• 한국식품위생안전성학회지
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• 제33권1호
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• pp.65-70
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• 2018

L. monocytogenes는 Listeriosis를 일으키는 중요한 식중독 균으로 현재 국내 식품공전에서는 증균배양을 기초로 검출하며, 규격은 불검출로 관리하고 있다. 그러나 Listeria종 간의 혼합오염시 증균 과정에서 경쟁생육이 존재하여 L. monocytogenes 위음성의 가능성이 있다고 보고되고 있다. 국내 식품공전은 L. monocytogenes 증균을 위한 1차 배지로 규정되어 있으나 LEB 배지에서의 Listeria 종 간의 생육 연구는 보고된 바 없다. 본 연구는 식품에서 주로 검출되는 Listeria 속 4종(L. monocytogenes, L. innocua, L. ivanovii, L. seeligeri)을 LEB배지에 혼합배양하며 증균과정에서 생육의 차이가 존재하는 것을 확인하였다. 특히, L. innocua에 의해 L. monocytogenes의 생육이 저해되며, L monocytogenes가 L. innocua보다 초기균수가 2.0 log CFU/mL 이상 오염이 되어있어야지만 L. innocua보다 생육이 잘 되는 것을 확인하였다. Listeria 종 간의 혼합오염이 있을 경우 현재 검출법으로는 L. monocytogenes의 검출이 어려울 수 있다고 판단된다. 따라서 L. monocytogenes 검출율을 높이는 새로운 증균배지 개발의 필요성을 확인하였다. 향후 본 연구는 L. monocytogenes 검출률을 높여 국내 식품의 식품 안전에 기여 할 수 있으며 국내 식품 관리 규격 개정 시 기초가 되는 참고 자료로 활용 할 수 있을 것으로 생각된다.

• 대한수의학회지
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• 제43권2호
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• pp.231-237
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• 2003

Listeria (L.) monocytogenes in samples could not be detected occasioally by faster growth of other Listeria spp. especially L. innocua. The aim of this study was to develop the differential media and multiplex polymerase chain reaction (PCR) assays for the rapid detection of L. monocytogenes. L. monocytogenes colonies were characterized by their ${\beta}$-hemolysis with fluorescence under 366 nm UV light on the Listeria hemolysis agar (LHA). L. innocua, a species commonly present in foods, did not produce ${\beta}$-hemolysis on LHA. Therefore, one or more colonies of L. monocytogenes were easily distinguished from large populations of L. innocua. The multiplex PCR assays were developed to distinguish from L. monocytogenes and other Listeria spp. with two pairs of primers. The primers were designed in 16S rRNA and listeriolysin O gene for specific amplification of all members of the genus Listeria and L. monocytogenes, respectively. The multiplex PCR assays produced 560 and 938 bp products in L. monocytogenes; only 938 bp products in the genus Listeria. The multiplex PCR assays could detect as little as 50 pg of L monocytogenes DNA. These results indicated that the differential media and multiplex PCR assays might be useful diagnostic tools for the rapid detection of L. monocytogenes.

• 한국식품영양과학회지
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• 제23권2호
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• pp.293-297
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• 1994

The effect of organic acids on growth and heat resistance of Listeria monocytogenes Scott A were investigated. The growth of L. monocytogenes was inhibited in Tryptic Soy Broth(TSB) with 0.1 or 0.2% of acetic , tartic , propionic , citric and lactic acid at 35$^{\circ}C$, respectively. The growth of l. Monocytogenes did not occur in TSB with 0.2% of acetic acid or propionic acid during 48h of incubation. The heat resistance of L.monocytogenes was affected by kind of organic acid, ph and heating substrate. L.monocytogenes showed more heat resistant in TSB with various organic acids than in 0.1M sodium phosphate with the same organic acids. Heat resistance decreased as pH of heating substrate decreased . Surface-adherent microcolony was more heat resistant than planktonic cell of L. monocytogenes. Propionic and lactic acids more affected on heat resistance of L.monocytogenes than acetic , tartaric and citric acids.

