• Title/Summary/Keyword: Listeria spp

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Simultaneous Detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in Low-fatted Milk by Multiplex PCR

  • Kim, Ji-Hyun;Rhim, Seong-Ryul;Kim, Kee-Tae;Paik, Hyun-Dong;Lee, Joo-Yeon
    • Food Science of Animal Resources
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    • v.34 no.5
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    • pp.717-723
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    • 2014
  • A rapid and specific PCR assay for the simultaneous detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in foods was developed to reduce the detection time and to increase sensitivity. Multiplex PCR developed in this study produced only actA, fliC, hbl, invA, ileS amplicons, but did not produce any non-specific amplicon. The primer sets successfully amplified the target genes in the multiplex PCR without any non-specific or additional bands on the other strains. The multiplex PCR assays also amplified some target genes from five pathogens, and multiplex amplification was obtained from as little as 1 pg of DNA. According to the results from the sensitivity evaluation, the multiplex PCR developed in this study detected 10 cells/mL of the pathogens inoculated in milk samples, respectively. The results suggested that multiplex PCR was an effective assay demonstrating high specificity for the simultaneous detection of five target pathogens in food system.

Characterization of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju

  • Lim, Sung-Mee;Park, Mi-Yeon;Chang, Dong-Suck
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.49-57
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    • 2005
  • Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.

Effect of Applying Pretreatment Methods before cooking for decreasing the Microbiological Hazard of Cooked Dried fish in Foodservice establishments (건어물을 이용한 조리음식의 미생물학적 위해 감소를 위한 조리 전처리 적용 효과)

  • 배현주;이재학;오세인
    • Korean journal of food and cookery science
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    • v.19 no.5
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    • pp.555-561
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    • 2003
  • The purpose of this study was to analyze the microbiological hazards of dried fish (Jwieochae, Ojingeochae and Bugeochae), and to apply pretreatment methods to increase the safety of cooked dried fish within foodservice establishments. Microbiological inspections were conducted on Total Plate Count, Coliforms, E. coli, E. coli O157:H7, Staphylococcus aureus, Salmonella spp. and Listeria monocytogenes. The study results are summarized as follows. According to the Hazard analysis, there are many problems showing high numbers in terms of Total Plate Count and Coliforms, which were both well over acceptable standard levels. E. coli and Staphylococcus aureus were detected at certain foodservice establishments, while E. coli O157:H7, Salmonella spp. and Listeria monocytogenes were not detected at all. By applying various pretreatment methods, such as washing, blanching, pan frying and microwave heating, the levels of microbiological hazards were able to be controlled and lowered. Blanching was the most effective method, followed by panfrying, microwave heating and washing. The Total Plate Counts gradually decreased with increasing number of times washed and seconds panfried. From these results, it is concluded that to guarantee food safety, cooked dried fish raw materials should be kept hygienically and appropriate pretreatment methods applied before cooking.

Multiplex PCR Assay from Colon Biopsy Specimens in Acute Infectious Colitis Patients

  • Lee, Gyu-Sang;Lim, Kwan-Hun;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.13 no.1
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    • pp.71-73
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    • 2007
  • For the rapid detection of objective pathogenic bacteria from colon biopsy specimens, multiplex PCR (polymerase chain reaction) method was developed. The major objective bacteria in this study were Shiga-like toxin producing E. coli O157:H7, Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, Shigella spp. Salmonella spp. and Yersinia spp.. To detect simultaneously 7 kinds of pathogenic bacteria in single reaction tube, multiplex PCR system was executed using 6 sets of primers used in single PCR system for the respective bacteria. The results in this research might be applied for the detection of pathogenic bacteria form colon biopsy samples.

