• Journal of Microbiology and Biotechnology
• /
• 제27권7호
• /
• pp.1216-1222
• /
• 2017

To develop starters for the production of functional foods or materials, lactic acid bacteria producing ${\gamma}-aminobutyric$ acid (GABA) were screened from jeotgals, Korean fermented seafoods. One isolate producing a high amount of GABA from monosodium $\text\tiny{L}$-glutamate (MSG) was identified as Enterococcus avium by 16S rRNA gene sequencing. E. avium M5 produced $18.47{\pm}1.26mg/ml$ GABA when incubated for 48 h at $37^{\circ}C$ in MRS broth with MSG (3% (w/v)). A gadB gene encoding glutamate decarboxylase (GAD) was cloned and overexpressed in E. coli BL21 (DE3) using the pET26b (+) expression vector. Recombinant GAD was purified through a Ni-NTA column and the size was estimated to be 53 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 4.5 and $55^{\circ}C$and the activity was dependent on pyridoxal 5'-phosphate. The $K_m$ and $V_{max}$ values of GAD were $3.26{\pm}0.21mM$ and $0.0120{\pm}0.0001mM/min$, respectively, when MSG was used as a substrate. Enterococcus avium M5 secretes a lot of GABA when grown on MRS with MSG, and the strain is useful for the production of fermented foods containing a high amount of GABA.

• Journal of Microbiology and Biotechnology
• /
• 제28권6호
• /
• pp.946-956
• /
• 2018

The gut microbiota of aquatic animals plays a crucial role in host health through nutrient acquisition and outcompetition of pathogens. In this study, on the basis of the high-throughput sequencing of 16S rRNA gene amplicons, we examined the bacterial communities in the gut of freshwater shrimp (Macrobrachium nipponense) and in their living environments (sediment and pond water) and analyzed the effects of abiotic and biotic factors on the shrimp gut bacterial communities. High bacterial heterogeneity was observed in the freshwater shrimp gut samples, and the result indicated that both the surrounding bacterial community and water quality factors (particularly dissolved oxygen and temperature) could affect the shrimp gut bacterial community. Despite the observed heterogeneity, 57 genera, constituting 38-99% of the total genera in each of the 40 shrimp gut samples, were identified as the main bacterial population in the gut of M. nipponense. In addition, a high diversity and abundance of lactic acid bacteria (26 genera), which could play significant roles in the digestion process in shrimp, were observed in the shrimp gut samples. Overall, this study provides insights into the gut bacterial communities of freshwater shrimp and basic information for shrimp farming regarding the application of probiotics and disease prevention.

• Journal of Microbiology and Biotechnology
• /
• 제30권11호
• /
• pp.1640-1650
• /
• 2020

Colorectal cancer (CRC) is the leading cause of common malignant neoplasm worldwide. Many studies have analyzed compositions of gut microbiota associated with various diseases such as inflammatory bowel diseases (IBD) and colon cancer. One of the most representative bacteria involved in CRC is enterotoxigenic Bacteroides fragilis (ETBF), a species belonging to phylum Bacteroidetes. We used ETBF colonized mice with azoxymethane (AOM)/dextran sulphate sodium (DSS) and zerumbone, a compound with anti-bacterial effect, to determine whether zerumbone could restore intestinal microbiota composition. Four experimental groups of mice were used: sham, ETBF colonized AOM/DSS group, ETBF colonized AOM/DSS group zerumbone 60 mg kg-1 (ETBF/AOM/DSS + Z (60)), and only zerumbone (60 mg kg^-1)-treated group. We performed reversible dye terminators-based analysis of 16S rRNA gene region V3-V4 for group comparison. Microbiota compositions of ETBF/AOM/DSS + Z (60) group and ETBF colonized AOM/DSS group not given zerumbone were significantly different. There were more Bacteroides in ETBF/AOM/DSS + Z (60) group than those in ETBF colonized AOM/DSS group, suggesting that B. fragilis could be a normal flora activated by zerumbone. In addition, based on linear discriminant analysis of effect size (LEfSe) analysis, microbial diversity decreased significantly in the ETBF colonized AOM/DSS group. However, after given zerumbone, the taxonomic relative abundance was increased. These findings suggest that zerumbone not only influenced the microbial diversity and richness, but also could be helpful for enhancing the balance of gut microbial composition. In this work, we demonstrate that zerumbone could restore the composition of intestinal microbiota.

