• Title/Summary/Keyword: Pathogenic E. coli O-157

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Molecular characterization and functionality of rumen-derived extracellular vesicles using a Caenorhabditis elegans animal model

  • Hyejin Choi;Daye Mun;Sangdon Ryu;Min-jin Kwak;Bum-Keun Kim;Dong-Jun Park;Sangnam Oh;Younghoon Kim
    • Journal of Animal Science and Technology
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    • v.65 no.3
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    • pp.652-663
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    • 2023
  • The rumen fluids contain a wide range of bacteria, protozoa, fungi, and viruses. The various ruminal microorganisms in the rumen provide nutrients by fermenting the forage they eat. During metabolic processes, microorganisms present in the rumen release diverse vesicles during the fermentation process. Therefore, in this study, we confirmed the function of rumen extracellular vesicles (EVs) and their interaction with the host. We confirmed the structure of the rumen EVs by transmission electron microscope (TEM) and the size of the particles using nanoparticle tracking analysis (NTA). Rumen EVs range in size from 100 nm to 400 nm and are composed of microvesicles, microparticles, and ectosomes. Using the Caenorhabditis elegans smart animal model, we verified the interaction between the host and rumen EVs. Exposure of C. elegans to rumen EVs did not significantly enhance longevity, whereas exposure to the pathogenic bacteria Escherichia coli O157:H7 and Staphylococcus aureus significantly increased lifespan. Furthermore, transcriptome analysis showed gene expression alterations in C. elegans exposed to rumen EVs, with significant changes in the metabolic pathway, fatty acid degradation, and biosynthesis of cofactors. Our study describes the effect of rumen EV interactions with the host and provides novel insights for discovering biotherapeutic agents in the animal industry.

Microbiological Hazard Analysis for HACCP System Application to Vinegared Pickle Radishes (식초절임 무의 HACCP 시스템 적용을 위한 미생물학적 위해 분석)

  • Kwon, Sang-Chul
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.69-74
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    • 2013
  • This study has been performed for 150 days from February 1 - June 31, 2012 aiming at analyzing biologically hazardous factors in order to develop HACCP system for the vinegared pickle radishes. A process chart was prepared as shown on Fig. 1 by referring to manufacturing process of manufacturer of general vinegared pickle radishes regarding process of raw agricultural products of vinegared pickle radishes, used water, warehousing of additives and packing material, storage, careful selection, washing, peeling off, cutting, sorting out, stuffing (filling), internal packing, metal detection, external packing, storage and consignment (delivery). As a result of measuring Coliform group, Staphylococcus aureus, Salmonella spp., Bacillus cereus, Listeria Monocytogenes, E. coli O157:H7, Clostridium perfringens, Yeast and Mold before and after washing raw radishes, Bacillus cereus was $5.00{\times}10$ CFU/g before washing but it was not detected after washing and Yeast and Mold was $3.80{\times}10^2$ CFU/g before washing but it was reduced to 10 CFU/g after washing and other pathogenic bacteria was not detected. As a result of testing microorganism variation depending on pH (2-5) of seasoning fluid (condiment), pH 3-4 was determined as pH of seasoning fluid as all the bacteria was not detected in pH3-4. As a result of testing air-borne bacteria (number of general bacteria, colon bacillus, fungus) depending on each workplace, number of microorganism of internal packing room, seasoning fluid processing room, washing room and storage room was detected to be 10 CFU/Plate, 2 CFU/Plate, 60 CFU/Plate and 20 CFU/Plate, respectively. As a result of testing palm condition of workers, as number of general bacteria and colon bacillus was represented to be high as 346 $CFU/Cm^2$ and 23 $CFU/Cm^2$, respectively, an education and training for individual sanitation control was considered to be required. As a result of inspecting surface pollution level of manufacturing facility and devices, colon bacillus was not detected in all the specimen but general bacteria was most dominantly detected in PP Packing machine and Siuping machine (PE Bulk) as $4.2{\times}10^3CFU/Cm^2$, $2.6{\times}10^3CFU/Cm^2$, respectively. As a result of analyzing above hazardous factors, processing process of seasoning fluid where pathogenic bacteria may be prevented, reduced or removed is required to be controlled by CCP-B (Biological) and threshold level (critical control point) was set at pH 3-4. Therefore, it is considered that thorough HACCP control plan including control criteria (point) of seasoning fluid processing process, countermeasures in case of its deviation, its verification method, education/training and record control would be required.

