• Title/Summary/Keyword: bacterial culture

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Probiotic Characterization of Acid- and Bile-tolerant Lactobacillus salivarius subsp. salivarius from Korean Faeces

  • Bae, H.C.;Nam, M.S.;Lee, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1798-1807
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    • 2002
  • This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

Inhibition of LPS-induced nitric oxide production by transduced Tat-arginine deiminase fusion protein in Raw 264.7 cells

  • Lee, Min-Jung;Kim, Dae-Won;Lee, Yeom-Pyo;Jeong, Hoon-Jae;Kang, Hye-Won;Shin, Min-Jae;Sohn, Eun-Jeong;Kim, Mi-Jin;Jang, Sang-Ho;Kang, Tae-Cheon;Won, Moo-Ho;Min, Bon-Hong;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.42 no.5
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    • pp.286-292
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    • 2009
  • Arginine deiminase (ADI), an arginine-degrading enzyme, has anti-proliferative and anti-tumor activities and is capable of inhibiting the production of nitric oxide (NO). Modulation of nitric oxide (NO) production is considered a promising approach for the treatment of various diseases including cancer, inflammation and neuronal disorders. In this study, an ADI gene was fused with an HIV-1 Tat peptide in a bacterial expression vector to produce an genetic in-frame Tat-ADI fusion protein. When added exogenously to the culture media, the expressed and purified Tat-ADI fusion proteins were efficiently transduced into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced Tat-ADI fusion proteins markedly increased cell viability in cells treated with lipopolysaccharide (LPS). This increase in viability was mediated by an inhibition of NO production. These results suggest that this Tat-ADI fusion protein can be used in protein therapies of NO-related disorders such as cancer, inflammation and neuronal diseases.

Analysis of Microbial Community During the Anaerobic Dechlorination of Tetrachloroethylene (PCE) in Stream of Gimpo and Inchon Areas (경기도 김포, 인천 서구지역 소하천의 PCE 탈염소화 군집의 선별 및 다양성 분석)

  • Kim, Byung-Hyuk;Baek, Kyung-Hwa;Cho, Dea-Hyun;Sung, Youl-Boong;Ahn, Chi-Yong;Oh, Hee-Mock;Koh, Sung-Cheol;Kim, Hee-Sik
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.140-147
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    • 2009
  • In this study, anaerobic enrichment cultivation was performed with the sediments from the Gimpo and Inchon areas. Lactate as an electron donor and PCE as an electron acceptor was injected into the serum bottle with an anaerobic medium. After the incubation of 8 weeks, the reductive dechlorination of PCE was observed in 7 sites among 16 sites (43%). Three enrichment cultures showed completely dechlorination of PCE to ethene, while four enrichment culture showed transformation of PCE to cis-DCE. The bacterial community structure was analyzed by PCR-DGGE. Dechlorinating bacteria were detected by species-specific primers. The dominant species in seven anaerobic enrichments were found to belong to the genus of Dehalococcoides sp. and Geobacter sp., and Dehalobacter sp.

Purification and Characterization of a Fibrinolytic Enzyme Produced by Bacillus amyloliquefaciens HC188 (Bacillus amyloliquefaciens HC188이 생산하는 혈전분해 효소의 정제 및 특성)

  • Shin, So Hee;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.33-43
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    • 2013
  • A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

The Endophytic Bacteria Bacillus velezensis Lle-9, Isolated from Lilium leucanthum, Harbors Antifungal Activity and Plant Growth-Promoting Effects

