• Korean Journal of Microbiology
• /
• v.55 no.1
• /
• pp.77-79
• /
• 2019

Here we report the genome sequence of Veillonella atypica strain KHUD-V1 isolated from subgingival dental plaque of Korean chronic periodontitis patients. Unlike other V. atypica strains, KHUD-V1 carries two prophage regions and prophage remnants, as well as several genes homologous to prophage-associated virulence factors, such as virulence-associated protein E, a Clp protease, and a toxin-antitoxin system. The isolate and its genome sequence obtained here will aid to understand the diversity of the genome architecture of Veillonella within an evolutionary framework and the role of prophages that contribute to the genetic diversity as well as the virulence of V. atypica.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2019.10a
• /
• pp.8-8
• /
• 2019

The stringent response (SR) is characterized as a bacterial defense mechanism in response to various growth-inhibiting stresses. It is activated by accumulation of a small nucleotide regulator, (p)ppGpp, and induces global changes in bacterial transcription and translation. Recent work from our group has shown that (p)ppGpp plays a critical role in virulence and survival in Vibrio cholerae. The genes, relA and relV, are involved in the production of (p)ppGpp, while the spoT gene encodes an enzyme that hydrolyzes it in V. cholerae. A mutant strain defective in (p)ppGpp production (i.e. ${\Delta}relA{\Delta}relV{\Delta}spoT$ mutant) lost the ability to produce cholera toxin (CT) and lost their viability due to uncontrolled production of organic acids, when grown with extra glucose. In contrast, the ${\Delta}relA{\Delta}spoT$ mutant, a (p)ppGpp overproducer strain, produced enhanced level of CT and exhibited better growth in glucose supplemented media via glucose metabolic switch from organic fermentation to acetoin, a neutral fermentation end product, fermentation. These findings indicates that (p)ppGpp, in addition to its well-known role as a SR mediator, positively regulates CT production and maintenance of growth fitness in V. cholerae. This implicates SR as a promising drug target, inhibition of which may possibly downregulate V. cholerae virulence and survival fitness. Therefore, we screened a chemical library and identified a compound that induces medium acidification (termed iMAC) and thereby loss of wild type V. cholerae viability under glucose-rich conditions. Further, we present a potential mechanism by which the compound inhibits (p)ppGpp accumulation. Together, these results indicate that iMAC treatment causes V. cholerae cells to produce significantly less (p)ppGpp, an important regulator of the bacterial virulence and survival response, and further suggesting that it has a therapeutic potential to be developed as a novel antibacterial agent against cholera.

• Korean Journal of Veterinary Service
• /
• v.42 no.2
• /
• pp.93-112
• /
• 2019

Calf diarrhea is a common disease in young claves and is still a major cause of productivity and economic loss in livestock farms. Fecal samples from Korean native cattle (n=100) with diarrhea from 64 farms in Gwangju area, Korea from september 2017 to December 2018 were examined for shedding of important protozoan parasitic, viral and bacterial pathogens using culture, rapid test kit and PCR methods. Of 57 (89.1%) of the 64 Korean native cattle farms examined had samples infected with at least one of the investigated pathogens. Among 100 fecal samples, 88 samples were positive for at least one the twelve pathogens and 51 samples were simultaneously positive for two or more pathogens by culture and PCR assay. Bovine group A rotavirus (BRV) was the most common pathogen, found in 43/100 (43.0%) samples on 32/64 (50.0%) farms. Subsequently, kobuvirus (30.0%), pathogenic E. coli (29.0%), bovine parvovirus (17.0%), Giardia spp. (13.0%), Eimeria spp. (10.0%), Clostridium perfringens type A (8.0%), bovine torovirus (8.0%), bovine viral diarrhea virus (6.0%), bovine coronavirus (5.0%), bovine norovirus (2.0%) and Cryptosporidium spp. (2.0%) were detected. Nebovirus, kırklareli virus, bovine adenovirus, Salmonella spp. and intestinal parasites were not detected. Of the 72 calves sampled in this age group, 64 (88.9%) samples were positive for at least one enteropathogen. BRV was identified in 34/72 (47.2%) samples from 27/48 (56.3%) farms. Subsequently, pathogenic E. coli (30.6%), kobuvirus (29.2%), BPaV (22.2%), Giardia spp. (15.3%), Eimeria spp. (9.7%), BVDV (6.9%), Cl. perfringens type A (6.9%), BCoV (4.6%) and Cryptosporidium spp. (2.8%) were detected in fecal samples. A total of ninety-six strains of E. coli were isolated from one hundred fecal samples collected from Korean native cattle with diarrhea. The presence of stx1, stx2, eaeA, LT, STa, STb, ehxA, saa, F4, F5(K99), F6, F17, F18 and F41 genes in the isolates was investigated by PCR. Out of ninety-six E. coli isolates screened for specific genes, 30 strains E. coli were identified to harbor shiga toxin-producing E. coli (STEC) 7 (7.3%), enterohemorrhagic E. coli (EHEC) 8 (8.3%), enteropathogenic E. coli (EPEC) 6 (6.3%), enterotoxigenic E. coli (ETEC) 2 (2.1%) and STEC/ETEC hybrid 7 (7.3%). This study provides epidemiological estimates of the prevalence of Korean native cattle's enteropathogens in Gwangju area, Korea, which would be used for cattle farmers and veterinarians to select appropriate therapeutic method.

• Journal of Food Hygiene and Safety
• /
• v.38 no.4
• /
• pp.217-227
• /
• 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.

• Korean Journal of Ecology and Environment
• /
• v.50 no.1
• /
• pp.137-147
• /
• 2017

The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.