A Study on Dose-Response Models for Foodborne Disease Pathogens (주요 식중독 원인 미생물들에 대한 용량-반응 모델 연구)

  • Park, Myoung Su;Cho, June Ill;Lee, Soon Ho;Bahk, Gyung Jin
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.299-304
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    • 2014
  • The dose-response models are important for the quantitative microbiological risk assessment (QMRA) because they would enable prediction of infection risk to humans from foodborne pathogens. In this study, we performed a comprehensive literature review and meta-analysis to better quantify this association. The meta-analysis applied a final selection of 193 published papers for total 43 species foodborne disease pathogens (bacteria 26, virus 9, and parasite 8 species) which were identified and classified based on the dose-response models related to QMRA studies from PubMed, ScienceDirect database and internet websites during 1980-2012. The main search keywords used the combination "food", "foodborne disease pathogen", "dose-response model", and "quantitative microbiological risk assessment". The appropriate dose-response models for Campylobacter jejuni, pathogenic E. coli O157:H7 (EHEC / EPEC / ETEC), Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio parahaemolyticus, Vibrio cholera, Rota virus, and Cryptosporidium pavum were beta-poisson (${\alpha}=0.15$, ${\beta}=7.59$, fi = 0.72), beta-poisson (${\alpha}=0.49$, ${\beta}=1.81{\times}10^5$, fi = 0.67) / beta-poisson (${\alpha}=0.22$, ${\beta}=8.70{\times}10^3$, fi = 0.40) / beta-poisson (${\alpha}=0.18$, ${\beta}=8.60{\times}10^7$, fi = 0.60), exponential (r=$1.18{\times}10^{-10}$, fi = 0.14), beta-poisson (${\alpha}=0.11$, ${\beta}=6,097$, fi = 0.09), beta-poisson (${\alpha}=0.21$, ${\beta}=1,120$, fi = 0.15), exponential ($r=7.64{\times}10^{-8}$, fi = 1.00), betapoisson (${\alpha}=0.17$, ${\beta}=1.18{\times}10^5$, fi = 1.00), beta-poisson (${\alpha}=0.25$, ${\beta}=16.2$, fi = 0.57), exponential ($r=1.73{\times}10{-2}$, fi = 1.00), and exponential ($r=1.73{\times}10^{-2}$, fi = 0.17), respectively. Therefore, these results provide the preliminary data necessary for the development of foodborne pathogens QMRA.

Sterilization of Rapeseed Sprouts by Intense Pulsed Light Treatment (고강도 광원을 이용한 새싹 채소의 살균)

  • Park, Heeran;Cha, Gyung-Hee;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.48 no.1
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    • pp.36-41
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    • 2016
  • In this study, the effects of intense pulsed light (IPL) treatment on microbial inactivation and quality in rapeseed sprouts were investigated. Untreated rapeseed sprouts exhibit a high level of total aerobic bacteria (TAB) ($1.2{\times}10^7CFU/g$), coliform bacteria (coliform) ($3.3{\times}10^6CFU/g$), and pathogenic E. coli (PE) ($2.1{\times}10^5CFU/g$). The microorganisms found on rapeseed sprouts decreased with exposure to increasing light intensity and treatment time. The greatest reduction in microbial content was observed with a treatment of 1000 V, 5 pps for 10 min, where TAB, coliform, and PE levels decreased to 1.0 log CFU/g, 1.6 log CFU/g, and 1.8 log CFU/g, respectively. In agreement with these data, the microbial inactivation rate increased with the increase in the distance between the light source and the samples during IPL treatment. After IPL treatment of rapeseed sprouts, water content and vitamin C content decreased.

Biological and Chemical Hazards Factor Analysis for CCP(Critical Control Point) in Fried Process of Fried Noodles (유탕면류의 유탕공정 중 중요관리점(CCP)을 위한 미생물학적, 화학적 위해요소분석)

  • Kwon, Sang-Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.8
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    • pp.3578-3585
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    • 2012
  • The purpose of this study was to establish the critical limit at CCP (Critical Control Point) of HACCP (Hazard Analysis Critical Control Point) system for instant noodle and it was conducted at P company in Ichen(Gyeonggi-do), Korea. According to the CCP, Fried process were experimented to removal and decrease of microbiological and chemical hazards by measuring of each temperature and times. As a result, the standard plate count and pathogenic microorganism were not detected by fried processing (Temperature : $145{\pm}10^{\circ}C$, Time : $75{\pm}30$ sec). The acid value of chemical hazards produced by fried processing was able to manage, showed lower (0.2) than the legal limit (0.6). Air-borne bacterial examination results detected(3 CFU/mL, 3 CFU/mL) in the Frying Room and Steam Room. Therefore, the CCP-BC of fried process would be a great alternative to prevent and remove hazard analysis, such as general and pathogenic microorganism (E. coli O157:H7, B. cereus, Listeria monocytogenes, Salmonella spp, Sthaph. aureus etc), chemical hazard analysis. In conclusion, it suggested that HACCP plan was necessary for management standard and systematic approach in establishement of critical limit, solving the problem, method of verification, education and records management by fried processing.

