• Korean Journal of Environmental Biology
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• v.29 no.3
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• pp.154-161
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• 2011

The species diversity of Betaproteobacteria in the Sumunmulbengdui Wetland Area of Jeju Island was studied using culture based techniques, and candidates for novel taxa were screened. Twenty two novel bacterial strains belonging to Betaproteobacteria were isolated, which could be assigned to 16 genera of 4 families, namely Burkholderiaceae (3 strains), Comamonadaceae (8 strains), Oxalobacteraceae (5 strains), Neisseriaceae (5 strains), and an unassigned group belonging to Burkholderiales (1 strain) based 16S rRNA gene sequences. The genus Chromobacterium contained three candidates of novel species, and each of the genera Burkholderia, Comamonas, Pelomonas and Herbaspirillum contained two candidates respectively. Through the analysis of membrane fatty acid profiles and physiological properties using API 20NE as well as morphological and cultural properties, each of the isolates was found to form potentially novel species. Brief description of 22 potential candidates for new species or subspecies is given accordingly.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.9
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• pp.1340-1349
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• 2017

Objective: This study aimed to evaluate the microbiological and cellular milk profile for the diagnosis of subclinical mastitis in female buffaloes and to assess risk factors for predisposition of the disease. Methods: Analyses were carried out by standard plate count (SPC), identification of species and antibiotic resistance, somatic cell count (SCC), electrical electrical conductivity of milk (ECM), and lactoferrin content in milk. Teat cups were swabbed to evaluate risk factors, observing hyperkeratosis, milking vacuum pressure and cleanliness of the site. Hence, 30 female buffaloes were randomly selected (15 from a group in early lactation and 15 in late lactation). Results: The most common bacteria in the microbiological examination were Staphylococcus spp., Streptococcus spp. and Corynebacterium sp. In the antibiotic sensitivity test, 10 (58.82%) of the 17 antibiotics tested were sensitive to all isolates, and resistant bacteria were Streptococcus uberis, Streptococcus dysgalactiae, Streptococcus haemolyticus, and Escherichia coli. It was observed that positive samples in the microbiological examination showed total bacterial count between $9.10{\times}10^3$ to $6.94{\times}10^6$ colony forming units/mL, SCC between 42,000 to 4,320,000 cells/mL and ECM ranging from 1.85 to 7.40 mS/cm. It was also found that the teat cups had high microbial counts indicating poor hygiene, and even faults in the cleanliness of the animals' waiting room were observed. It is concluded that values of SCC above 537,000 cells/mL and ECM above 3.0 mS/mL are indications of mammary gland infection for this herd; however, the association of these values with a microbiological analysis is necessary to more accurately evaluate the health status of mammary glands with subclinical mastitis. Conclusion: Through phenotypic characterization of bacteria involved in the samples, the genera Staphylococcus spp., Streptococcus spp., and Corynebacterimum bovis were the most prevalent in this study. Faults in environment and equipment hygienization are factors that are directly associated with mastitis.

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

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.125-132
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• 2009

In order to develop probiotics which may be a viable alternative of antibiotic use in pig industry, five bacterial strains (Lactobacillus sp. HN 52, 92, 98, 235 and AP 116) possessing antimicrobial properties were selected from 500 strains isolates of pig intestines. The bacteriocin produced by Lactobacillus sp. HN 235 displayed a relative broad spectrum of inhibitory activity against all Enterococcus strains, Pseudomonas aeruginosa, Listeria monocytogenes and Clostridium perfringens using the spot-on-lawn method. The production of antimicrobial substance started in the middle of the exponential growth phase, reached maximum levels (6,400 AU/mL) in the stationary phase, and then declined. Bacteriocin activity remained unchanged after 30 min of heat treatment at $95^{\circ}C$ and stable from pH 2.0 to 10 for 1 h, or exposure to organic solvents; however, it diminished after treatment with proteolytic enzymes. The molecular weight of the bacteriocin was about 5 kDa according to a tricine SDS-PAGE analysis.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.775-779
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• 2003

