• 제목/요약/키워드: pulsed field gel electrophoresis

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Characterization of Antimicrobial Resistance Patterns and Integrons of Nontyphoid Salmonella Isolates from Infants in Seoul (서울지역 소아에서 분리된 Nontyphoid Salmonella의 항생제 내성과 Integron의 특징)

  • Jin, Young-Hee;Kim, Jin-Ah;Jung, Ji-Hun;Jeon, Soo-Jin;Lee, Jae-Kyoo;Oh, Young-Hee;Han, Ki-Young;Lee, Young-Ki
    • Korean Journal of Microbiology
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    • 제46권4호
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    • pp.326-333
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    • 2010
  • A total of 105 nontyphoid Salmonella isolated from infants in Seoul from 2003 to 2009 was investigated for their serotype, antimicrobial resistance, characterization of integron, and the patterns of Pulsed-field gel electrophoresis (PFGE). Eighteen serotypes were detected in 105 isolates, and the two most common serotypes were S. Enteritidis (47.6%) and Montevideo (15.2%). Among the Salmonella serovars, a high level of antimicrobial resistance was found to ampicilin (60%), tetracycline (46.7%), streptomycin (35.2%) and nalidixic acid (28.6%). In the multi-drug resistance patterns, the predominant patterns were only nalidixic acid (15.7%), ampicillin-ampicillin/sulbactam-tetracycline (14.5%), and ampicillin-streptomycin-chloramphenicol-tetracycline (10.8%). PCR and DNA sequencing analysis revealed the presence of class 1 integron in 20 isolates (19%). Of the class 1 integron positive isolates 20% harboured the integron-associated gene cassettes : aadA2, blaP1, dfr17-aadA5, dfrA12-aadA2, and aadA7. PFGE was carried out to examine the genetic relatedness among S. Enteritidis isolates. Except for three strains, fifty strains were divided by three pulsotypes.

Bacterial Contamination Reduction of Minimally Processed Agricultural Products using Antibacterial Foods and Molecular Biological Analysis (항균성 식품을 이용한 간편섭취 농산물 미생물오염의 감소 및 분자생물학적 분석)

  • Kim, Jin-Ah;Lee, Sung-Deuk;Hwang, Kwang-Ho;Song, Mi-Ok;Park, Jung-Eun;Kim, Da-Mi;Chung, Ae-Hee;Oh, Young-Hee
    • Journal of Food Hygiene and Safety
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    • 제31권1호
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    • pp.8-14
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    • 2016
  • The purposes of this study are to confirm ways to reduce bacteria of minimally processed agricultural products, using antibacterial foods that are easily available in home and to improve bacterial hygienic condition of them. We chose garlic-allicin, ginger-gingerol, green tea-catechin, cinnamon-cinnamic aldehyde, wasabi-allyl isothiocyanate as antibacterial foods and their unique antibiotic materials. We confirmed the better washing effect when these antibiotic extracts were used, compared to washing effect by only distilled water. Their antibiosis was proved by statistical processing. PFGE (Pulsed Field Gel Electrophoresis) of Bacillus cereus shows continuous contamination probability of minimally processed vegetables by same product suppliers and the necessity of systematic measures against bacterial contamination.

Molecular Characteristics of Extended-Spectrum β-Lactamase Genes in Clinical Isolates of Escherichia coli and Klebsiella pneumoniae (임상검체에서 분리된 Escherichia coli와 Klebsiella pneumoniae의 Extended-Spectrum β-Lactamase 유전자형 및 분자유전학적 특성)

  • Chung, Kyung-Suk
    • Korean Journal of Clinical Laboratory Science
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    • 제38권1호
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    • pp.26-33
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    • 2006
  • Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

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Prevalence and Genetic Characteristics of Meatborne Listeria monocytogenes Isolates from Livestock Farms in Korea

