• Fisheries and Aquatic Sciences
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• v.22 no.2
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• pp.5.1-5.9
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• 2019

Background: Live fish import may lead to the unintended introduction of pathogens. We examined the monthly distribution of microbial pathogens in ornamental finfish imported into South Korea over a 6-month period. Results: Vibrio alginolyticus was detected in one lemon damsel in June and July; V. vulnificus was detected in one lemon damsel, one caerulean damsel, and one pearl-spot chromis and one ocellaris clownfish in July, April, and May, respectively; Photobacterium damselae was detected in one ocellaris clownfish and one caerulean damsel in June and July, respectively; V. anguillarum was detected in one pearl-spot chromis in February; V. harveyi was detected in one ocellaris clownfish and two mandarin fish in February and April, respectively; Yersinia ruckeri was detected in a pearlscale goldfish group in June and July and in two colored carp groups in July; and Lactococcus garvieae was detected in a lemon damsel group and a sutchi catfish group in July and May, respectively. European catfish virus, the only viral pathogen detected, was found in two sutchi catfish groups in May. Conclusion: This study is the first to identify pathogenic species and the presence or absence of pathogens (non-quarantine diseases) in imported ornamental finfish. These results demonstrate that various pathogens with the potential to harm indigenous fish populations can accompany ornamental finfish imported into South Korea.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.43-50
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• 2021

A newly cloned 4-α-glucanotransferase (αGT) from Deinococcus geothermalis and two typical bacterial αGTs from Thermus scotoductus and Escherichia coli (MalQ) were investigated. Among 4 types of catalysis, the cyclization activity of αGTs that produces cycloamylose (CA), a valuable carbohydrate making inclusion complexes, was intensively studied. The new αGT, DgαGT, showed close protein sequence to the αGT from T. scotoductus (TsαGT). MalQ was clearly separated from the other two αGTs in the phylogenetic and the conserved regions analyses. The reaction velocities of disproportionation, cyclization, coupling, and hydrolysis of three αGTs were determined. Intriguingly, MalQ exhibited more than 100-fold lower cyclization activity than the others. To lesser extent, the disproportionation activity of MalQ was relatively low. DgαGT and TsαGT showed similar kinetics results, but TsαGT had nearly 10-fold lower hydrolysis activity than DgαGT. Due to the very low cyclizing activity of MalQ, DgαGT and TsαGT were selected for further analyses. When amylose was treated with DgαGT or TsαGT, CA with a broad DP range was generated immediately. The DP distribution of CA had a bimodal shape (DP 7 and 27 as peaks) for the both enzymes, but larger DPs of CA quickly decreased in the DgαGT. Cyclomaltopentaose, a rare cyclic sugar, was produced at early reaction stage and accumulated as the reactions went on in the both enzymes, but the increase was more profound in the TsαGT. Taken together, we clearly demonstrated the catalytic differences between αGT groups from thermophilic and pathogenic bacteria that and showed that αGTs play different roles depending on their lifestyle.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.255-269
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• 2022

Actinomycetes isolated from marine habitats represent a promising source of bioactive substances. Here, we report on the isolation, identification, productivity enhancement and application of the bioactive compounds of Streptomyces qinglanensis H4. Eighteen marine actinomycetes were isolated and tested for resistance to seven bacterial diseases. Using 16S rRNA sequencing analysis (GenBank accession number MW563772), the most powerful isolate was identified as S. qinglanensis. Although the strain produced active compound(s) against a number of Gram-negative and Gram-positive bacteria, it failed to inhibit pathogenic fungi. The obtained inhibition zones were 22.0 ± 1.5, 20.0 ± 1, 16.0 ± 1, 12.0 ± 1, 22.0 ± 1 and 24.0 ± 1 mm against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 19404, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa ATCC 9027, Candida albicans ATCC 10231 and Staphylococcus aureus ATCC6538, respectively. To maximize bioactive compound synthesis, the Plackett-Burman design was used. The productivity increased up to 0.93-fold, when S. qinglanensis was grown in optimized medium composed of: (g/l) starch 30; KNO₃ 0.5; K₂HPO₄ 0.25; MgSO₄ 0.25; FeSO₄·7H₂O, 0.01; sea water concentration (%) 100; pH 8.0, and an incubation period of 9 days. Moreover, the anticancer activity of S. qinglanensis was tested against two different cell lines: HepG2 and CACO. The inhibition activities were 42.96 and 57.14%, respectively. Our findings suggest that the marine S. qinglanensis strain, which grows well on tailored medium, might be a source of bioactive substances for healthcare companies.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.329-338
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• 2023