Cultivation environment for effective production of shrimp in shrimp aquaculture, Institute of Marine & Fishery, Soonchunhyang University located at Tae-An, Chung-Nam Province, with the ultimate aim of probiotics development, was monitored. Several environmental factors including dissolved oxygen, water body temperature, pH, salinity, $NH_4-N$, $NO_2-N$, $NO_3-N$, chlorophyll a affecting on the growth of shrimp were examined. Concentrations of $NH_4-N$, $NO_2-N$, and $NO_3-N$ dissolved in water samples were determined to 0.024-0.034 mg/L, 0.03-0.02, and 0.004-0.009 mg/L, respectively. Chlorophyll a content was examined in the range of 0.002-0.118 $ug/m^3$. In order to understand the distribution of different bacteria in water samples collected in shrimp aquaculture, bacteria were isolated and enumerated on the marine agar plates. Total number of bacteria were increased to approximately $6.5\;{\times}\;10^4$. Thirteen predomonant bacteria were isolated and identified. As the results of BIOLOG test of the isolates, these bacteria were identified as Corynebacterium nirilophilus, Clavibacter agropyri, Sphingomonas adhaesiva, Brevundimonas vesicularis, Vibrio parahaemolyticus, Pseudomonas bathycetes, Vibrio tubiashii, Sphingomonas macrogoltabidus, Rhodococcus, rhodochrous, Burkholderia glumae, Corynebacterium urealyticus, Rhodococcus fascians, Psychrobacter immobilis, respectively. Further work will stimulate the elucidation of pathogenicity, corresponding bacteria related to environment and probiotics, providing good information for effective production of shrimp.

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.519-532
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• 1999

Helicobacter pylori is a causative agent of type B gastritis and plays a central role in the pathogenesis of gastroduodenal ulcer and gastric cancer. To elucidate the host-parasite relationship of the H. pylori infection on the basis of molecular biology, we tried to evaluate the genomic diversity of H. pylori. An ordered overlapping bacterial artificial chromosome (BAC) library of a Korean isolate, H. pylori 51 was constructed to set up a genomic map. A circular physical map was constructed by aligning ApaI, NotI and SfiI-digested chromosomal DNA. When the physical map of H. pylori 51 was compared to that of unrelated strain, H. pylori 26695, completely different restriction patterns were shown. Fifteen known genes were mapped on the chromosome of H. pylori 51 and the genetic map was compared with those of strain 26695 and J99, of which the entire genomic sequences were reported. There were some variability in the gene location as well as gene order among three strains. For further analysis on the genomic diversity of H. pylori, when comparing the genomic structure of 150 H. pylori Korean isolates with one another, genomic macrodiversity of H. pylori was characterized by several features: whether or not susceptible to restriction digestion of the chromsome, variation in chromosomal restriction fingerprint and/or high frequency of gene rearrangement. We also examined the extent of allelic variation in nucleotide or deduced amino acid sequences at the individual gene level. fucT, cagA and vacA were confirmed to carry regions of high variation in nucleotide sequence among strains. The plasticity zone and strain-specific genes of H. pylori 51 were analyzed and compared with the former two genomic sequences. It should be noted that the H. pylori 51-specific sequences were dispersed on the chromosome, not congregated in the plasticity zone unlike J99- or 26695-specific genes, suggesting the high frequency of gene rearrangement in H. pylori genome. The genome of H. pylori 51 shows differences in the overall genomic organization, gene order, and even in the nucleotide sequences among the H. pylori strains, which are far greater than the differences reported on the genomic diversity of H. pylori.