  • Oh, Hyemin;Kim, Sejeong;Lee, Soomin;Lee, Heeyoung;Ha, Jimyeong;Lee, Jeeyeon;Choi, Yukyung;Choi, Kyoung-Hee;Yoon, Yohan
    • Food Science of Animal Resources
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    • 제36권6호
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    • pp.779-786
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    • 2016
  • This study aimed to evaluate the prevalence of Listeria monocytogenes on livestock farms in Korea and determine their serotypes and genetic correlations. Twenty-five livestock farms in Korea (central: 15, south west: 7, south east: 3) were visited 2-3 times, and 2,018 samples (feces: 677, soil: 680, silage: 647, sludge: 14) were collected. Samples were enriched in LEB (Listeria enrichment broth) and Fraser broth media, and then plated on Palcam agar. The isolates were identified by PCR and 16S rRNA gene sequencing. Then, the sero-types, presence of virulence genes (actA, inlA, inlB, plcB, and hlyA), and antibiotic resistance were determined. Genetic correlations among the isolates were evaluated by analyzing the restriction digest pattern with AscI. Of the 2,018 samples, only 3 (0.15%) soil samples (FI-1-FI-3) from 1 farm in the south east region were positive for L. monocytogenes. Based on biochemical tests and multiplex PCR, the serotype of the isolates were 4ab (FI-1 and FI-3) and 3a (FI-2), which are not common in foodborne L. monocytogenes. The 3a sero-type isolate was positive for all tested virulence genes, whereas the 4ab serotype isolates were only positive for hlyA, actA, and inlA. The isolates were resistant to all 12 tested antibiotics, especially FI-3. The genetic correlations among the isolates were 100% for those of the same serotype and 26.3% for those of different serotypes. These results indicate that the prevalence of L. monocytogenes on livestock farms in Korea is low; however, the isolates are pathogenic and antibiotic resistant.

Serotypes and genotypes of Salmonella isolates from slaughtered pigs (도축돈에서 분리된 살모넬라의 혈청형 및 유전형)

  • Choi, Won-Zong;Jung, Ji-Hun;Won, Ho-Keun;Kang, Zheng-Wu;Hahn, Tae-Wook
    • Korean Journal of Veterinary Service
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    • 제31권1호
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    • pp.1-16
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    • 2008
  • Salmonella infections cause the disease in pigs but also some zoonotic Salmonella serotypes can be transmitted to human through swine products, resulting in food poisoning. The objective of this study was to investigate the bacteriological prevalence and detection of invA gene using Salmonella specific polymerase chain reaction (PCR), the epidemiological characteristics related to Salmonella strains cultured from pig samples in Gangwon areas using serotyping, random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE) methods. During the period of November 2001 through April 2002, 1,174 ileocecal lymph node were collected from the slaughtered pigs raised in 38 farms located in Gangwon province. The samples were submerged in boiling water and macerated in saline and lymph node homogenates were inoculated into Tetrathionate broth with iodine (TTB, Difco, 0.5% iodine was added) for enrichment growth. Then additional tests were performed using several mediums, and suspects were identified by API 20E kit (BioMerieux) and PCR. Of total 1,174 samples from 38 farms, 44 (3.7%) were isolated as Salmonella spp from 13 farms (34.2%). Of 44 isolates, 31 were in Yangyang region, followed by 9 in Goseong, 2 in both Gangneung and Sokcho. However, there was no difference in regional isolation frequency. All isolates have a 521bp amplified product in Salmonella specific PCR with primer invA which encodes in proteins for invasion of epithelial cells. Of 44 recovered serotypes, 23 (52.3%) were S Eingedi, 10 (22.7%) S Schwarzengrund, 9 (20.5%) S Typhimurium, and 2 (4.5%) S Mbandaka. In RAPD analysis, there appeared to be unique bands distinguishing each serotype, although similarities exist between the different serotypes. Four serotypes of 44 Salmonella isolates appeared to fall into 14 different RAPD types. In PFGE analysis, 9 S Typhimurium were tested with XbaI enzyme and SpeI enzyme. The combination of results obtained with two enzymes subdivided the 9 S Typhimurium into 4 PFGE types.

Molecular Characterization of Vibrio parahaemolyticus Isolates from Seawater, Fish Tanks, and Distributed Fishery Products in Jeju (제주지역 해수, 수족관물, 유통수산물에서 분리된 장염비브리오균의 분자생물학적 특성)

