• Title/Summary/Keyword: cpe gene

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Toxin Gene Typing, DNA Fingerprinting, and Antibiogram of Clostridium perfringens Isolated from Livestock Products

  • Lee, Seung-Bae;Choi, Suk-Ho
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.394-401
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    • 2006
  • Forty Clostridium perfringens isolates were obtained from twelve animal products, following the examination of eighty six beef, pork, broiler chicken and salami meat products, and eleven milk powder products. There were 21 isolates from salami stored at $25^{\circ}C$, 3 isolates from pork, 4 isolates from beef, 9 isolates from broiler chicken, and 3 isolates from milk powder. Only the cpa gene encoding a toxin among the 5 toxin genes tested (cpa, cpb, etx, iap, and cpe) was detected in all forty isolates, suggesting contamination with C. perfringens type A. DNA fingerprinting analysis using PCR of the tRNA intergenic spacer (tDNA-PCR) and the 16S-23S internal transcribed spacer (ITS-PCR), and randomly amplified polymorphic DNA (RAPD) analysis were attempted to differentiate the isolates. RAPD analysis was the most discriminating method among the three PCR analyses. Isolates from the same products tended to show similar RAPD patterns. Antimicrobial susceptibility tests showed that some isolates from broiler chickens had the same antibiogram with multiple resistance to streptomycin, colistin, and ciprofloxacin. Antibiograms were similar between isolates from the same livestock products, but differed considerably between the products.

Toxin Genes and Antimicrobial Resistance of Clostridium perfringens Strains Isolated from Commercial Jeotgals (시판 젓갈에서 분리한 Clostridium perfringens의 독소 유전자 및 항균제 내성 분석)

  • Shin-Hye Lee;Kwon-Sam Park
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.6
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    • pp.826-832
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    • 2023
  • Clostridium perfringens causes diarrhea and other diseases in humans and animals. We investigated the prevalence, toxin gene profiles, and antimicrobial resistance of C. perfringens isolated from commercial jeotgal sample. C. perfringens was isolated from 11 of 22 commercial jeotgals. All C. perfringens strains were positive for the alpha toxin gene, but not for the beta, epsilon, iota, CPE or NetB toxin genes; therefore, all strains were identified as type A C. perfringens. However, the beta2 toxin gene was identified in 54.5% of isolates. Disk diffusion susceptibility tests showed that most isolates were resistant to kanamycin (90.9%), nalidixic acid (72.7%), oxacillin (54.5%), erythromycin (27.3%), ciprofloxacin (9.1%) and clindamycin (9.1%). However, all strains were susceptible to 14 other antimicrobial including amoxicillin, ampicillin, and chloramphenicol. The average minimum inhibitory concentrations against C. perfringens of clindamycin, kanamycin, and nalidixic acid were 128.0, 128.0, and 54.0 ㎍/mL, respectively. These results provide new insight into the necessity for sanitation of commercial jeotgal, and provide evidence to help reduce the risk of contamination with antimicrobial-resistant bacteria.

Cytotoxic Effects of an Oncolytic Adenoviral Vector AdLPCDIRESE1A in Hepatocellular Carcinoma Cells (암세포 용해성 AdLPCDIRESE1A 벡터의 간암 세포독성효과)

  • Chung, In-Jae
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.75-79
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    • 2011
  • The replication competent adenoviral vector (AV), AdLPCDIRESE1A was generated and reported previously to have cytotoxic effects in some cell lines. In AdLPCDIRESE1A, the expression of cytosine deaminse (CD) and E1A genes are under the control of tumor-specific L-plastin promoter. CD enzyme can deaminate the nontoxic prodrug 5-fluorocytosine (5-FC) to the toxic 5-fluorouracil (5-FU). E1A gene is essential for viral replication. Primary liver cancer, most of which is hepatocellular carcinoma (HCC), is the third common leading cancer in Korea. Thus, we have conducted in vitro preclinical study to evaluate effectiveness of AdLPCDIRESE1A on HCC. The efficacy of cytotoxicity was measured by generation of cytopathic effect (CPE) and cell counting. We infected HepG2 cells with various MOI of vector alone or concurrent with 5-FC. Exposure of cells to AdLPCDIRESE1A generated a significant cytotoxic effect as compared to the control. Almost 83% of the cell had manifested the characteristic cytotoxic effect on day 9 after infection of cells with 10 MOI of vector. We also observed the additive cytotoxic effects when AdLPCDIRESE1A vector had been coadministrated with 5-FC. The results suggest that the use of AdLPCDIRESE1A/5FC may be value in treatment of liver cancer. Further animal studies are needed for clinical trial.

Inhibition of Human Cytomegalovirus Replication using Peptide Nucleic Acids with Polyethylenimine

  • Eum, Jin-Seong;Park, Young-Doo;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2010.10a
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    • pp.660-662
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    • 2010
  • To control replication of human cytomegalovirus (HCMV) effectively, inhibitors of peptide nucleic acids (PNA) with a gene delivery agent, PEI (polyethylenimine) against HCMV were applied. The transfection of these PNA inhibitors with PEI agent into host cells showed synergic inhibition effect of HCMV replication. These inhibition effect was confirmed by methods of RT-PCR, CPE, real-time-PCR, and Western blot.

