• 대한의생명과학회지
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• 제25권1호
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• pp.40-53
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• 2019

Of the Acinetobacter spp., A. baumannii (genospecies 2) is the most clinically significant in terms of hospital-acquired infections worldwide. It is difficult to perform Acinetobacter-related taxonomy using phenotypic characteristics and routine laboratory methods owing to clusters of closely related species. The ability to accurately identify Acinetobacter spp. is clinically important because antimicrobial susceptibility and clinical relevance differs significantly among the different genospecies. Based on the medical importance of pathogenic Acinetobacter spp., the distribution and characterization of Acinetobacter spp. isolates from 123 clinical samples was determined in the current study using four typically applied bacterial identification methods; partial rpoB gene sequencing, amplified rRNA gene restriction analysis (ARDRA) of the intergenic transcribed spacer (ITS) region of the 16~23S rRNA, the $VITEK^{(R)}$ 2 system (an automated microbial identification system) and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). A. baumannii isolates (74.8%, 92/123) were the most common species, A. nosocomialis (10.6%, 13/123) and A. pittii isolates (7.5%, 9/123) were second and third most common strains of the A. calcoaceticus-A. baumannii (ACB) complex, respectively. A. soli (5.0%, 6/123) was the most common species of the non-ACB complex. RpoB gene sequencing and ARDRA of the ITS region were demonstrated to lead to more accurate species identification than the other methods of analysis used in this study. These results suggest that the use of rpoB genotyping and ARDRA of the ITS region is useful for the species-level identification of Acinetobacter isolates.

• Asian-Australasian Journal of Animal Sciences
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• 제30권4호
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• pp.495-504
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• 2017

Objective: Manipulating the fermentation to improve the performance of the ruminant has attracted the attention of both farmers and animal scientists. Propionate salt supplementation in the diet could disturb the concentration of propionate and total volatile fatty acids in the rumen. This study was conducted to evaluate the effect of calcium propionate supplementation on the ruminal bacterial community composition in finishing bulls. Methods: Eight finishing bulls were randomly assigned to control group (CONT) and calcium propionate supplementation (PROP) feeding group, with four head per group. The control group was fed normal the total mixed ration (TMR) finishing diet, and PROP group was fed TMR supplemented with 200 g/d calcium propionate. At the end of the 51-day feeding trial, all bulls were slaughtered and rumen fluid was collected from each of the animals. Results: Propionate supplementation had no influence the rumen fermentation parameters (p>0.05). Ruminal bacterial community composition was analyzed by sequencing of hypervariable V3 regions of the 16S rRNA gene. The most abundant phyla were the Firmicutes (60.68%) and Bacteroidetes (23.67%), followed by Tenericutes (4.95%) and TM7 (3.39%). The predominant genera included Succiniclasticum (9.43%), Butyrivibrio (3.74%), Ruminococcus (3.46%) and Prevotella (2.86%). Bacterial community composition in the two groups were highly similar, except the abundance of Tenericutes declined along with the calcium propionate supplementation (p = 0.0078). Conclusion: These data suggest that the ruminal bacterial community composition is nearly unchanged by propionate supplementation in finishing bulls.

• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.551-560
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• 2018

Bellflower root (Platycodon grandiflorum), which belongs to the Campanulaceae family, is a perennial grass that grows naturally in Korea, northeastern China, and Japan. Bellflower is widely consumed as both food and medicine owing to its high nutritional value and potential therapeutic effects. Since foodborne disease outbreaks often come from vegetables, understanding the public health risk of microorganisms on fresh vegetables is pivotal to predict and prevent foodborne disease outbreaks. We investigated the microbial communities on the bellflower root (n = 10). 16S rRNA gene amplicon sequencing targeting the V6-V9 regions of 16S rRNA genes was conducted via the 454-Titanium platform. The sequence quality was checked and phylogenetic assessments were performed using the RDP classifier implemented in QIIME with a bootstrap cutoff of 80%. Principal coordinate analysis was performed using the weighted Fast UniFrac distance. The average number of sequence reads generated per sample was 67,192 sequences. At the phylum level, bacterial communities from the bellflower root were composed primarily of Proteobacteria, Firmicutes, and Actinobacteria in March and September samples. Genera Serratia, Pseudomonas, and Pantoea comprised more than 54% of the total bellflower root bacteria. Principal coordinate analysis plots demonstrated that the microbial community of bellflower root in March samples was different from those in September samples. Potential pathogenic genera, such as Pantoea, were detected in bellflower root samples. Even though further studies will be required to determine if these species are associated with foodborne illness, our results indicate that the 16S rRNA gene-based sequencing approach can be used to detect pathogenic bacteria on fresh vegetables.