• Korean Journal of Veterinary Research
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• v.47 no.3
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• pp.291-298
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• 2007

Olive flounder, Paralichthys olivaceus is one of the most important cultured fish in Korea, its farming has been negatively impacted by viral, bacterial and parasitic diseases. Streptococcal infection was considered as a serious problem because of significant economic losses in olive flounder farm industry. The development and evaluation of vaccine for protection against infection by this agent were required. We evaluated the safety and efficacy of ${\beta}$-hemolytic Streptococcus (S.) iniae vaccine on olive flounder Three hundreds of flounders (weight $119.8{\pm}20.7g$, body length $22.6{\pm}1.4cm$) were reared in 0.5 tons aquaria in land-marine tank system. Seawater was provided from the sea of Inchon in Korea, and water temperature was set to $22^{\circ}C$ and $25^{\circ}C$ in the vaccination and challenge test, respectively. We used the formalin-inactivated ${\beta}$-hemolytic S. iniae (F2K) vaccine (M VAC INIAE; Kyoritsu seiyaku, Japan) originated in Japan. The vaccine was intraperitoneally administered to fish. Both of vaccinated group and control group were challenged with intraperitoneally injection by virulent S. iniae SI-36 isolates with $1.0{\times}10^7CFU/fish$ at 3 weeks after vaccination. Difference on mortality of control and vaccinated group (90.0 and 15.0%, 76.5 and 8.0% respectively) at two trials were found significant (p<0.05), and relative percent survival were 83.4% and 89.5%, respectively. The dead fishes were showed dark pigmentation of skin, abdominal extension, hemorrhagic ascites, and liver necrosis, and isolated the S. iniae strain from ascites, liver and kidney. We confirmed the safety and efficacy of ${\beta}$-hemolytic S. iniae vaccine by determinations of the optimal management condition and artificial challenge test in olive flounder.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.144-148
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• 1998

Soil microorganisms including bacteria, fungi, and actinomycetes were seasonally isolated at depths (0.5~2, $10{\pm}1$, $50{\pm}1cm$) of field. The frequency of microbial isolates was employed for the determination of microbial population (CFU/g dry soil) and distribution ratio (%) in soil. Both bacteria (24-fold) and actinomycetes (7-fold) exhibited the biggest change at the depth of $50{\pm}1cm$, whereas fungi showed the maximum (13-fold) at $10{\pm}1cm$. On the whole, the bacterial population was high in spring soil, fungi in winter, and actinomycetes in autumn. Soil microorganisms also exhibited the seasonal variation on their distribution ratio (%). The maximum distribution ratio (85.7%) of bacteria was observed at the depth of $50{\pm}1cm$ in spring, whereas bacteria showed the minimum (35.2%) at the depth of $10{\pm}1cm$ in spring. The maximum distribution ratio (23.0%) of fungi was found at the depth of $50{\pm}1cm$ in spring, whereas its minimum (0.5%) at the depth of $10{\pm}1cm$ in spring. Actinomycetes exhibited the maximum distribution ratio (45.2%) at the depth of $10{\pm}1cm$ in spring, whereas its minimum (12.2%) was showed at the depth of $50{\pm}1cm$ in spring.

• Journal of fish pathology
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• v.20 no.2
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• pp.99-108
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• 2007

Vibrio harveyi, one of the major causal agent of vibriosis, affects a diverse range of marine vertebrates and invertebrates over a wide geographical area. The organism is synonymous with Vibrio carchariae, which is also known as a fish pathogen. The aims of this study were to investigate the characteristics of the pathogenic non-luminous V. harveyi and the luminous V. harveyi. And V. harveyi isolates were examined the pathogenicity to the black rockfish, Sebastes schlegeli. Both strains of V. harveyi showed haemolytic activity, and the survival rate of non-luminous V. harveyi FR 2 was higher than other strains in the skin, gut mucus and fresh serum of olive flounder, Paralichthys olivaceus and black rockfish, Sebastes schlegeli, respectively. The virulence of non-luminous V. harveyi FR 2 was higher than that of luminous V. harveyi VIB 391 in the intraperitoneally infected black rockfish. In conclusion, the present study revealed that the pathogenicity of V. harveyi FR 2 isolated from marine fish was higher than that of V. harveyi VIB 391 isolated from shrimp for black rockfish. It was suggested that the pathogenicity of V. harveyi on the black rockfish was related with bacterial luminescense.

• The Korean Journal of Pesticide Science
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• v.10 no.1
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• pp.56-65
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• 2006

An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.