  • Man Jae Cho;Eunok Kang;Ye-seul Heo;Eun A Koh
    • Journal of Food Hygiene and Safety
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    • 제38권4호
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    • pp.246-254
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    • 2023
  • V. parahaemolyticus causes waterborne and foodborne disease such as acute diarrhea. In this study, V. parahaemolyticus isolates from seawater, fish tanks, and distributed fishery products in Jeju were investigated for potential toxin or species-specific genes (tdh, trh, tlh, and toxR) using RT-PCR and their genetic characteristics were analyzed using Pulsed-field gel electrophoresis (PFGE). Overall, V. parahaemolyticus of 90 strains (36.7%), including 33 strains from seawater, 8 strains from fish tanks, and 50 strains from fishery products, were isolated from 245 samples. All V. parahaemolyticus strains did not detect the tdh gene, whereas all strains detected tlh or toxR genes. In addition, trh genes were detected in 3 strains from seawater and 1 strain from fishery products. Monthly quantitative testing of seawater revealed that V. parahaemolyticus was positively correlated with water temperature. The 90 strains of V. parahemolyticus obtained in this study showed by gene homology between types, ranging from 64.0-97.3%. Among these, thirteen types showed 100% homology between genes. These results indicate that continuous monitoring is needed to facilitate food poisoning epidemiological investigations because some isolated V. parahaemolyticus strains harbored toxin genes and V. parahaemolyticus strains isolated from seawater, fish tanks, and distributed fishery products showed genetic similarity.

Prevalence of Toxin Genes and Antibiotic Resistance Profiles of Vibrio vulnificus strains isolated from Jeju Island (제주도에서 분리된 비브리오패혈증균의 독소 유전자 분포 및 항생제 내성)

  • Eunok Kang;Man Jae Cho;Ye-Seul Heo;Eun A Koh
    • Journal of Food Hygiene and Safety
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    • 제38권5호
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    • pp.381-389
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    • 2023
  • Vibrio vulnificus, the most fatal waterborne and foodborne pathogens of 50% fatality rate in the world, is common in seawater and occurs particularly in warmer months. In this study, we investigated the toxin genes using reverse transcription-polymerase chain reaction (RT-PCR), antibiotic resistance status using Vitek, and genetic characteristics using pulsed-field gel electrophoresis (PFGE) of different V. vulnificus strains isolated from the Jeju Island seawater, distribution fishery products, and fish tanks. We examined a total of 487 samples and isolated a total of 46 strains (including overlapping strains) of V. vulnificus, 44 strains from seawater and 1 strain each from fishery products and fish tank. We detected toxin gene vvhA in all 46 strains and rtxA, viu in 8 strains (17.4%) and 9 strains (19.6%) strains, respectively. Antibiotic resistance tests indicated 100% resistance to cefoxitin antibiotics. The PFGE analysis of the 46 strains identified a total of 6 types showed 100% homology and the degree of similarity was 81.3-98.0%; however, there were no similarity between the regions and samples. These results indicate that V. vulnificus isolated from the seawater, fishery products, and fish tanks should be continuously monitored as cases of food poisoning caused by V. vulnificus with toxin genes have been reported in Jeju Island.

Serovars and Genetic Characteristic of Salmonella spp. Isolates from Jeju Island, South Korea (제주도에서 분리된 살모넬라균의 혈청형 및 유전학적 특성)

  • Eunok Kang;Man Jae Cho;Chang Hui Yang
    • Journal of Food Hygiene and Safety
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    • 제39권2호
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    • pp.134-151
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    • 2024
  • Salmonella spp. is among the most important water-borne and food-borne pathogens and is one of the most common causes of human gastroenteritis and diarrheal diseases globally. In this study, Salmonella spp. isolated from food, environmental samples, and patients with food poisoning or diarrhea were investigated Salmonella serovars, antibiotic resistance using Vitek2, and genetic characteristics through pulsed-field gel electrophoresis (PFGE). Salmonella spp. of 339 strains, including 26 strains from food or environmental samples and 313 strains from patients, were isolated from Jeju Island of South Korea between 2020 and 2023. The monthly number of isolated Salmonella spp. gradually increased from March, with the highest number being in August. No significant differences in Salmonella spp. isolated from patients according to gender was observed. However, Salmonella spp. was most frequently isolated from people aged 70 years or older and least frequently isolated from those between ages 10 and 19 years. Salmonella spp. isolated from food or environmental samples were distributed among eight different serovars and the main serovars were identified in the order of S. Bareilly (26.9%), S. Rissen (23.1%), and S. Thompson (19.3%). Salmonella spp. isolated from patients were distributed among 27 different serovars and the main serovars were identified in the order of S. Bareilly (31.0%), S. Typhimurium (24.6%), and S. Enteritidis (11.5%). The main cause serovars of Salmonella spp. outbreaks are S. Bareilly, S. Enteritidis, S. Thompson. Antibiotic resistance tests indicated resistance to various antibiotics and some Salmonella spp. exhibited multidrug resistance. Salmonella spp. showed various genetic correlations among the 17 serovars. These results indicate that they can be used as basic data for epidemiological investigations by predicting the appearance of Salmonella spp. and providing a scientific basis.