• 한국식품과학회지
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• 제26권5호
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• pp.545-551
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• 1994

식용 가능성이 있는 식물 49종의 열매, 줄기, 잎, 껍질 혹은 뿌리를 75% ethanol을 용매로 추출물을 얻고 이를 Listeria monocytogenes ATCC 19111, L. monocytogenes ATCC 19112, L. monocytogenes ATCC 19113, L. monocytogenes ATCC 19114 및 L. monocytogenes ATCC 15313을 대상으로 증식저해 정도를 disk method로 검색하고 효과가 있는 것은 액체 배지에 농도별로 첨가, 그 효과를 비교하였다. L. monocytogenes ATCC 19111에 비교적 높을 항균성을 보인 식물의 ethanol 추출물은 털진득찰, 뽕나무이고 L. monocytogenes ATCC 19112와 ATCC 19114에는 뽕나무, ATCC 19113에는 고삼, 뽕나무, ATCC 15313에는 뽕나무, 꿀풀, 회향 등 이었다 이들 식물 추출물중 광범위하게 항균성을 나타내는 뽕나무껍질 추출물은 실험한 L. monocytogenes 5종 모두에 대하여 500ppm 첨가 수준에서 대부분 완전 증식억제 현상을 보였다.

• 한국식품과학회지
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• 제31권5호
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• pp.1324-1329
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• 1999

국내 유통중인 냉동만두와 피자 중 L. monocytogenes의 분포와 분리균의 특성을 조사하기 위하여 1998년 8월부터 11월에 걸쳐 시료 72개를 구입하여 실험하였다. 전체 냉동식품 중 9.7%(7개)에서 L. monocytogenes가 분리되었으며 냉동만두 중 11.1%,피자 중 5.6%가 오염되어 있었다. USDA와 FDA방법. 그리고 modified cold enrichment 방법 중 USDA방법이 가장 L. monocytogenes의 분리율이 높았으며 분리된 L. monocytogenes 혈청형은 4였다. PCR실험결과 CAMP test와 API Listeria kit를 사용하여 분리된 균주가 L. monocytogenes임이 확인되었다. USDA방법과 PCR을 이용하여 L. monocytogenes를 분리하고 확인하는데 3-4일 정도의 시간이 소요되어 시간을 단축시킬 수 있는 분리 및 확인방법의 개발이 필요하리라 사료된다. 분리된 L. monocytogenes의 내열성을 조사한 결과 tryptic soy broth에서 D값이 $60^{\circ}C$에서 49.2초, $65^{\circ}C$에서 8.8초였으며 냉동식품의 안전한 사용을 위하여 L. monocytogenes의 분리율이 높은 식품에서 D값의 측정이 필요하다고 사료된다.

• 한국미생물·생명공학회지
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• 제25권5호
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• pp.442-447
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• 1997

To development food preservative, antimicrobial activities of Schizandra chinensis (SC) against Listeria monocytogenes Scott A, L. monocytogenes Brie I and L. monocytogenes ATCC 19111 were investigated. The growth of L. monocytogenes Scott A, L. monocytogenes Brie I and L. monocytogenes ATCC 19111 was inhibited apparently in Tryptic Soy Broth (TSB) containing 1% SC at 35$\circ$C and it was found that these had antibacterial effects against a broad spectrum of pathogenic bacteria such as S. aureus ATCC 29737, B. subtilis KCTC 1021, E. coli ATCC 11775. The growth of L. monocytogenes was also inhibited about 3 to 5 log$_{10}$ cycle by 0.1% of three fractions of the alcohol extract such as ether, ethyl acetate and butanol. Acidic, weakly acid and neutral fraction of ether fraction showed inhibitory properties against L. monocytogenes.

