• Title/Summary/Keyword: Shigella spp.

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Simultaneous Detection of Food-borne Pathogenic Bacteria in Ready-to-eat Kimbab Using Multiplex PCR Method

  • Cho, Kye-Man;Kambiranda, Devaiah M;Kim, Seong-Weon;Math, Renukaradhya K;Lim, Woo-Jin;Hong, Su-Young;Yun, Han-Dae
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1240-1245
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    • 2008
  • Kimbab is the most popular ready-to-eat (RTE) food in Korea. A rapid detection method based on multiplex PCR technique was developed for detection of major food-borne pathogens like Salmonella spp., Shigella spp., Bacillus cereus, Listeria monocytongenes, and Staphylococcus aureus. Specific bands were obtained as 108 bp (Sau, S. aureus), 284 bp (Sal, S. enterica, S. enteritids, and S. typhmurium), 404 bp (Lmo, L. monocytogenes), 475 bp (Bce, B. cereus), and 600 bp (Shi, S. flexineri and S. sonnei). Visible cell numbers varied from 4.14-5.03, 3.61-4.47, and 4.10-5.11 log CFU/g in randomly collected June, July, and August samples, respectively. Among the 30 kimbab samples obtained 83.3% samples were contaminated and 16.7% samples were free from contamination. The highest rate of contamination was with S. aureus (56.7%) followed by B. cereus (43.3%), Salmonella spp. (36.7%), Shigella spp. (13.3%), and L. monocytogenes (6.7%). The identification of the pathogenic species could be faster using one polymerase chain reaction (PCR) and the ability to test for food-borne pathogenic species in kimbab will save time and increase the ability to assure its quality.

Studies on Antibiotic Resistance and Growth Characteristics of Shigella Sonnei Isolated from Patients of Shigellosis (이질환자에서 분리한 Shigella Spp.의 항생제 내성과 성장특성)

  • 이영희;박나영;이신호
    • Journal of Food Hygiene and Safety
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    • v.16 no.1
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    • pp.27-32
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    • 2001
  • Antibiotic resistance of thirty strains of Shigella sonnei isolatedfrom patient of Shigellosis outbreke at Young Cheon area in 1998 was tested. Twenty-seven strains were resistant to Tr(Trimethoprim-Sulfamethoxazol) and Shigella sonnei SG-48 was resistant to Tr(Trimethopirm-Sulfamethoxazol), Ap(Ampicillin), Cp(Cephalothin) and Pi(Piperacillin). Shigella sonnei SG-49, SG-66, and SG-73 were senstive to all tested antibiotics. Physiological charactristics of isolated Shigella sonnei SG-48, SG-49, SG-57, and SG-73 such as effect of pH, NaCl concentration and temperature on the growth, survival in adverse condition and heat resistance were investigated Growth of the strains were inhibited at pH 4 and pH 9. All strains were grown in Tryptic soy broth containing 6% of NaCl but inhibited in TSB containing 9% of NaCl except Shigella sonnei SG-73 after incubation for 18hrs at 37$^{\circ}C$. Selected strains grew during storage at 10 but did not grow at 4. The strains were survived in 1% pepton solution for 15 days at 37$^{\circ}C$. Viable cell of selected strains were decreased 45 log cycle after heat treatment for 30 mins at 6$0^{\circ}C$ but did not detect by heat treatment for 5 mins at 7$0^{\circ}C$.

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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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Waterborne Pathogens Identification in Public Bathroom by PCR-Reverse Blot Hybridization Assay

  • Choi, Seung-Gu;Song, Woon-Heung;Lee, Jae-Sang;Yang, Byoung-Seon;Choi, Myeong-Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.3
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    • pp.120-123
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    • 2011
  • A total of 30 water samples were collected from 30 different public baths in Seoul, Korea. Contamination of public bath water by waterborne pathogens can cause disease outbreaks and contribute to increase background rates of disease. Pathogens in water was filtered by nitrocellulose membrane with $0.45{\mu}m$ pore size. The membrane filters were analyzed by both cultivation and polymerase chain reaction (PCR) amplification of partial 16S rRNA gene. Various microorganisms including 4 Escherichia coli/Shigella spp. 1 Salmonella spp. 3 Pseudomonas aeruginosa and 2 Mycobacterium spp. were identified by reverse blot hybridization assay (REBA). PCR-REBA was able to identify many bacterial genera in one assay. Our results suggest that appropriate hygiene practice and continuous monitoring is needed for reducing health risk associated with public bath houses.