  • Khan, Mohammad Sayyar;Gao, Junlian;Chen, Xuqing;Zhang, Mingfang;Yang, Fengping;Du, Yunpeng;Moe, The Su;Munir, Iqbal;Xue, Jing;Zhang, Xiuhai
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.668-680
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    • 2020
  • Bacillus velezensis is an important plant growth-promoting rhizobacterium with immense potential in agriculture development. In the present study, Bacillus velezensis Lle-9 was isolated from the bulbs of Lilium leucanthum. The isolated strain showed antifungal activities against plant pathogens like Botryosphaeria dothidea, Fusarium oxysporum, Botrytis cinerea and Fusarium fujikuroi. The highest percentage of growth inhibition i.e., 68.56±2.35% was observed against Fusarium oxysporum followed by 63.12 ± 2.83%, 61.67 ± 3.39% and 55.82 ± 2.76% against Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi, respectively. The ethyl acetate fraction revealed a number of bioactive compounds and several were identified as antimicrobial agents such as diketopiperazines, cyclo-peptides, linear peptides, latrunculin A, 5α-hydroxy-6-ketocholesterol, (R)-S-lactoylglutathione, triamterene, rubiadin, moxifloxacin, 9-hydroxy-5Z,7E,11Z,14Z-eicosatetraenoic acid, D-erythro-C18-Sphingosine, citrinin, and 2-arachidonoyllysophosphatidylcholine. The presence of these antimicrobial compounds in the bacterial culture might have contributed to the antifungal activities of the isolated B. velezensis Lle-9. The strain showed plant growth-promoting traits such as production of organic acids, ACC deaminase, indole-3-acetic acid (IAA), siderophores, and nitrogen fixation and phosphate solubilization. IAA production was accelerated with application of exogenous tryptophan concentrations in the medium. Further, the lily plants upon inoculation with Lle-9 exhibited improved vegetative growth, more flowering shoots and longer roots than control plants under greenhouse condition. The isolated B. velezensis strain Lle-9 possessed broad-spectrum antifungal activities and multiple plant growth-promoting traits and thus may play an important role in promoting sustainable agriculture. This strain could be developed and applied in field experiments in order to promote plant growth and control disease pathogens.

Biological Control of Sclerotinia sclerotiorum in Lettuce Using Antagonistic Bacteria (길항세균을 이용한 상추 균핵병의 생물학적 방제)

  • Chon, Bong-Goan;Park, Suji;Kim, Jin-Won
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.12-20
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    • 2013
  • To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

Isolation of Anaerobic Bacteria from Clinical Specimens in Veterinary Medical Teaching Hospital (대학 동물병원 임상 검체로부터 분리된 혐기성 세균과 항생제 감수성 양상)

  • Park, Se-won;Chung, Jin-young;Hwang, Cheol-yong;Youn, Hwa-young;Han, Hong-ryul
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.15-19
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    • 2004
  • The emergence of bacterial resistance to antibiotics during therapy is a matter of great problem in clinical medicine. This may be because many veterinarians have used inappropriate antibiotics without bacteriological culture. Therefore, the purpose of this study is to determine isolation of anaerobic bacteria as pathogens from veterinary clinical specimens as well as susceptibility pattern for choosing antibiotics. Various anaerobic bacteria were isolated from clinical specimens of dogs, cats, rabbits at Veterinary Medical Teaching Hospital of Seoul National University from May 2001 to October 2002. The total number of isolated anaerobic bacteria was 13 isolates; Bacteroides spp. (3 isolates), Fusobacterium spp. (2 isolates), Peptostreptococcus spp. (2 isolates), Porphyromonas gingivalis (2 isolates), Prevotella spp. (3 isolates), and Propionibacterium acnes (1 isolate). For evaluating the antibiotic susceptibility patterns of the isolates disk diffusion method was used. All isolates were susceptable to all tested antibiotics except only one Fusobacterium varium was resistant to norfloxacin.

Outcome of High Dose AmpicillinSulbactam and Colistin Combination Therapy for Treating VentilatorAssociated Pneumonia Caused by Carbapenem-Resistant Acinetobacter baumannii: a Pilot Study (Carbapenem내성 Acinetobacter baumannii로 인한 인공호흡기연관 폐렴 환아에서 고용량 Ampicillin-Sulbactam 과 Colistin 항균제 병합요법의 치료적 예후: 예비 연구)

  • Jeong, Seong Hee;Kim, Young A;Choi, Go-eun;Park, Su Eun
    • Pediatric Infection and Vaccine
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    • v.27 no.1
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    • pp.45-52
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    • 2020
  • Purpose: This pilot study aimed to evaluate the efficacy of high dose ampicillin-sulbactam and colistin combination therapy for ventilator-associated pneumonia (VAP) caused by carbapenem-resistant Acinetobacter baumannii (CRAB) in the pediatric intensive care unit of Pusan National University Children's Hospital. Methods: We retrospectively reviewed 17 pediatric patients with VAP caused by CRAB from June 2017 to August 2018. Ten (58.8%) patients were treated with high dose ampicillin-sulbactam and colistin combination therapy (combination therapy group), whereas 7 were treated with colistin only or with various combinations with or without colistin (other antibiotics group). Clinical and bacteriological outcomes were compared between the groups. Results: The mean duration of fever after antibiotic use was 1.30±1.70 days in the combination therapy group and 1.71±1.49 days in the other antibiotics group. The mean duration of days for negative conversion of endotracheal aspirate bacterial culture after antibiotic therapy was 3.40±1.71 days in the combination therapy group and 11.80±8.86 days in the other antibiotics group. The mortality rate within 30 days of antibiotic therapy was 1/10 (10%) in the combination therapy group and 3/7 (42.9%) in the other antibiotics group. Conclusions: High dose ampicillin-sulbactam and colistin combination therapy as early antibiotic treatment in VAP caused by CRAB in children could improve clinical outcomes.