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.

• IMMUNE NETWORK
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• v.23 no.4
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• pp.31.1-31.18
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• 2023

Evidence suggests that the human respiratory tract, as with the gastrointestinal tract, has evolved to its current state in association with commensal microbes. However, little is known about how the airway microbiome affects the development of airway immune system. Here, we uncover a previously unidentified mode of interaction between host airway immunity and a unique strain (AIT01) of Staphylococcus epidermidis, a predominant species of the nasal microbiome. Intranasal administration of AIT01 increased the population of neutrophils and monocytes in mouse lungs. The recruitment of these immune cells resulted in the protection of the murine host against infection by Pseudomonas aeruginosa, a pathogenic bacterium. Interestingly, an AIT01-secreted protein identified as GAPDH, a well-known bacterial moonlighting protein, mediated this protective effect. Intranasal delivery of the purified GAPDH conferred significant resistance against other Gram-negative pathogens (Klebsiella pneumoniae and Acinetobacter baumannii) and influenza A virus. Our findings demonstrate the potential of a native nasal microbe and its secretory protein to enhance innate immune defense against airway infections. These results offer a promising preventive measure, particularly relevant in the context of global pandemics.

• Pediatric Infection and Vaccine
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• v.24 no.2
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• pp.95-101
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• 2017

Purpose: Acute bacterial gastroenteritis (ABG) can cause more severe symptoms than acute viral gastroenteritis in children. This study was aimed at determining the etiologic trends and to examine the clinical characteristics of ABG in children. Methods: We sent stool samples from the children with acute gastroenteritis who were treated at a secondary hospital located in Seoul, Korea between January 2011 and December 2014 to Seoul Metropolitan Government Research Institute of Public Health and Environment to find the causative organisms. Clinical characteristics of patient were analyzed through a medical records review. Results: Out of 664 stool samples, 183 (27.6%) yielded bacterial pathogens. Staphylococcus aureus was the most common bacterial pathogen, found in 72 cases (39.3%), even though it was only tested for since 2012. The monthly isolation rate was the highest (24.6%) in August. The isolation rate of Campylobacter spp. by patient's age group was high (16.7%) in the 12- to 18-year-age group (P=0.04). In patients with bloody stool, Campylobacter spp. was the most commonly isolated (31.0%, P=0.04). When comparing C-reactive protein, the Salmonella spp.- or Campylobacter spp.-isolated group showed higher values than the S. aureus- or pathogenic Escherichia coli-isolated group ($5.7{\pm}0.6mg/dL$ vs. $2.1{\pm}0.3mg/dL$, P<0.01). Conclusions: S. aureus, Salmonella spp., pathogenic E. coli, and Campylobacter spp. were important pathogens of ABG among children. Considering the differences in pathogens found according to age, a clinical symptom and inflammation index might be helpful in assuming the causative organism.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.255-261
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• 2006

