• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.710-716
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• 2021

A risk analysis of Shiga toxin (Stx)-encoding bacteriophage was carried out by confirming the transduction phage to non-Stx-producing Escherichia coli (STEC) and subsequent expression of the Shiga toxin genes. The virulence factor stx1 was identified in five phages, and both stx1 and stx2 were found in four phages from a total of 19 phage isolates with seven non-O157 STEC strains. The four phages, designated as ϕNOEC41, ϕNOEC46, ϕNOEC47, and ϕNOEC49, belonged morphologically to the Myoviridae family. The stabilities of these phages to temperature, pH, ethanol, and NaClO were high with some variabilities among the phages. The infection of five non-STEC strains by nine Stx-encoding phages occurred at a rate of approximately 40%. Non-STEC strains were transduced by Stx-encoding phage to become lysogenic strains, and seven convertant strains had stx1 and/or stx2 genes. Only the stx1 gene was transferred to the receptor strains without any deletion. Gene expression of a convertant having both stx1 and stx2 genes was confirmed to be up to 32 times higher for Stx1 in 6% NaCl osmotic media and twice for Stx2 in 4% NaCl media, compared with expression in low-salt environments. Therefore, a new risk might arise from the transfer of pathogenic genes from Stx-encoding phages to otherwise harmless hosts. Without adequate sterilization of food exposed to various environments, there is a possibility that the toxicity of the phages might increase.

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.35-41
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• 1982

Enterotoxigenk E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-labile enterotoxin is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a marker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. Therefore knowledge about the heat-labile enterotoxin is essential not only for understanding the pathogenesis but also for the diagnosis of the diarrhea. However the in-vitro heat-labile enterotoxin production is reported to be greatly affected by the cultural condition. In this regards, this study was designed to know the optimal conditions for the production of the heat-labile enterotoxin by assaying the permeability factor in the 18 hours culture supernatant of E. coli 08K25(B2) H9 and of E. coli 015 H11. Results obtained were summerized as follows: 1. Amounts of heat-labile enterotoxin produced were greater at initial pH 8.5 than at 7.0 of CYES-2 broth culture. However, the bacterial growth itself was more abundant at 7.0 than at 8.5. 2. Heat-labile enterotoxin per unit volume of culture supernatant was greater at shaking culture than at standing culture condition, but ratio of the enterotoxin produced over the unit mass of E. coli calculated was greater at standing culture than shaking culture condition, indicating that the greater yields of the toxin produced at shaking culture was due to increase in E. coli cell mass compared to the standing culture condition: 3. The enterotoxin produced in the lincomycin(128 microgram/ml) supplemented media was 5 or 11 times greater on the basis of enterotoxin per unit mass of E. coli, compared to the lincomycin-non-supplemented media, indicating that lincomycin itself increases the enterotoxin production. 4. Treatment of 18 hours culture of E. coli with polymyxin B(0.2 mg/ml) for 1 hour increased the yields of enterotoxin amounting to 2 or 5 times of the non-treated control cultures.

• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.147-155
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• 1991

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

• Korean Journal of Soil Science and Fertilizer
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• v.46 no.6
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• pp.647-651
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• 2013

Animal manure compost is a commonly used fertilizer in organic vegetable and fruit production in Korea. However, livestock manure compost produced from animal feces can contain a lot of the non-pathogenic and pathogenic bacteria. Of particular concern are bacteria causing human food-borne illness such as Escherichia coli O157:H7 and Listeria monocytogenes. The objective of this study was to investigate effect of temperature on survival of E. coli O157:H7 and L. monocytogenes in livestock manure compost. Commercial livestock manure compost (manure 60%, sawdust 40%) was inoculated with E. coli O157:H7 and L. monocytogenes. Compost was incubated at four different temperatures (10, 25, 35, and $55^{\circ}C$) for 20 weeks. Samples were taken every week during incubation depending on the given conditions. E. coli O157:H7 persisted for up to 1 day in livestock manure compost at $55^{\circ}C$, over 140 days at $10^{\circ}C$, 140 days at $25^{\circ}C$, and 120 days at $35^{\circ}C$, respectively. L. monocytogenes persisted for up to 1 day in livestock manure compost at $55^{\circ}C$ and 140 days at $10^{\circ}C$, 70 days at $25^{\circ}C$, and 40 days at $35^{\circ}C$, respectively. The results indicated that E. coli O157:H7 and L. monocytogenes persisted longer under low temperature condition. E. coli O157:H7 survived longer than L. monocytogenes at three different temperatures (10, 25, and $35^{\circ}C$). The results are being used to develop guidelines on the management of manure to reduce the risks of E. coli O157:H7 and L. monocytogenes transmission to foods produced in the presence of animal waste.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.6
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• pp.617-624
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• 2019

Dielectric barrier discharge (DBD) plasma is one of the promising next generation non-thermal technologies for food sterilization. The present study investigated the effects of DBD plasma on the reduction of most common food-borne pathogenic bacteria (Staphylococcus aureus, Bacillus cereus, Vibrio parahaemolyticus, Salmonella enterica) and sanitary indicative bacteria (Escherichia coli) in the suspension (initial inoculum of approx. 9 log CFU/mL). The bacterial counts were significantly (P<0.05) reduced with the increase in the treatment time (1-30 min) of DBD plasma in the suspension. The D-values (time for 90% reduction) of DBD plasma by first-order kinetics for S. aureus, B. cereus, V. parahaemolyticus, S. enterica, and E. coli were 17.76, 19.96, 32.89, 21.55, and 15.24 min, respectively (R²>0.90). These results specifically showed that 30 min of DBD plasma treatment in > 90% reduction of seafood-borne pathogenic and sanitary indicative bacteria. This suspension study may provide the basic data for use in seafood processing and distribution.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1339-1348
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• 2015

Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

• Food Science of Animal Resources
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• v.38 no.4
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• pp.727-736
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• 2018

In this study, an immuno-magnetic bead (IMB)-based assay was developed to simultaneously detect Escherichia coli, Staphylococcus aureus, and Salmonella spp. and was tested in four animal-derived foods: beef, ham, egg, and ricotta cheese. The IMB-based assay exhibited good specificity by binding to five E. coli serotypes [capture efficiency (CE) average (avg.) 90.4%], five S. aureus strains (CE avg. 91.4%), and five Salmonella serotypes (CE avg. 95.4%) but not binding to non-target bacteria (CE<10%). Furthermore, the assay detected all three pathogens with a detection limit of 10 CFU/g without the need for enrichment or additional platforms. Since the results demonstrated that the IMB-based assay can effectively separate and enrich target bacteria from a variety of animal-derived food matrixes, the assay exhibits good specificity for potential use in providing rapid, immunological, presumptive identification of pathogenic bacteria.

• Journal of Food Hygiene and Safety
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• v.18 no.3
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• pp.118-124
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• 2003

For the rapid detection of various pathogenic microorganisms from food sample, various kinds of kits have been developed and commercially available in the markets. With the advantages of speed, accuracy and easiness, the market of these kits has gradually increased for the QC and QA field of food company as well as testing facilities or laboratories. In this study, the characteristics such as the detection limit and the sensitivity of immunochromatographic type of rapid detection kit (Donga Co, Korea, D-kit) for E. coli 0157:H7 developed by monoclonal antibody were examined and also the possibility of application of the kit to food samples was evaluated. The reference kits used for comparison study were Reveal E. coli 0157:H7 (Neogen Co., USA, R-kit) and VIP EHEC kit (Biocontrol Inc., USA, V-kit) occupying major market share. In the detection limit test with the E. coli 0157:H7 reference, both R-kit and D-kit showed a distinct positive reaction in $10^4$/ml and weak positive reaction in $10^3$/ml, whereas V-kit showed a same reaction in 105/ml. Also, it was identified that the culture treated with heat showed more sensitivity than no heat treated culture. The sensitivity test was conducted against 22 isolates of E. coli 0157:H7, 7 strains of non-O157:H7 verotoxin-producing E. coli, 40 strains of E. coli with different O and H antigen type, and 38 strains of non-E. coli Enterobacteriaceae, and all of the test strains except three were showed exactly three were showed exactly the same reaction against three kinds of the tested kits. All the three kinds of kits showed a positive reaction against E. coli O157:H19, E. coli O148:H18 and Salmonella galinarium. We suppose that there might be a similarity in serological property between these three strains and O157:H7. From the test results, it can be concluded that there is (was) no difference between the D-kit developed in this study and R-kit or V-kit based on the detection limit and sensitivity.

• Journal of Animal Science and Technology
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• v.53 no.1
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• pp.67-74
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• 2011

The aim of this study was to understand the management level of pathogenic bacteria in HACCP system implemented animal farms. Microbial samples were collected from manure, floor, compost depot, manure on belt, low milk tank, dust in laying house and egg collector in HACCP system implemented Korean beef cattle, dairy cattle, swine, and laying Hens farms. O157, O111 and O26 strains of E. coli were not detected in HACCP system implemented Korean beef cattle farm. The detection rate of E. coli from manure and floor in HACCP system implemented cattle farms (Korean beef cattle and dairy farm) was lower than those of non-HACCP system implemented cattle farm. Salmonella spp. was detected in HACCP system implemented cattle farms (Korean beef cattle and dairy farm). Compared with pervious studies, lower detection rate of Salmonella spp. at floor and compost depot in HACCP system implemented swine and commercial layer farms were indicated. In conclusion, implementation of HACCP system in animal farms would enhance the management level of biological hazard compare to normal animal farms.

• Journal of Microbiology and Biotechnology
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• v.29 no.9
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• pp.1401-1411
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• 2019

Mycobacterial cell walls comprise thick and diverse lipids and glycolipids that act as a permeability barrier to antibiotics or other chemical agents. The use of OH radicals from a non-thermal plasma jet (NTPJ) for the inactivation of mycobacteria in aqueous solution was adopted as a novel approach. Addition of water vapor in a nitrogen plasma jet generated OH radicals, which converted to hydrogen peroxide ($H_2O_2$) that inactivated non-pathogenic Mycobacterium smegmatis and pathogenic Mycobacterium tuberculosis H37Rv. A stable plasma plume was obtained from a nitrogen plasma jet with 1.91 W of power, killing Escherichia coli and mycobacteria effectively, whereas addition of catalase decreased the effects of the former. Mycobacteria were more resistant than E. coli to NTPJ treatment. Plasma treatment enhanced intracellular ROS production and upregulation of genes related to ROS stress responses (thiolrelated oxidoreductases, such as SseA and DoxX, and ferric uptake regulator furA). Morphological changes of M. smegmatis and M. tuberculosis H37Rv were observed after 5 min treatment with $N_2+H_2O$ plasma, but not of pre-incubated sample with catalase. This finding indicates that the bactericidal efficacy of NTPJ is related to the toxicity of OH and $H_2O_2$ radicals in cells. Therefore, our study suggests that NTPJ treatment may effectively control pulmonary infections caused by M. tuberculosis and nontuberculous mycobacteria (NTM) such as M. avium or M. abscessus in water.