• 한국축산식품학회지
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• 제33권4호
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• pp.481-486
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• 2013

This study evaluates the effects of fat contents on the thermal resistance, antibiotic sensitivity, and Caco-2 cell invasion of Listeria monocytogenes. Ten strain mixture of L. monocytogenes in milk (0, 1, and 4% fat) and pork sausage patties (10, 20, and 30% fat) were exposed to $63^{\circ}C$. To evaluate effects of fat on the antibiotic sensitivity of L. monocytogenes, the L. monocytogenes strains NCCP10811 (most antibiotic resistant to streptomycin) and NCCP10943 (most antibiotic sensitive to streptomycin) were exposed to different fat contents in milk and pork sausage patties, and L. monocytogenes from the foods were used for antibiotic sensitivity assays. The most invasive L. monocytogenes strains (NCCP10943) was exposed to different fat contents in milk or pork sausage patties, and L. monocytogenes from the foods were used for the Caco-2 cell invasion assays. The reductions of L. monocytogenes populations were not generally influenced by fat contents. The L. monocytogenes subjected to milk fat had increased sensitivities (p<0.05) due to some antibiotics. In addition, Caco-2 cell invasion efficiency of L. monocytogenes NCCP10943 increased (p<0.05) as fat contents increased. These results indicated that higher fat contents may be related to L. monocytogenes invasions and heat resistances in pork sausage patties, but the relationship between fat and antibiotic sensitivity varied according to antibiotics, strains, and fat contents.

• 한국축산식품학회지
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• 제33권3호
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• pp.395-402
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• 2013

Sodium chloride is used to improve various properties of processed meat products, e.g., taste, preservation, water binding capacity, texture, meat batter viscosity, safety, and flavor; however, many studies have shown that sodium chloride increases the resistance of many foodborne pathogens to heat and acid. Listeria monocytogenes has been isolated from various readyto- eat (RTE) meat and dairy products formulated with sodium chloride; therefore, the objective of this paper was to review the effects of sodium chloride on the physiological characteristics of L. monocytogenes. The exposure of L. monocytogenes to sodium chloride may increase biofilm formation on foods or food contact surfaces, virulence gene transcription, invasion of Caco-2 cells, and bacteriocin production, depending on L. monocytogenes strain and serotype as well as sodium chloride concentration. When L. monocytogenes cells were exposed to sodium chloride, their resistance to UV-C irradiation and freezing temperatures increased, but sodium chloride had no effect on their resistance to gamma irradiation. The morphological properties of L. monocytogenes, especially cell elongation and filament formation, also change in response to sodium chloride. These findings indicate that sodium chloride affects various physiological responses of L. monocytogenes and thus, the effect of sodium chloride on L. monocytogenes in RTE meat and dairy products needs to be considered with respect to food safety. Moreover, further studies of microbial risk assessment should be conducted to suggest an appropriate sodium chloride concentration in animal origin foods.

• 대한수의학회지
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• 제36권1호
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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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• 제19권3호
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• pp.238-249
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• 2009

The genus Listeria consists of six closely related species and forms three phylogenetic groups: L. monocytogenes-L. innocua, L. ivanovii-L. seeligeri-L. welshimeri, and L. grayi. In this report, we attempted to examine the evolutionary relationship in the L. monocytogenes-L. innocua group by probing the nucleotide sequences of 23S rRNA and 16S rRNA, and the gene clusters lmo0029-lmo0042, ascB-dapE, rplS-infC, and prs-ldh in L. monocytogenes serovars 1/2a, 4a, and 4b, and L. innocua. Additionally, we assessed the status of L. monocytogenes-specific inlA and inlB genes and 10 L. innocua-specific genes in these species/serovars, together with phenotypic characterization by using in vivo and in vitro procedures. The results indicate that L. monocytogenes serovar 4a strains are genetically similar to L. innocua in the lmo0035-lmo0042, ascB-dapE, and rplS-infC regions and also possess L. innocua-specific genes lin0372 and lin1073. Furthermore, both L. monocytogenes serovar 4a and L. innocua exhibit impaired intercellular spread ability and negligible pathogenicity in mouse model. On the other hand, despite resembling L. monocytogenes serovars 1/2a and 4b in having a nearly identical virulence gene cluster, and inlA and inlB genes, these serovar 4a strains differ from serovars 1/2a and 4b by harboring notably altered actA and plcB genes, displaying strong phospholipase activity and subdued in vivo and in vitro virulence. Thus, by possessing many genes common to L. monocytogenes serovars 1/2a and 4b, and sharing many similar gene deletions with L. innocua, L. monocytogenes serovar 4a represents a possible evolutionary intermediate between L. monocytogenes serovars 1/2a and 4b and L. innocua.