• 한국수산과학회지
• /
• 제44권6호
• /
• pp.605-611
• /
• 2011

This study examined the suitability of characteristics of potential strains of probiotic bacteria. Among 25 lactic acid bacteria isolated from Korean traditional fermented food, the Hard Clam Meretrix meretrix Shikhae, the SH-10 strain, which exhibited superior resistance to low pH and bile salts, was selected as a potential probiotic bacteria. By examining carbohydrate utilization, morphological properties, and the 16S rRNA gene sequence, the SH-10 strain was identified as Pediococcus pentosaceus (hereafter, P. pentosaceus SH-10). P. pentosaceus SH-10 was resistant to amikacin, cefotetan, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, and vancomycin. Tests of antimicrobial activities against pathogens such as Bacillus cereus, Listeria monocytogenes, Salmonella choleraesuis, and Staphylococcus aureus, indicated that P. pentosaceus SH-10 inhibited the growth of pathogenic bacteria. These results suggest that P. pentosaceus SH-10 can be developed as a probiotic bacteria.

• 대한화학회지
• /
• 제47권2호
• /
• pp.127-132
• /
• 2003

소의 타일레리아병 진단을 위해 DNA 추출과정 없이 전혈에서 바로 중합효소연쇄반응(PCR)을 통하여 T. buffeli(buffeli/orientalis/sergenti)의 16S rRNA 유전자 단편을 증폭시킨 뒤, 증폭된 DNA를 전기영동법으로 분석하는 방법을 개발하였다. 특이유전자 단편의 증폭을 위해 formamide를 사용하여 혈액세포를 용해시켰으며, 단백질의 응고를 줄이기 위해 낮은 반응온도를 사용하는 FoLT(Formamide Low Temp.) PCR법을 이용하였다. 전혈 100-200 nL를 바로 PCR 증폭에 사용하였으며, PCR 산물(816-bp DNA)은 전기영동법으로 분석하였다. 본 결과는 T. buffeli에 감염된 소의 혈액으로부터 정제된 DNA를 사용하여 얻은 실험결과와 잘 일치하였다.

• 한국동물위생학회지
• /
• 제35권2호
• /
• pp.99-104
• /
• 2012

Otitis externa (OE) is a frequent disease in the ear canals of dogs. To identify the pathogens causing OE in dogs and to determine their antimicrobial resistances, specimens were collected from animal hospitals in Daejeon. The isolates were examined by morphological and biochemical tests, 16S rRNA analysis and antimicrobial susceptibility tests. We analyzed correlation between the isolated pathogens and external factors of dogs such as breed, age, gender, ear mite, hair in ears and experience with antibiotic therapy. Thirty three strains of bacteria were isolated from 26 of the 68 heads of dogs with OE. The most isolated bacteria were Enterococcus faecalis (E. faecalis) followed by Staphylococcus aureus (Sta. aureus), Sta. pseudointermedius, E. faecium, E. avium and Streptococcus canis (Strep. canis) in order of frequency of occurrence. Isolation frequency of Enterococcus spp. and Staphylococcus spp. were 51.5% and 45.5%, respectively. E. faecalis and E. faecium isolates showed VanB phenotype, which is resistant to vancomycin but sensitive to teicoplanin were 58% and 25%, respectively. Nine isolates among total twelve isolates of E. faecalis were isolated from the dogs treated with antibiotics. There was no methicillin-resistant Sta. aureus (MRSA), but were MR-Sta. pseudointermedius (MRSP) (57.1%) and vancomycin-resistant (VR)-Sta. pseudointermedius (14.3%) (VRSP) showing VanB phenotype. However, vanA, vanB and vanC genes were not detected in VR isolates from the dogs. Taken together, VR-Enterococcus spp. (VRE) is one of the major pathogens in domestic animals, as well as community-and hospital-acquired infection.

