• Korean Journal of Veterinary Service
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• v.46 no.4
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• pp.325-334
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• 2023

Edema disease (ED) in pigs is enterotoxemia caused by Shiga toxin type 2e (Stx2e)-producing Escherichia coli (STEC) and frequently occurs in young piglets. Therefore, ED causes enormous economic losses in pig farms. In this study, a modified Stx2e (mStx2e) antigen was expressed and purified using commercial E. coli expression system. Monoclonal antibody was serviced by Ynto Ab Inc., using Phage Display Technique. Anti-Stx2e antibodies in piglets were measured by indirect ELISA using mStx2e antigens. Naive Stx2e in piglets were detected by Sandwich ELISA using Stx2e-monoclonal antibodies and commercial Stx2e-polyclonal antibodies. Among 3,480 piglets, anti-Stx2e antibodies were observed in 2,573 piglets. The 49.4% among 830 piglet serum samples possessed 0.625 ㎍/mL or more of Stx2e proteins. The 18.3% of 830 sera had 0.313 ㎍/mL of Stx2e proteins. The 32.3% of 830 samples held 0.156 ㎍/mL or less of Stx2e proteins. These results show that indirect ELISA using mStx2e antigen and Sandwich ELISA using Stx2e-monoclonal and polyclonal antibodies can be useful to detect ED in piglets.

• 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.

• 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.

• Korean Journal of Veterinary Research
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• v.44 no.2
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• pp.251-257
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• 2004

This study was designed to determine optimal condition for expression of cloned Shigatoxin2e(Stx2e) gene from transformed E. coli PED18, to compare the cytotoxicity titer between cloned Stx2e and Stx2e from original strain, and to confirm of receptor binding affinity of Stx2e for use of development of receptor binding ELISA to detect of Stx2e. The optimum composition of medium for expression of Stx2e gene in E.coli host-vector system was definded as the medium containing 0.5% glucose and 0.5 mM IPTG. The cytotoxicity titer of expressed Stx2e for Vero cell was 1000 fold higher than that of Stx2e from original strain AY93258. The binding affinity of Stx2e to receptor globotetraosyl ceramide($Gb_4$) was confirmed by immunobloting.

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.316-321
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• 2014

The aim of study was to investigate the virulence profile of Escherichia coli O157:H7 bacteriophages isolated from sewage and livestock stools. Among 23 E. coli O157:H7 bacteriophages, 14 strains were isolated from sewage and 9 were from animal stools collected from 10 livestock farms in Korea. For each bacteriophage DNA sample, the presence of stx1, stx2, eae, aafII, ial, elt, estI, estII, astA, afa, and cnf was examined by polymerase chain reaction. The detection rate of eae, stx2, estI, astA, and ial was 100%, 69.6%, 13.0%, 13.0%, 8.7%, respectively. While all E. coli O157:H7 bacteriophages isolated from stools carried eae+stx2, stx2+eae, eae+astA, eae, stx2+eae+estI, eae+estI, stx2+eae+ial, and eae+ial were observed in bacteriophages isolated from sewage. As several plasmid-carrying virulence factors (estI, astA, and ial) were found in E. coli O157:H7 bacteriophages obtained from sewage and stools, the microbial safety of bacteriophages should be investigated in further study.

• Korean Journal of Veterinary Service
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• v.32 no.3
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• pp.241-249
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• 2009

Shiga toxin (Stx)-producing Escherichia coli (STEC) strains can cause broad spectrum of human disease, including diarrhea, hemorrhagic colitis, and the life-threatening hemolytic uremic colitis (HUS). We examined 868 samples was taken from bovine feces and carcass from January to December 2008 in Seoul. Twenty two (9.5%) shiga toxin -producing Escherichia coli were isolated from the 230 of bovine feces, and two (0.31%) were isolated from the 638 of carcasses. Serotype of E. coli isolates were O157 (10, 41.6%), O26 (10, 41.6%), O111 (1, 4.2%) and UT (3, 12.6%). In PCR, the isolates displayed three different stx gene combination (stx1 [2, 8.4%]), stx2 [3, 12.6%] and stx1 and stx2 [19,87.5%]). The eaeA and hlyA gene were found in 11 (45.8%) of the 24 strains. Saa gene was present only one strains (4.2%). Toxin typing using reverse passive latex agglutination test showed the same result in VT 1. But it showed different result in VT 2. In antimicrobial susceptibility test, all isolates were sensitive to amikacin, amoxicillin/clavulanic acid, ciprofloxacin and colistin. Eighteen strains (75.0%) of 24 isolates showed the multi-resistant patterns with over 3 drugs. PFGE was performed after the genomic DNA of twenty four isolates was digested with Xba I. the 24 isolates showed 7 (A~G) PFGE type.