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Growth Inhibition Profile of an Antibacterial Entity from Paenibacillus DY1 Isolated from Korean Soil against Multidrug Resistant Enteric Bacterial Strains and Its Characterization

  • Shin, Eun-Seok;Kwon, Sun-Il;Yoo, Kwan-Hee
    • Biomedical Science Letters
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    • v.13 no.1
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    • pp.47-53
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    • 2007
  • Due to wide abuse of antibiotics both in human and livestock use, the advent and spread of multidrug resistant (MDR) pathogens becomes a serious health problem all over the world. Since the development of new antibiotics is at a standstill in pharmaceutical industry, the choice of therapeutic antibiotics is getting narrower. In this study, in an effort to search new antibiotics, the antimicrobial activity of Paenibacillus DY1 isolated from Korean soil was characterized on its growth inhibition spectrum against various health threatening MDR strains, with its stability and chemical structure. Extracellular culture filtrate of Paenibacillus DY1 effectively inhibits the growth of all the tested MDR enteropathogenic Eshcherichia coli, enterohemolytic E. coli, and enterotoxigenic E. coli strains, at a similar level to that on the nonresistant control E. coli strains. It showed significant growth inhibition effect against the causative agents of class one legal communicable disease, MDR Salmonella typhi, MDR Salmonella paratyphi A, food poisoning bacteria, MDR Salmonella typhimurium, and other MDR Salmonella spp. The growth of all of 10 different MDR Shigella spp. strains and 6 different Vibrio spp. strains tested was also inhibited. The antimicrobial activity of Paenibacillus DY1 was well preserved after heat treatment, and was also stable in both alkaline and acidic environment. The antimicrobial activity was partially purified with Diaion HP20 column and TLC. By NMR study, the putative structure of the activity was postulated as an alkane having hydroxyl groups.

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The Shigella Flexneri Effector OspG Interferes with Innate Immune Responses by Targeting Ubiquitin-Conjugating Enzymes

  • Kim, Dong-Wook
    • Proceedings of the PSK Conference
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    • 2005.11a
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    • pp.231-232
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    • 2005
  • Bacteria of Shigella spp. are responsible for shigellosis in humans, a disease characterized by destruction of the colonic epithelium that is induced by the inflammatory response elicited by invasive bacteria. They use a type III secretion system injecting effector proteins into host cells to induce their entry into epithelial cells and triggers apoptosis in macrophages. We present evidence that the effector OspG is a protein kinase that binds various ubiquitinylated ubiquitin-conjugating enzymes (E2s) and blocks degradation of phospho-$I{\kappa}B{\alpha}$ induced upon entry of bacteria into epithelial cells. Transfection experiments confirmed that OspG interferes with the $NF-{\kappa}B$ activation patway by preventing phospho-$I{\kappa}B{\alpha}$ degradation, suggesting that OspG inactivates a component of the $SCF^{{\beta}-TrCP}$ ubiquitin ligase complex (E3) involved in phospho-$I{\kappa}B{\alpha}$ ubiquitination. Upon infection of ileal loops in rabbits, the ospG mutant induced a stronger inflammatory response compared with the wild-type strain, indicating that OspG down-regulates the host innate response induced by invasive bacteria.

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Investigation for the Infectious Diarrhea by Pathogenic Microorganism from Hospitals in ChungNam Province in 2004 (2004년 충남지역 의료기관의 설사환자 가검물에서 분리된 병원성미생물 감염실태에 관한 조사연구)

  • Kim, Woo-Sik;Song, Nak-Soo;Sung, Si-Yeoul;Cha, Yeun-Tae;Seo, Woo-Sung;Lee, Moo-Sik;Kim, Keon-Yeop;Na, Bak-Ju
    • Journal of agricultural medicine and community health
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    • v.30 no.2
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    • pp.137-149
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    • 2005
  • Objectives and Methods: This study was performed to investigate the etiologic bacterial, viral and protozoal organisms for the diarrhea from hospitals in Chungnam area from January to December in 2004. Total of 787 fecal samples were collected and examined. Results and Conclusions: In test for enteropathogenic bacteria, total of 79 cases out of 787 samples from hospitals showed positive isolation. Among 79 positive samples, 27 cases were confirmed as Salmonella spp.. 20 cases as pathogenic E. coli, 18 cases as Clostridium perfringens, 6 cases as Staphylococcus aureus, 4 cases as Shigella spp. and 4 cases as Vibrio parahaemolyticus. In test for enteropathogenic virus, 190 cases out of 787 samples from hospitals showed positive reaction. Among 190 samples, 115 cases were confirmed as rotavirus, 55 cases as norovirus, 5 case as astrovirus, 4 case as rotavirus & norovirus, 3 cases as adenovirus, 2 case as rotavirus & astrovirus. In test for enteropathogenic protozoa, 6 cases out of 787 samples from hospitals showed positive result. Among 6 samples, 5 cases were confirmed as Entamoeba histolytica and 1 cases as Giardia lamblia. When we classified the positive results by the age of the patients, the highest isolation rate was noted in a group of age under 10 and over 60 for bacterial, viral and protozoal pathogens. Especially, patient below age of 5 showed high positive rate. When we classified the positive results by the time, pageathogenic bacteria were isolated throughout the year, and the highest frequency was noted in August. On the other hand, pathogenic viruses were detected more frequently during the colder season from December to April. Antimicrobial susceptibility test for the isolated bacteria resulted as follows; Salmonella strains showed high drug resistance rates against ampicillin, chloramphenicol, tetracycline, nalidixic acid, ticarcillin. Shigella strains showed high drug resistance rates against ampicillin, trimethoprim/sulfamethoxazole, chloramphenicol, tetracycline, ampicillin/sulbactam, ticarcillin. Pathogenic E. coli strains showed high drug resistance rates against ampicillin, cephalothin, gentamicin, tetracycline, nalidixic acid, ampicillin/sulbactam, ticarcillin.