Analysis of the Causes of a Large Food Poisoning Outbreak Attributable to Bacillus cereus (Bacillus cereus에 의한 대규모 집단식중독 원인 분석)

  • Hyunah Lee;Youngeun Ko;Dayeon Lee;KyungA Yun;Hyeonjeung Kim;Ok Kim;Junhyuk Park
    • Journal of Food Hygiene and Safety
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    • 제39권2호
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    • pp.102-108
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    • 2024
  • This study was performed to establish the epidemiological features of a food poisoning outbreak that occurred in the cafeteria of a company in Chungcheongnam-do Province, Korea, in October 2020, and to recommend measures to prevent similar outbreaks. Twenty-one patients with acute gastroenteritis, three food handlers, seven cooking utensils, and 12 preserved food samples were subjected to viral and bacterial analyses based on procedures described in the "Manual for Detection of Foodborne Pathogens at Outbreaks". Among 135 individuals who had been served the meals, 21 (15.6%) showed symptoms of nausea and vomiting within an hour of consuming the food. Bacillus cereus were isolated from 11 (52.4%) of the 21 patients, one food service employee, one item of cooking ware, and 12 preserved food samples. In addition, we confirmed the toxin genes CER, nheA, and entFM from the isolated B. cereus strains. Pulsed-field gel electrophoresis results indicated that all of the isolated B. cereus strains were closely related, with the exception of strains obtained from one patient and one sample of preserved food. These findings provide evidence to indicate that the isolated B. cereus originated from preserved foods and an unhygienic eating environment. This outbreak highlights that the provision of food in non-commercial food systems must be thoroughly managed. In addition, it emphasizes the necessity for the correct and timely identification of causal pathogens for tracing the cause of food poisoning outbreaks, and the need to preserve food under appropriate conditions. To prevent similar cases of food poisoning, it is necessary to investigate cases based on an epidemiological approach and share the findings.

Outbreak of Shigellosis Occurred in a Preschool and Two Elemetary Schools in Mapo-Gu, Seoul (서울시 마포구 관내 어린이집 및 초등학교에서 집단 발병한 세균성 이질)

  • Park, Tae Su;Lee, Ho Jun;Kim, Su Yeon;Lee, Dong Woo;Kim, Jae Yoon;Baik, Ji Na;Park, Yu Mi;Park, Mi Sun;Lee, Bok Kwon
    • Pediatric Infection and Vaccine
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    • 제13권1호
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    • pp.71-77
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    • 2006
  • Purpose : Shigellosis is still an important cause of acute food-borne diarrheal diseases throughout the world. We investigated the transmission routes and clinical course through the outbreak of shigellosis in Mapo-Gu, Seoul. Methods : From October 7th to November 19th in 2005, Mapo-Gu public health center had referred 32 patients with confirmed or suspected shigellosis to the National Medical Center. We had investigated source of infection, epidemiology, laboratory findings, and clinical course of the cases occurred during this outbreak. Results : Among 32 patients, 24 patients had been confirmed with shigellosis, 8 patients had been diagnosed with suspected shigellosis. They ranged in age from 5 months to 12 years old and their mean age was 6.5 years. The clinical manifestations were as follows; diarrhea, fever, abdominal pain and asymptomatic condition. Symptoms had sustained for 3.7 days on the average. S. sonnei were cultured by rectal swab and founded to be resistant to ampicillin and TMP/SMX except to 3rd generation cephalosporin. After treatment with antibiotics such as cefixime and ceftriaxone or imipenem and conservative treatment with electrolyte and fluid replacement for 5~7 days, Stool cultures of the rectal swab grew no Shigella in these cases except 3 cases. Conclusion : An outbreak of shigellosis had occurred in a preschool and elementary school children. From the same results of antimicrobial susceptibility and pulsed-field gel electrophoresis patterns in this study, we suggest that the outbreak of shigellosis in this report had been originated from a single strain. According to all negative results about suspected food and water cultures, we couldn't find out source of infection. Through materials offerred by Mapo-Gu Public Health Center, we presumed the trasmission routes probably were person-to-person.

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