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A Study of the Distribution of Listeria spp. in Fresh Agricultural Products Distributed in the Busan Area, the Republic of Korea (부산지역에서 유통되는 신선농산물 중 리스테리아균 분포에 관한 연구)

  • Youn-ju Ok;Young-hee Kwon;Hye-sun Hwang;Ye-jee Byun;Ji-young Park;Byung-jun Kim
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.9-15
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    • 2024
  • This study was performed to survey the distribution of Listeria spp. in fresh agricultural products in the Busan area, the Republic of Korea, from January to November 2022. We investigated the pathogenicity and epidemiological relationships by tracing isolated strains using polymerase chain reaction and pulsed-field gel electro-phoresis (PFGE) methods. Forty cases of Listeria spp. were detected in the 210 samples of fresh agricultural products analyzed. Four species, Listeria rocourtiae, L. innocua, L. grayi, and L. monocytogenes were detected only in green vegetables (lettuce, perilla leaps) and the others (L. innocua, L. monocytogenes, and L. grayi) were detected in enoki and oyster mushrooms. L. innocua was detected in 22 samples and L. grayi in six samples. L. monocytogenes, which causes foodborne diseases, was only detected in enoki mushrooms and the strains that were isolated had genes responsible for the pathogenicity of listeriosis (iap, prfA, inlA, inlC, inlJ, and hly). To investigate the genetic similarity and contamination route of L. monocytogenes, serotyping and PFGE were conducted for 12 strains isolated from fresh agricultural (10 strains) and poultry (2 strains) products distributed at a market in the Busan area. Two serotypes (1/2a, 1/2b) were detected in strains isolated from the agricultural and poultry products, but serotype 1/2b was only detected in strains isolated from agricultural products. PFGE analysis showed index of similarity values of 45.7 to 100% and the same patterns were represented in isolates from some enoki mushrooms. These isolates had the same serotypes and showed significant epidemiological relationships.

Microbiological Investigation of the Frozen-Raw Sliced Fishes for Sushi Manufacturing (생선초밥용 냉동횟감의 미생물학적 조사)

  • Jeon, Eun Bi;Kim, Ji Yoon;Son, Min Gyu;Park, Shin Young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.2
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    • pp.224-230
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    • 2021
  • This study assessed the contamination levels of total aerobic bacteria, fungi, coliforms, Escherichia coli, and Staphylococcus aureus, and qualitative analysis of Bacillus cereus, Salmonella spp., Listeria spp., and Vibrio spp. in four frozen-raw sliced fishes (cuttlefish, flatfish, salmon, and shrimp) for sushi production. The total aerobic bacteria, fungi, and coliforms were 2.95-3.38, 1.96-2.88, and 0.92-1.29 log CFU/g, respectively. In particular, shrimp was highly contaminated with total aerobic bacteria (3.38 log CFU/g) and fungi (2.88 log CFU/g). Over 3 log CFU/g of total aerobic bacteria was also detected in cuttlefish, flatfish, and salmon. Less than 1-2 log CFU/g of E. coli was detected in all frozen samples. S. aureus was detected at 2.25-3.13 log CFU/g in most samples. B. cereus was qualitatively detected at 25% in most samples, except for salmon (0%). Salmonella spp., L. monocytogenes, and Vibrio spp. were qualitatively detected at 25-50% of all four samples. The microbial contamination levels determined in the current study may be potentially used as basic data to perform microbial risk assessments of frozen-raw sliced fishes.

An Analysis Microbiological Contamination on the Facilities and Utilities of Japanese Restaurants (일식 레스토랑 설비와 조리기구의 미생물 오염 실태 분석)

  • Kim, Gi-Young;Park, Gye-Young;Chae, Myoung-Hee
    • Culinary science and hospitality research
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    • v.15 no.4
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    • pp.9-17
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    • 2009
  • The microbiological examinations was conducted for the hygienic evaluation on ten Japanese restaurants during summer season in Gwangju, Korea. Total two hundreds swabbed samples using sponge were collected from the surface of facilities and utensils at restaurants. The number of total microorganism, coliform, E. coli, Staphylococcus, Salmonella, and Listeria were measured. The results demonstrated that most swabbed samples were contaminated with microorganisms and coliforms. The degree of contamination depended on the sample sites. The severely contaminated sites were floor, trench, and working table for the fish and total counts of those samples were over $10^4$ CFU/100 $cm^2$. The coliforms were detected on the floor, trench, and wood cutting board over $10^3$ CFU/100 $cm^2$. Moreover, coliforms was detected on the towel, dish, and working place for fish and vegetables. The E. coli was detected on the floor and trench of several restaurants. The Staphylococcus spp. was detected on the cutting board, towel, floor and trench of several restaurants. The Salmonella spp. and Listeria spp. were not detected all over the samples. From those results showed that sanitation management and disinfection were required on the Japanese restaurants.