An Investigation of the Hazards Associated with Cucumber and Hot Pepper Cultivation Areas to Establish a Good Agricultural Practices (GAP) Model (오이와 고추생산 환경에서의 GAP 모델 개발을 위한 위해요소 조사)

  • Shim, Won-Bo;Lee, Chae-Won;Jeong, Myeong-Jin;Kim, Jeong-Sook;Ryu, Jae-Gee;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.108-114
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    • 2014
  • To analyze the hazards associated with cucumber and hot pepper cultivation areas, a total of 72 samples were obtained and tested to detect the presence of biological (sanitary indicative, pathogenic bacteria and fungi) and chemical hazards (heavy metals and pesticide residues). The levels of sanitary indicative bacteria (aerobic plate counts and coliforms) and fungi were ND-7.2 and ND-4.8 log CFU/(g, mL, hand, or $100cm^2$) in cucumber cultivation areas, and ND-6.8 and 0.4-5.3 log CFU/(g, mL, hand, or $100cm^2$) in hot pepper cultivation areas. More specifically, the soil of hot pepper cultivation areas was contaminated with coliforms at a maximum level of 5.6 log CFU/g. Staphylococcus aureus was detected only in glove samples at a level of 1.4 log CFU/$100cm^2$ and Bacillus cereus was detected in the majority of samples at a level of ND-4.8 log CFU/(g, mL, hand, or $100cm^2$). Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella spp. were not detected. Heavy metal (Zn, Cu, Ni, Pb, and Hg) chemical hazards were detected at levels lower than the regulation limit. Residual insecticides were not detected in cucumbers; however, hexaconazole was detected at a level of 0.016 mg/kg (maximum residue limit: 0.3 mg/kg) in hot peppers.

Highly Sensitive Detection of Pathogenic Bacteria Using PDMS Micro Chip Containing Glass Bead (유리비드를 포함한 PDMS 마이크로칩을 이용한 고감도 감염성 병원균 측정에 관한 연구)

  • Won, Ji-Yeong;Min, Jun-Hong
    • KSBB Journal
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    • v.24 no.5
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    • pp.432-438
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    • 2009
  • Here, we demonstrated simple nucleic acid, RNA, concentration method using polymer micro chip containing glass bead ($100\;{\mu}m$). Polymer micro chip was fabricated by PDMS ($1.5\;cm\;{\times}\;1.5\;cm$, $100\;{\mu}m$ in the height) including pillar structure ($160\;{\mu}m\;(I)\;{\times}\;80\;{\mu}m\;(w)\;{\times}\;100\;{\mu}m\;(h)$, gap size $50\;{\mu}m$) for blocking micro bead. RNA could be adsorbed on micro glass bead at low pH by hydrogen bonding whereas RNA was released at high pH by electrostatic force between silica surface and RNA. Amount of glass beads and flow rate were optimized in aspects of adsorption and desorption of RNA. Adsorption and desorption rate was measured with real time PCR. This concentrated RNA was applied to amplification micro chip in which NASBA (Nucleic Acid Sequence Based Amplification) was performed. As a result, E.coli O157 : H7 in the concentration of 10 c.f.u./10 mL was successfully detected by these serial processes (concentration and amplification) with polymer micro chips. It implies this simple concentration method using polymer micro chip can be directly applied to ultra sensitive method to measure viable bacteria and virus in clinical samples as well as environmental samples.

Physiological Activities of Fermented Garlic Broth during Fermentation (발효기간에 따른 마늘 발효액의 기능성)