Experimental animals have been used to biological and medical purposes and the animals must be, for these purposes, healthy and clean to microbial infection. However, the animals can be easily exposed to pathogenic microorganism via several routes. Of the routes, environmental conditions are the most important factors to keep the animals healthy and clean, especially air condition. Monitoring of air-condition has been required to keep the animal healthy and clean. However, any guideline is not available for experimental conditions with pigs. Therefore, the sampling times and points were compared in different conditions to establish an optimal protocol for monitoring of air borne bacteria. Tryptic soy agar(TSA), blood agar containing 5% defibrinated sheep blood and Sabraud dextrose agar(SDA) were used as media to capture total bacteria, pathogenic bacteria and fungi, respectively. Two methods, compulsive capture using an air-sampler and capturing fall-down bacteria were used to capture the microorganisms in the air. The points and time of capturing were different at each experiment. Air borne microorganisms were captured at three and five points in the open and closed equipments, respectively. Air was collected using an air-sampler for 1 min and 5 min and the agar plates as open status were left from 30 min to 2hr. At first, we monitored an experimental laboratory which dealt with several pathogenic bacteria and then, a protocol obtained from the investigation was applied to open or close experimental conditions with pigs. Number of bacteria was high from 10:00 to 15:00, especially on 13:30-15:30 but sharply decreased after 17:00. The tendency of the number of bacteria was similar between two methods even though the absolute number was higher with air sampler. Critical difference in the number of cells was observed at 5 min with air sampler and 2 hr with fall-down capturing method. However, 1 min with air sampler and 1 hr with fall-down capturing were the best condition to identify bacterial species collected from the air. Number of bacteria were different depending on the sampling points in closed condition but not in opened condition. Based on our results, a guide-line was suggested for screening air-borne microorganism in the experimental conditions with pigs.

• Journal of Veterinary Clinics
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• v.31 no.6
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• pp.502-506
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• 2014

Recently nano particles have proven for wide array of bioactive properties. In the present study, antibacterial properties of chitosan silver nano composites (CAgNCs) were investigated against fish pathogenic Edwardsiella tarda. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs against E. tarda were $25{\mu}g/mL$ and $125{\mu}g/mL$, respectively. The field emission scanning electron microscope (FE-SEM) image of CAgNCs treated E. tarda showed the strongly damaged bacteria cells than non-treated bacteria. Furthermore, treatment of CAgNCs induced the level of intracellular reactive oxygen species (ROS) in E. tarda cells in concentration and time dependent manner suggesting that it may generate oxidative stress leading to bacterial cell death. In addition, MTT assay results showed that the lowest cell viability at $100{\mu}g/mL$ of CAgNCs treated E. tarda. Overall results of this study suggest that CAgNCs is a potential antibacterial agent to control pathogenic bacteria.

• Korean Journal of Veterinary Service
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• v.39 no.4
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• pp.211-219
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• 2016

For determining the prevalence of major enteric pathogens, clinical examination and etiological diagnosis were carried out on 75 Korean pig farms. Enteric disease-suspected signs were observed in 90.7% of the farms and the incidence and severity were higher in younger age groups of the pigs. Five of seven pathogens were detected in 375 fecal samples collected from the 75 farms, and the farm-level prevalence of porcine rotavirus group A (PoRVA), pathogenic Escherichia (E.) coli, Lawsonia (L.) intracelluraris, Salmonella spp., and Brachyspira (B.) hyodysenteriae was 54.7%, 54.7%, 16.0%, 10.7% and 2.7%, respectively. PoRVA was extensively infected in suckling and weaning pig groups. The prevalence of pathogenic E. coli was highest in suckling period, and after the period, it exhibited a tendency to decrease. Salmonella spp. and L. intracelluraris were detected in all feeding groups of pigs in a ratio of 1.3~6.7%. B. hyodysenteriae was detected in 1.3~2.7% of growing and fattening pig groups but not detected in suckling and weaning pig groups. At least one or more pathogens were detected in 30.1% of 375 fecal samples. Among these, 25.0% or 5.1% of cases were single or mixed infection. Enteric disease signs of the pigs were significantly co-related with the detection of PoRVA, pathogenic E. coli or Salmonella spp. (P<0.01) but not with L. intracelluraris or B. hyodysenteriae (P>0.05). Conclusively, it will be expected that these data obtained in this study are very useful for subsequent studies and prevention strategies for swine enteric disease in Korean pig farms.

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