• Korean Journal of Clinical Laboratory Science
• /
• v.48 no.2
• /
• pp.82-87
• /
• 2016

Enterotoxigenic Bacteroides fragilis (ETBF) produces enterotoxins known to be a virulence factor. Three isotypes of the B. fragilis toxin (BFT) gene have been identified: bft-1, bft-2, and bft-3. We investigated the presence of bft isotypes in clinical B. fragilis isolates and the antimicrobial resistance of BFT-negative and BFT-positive isolates. Overall, 537 B. fragilis isolates were collected from extraintestinal specimens over 8 years (2006~2013) from a university hospital in Korea. Samples were analyzed by multiplex PCR to identify the bft gene isotypes. Additionally, the antimicrobial susceptibility of 107 B. fragilis isolates (74 BFT-negative and 33 BFT-positive) was examined by the CLSI agar dilution method. PCR revealed a total bft gene detection rate of 30%, while 33% and 29% of blood and other extraintestinal isolates contained the gene, respectively. Among ETBF isolates, the most common isotype was bft-1 gene, followed by bft-2 and bft-3 (bft-1 77%, bft-2 14%, bft-3 9%). Resistance rates (%) for BFT-negative and positive isolates differed in response to various antimicrobial agents, with 3%, 5%, 1% and 38% of BFT-negative isolates and 3%, 6%, 3% an 42% of BFT-positive isolates being resistant to piperacillin-tazobactam, cefoxitin, imipenem, and clindamycin, respectively. Interestingly, neither BFT-negative nor positive isolates showed antimicrobial resistance to chloramphenicol and metronidazole. Overall, the proportion of ETBF from blood was similar to that of other extraintestinal sites and the bft-1 gene was the predominant isotype. Higher antimicrobial resistance rates were found in BFT-positive isolates than BFT-negative isolates, but these differences were not statistically significant.

• Korean Journal of Microbiology
• /
• v.51 no.3
• /
• pp.300-307
• /
• 2015

For selecting Bacillus strains producing high-quality Cheonggukjang, 8 strains were isolated from the different Cheonggukjang samples. Seven of them exhibited the highest 16S rRNA gene sequence similarity value of over 99.9% to Bacillus subtilis subsp. subtilis and one of them showed the similarity to B. licheniformis. All the strains showed positive activities for amylase, cellulase, protease and lipase, and 6 strains are positive for fibrinolytic activity. To confirm the safety of the strains isolated from the samples of Cheonggukjang which are manufactured by traditional method, strains were analyzed for the presence of seven toxin genes of Bacillus cereus and results were found negative. And 7 strains did not produce at all or merely produce both histamine and tyramine, the representative biogenic amines. Biogenic amine degradation analysis by HPLC revealed that, most of them exhibited tyramine degradation activity. For Cheonggukjang fermented by artificial inoculation of selected strains, fermentation property, sensory test, volatile basic nitrogen production and metabolic profiles by $^1H-NMR$ were tested. Seven strains were confirmed to make high-quality Cheonggukjang.

• Korean Journal of Food Science and Technology
• /
• v.47 no.1
• /
• pp.126-131
• /
• 2015

Pathogenic Escherichia coli is recognized as an important cause of diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome worldwide. This study was conducted to investigate the prevalence E. coli contamination in the Korean traditional food bibimbap. E. coli were isolated from 84 of 1142 (7.3%) bibimbap investigated from 2005 to 2011. Antibiotic resistance profiling demonstrated that 6 of the 84 isolates (7.2%) showed multiple drug resistance. Fifteen virulence genes specific for pathogenic E. coli such as Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli (EAEC) were examined by multiplex PCR for mixed bacterial cultures derived from bibimbap samples. The EPEC virulence gene (ent) was detected in 5 strains (5.9%), while ETEC, EAEC, and EIEC were not detected. STEC serotypes O103 (1.2%), O91 (1.2%), and O128 (6.0%) were found, but other serogroups such as O26, O157, O145, O111 and O121 were not detecded. Automated Repetitive-Sequence-Based PCR analysis showed different patterns.

• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.4
• /
• pp.430-436
• /
• 2010

The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

• Journal of Preventive Medicine and Public Health
• /
• v.36 no.1
• /
• pp.77-84
• /
• 2003

Objectives : Two related cases of Hemolytic-Uremic Syndrome (HUS) were reported to the Korea National Institute of Health in May, 2001. Shiga toxin 2 genes were detected in both stool samples. We suspected an enterohemorrhagic Escherichia coli (EHEC) infection as the cause of the HUS, and conducted an investigation to find the source of the infection and its route of transmission. Methods : We peformed case investigations on these two related HUS cases, and obtained interviews and rectal swabs form the family members and other close contacts. Additionally, we peformed rectal swabs on the cattle raised by the household of the index patient. Results : We found a 20 month old index patient and a 6 year-old cousin had developed HUS, where there had been a 2 day history of contact with the index, and bacteriological examinations for these two patients revealed, indistinguishably, the same E. coli O171. The grandmother of the index patient was found to be asymptomatic, but E. coli O26 was isolated. We also found a probable case in the mother of the cousin. She reported a history of contact with the index, and developed bloody diarrhea of 3 days duration. The test results for the cattle revealed E. coli O26 in one cow, and E. coli O26 and O55 in another. E. coli O26, which was isolated in both cows and the grandmother of the index, were indistinguishably the same. Conclusions : We found that the E. coli O26 in the grandmother had originated from the cows, and that the E. coli O171 found in the index patient had been transmitted to the cousin through person-to-person contact.