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Development of a Panel of Multiplex Real-Time Polymerase Chain Reaction Assays for Simultaneous Detection of Canine Enteric Bacterial Pathogens (개의 장내 병원균의 동시 검출을 위한 다중 실시간 중합효소연쇄반응분석 패널개발)

  • Jang, Hye-Jin;Han, Jae-Ik;Kang, Hyo-Min;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.154-157
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    • 2015
  • A major cause of diarrhea in a dog is an infection with bacteria which include Salmonella spp., Campylobacter (C.) spp., and Clostridium (Cl.) spp.. It is fastidious to identify these bacteria by the culture. The purpose of this experiment is to devise the method for detecting Cl. perfringens, C. jejuni, C. coli, and Salmonella spp. with rapid and high sensitivity. The fecal samples collected from 71 normal and 66 diarrheic dog feces were used to compare the prevalence of the enteric pathogens and to develop a multiplex real-time polymerase chain reaction (PCR) assay for clinical use. Detection of Cl. perfringens, C. coli, and C. jejuni in diarrhea feces was higher than normal feces. A developed multiplex real-time PCR is useful for determining the presence and quantity of pathogen-specific or other unique sequences with in a fecal sample.

Isolation and Identification of Pathogenic Bacteria from Spinach (시금치로부터 병원성세균의 분리 및 동정)

  • Kim, Hye-Jung;Kim, Young-Hoon;Lee, Dong-Sun;Paik, Hyun-Dong
    • 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.

Changes of Indicator Microorganisms and Pathogenic Bacteria in Spinach during Cook-Chill Process (시금치의 cook-chill 가공 중 오염지표균 및 병원성세균의 변화)

  • Kim, Hye-Jung;Park, Jae-Kap;Lee, Dong-Sun;Paik, Hyun-Dong
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.927-930
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    • 2002
  • Spinach minimally processed using cook-chill and sous vide techniques was vacuum-packed in low gas permeable plastic film, pasteurized at $70^{\circ}C$ for 2 min, cooled rapidly at $3^{\circ}C$, and stored at 3 and $10^{\circ}C$. Contents of mesophilic bacteria, psychrophilic bacteria, anaerobic bacteria, spore-forming bacteria, total coliforms, yeast and molds, fecal Streptococcus, and Enterobacteriacea were measared to identify the degree of food contamination. Number of mesophilic bacteria, detected at $2.2{\times}10^8\;cfu/g$ in raw spinish, decreased to about $6.0{\times}10^3\;cfu/g$ after cook-chill process. During the storage at 3 or $10^{\circ}C$, levels of mesophilic, psychrophilic and anaerobic bacteria increased, whereas total coliforms, yeast and molds, fecal Streptococcus, and Enterobacteriacea were not detected. Twelve strains of Aeromonas hydphila, Escherichia coli O157:H7, Plesiomonas shigelloides, Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Yersinia enterocolitica, Bacillus cereus, Campylococcus spp., Clostridium perfringens, Listeria monocytogenes, and Staphylococcus aureus were examined for detecting the presence of pathogenic bacteria in spinach. B. cereus and C. perfringens were isolated from raw, washed, and cook-chilled spinach, whereas A. hydrophila was isolated only from washed spinach. S. aureus was isolated from raw and washed spinach, but not from cook-chilled spinach. Other pathogenic organisms were not detected in raw, washed, and cook-chilled spinach.