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Hygienic Quality of Beef and Distribution of Pathogens during Cut-Meat Processing (식육의 처리 단계별 미생물 오염실태와 병원성 미생물의 분포)

  • 오영숙;이신호
    • Journal of Food Hygiene and Safety
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    • v.16 no.2
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    • pp.96-102
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    • 2001
  • Bacteriological quality of beef carcass and distributions of pathogens in beef processing environments were investigated to improve the hygienic quality of fresh beef. Total bacterial contamination of carcass surface in slaughtering process and cutting board in cut-meat process showed 10$^{5}$ -10$^{6}$ CFU/$\textrm{cm}^2$ and 10$^{5}$ CFU/$\textrm{cm}^2$ in summer, respectively. The viable bacterial count of cotton glove was similar to that of cutting board during and entire period of year. Microbial contamination of carcass surface, cutting board, cotton glove and deboned meat showed the highest in summer and the lowest in winter during the year. Escherichia coli O157, Pseudomonas aeruginosa, Klebsiella. ornithinolytica, Staphylococcus aureus, E coli, Tatumella. ptyseos, Serratia odorifera, Aero-monas sobria, Enterobacter cloacae and Flavimonas oryzihabitans were isolated from carcass surface during slaughter treatments. S. aureus, Listeria grayi and L. monocytogenes were isolated from cutting board and L. grayi, Erwinia spp. Salmonella app. and S. aureus were isolated from cotton glove in cut-meat process environments. Citrobacter freundii; L. monocytogenes; and S. aureus were isolated from deboned meat.

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Antimicrobial Activity against Food-hazardous Microorganisms, Dermatophytes, and Pytopathogens and Antioxidative Activity of Sancho Oil (식품위해성균, 피부사상균 및 식물성 병원균에 대한 산초유의 항균 활성 및 항산화 활성)

  • Kim, Hak Gon;Kang, Seung Mi;Yong, Seong Hyeon;Seol, Yu Won;Choi, Eun Ji;Park, Jun Ho;Yu, Chan Yeol;Solomon, Tamirat;Choi, Myung Suk
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.1
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    • pp.38-46
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    • 2020
  • Background: Although Sancho (Zanthoxylum schinifolium Siebold & Zucc) oil has traditionally been used for its antibiotics properties, there is currently a lack of scientific evidence regarding its biological activities. In this study, we investigated the antimicrobial and antioxidant activities of Sancho oil against food-hazardous microorganisms, phytopathogens, and dermatophytes. Methods and Results: We investiated the antimicrobial activity of Sancho oil against 11 food-hazardous microorganisms, nine phytopathogens, and six dermatophytes. The Sancho oil was found to show the strongest antibacterial activity against Shigella flexneri and Listeria spp. Sancho oil also showed high antifungal activity against plant pathogens, particularly Fusarium oxysporum, and showed antimicrobial activity against dermatophytes such as Trichophyton rubrum, Microsporum canis and Candida albicans. The antioxidant activity of Sancho oil was measured using the DPPH method, and was found to be stronger than that of unrefined oil. Moreover, this activity increased with increasing oil concentration. Conclusions: We found that Sancho oil showed differing antimicrobial activities against food-hazardous microorganisms, dermatophytes, and plant pathogens. The antimicrobial activity spectrum of Sancho oil was not broad and varied among microbial strains. On the basis of our findings, we consider that Sancho oil could be used an antibacterial material for food-borne S. flexneri and Listeria spp., a biopesticide for Fusarium spp., and a treatment for dermatophytes such as T. rubrum.

Improvement of polymerase chain reaction methods for rapid detection of Listeria monocytogenes in raw milk (원유로부터 Listeria monocytogenes의 신속검색을 위한 종합효소 연쇄반응법의 개선)

  • Yi, Chul-hyun;Son, Won-geun;Kang, Ho-jo
    • 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.

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