• KSBB Journal
• /
• 제28권1호
• /
• pp.18-23
• /
• 2013

The purpose of this work was to investigate the several functional characteristics of Lactobacillus sakei JK-17 isolated from long-term fermented kimchi, Muk Eun Ji. Initially, phylogenetic analysis using 16S rRNA sequencing was performed to identify the isolate JK-17, and the strain could be assigned to Lactobacillus sakei and designated as L. sakei JK-17. The strain was registered in GenBank as [JX841311]. The changes of bacterial growth and residual organic acids were monitored and HPLC was used to measure quantitatively two organic acids, lactic acid and acetic acid, produced in the culture during 84 hours of incubation. During the incubation period, several functional characteristics of L. sakei JK-17 were examined. L. sakei JK-17 culture depleted nitrite concentration 94.75%. Antioxidant activity of cultural supernatants of L. sakei JK-17 was approx. 53.8%, and ${\beta}$-galactosidase activities were 0.243 units/mL at pH 7.0 and 0.387 units/mL at pH 4.1, respectively. The antibacterial activities against food-poisoning causing bacteria were examined with 20-fold concentrated culture supernatants from L. sakei JK-17 and the antibacterial effects were clearly observed against all bacteria tested in this work.

• 한국응용곤충학회지
• /
• 제51권4호
• /
• pp.377-382
• /
• 2012

경기지역에서 담배가루이의 발생 및 약제반응을 조사하였다. 담배가루이는 13개 시군에서 발생되었으며, 유전자(16S rRNA, MtCOI) 분석결과, 고양시 일부지역은 biotype B와 Q가 혼재되었으나, 그 밖의 지역에서 2005부터 국내 발생이 보고되었던 biotype Q로 나타났다. 채집곤충 및 기주식물의 total DNA에 대해 PCR을 통한 염기서열 분석과 southern hybridization 분석 결과 모든 지역에서 Tomato Yellow Leaf Curl Virus(TYLCV)는 검출되지 않았다. 5개 지역에서 채집한 6 개체군의 담배가루이를 실내사육하면서 유묘검정법으로 성충에 대한 살충제 감수성을 검정한 결과, 지역별로 다소 차이는 있었지만 네오니코티노이드계 약제의 경우 biotype B에게 높은 살충활성을 나타낸 반면 biotype Q에게는 상대적으로 낮은 살충활성을 나타내었다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제29권1호
• /
• pp.48-55
• /
• 2003

The purpose of this study was to isolate and identify the bacteria in osteomyelitis lesion of 3 patients. Two lesions were due to the post-infection after extraction. The other was resulted from mal-fixation of both sides of mandibular angles. Pus samples were collected by needle aspiration from the lesion and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, $CO_2$, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene cloning and nucleotide sequencing method. Our results showed that Streptococci species was predominantly isolated in both lesions of extraction socket. Only one species (Proteus vulagris) was detected in lesion of mandibular angle. This study was not sufficient to identify the causative bacteria in those osteomyelitis. However, our data may be offered the clue to solve the problem.

• 한국균학회지
• /
• 제44권3호
• /
• pp.171-175
• /
• 2016

Pseudomonas agarici에 의해 발생하는 세균성회색무늬병은 양송이 재배에서 문제가 되는 대표적인 병해이다. 본 연구에서는 세균성회색무늬병의 생물학적 방제법에 이용할 수 있는 길항미생물의 항균활성과 선발된 길항미생물에 대해 폿트 수준의 생물검정 실험을 실시하였다. 재배중인 양송이 배지에서 세균성회색무늬병 병원균을 강하게 억제하는 길항세균 HC12를 선발하였으며, 생리 생화학적 실험과 유전적 실험결과 HC12균주는 Alcaligenes sp.로 동정되었다. Alcaligenes sp. HC12를 양송이에 처리한 결과 63%의 방제효과를 보였다. 따라서 Alcaligenes sp. HC12가 양송이버섯 세균성회색무늬병 방제를 위해 합성농약을 대체할 수 있는 친환경 방제제가 될 수 있을 것으로 생각된다.