  • Jung, Kyung-Ae;Park, Chan-Sung
    • Food Science and Preservation
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    • v.19 no.3
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    • pp.406-412
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    • 2012
  • The purpose of this research is to develop functional food with garlics(Allium sativum var. pekinense) as a healthy food. Fermented garlic broth(FGB)s were prepared with whole bulb of garlics preserved in sugar and sugar syrup, then fermented and aged at room temperature for 36 months. Biological activities of FGBs were tested antibacterial, antioxidative, fibrinolytic activities and analyzed for polyphenol contents. The total polyphenol contents of FGBs in 12~36 month fermented broth(870~885 mg/100 mL of broth) had significantly higher than those of 1~6 months fermented broths(p<0.001). The electron donating abilities(EDAs) and SOD-like activities of 24~36 month fermented broth had significantly higher than those of 1~6 months fermented broths(p<0.05). FGBs had shown strong antibacterial activities against four kinds of pathogenic bacteria(L. monocytogenes, S. aureus, E. coli O157:H7, and Sal. typhimurium). The fibrinolytic activities of 24~36 months fermented broth had more than twice of the fibrinolytic activity of plasmin. FGBs had increasing activities in antibacterial, antioxidative and fibrinolytic activity as the progress of fermentation period. FGBs can be used as natural antioxidant to prevent oxidative damage on normal cells probably because of their antibacterial, antioxidative and fibrinolytic activities.

Detection of Pathogenic Yersinia Enterocolitica in Drinking Water and Vegetables by Mutiplex-PCR (Multiplex-PCR에 의한 먹는샘물 및 야채류로부터의 병원성 Yersinia enterocolitica의 신속검출)

  • 이택수;박부길;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.1
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    • pp.35-41
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    • 2003
  • The study was conducted to develope a rapid method for the detection of Yersinia enterocolitica in spring water and vegetables via multiplex polymerase chain reaction (PCR) technique using ail, yst, uirF and subgenus-specific Y16S primers. Specificity and sensitivity of multiplex PCR and application of best primers for the detection of Y. enterocolitica from spring water and vegetables were investigeted. Y. enterocolitica ATCC 27729 strains gave 356 bP and 200 bp (Y16S) and 134 bp (yst) bands. but Y. enterocolitica ATCC 9610 and ATCC 23715 strains gave 200 bp and 134 bp bands.In the meanwhile, non-pathogenic Yersinia species, such as Y. frederikseni, Y. inter-media, Y. kristenseni and Y. pseudotuberculosis gave only single 200 bp band, and other bacteria including Escherichia coli O157:H7 ATCC 25392, Shigella dysenteri. Staphylococcu aureus ATCC 25923 and Listeria mo-nocytogenes ATCC 19111 did not show any bands. Among primers, yst and Y16S primer showed the best sensitivity. Seven CFU/mL Y. enterocolitica cells could be detected with yst and Y16S primers and the sensitivity was significantly improved by the further 2nd PCR after 38 cycles of first PCR amplication. Spring water, cabbage and mushroom were inoculated with Y. enterocolitica to determine the sensitivity of multiplex-PCR for the rapid detection of Y. enterocolitica. Multiplex-PCR assay could detect 7 or 70 cells in spring water and vegetables using whole cell lysate with repeating PCR amplication.

Hazard Analysis for the Cultivation Stage of Strawberry Farms for Securing Preliminary Data to Establish the Good Agricultural Practices (농산물우수관리제도 확립의 기초자료 확보를 위한 딸기농장 재배단계의 위해요소 분석)

  • Lee, Chi-Yeop;Lee, Won-Gyeong;Song, Jeong-Eon;Kim, Kyeong-Yeol;Shim, Won-Bo;Yoon, Yo-Han;Kim, Yun-Shik;Chung, Duck-Hwa
    • Journal of agriculture & life science
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    • v.46 no.3
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    • pp.97-108
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    • 2012
  • Physical, chemical and biological hazards of strawberry farms at the cultivation stage were analyzed to establish the GAP(Good Agricultural Practice) system. Samples were collected from the plants, cultivation environments(water, soil and air), and personal hygiene (hand, glove, and clothes) of three strawberry farms(A, B, and C) and were tested to analyze physical, chemical (heavy metals and pesticide residues), and biological(sanitary indications and foodborne pathogens) hazards. Physical hazards such as insects and pieces of metal and glass were found in the strawberry farms and can be potential bow for strawberry products. Heavy metal and pesticide residue as chemical hazards were detected at levels lower than the regulation limit. In case of biological hazards, total bacteria and coliform were detected at the levels of 1.6~7.3 and 1.3~5.6 log CFU/g, leaf, mL, hand or $100cm^2$. However, Escherichia coli was not detected in all samples. Bacillus cereus and Staphylococuus aureus were detected at levels of ${\leq}$ 1.1~6.1 log CFU and 4.7~5.4 log CFU/g, mL, hand or $100cm^2$, whereas Listeria monocytogenes, E. coli O157 and Salmonella spp. were not detected in all samples. This study demonstrates that various harzards were in strawberry farms at the growing stage. Therefore proper management such as GAP is needed to prevent the occurrence of food poisoning associated with the hazards revealed in this study.