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The Combined Anti-apoptotic Effect from Tamiflu and Pinoresinol of Forsythia fructus Extract Against Influenza Virus Infection (연교 추출물 Pinoresinol와 Tamiflu의 병용효과로부터 Influenza Virus 감염에 의한 세포사멸 억제효과)

  • Kim, Sang-Tae;Kim, Jang-Soo;Choe, Young-Uung;Kim, Young-Kyoon
    • Korean Journal of Pharmacognosy
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    • v.42 no.1
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    • pp.9-14
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    • 2011
  • The fruit body of Forsythiae Fructus (Oleaceae), a common Korean medical herb, is widely used in the treatment of cold and inflammation. In order to elucidate the action mechanism and the active principles from the plant against anti-influenza virus, the influenza virus hemagglutinin (HA) and neuraminidase (NA) gene RT-PCR and Viral Screening & Identification (VSI) assay were conducted, and the activity against viral replication was also investigated. Consequently, one active constituent, namely pinoresinol showed the in vitro antiviral principle using a cytopathic effect (CPE) reduction method, indicating pinoresinol possessed anti-influenza viral activity. Furthermore, combination of pinoresinol and Tamiflu exhibited higher activities than Tamiflu alone against influenza virus (H3N2) infection. The results suggested that combination of pinoresinol with Tamiflu could be a better candidate for an ant-H3N2 viral agent in the treatment of the influenza.

A survey on hygiene management for raw by-products of beef in Gwangju area, Korea (광주지역 생식용 소 부산물의 위생관리 실태 조사)

  • Kim, Ji-Yeon;Jang, Mi-Sun;Koh, Ba-Ra-Da;Ji, Tea-Kyung;Sung, Chang-Min;Park, Da-Hae;Kim, Hyun-Joong;Kim, Eun-Sun;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.36 no.3
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    • pp.209-216
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    • 2013
  • A total of 301 samples of bovine liver, spleen and omasum were collected from butchers and restaurants in Gwangju, Korea during 2012 and all samples were subjected to bacteriological examination and antibiotic residues. Also, this study was performed to survey the consciousness for hygiene of livestock workers who are handling bovine by-products in Gwangju. The detection rate of aerobic plate count (APC) was higher in summer than in other seasons in all by-products (P=0.000). The detection rate of E. coli count was lower in the liver than the spleen and omasum (P=0.000). Twenty four of the samples (8.0%) were contaminated with S. aureus while one spleen sample (0.3%) was contaminated with L. monocytogenes and finally 10 (3.3%) of the liver and omasum samples were contaminated with Cl. perfringens. Five of the twenty-four S. aureus isolates harbored enterotoxin gene. However, the cpe gene of Cl. perfingens was not detected among any of the 10 isolates. Antibiotic residues were not detected in the liver samples. The consciousness survey's results showed that most of them (58.8%) were safe.

Inhibition of Herpesvirus-6B RNA Replication by Short Interference RNAs

  • Yoon, Jong-Sub;Kim, Sun-Hwa;Shin, Min-Chul;Lee, Dong-Gun;Hong, Seong-Karp;Jung, Yong-Tae;Khang, In-Gu;Shin, Wan-Shik;Kim, Chun-Choo;Paik, Soon-Young
    • BMB Reports
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    • v.37 no.3
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    • pp.383-385
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    • 2004
  • RNA interference (RNAi) is a process of sequence-specific gene silencing, which is initiated by double-stranded RNA (dsRNA). RNAi may also serve as an antiviral system in vertebrates. This study describes the inhibition of herpesvirus-6B (HHV-6B) replication by short interference RNAs (siRNAs) that are targeted to the U38 sequence that encodes DNA polymerase. When virus-infected SupT1 cells were treated by siRNA, these cells blocked the cytopathic effect (CPE) and detected the HHV-6B antibody-negative in indirect immunofluorescence assays (IFA). Our result suggests that RNAi can efficiently block Herpesvirus-6B replication.

Evaluation of the virulence genes and Shiga toxin-producing abilities of Escherichia coli field isolates causing edema disease in pigs (국내 분리 돼지 부종병 대장균의 병원성 유전자 및 시가독소 생성 검증)

