objectives of this study were to investigate the microbial contamination level of domestic kitchen environments and to provide information to improve food safety in 50 domestic house kitchens located in Seoul, Incheon, and Gyeonggi-do. Dishcloth, chopping board, and refrigerator swabs were examined for the presence of coliforms, Salmonella spp., Campylobacter jejuni/coli, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus. The means and standard deviations of coliform counts for dishcloths was $4.8{\pm}1.84log\;CFU/100g$, chopping boards, and refrigerator drawers were $4.04{\pm}1.53$, $4.11{\pm}1.65log\;CFU/100cm^2$, respectively. Salmonella spp. and Campylobacter jejuni/coli were not detected in all samples. E. coli were detected in 3 on the dishcloths and 1 of 50 samples in the refrigerator drawer. Listeria monocytogenes was detected in the drawer of the refrigerator in 2 of 50 samples. In the case of Staphylococcus aureus, the detection on dishcloths, chopping boards, and drawers in refrigerators was 21, 12, and 14 of 50 samples, respectively. The results of microbiological tests of domestic kitchen utensils can be used to emphasize the importance of the sanitary conditions in domestic kitchen environments.

• Journal of Food Hygiene and Safety
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• v.22 no.4
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• pp.388-394
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• 2007

This study was conducted to evaluate the selective media listed in currently available Food Code in Korea. The 29 different types of media of five different types of foodborne bacteria including Salmonella spp., Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus were tested in the broth and food. The recovery test for five different types of foodborne bacteria was performed in the artificially inoculated into chicken, rice, pork and mackerel. There was no significant differences in isolation capabilities among twenty nine different types of isolation selective media for five different types of foodborne bacteria in broth condition, while there was significantly a little differences in isolation capabilities among those on foods (P<0.05). The higher number of foodborne pathogens were isolated from conventional selective media approved in Food Code than newly developed selective media such as chromogenic media. This results suggest that there was differences of selectivities among currently available isolation selective media in many countries and further studies are needed to be approved by Korean Food and Drug Administration.

• Journal of Food Hygiene and Safety
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• v.13 no.4
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• pp.388-393
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• 1998

Recently there has been an increasing amount of foreign livestock products distributed in the domestic market due to the market opening. Some vicious dealers sell the foreign beef in the trade name of the native beef during the final distribution step to arouse the social criticism frequently. In this report, we investigated a method to distinguish the native beef from the foreign one scientifically using the PCR-RAPD, a recent gene technique. Hygienical safety was also examined using a microbiological test for toxicity of Escherichia coli 0157:H7 and the food poisoning bacteria. The conditions of DNA amplification for the PCR analysis were $1{\times}Taq$ polymerase buffer, 1.5 mM $MgCl_2,\;50\;\mu\textrm{M}$ dNTP, 100 ng primers, 2.5 unit Taq polymerase and 5~20 ng template DNA, with the fmal volume of $50\;\mu\textrm{\ell}$. The size of the amplified product was detected mostly in the range of 0.5~2.0 kbp. The size of DNA, gene marking factor, which could be a criterion distinguishing the native beef from the foreign one, appeared approximately 1.2 kbp. The native beef was distinguished from the foreign beef with more than 90% of confidence by the gene marking factor. This method was expected to be useful in the breed discrimination between the native beef and the foreign one. The hygienical test results showed that, fortunately, neither Salmonella spp. and Listeria monocytogenes which form a principal cause of the food poisoning nor Enterohemorrhagic Escherichia coli : EHEC which have provoked a recent social disturbance, were detected at all.

• 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.