• Journal of the Society of Naval Architects of Korea
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• v.51 no.2
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• pp.130-137
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• 2014

As one of the largest shipbuilding company in the world, STX Offshore & Shipbuilding currently developed an inventory managing system for steel-plates, which is applied to their steel stock yard. In a traditional way to manage steel yard, almost every work has been done by manually. The manual steel-plate piling process caused some problems such as process delay due to piling errors and the uncertainty of work plan due to lack of information. To solve these problems, we developed an integrated inventory managing system based on real-time crane tracking system which automatically updates steel-plates' piling status. We built the integrated steel-plate database, developed several programs including steel-plate input program, real-time steel-plate monitoring program and steel-yard management program, and constructed hardware system for tracking magnetic cranes. As a result, a supervisor of steel-yard can manage the inventory of steel-plates efficiently and furthermore plan an efficient piling schedule and crane working schedule.

• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1460-1466
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• 2015

In this study, the prevalence of Shiga toxin-producing Escherichia coli (STEC) was investigated among raw meat or meat products from slaughterhouses and retail markets in South Korea, and their potential for antibiotic resistance and virulence was further analyzed. A total of 912 raw meats, including beef, pork, and chicken, were collected from 2008 to 2009. E. coli strains were frequently isolated in chicken meats (176/233, 75.9%), beef (102/217, 42.3%), and pork (109/235, 39.2%). Putative STEC isolates were further categorized, based on the presence or absence of the Shiga toxin (stx) genes, followed by standard O-serotyping. Polymerase chain reaction assays were used to detect the previously defined virulence genes in STEC, including Shiga toxins 1 and Shiga toxin 2 (stx1 and 2), enterohemolysin (ehxA), intimin (eaeA), STEC autoagglutination adhesion (saa), and subtilase cytotoxin (subAB). All carried both stx1 and eae genes, but none of them had the stx2, saa, or subAB genes. Six (50.0%) STEC isolates possessed the ehxA gene, which is known to be encoded by the 60-megadalton virulence plasmid. Our antibiogram profiling demonstrated that some STEC strains, particularly pork and chicken isolates, displayed a multiple drug-resistance phenotype. RPLA analysis revealed that all the stx1-positive STEC isolates produced Stx1 only at the undetectable level. Altogether, these results imply that the locus of enterocyte and effacement (LEE)-positive strains STEC are predominant among raw meats or meat products from slaughterhouses or retail markets in Korea.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.3
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• pp.364-372
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• 1995

On the outbreak of paralytic shellfish poisoning in April 1993 in most of shellfish harvesting areas in Jinhae Bay, Korea, to clarify the toxin production of causative organism Alexandrium species, 19 axenic clonal isolates established from the benthic resting cysts in three different stations of those culture grounds were subjected to PSP toxin analysis by HPLC. Individual toxin content per cell was highly variable among the strains isolated from a sampling area and originated from an individual cyst. Average toxin contents in those areas revealed higher values of 54-70 fmol/cell. Toxin profiles included C1/C2(epiGTX8/GTX8), GTX1/GTX4 and neoSTX as the major components, and GTX2/GTX3, GTX5, C4, dcSTX and STX as the minor or sporadic ones. neoSTX on the dominant toxins showed not only most diverse compositional changes comprising $5-54 mol\%$ ranges but also no detection on the half of the strains examined, which were implicated in arising of heterogeneity with a genetic trait within a geographical region. When average toxin composition was compared, carbamate toxins comprised large proportions of $57\%,\;54\%\;and\;67\%$ as total toxin in St. 1, St. 2 and St. 4, respectively. These results suggested that an extensive paralytic shellfish toxification in Jinhae Bay could be largely due to the production of highly potent carbamate toxins in the causative dinoflagellate Alexandrium species.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.2
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• pp.212-218
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• 2006

Changes of paralytic shellfish poison (PSP) contents, toxicity and toxin composition with pH and storing periods at different temperature in toxic blue mussel, Mytilus edulis, were tested by using fluorometric HPLC method. Toxicity at pH 3 was the highest as 14.1 MU/g $(100\%)$ and showed 12.9 MU/g $(92.1\%)$ at pH 5, 9.0 MU/g $(63.8\%)$ at pH 7, 3.6 MU/g $(25.5\%)$ at pH 9 and 0.8 MU/g $(5.7\%)$ at pH 10 which suggested PSP was unstable at alkaline conditions. The decrease in toxicity during storage days was depend on pH and temperature. The toxicity markedly decreased until during the first S day storage $(19.9\~65.3\%)$ at all pH (3, 5, 7, 9) and temperature (30, 5, $-20^{\circ}C$), but, slightly decreased after then till to 30 days. C group toxin (C1 and C2) was the major components and other toxins such as GTX 1,2,3,4, STX and dcSTX were detected. Among the 8 toxins, GTX1,4, dcSTX and STX were firstly decreased according to the decreasing the toxicity at all processing conditions. The toxicity in blue mussel (14.1 MU/g) were able to remove by heating over 10 minutes at pH higher than 7.