  • Seo, Byoung-Joo;Jeong, Chang-Gi;Kang, A-Rum;Cho, Ho-Seong;Kim, Won-Il
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.87-92
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    • 2016
  • Porcine edema disease (ED) is a communicable disease of pigs caused by infection with Shiga toxin (Stx)-producing Escherichia coli (STEC) which expresses F18 fimbriae and/or Stx type 2e (Stx2e). While STEC causes a severe illness including hemorrhagic colitis and hemolytic-uremic syndrome in humans, it induces damage to the vascular endothelium, which results in edema, hemorrhage, and microthrombosis, leading in high mortality in pigs. In the present study, we cultured Stx2e-producing E. coli field isolates from conventional pig farms that experienced sudden deaths previously with symptoms similar to porcine edema disease, which were further investigated with Shiga toxin profiles. A total of 43 strains were identified from the collected samples by F18 or Stx2e specific PCR. Based on the PCR, 42 isolates out of 43 isolates were proved to carry one of F18 or Stx2e genes and 14 isolates to carry both F18 and Stx2e genes. All of the 30 isolates that harbored Stx2e gene induced the cytopathic effect (CPE) in vero cells and especially, the isolate 150229 produced the highest level of Shiga toxin. Therefore, we identified the virulence genes (F18 and Stx2e) and demonstrated Shiga toxin-producing abilities from porcine edema disease causing E. coli filed isolates. These results suggested that one of the isolates could be a vaccine antigen candidate against STEC through further investigating to elicit an immune response.

Classical swine fever disease in Cheolwon

  • Park Yang-Soon;Shin Myung-Kyun;Chong Dong-Soo;Cheong Ki-Soo;Park Young-Nam;Choi Jee-Hee
    • Korean Journal of Veterinary Service
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    • v.27 no.4
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    • pp.345-354
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    • 2004
  • Two cases of classical swine fever (CSF) disease have broken out in Cheolwon (7 April, 2002). The suspected pig herds were huddled together because of high fever (over $40^{\circ}C$) and showed remarkable decrease of the leukocytes. The staggering gait related to posterior weakness, constipation and lethargy, hyperemia, hemorrhagic lesions (on the skin, muzzle, ears, limbs, tail and inner part of legs) and conjunctivitis with dirty streaks below the eyes were observed. The inflammation in the lung, infarction in the spleen, swelling and hemorrhage in lymph nodes, kidney, intestine, heart and cheese like purulent inflammation of the tonsil were observed. The ulcers of the colon were also detected. Several clinical and laboratory techniques including blood test, histo-pathological examinations, indirect fluorescent antibody (IFA) test and RT-PCR test were applied to diagnose the disease. Inoculation test on PK-15 cell was also performed. The necrosis of the lymphatic cells and infiltration of the vessel circumferential cells in the brain and lymph organs were commonly viewed. The proliferation of the glia cell (gliosis) in the lymph was particular. Cytopathogenic effect (CPE) and specific fluorescent-bright-green areas (with IFA) appeared in PK-15 cells inoculated with suspected blood plasma. The IFA test on the epithelial and mucous membrane cells of tonsil was positive. RT-PCR technique required more working hours and labor than other techniques in this examination but it was useful because of the sensitivity to the CSF viral gene.

Prevalence of Microbiological Contamination in the Ready-To-Eat Side Dishes Sold in Gyeongsangnam-do, South Korea (경남지역에서 유통되는 즉석 반찬류의 미생물 오염도 조사)

  • Ji-Yeon Um;Hye-Jeong Jang;Yeon-Ju Choi;So-Young Kim;Areum Jo;Min Young Kim;Jihee Ahn;Jea-Dong Kim
    • Journal of Food Hygiene and Safety
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    • v.38 no.4
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    • pp.217-227
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    • 2023
  • The consumption of ready-to-eat side dishes is rapidly growing in South Korea. These foods are particularly vulnerable to microbiological contamination as they are often cooked without any treatment, such as heating or stored at room temperature after cooking. Hence, in 2022, we analyzed the ready-to-eat side dishes sold in Gyeongsangnam-do, South Korea for microbiological contamination. We collected 100 samples from supermarkets in 7 cities, and then examined them for presence of food-borne pathogens and sanitary indicator bacteria. In the analysis of the food-borne pathogens, Bacillus cereus and Clostridium perfringens were isolated from 51 samples (51.0%) and 3 samples (3.0%), respectively. However, both quantitatively met the Korean Food Standards Codex. Genes of five different enterotoxins and one emetic toxin were analyzed from the 51 isolated B. cereus strains. We detected enterotoxin entFM (100.0%), nheA (94.1%), hblC (58.8%), cytK (56.9%), and bceT (41.2%) in 51 isolates, and emetic toxin gene, CER, in only one (2.0%) isolate. We did not detect C. perfringens toxin gene (cpe) that causes food poisoning in any one of the three C. perfringens isolates. In the case of sanitary indicator bacteria, Kimchi had the highest levels of total aerobic bacteria and coliforms, followed by Saengchae, Jeotgal, Jeolim, Namul, and Jorim, respectively. We counted total aerobic bacteria at two different storage temperatures (4℃ and 20℃) to determine the effect of storage temperature. When stored at 20℃, total aerobic bacteria count increased in most of the ready-to-eat side dishes, except for Jeotgal. This result conclusively shows the need for refrigerating the ready-to-eat side dishes after purchase. Further research is needed to assess the risk and safety of the ready-to-eat side dishes available in the market and determine appropriate safety management practices.