• 대한임상검사과학회지
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• 제52권3호
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• pp.202-213
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• 2020

미생물 연구에서 대량의 마이크로바이옴 데이터를 효율적으로 얻는 기술이 발전해왔지만, 마이크로바이옴 빅 데이터를 적절하게 분석하는 도구는 여전히 부족하다. 또한 빈약한 데이터베이스를 사용하여 미생물 군집을 분석하면 잘못된 결과를 초래할 수 있다. 따라서 본 연구는 대량의 미생물 데이터베이스 분석을 위한 적절한 방법을 설계하고자 하였다. 박테리아는 개인의 손끝과 개인 소지품(휴대 전화 및 랩탑 키보드)에서 수집되었다. 박테리아로부터 게놈 DNA를 추출하고 16S rRNA 유전자를 표적으로 하여 차세대 시퀀싱을 실시하였다. 손끝과 개인 소지품 간의 박테리아 매칭 비율의 정확성은 공식과 함께 환경 및 인간관련 데이터베이스를 사용하여 확인하였다. 적절한 분석을 설계하기 위해 다음 세가지 범주를 기준으로: 정성적 분석과 정량적 분석 비교, 성별에 관계없이 모든 참여자뿐만 아니라 동일 성별 참여자 내 비교, 환경(eDB) 및 인간 관련 데이터 베이스(hDB)를 이용하여 샘플간 비교하였다. 결과는 정성적 분석과 동일 성별 참가자 내에서의 비교 및 hDB의 사용이 비교적 정확한 결과를 제공하였다. 우리의 연구는 인간 유래 미생물을 사용하여 대량의 미생물학적 데이터를 포함하는 연구를 수행할 때 정확한 결과를 얻을 수 있는 분석 방법을 제공한다.

• Journal of Microbiology and Biotechnology
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• 제27권10호
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• pp.1778-1782
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• 2017

To identify and discriminate the bacterial species at the subspecific level, rep-PCR is a reliable genomic fingerprinting tool. Fourteen strains of bacteria were isolated from different food sources, identified as Leuconostoc citreum using 16S rRNA gene sequencing, and amplified using rep-primers (REP, ERIC, and $(GTG)_5$). Fingerprinting patterns generated bands in the range of 300-6,000 bp with REP, 150-6,000 bp with ERIC, and 200-1,700 bp with $(GTG)_5$ primers. In UPGMA dendrogram analysis, 14 strains were clustered into three clades (I, II, and III) with all the primers, thus differentiating them at the molecular level. The present study revealed the differentiation of L. citreum strains using rep-PCR.

• Mycobiology
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• 제42권2호
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• pp.158-166
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• 2014

In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant.

• Preventive Nutrition and Food Science
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• 제14권3호
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• pp.265-268
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• 2009

Three-hundred bacterial isolates from a natural cheese were screened for the production of bifidobacterial growth factor by whey fermentation. Based on this screen, two whey samples fermented by strains designated as CJNU 0421 and CJNU 0588 were found to effectively stimulate the growth of a bifidobacterial strain, Bifidobacterium longum FI10564, by 1.6$\sim$1.7 fold compared to a control, in which non-fermented whey medium was added. The two isolates were identified to be Lactobacillus casei (99% identity) by 16S rRNA gene sequencing and named Lactobacillus casei CJNU 0421 and CJNU 0588, respectively. The whey sample fermented by CJNU 0588 did not enhance the growth of other bacteria such as Escherichia coli and Listeria monocytogenes, suggesting that the whey fermentation metabolites from the isolate could be used for the selective stimulation of bifidobacteria.

• 한국토양비료학회지
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• 제50권6호
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• pp.598-605
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• 2017

To isolate rhizobacteria exhibiting antifungal activities for for five pathogenic fungi (Sclerotinia sclerotiorum, Fusarium solani, Collectotricum gloeosporides, Fusarium oxysporum, and Botrytis cinerea) which cause damage to Ginseng root in Ginseng grown fields, four soils were collected from Cheorlwon gun, in Korea. From 4 soils, a total of 160 bacterial strains were isolated by dilution plate method. Among 160 strains, 32 strains showed antifungal activities for one or more pathogens. From 32 strains, three strains exhibited antifungal activities for all pathogens. These are two Burkholderia cepacia (ATCC 25416 and ET 13) and one Paenibacillus polymyxa (ATCC 842). These potent antifungal strains showed high identities (99% using 16S-rRNA sequencing).

• KSBB Journal
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• 제30권5호
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• pp.239-244
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• 2015

Bacterial vaginosis (BV) is caused by microbial imbalance of the vaginal ecosystem and overgrowth of anaerobic bacteria. The antibiotic treatment often results in very high recurrence of BV because it disturbs the vaginal ecosystem. The high recurrence rates suggest a need for alternative therapeutic methods and probiotics are being recognized as alternative or additional treatment method for BV. The purpose of this study was to investigate how human vaginal isolates of Lactobacillus spp. inhibit the BV-associated pathogen Gardnerella vaginalis. Results show that selected strains significantly reduced the viability of G. vaginalis. Among these selected strains KLB410 and KLB416 were further selected based on acid/bile tolerance and identified through 16S rRNA gene sequencing being Lactobacillus plantarum. Further studies are underway to demonstrate that the selected strain can be applied as potential probiotics for recovering vaginal ecosystem.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.561-572
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• 2017

The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.