Type I immune-mediated polyarthritis with azathioprine therapy in a Shih-tzu dog

  • Jung, Dong-In;Park, Chul;Kang, Byeong-Teck;Kim, Ju-Won;Kim, Ha-Jung;Lim, Chae-Young;Ko, Ki-Jin;Lee, So-Young;Cho, Sue-Kyung;GU, Su-Hyun;Heo, Ra-Young;Park, Hyo-Jin;Jeon, Hyo-Won;Kim, Jung-Hyun;Han, Sung-Kuk;Yoon, Ah-Ram;Sung, Ju-Heon;Yoo, Jong-Hyun;Park, Hee-Myung
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.395-398
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    • 2006
  • A 2-month-old female Shih-tzu dog was referred because of lameness, exercise intolerance, depression, elbow and stifle joint swelling. Physical examination, complete blood counts, serum-chemistry, radiography, synovial fluid analysis, antinuclear antibody test, and rheumatoid factor measurement were initiated. On radiography, soft tissue swelling of elbow and stifle joints without erosiveness were founded. The results of synovial fluid analysis revealed severe neutrophilic pleocytosis (nondegenerative), decreased viscosity, increased turbidity, positive on mucin-clot test, and negative on bacterial culture. The results of rheumatoid factor measurement and antinuclear antibody test were negative and below 1 : 40, respectively. Based on all tests, we diagnosed this case as juvenile onset type I immune-mediated polyarthritis. Azathioprine (1 mg/kg body weight, per os q 24 h, for 4 weeks) was then administered and clinical signs improved gradually. Four weeks after azathioprine administration, clinical signs were disappeared. This report describes the clinical findings, imaging characteristics, synovial fluid findings, and other laboratory results of type I immune-mediated polyarthritis and successful management with azathioprine therapy.

Soybeans Fermented with Bacillus po/yfermenticus KJS-2 Protects Oplegnathus fasciatus from Iridovirus and Pathogenic Bacterial Infection (Bacillus polyfermenticus KJS-2에서 발효된 콩의 돌돔에 대한 이리도바이러스 및 병원성균에 대한 예방효과)

  • Kim, Kang-Min;Na, Hae-Choon;Park, Jung-Hee;Kang, Jae-Seon
    • Journal of Life Science
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    • v.19 no.6
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    • pp.720-727
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    • 2009
  • Lyophilized powder (BP2FS) of soybeans fermented with Bacillus polyfermenticus KJS-2 (B. polyfermenticus KJS-2) exhibited in vitro antibacterial activities against eight pathogenic bacteria. BP2FS was used as a fodder additive for Oplegnathus fasciatus (0. fasciatus) culture. One group (UFD) of O. fasciatus was fed a commercial fodder, while another group (FD) was fed the same fodder, but including BP2FS ($6{\times}10^{4}$ cfu $g^{-1}$ fodder), two times daily for 120 days. The mean body weight of the FD group (67.29${\pm}$12.62 g) was higher than that of the UFD group (56.56${\pm}$8.21 g) after 120 days. The survival rate of FD was 80% compared to 40% for the UFD group. Cumulative mortalities in the FD and UFD groups were 18.95% and 60.98% respectively. B. polyfermenticus KJS-2 was isolated from the intestines of the FD group and the number of viable colonies was estimated to be $1.04{\times}10^{4}$ cfu $g^{-1}$. Iridovirus and Vibrio vulnificus was detected in the organs of the UFD group but not in the FD group. All of the infected fish showed typical clinical symptoms of hemorrhage in their tail fins. Dissection of the infected internal organs revealed liver congestion and spleen enlargement - typical symptoms caused by iridovirus infection. These results clearly show that BP2FS is highly beneficial in preventing O. fasciatus from iridovirus infection.