• Title/Summary/Keyword: bacterial cell number

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Eradicating Bacterial Persister Cells with Substituted Indoles to Reduce Antibiotic Resistance (항생제 내성률 감소를 위한 퍼시스터 세포 박멸과 인돌의 기능)

  • Park, Garin;Song, Sooyeon
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.4
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    • pp.145-156
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    • 2021
  • Antibiotics are used in many sectors, including the dairy industry, to prevent bacterial infections in humans, animals, and plants. When bacterial cells are exposed to stressors, such as antibiotic exposure, a subpopulation of the cells becomes dormant. This helps the pathogen to revive and reconstitute its pathogenicity. Thus, eradicating the dormant cells may be an effective strategy to reduce the development of antibiotic resistance in bacteria caused by the abuse of antibiotics. In recent years, a large number of indole-related compounds have been reported to eradicate persister cells. In this review, we provide a summary of the mechanisms of persister cell formation and resuscitation, and the ability of indole and substituted indoles to eradicate persister cells.

Study on attachment of bacteria to tonsillar epithelial cell during acute tonsillitis (급성편도선염에서 편도상피세포의 세균부착성에 관한 연구)

  • 이흥만;정형목;최충식;이우섭;이상학;황순재
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.98-98
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    • 1993
  • To cause invasive disease, microorganism must attach firmly to the tonsillar epithelial cell. Once attached, the microorganisms can proliferated, form colonies and release extracillular toxins which can injure the underlying cells. The purpose of present study was to asertain whether or not there exist in vivo differences in bacterial attachment between patients with acute tonsillitis and healthy individuals as a control. This study was carried out on 20 patients suffering from acute tonsillitis and 20 healthy persons used as control. After scraping of the surface of tonsil, cellular mixture was stained with Acridine orange and the number of attached bacteria was calculated using a fluorescent microscope. The adherence rate was calculated as number of bacteria attached to each of 50 epithelial cells. simultaneously, we peformed conventional bacterial culture. Conclusively, the attachment of more than 10% bacteria to the tonsillar epithelial cell was significantly greater in acute tonsillitis group than control group, and there was a significant correlation between age and the number of the attached bacteria to the epithlium.

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The Role of AiiA, a Quorum-Quenching Enzyme from Bacillus thuringiensis, on the Rhizosphere Competence

  • Park, Su-Jin;Park, Sun-Yang;Ryu, Choong-Min;Park, Seung-Hwan;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1518-1521
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    • 2008
  • Bacteria sense their population density and coordinate the expression of target genes, including virulence factors in Gram-negative bacteria, by the N-acylhomoserine lactones (AHLs)-dependent quorum sensing (QS) mechanism. In contrast, several soil bacteria are able to interfere with QS by enzymatic degradation of AHLs, referred to as quorum quenching. A potent AHL-degrading enzyme, AiiA, from Bacillus thuringiensis has been reported to effectively attenuate the virulence of bacteria by quorum quenching. However, little is known about the role of AiiA in B. thuringiensis itself. In the present study, an aiiA-defective mutant was generated to investigate the role of AHA in rhizosphere competence in the root system of pepper. The aiiA mutant showed no detectable AHL¬-egrading activity and was less effective for suppression of soft-rot symptom caused by Erwinia carotovora on the potato slice. On the pepper root, the survival rate of the aiiA mutant significantly decreased over time compared with that of wild type. Interestingly, viable cell count analysis revealed that the bacterial number and composition of E. carotovora were not different between treatments of wild type and the aiiA mutant. These results provide evidence that AHA can play an important role in rhizosphere competentce of B. thuringiensis and bacterial quorum quenching to Gram-negative bacteria without changing bacterial number or composition.

Production of Bacillus thuringiensis Spore Using an Industrial Medium (산업용 배지를 이용한 Bacillus thuringiensis의 포지생산)

  • 최성호;강석권;유연우
    • KSBB Journal
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    • v.13 no.6
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    • pp.644-648
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    • 1998
  • In the production of a low cost bacterial insecticide, it is important to produce a high spore concentration using low price substrates. Experiments were carried out to investigate the effects of the addition of mineral salts and glucose, and of dissolved oxygen concentration on the cell growth and spore formation of Bacillus thuringiensis var aizawai using a cheap wheat and soybean meal in the batch culture. The maximum viable cell number was 1.2${\times}$109 CFU/mL at 12 hr culture and spore yield was 54.2% at 74 hr culture using an industrial medium containing 20 g/L wheat meal and 30 g/L soybean meal under 1.0 vvm aeration and 200 rpm agitation. The cell growth and the spore formation were not enhanced by the addition of mineral salts in industrial medium, whereas th addition of 10g/L glucose decreased the cell growth and spore formation. We could obtain a maximum viable cell number of 2.2${\times}$109 CFU/mL and spore number of 1.9${\times}$109 CFU/mL at the dissolved oxygen concentration of 60% of saturation. The spore concentration was enhanced approximately by 2 times as compared to the dissolved oxygen concentration of 50%. In the bench-scale culture, the maximum viable cell and spore number were 2.5${\times}$109 CFU/mL, and 2.2${\times}$109 CFU/mL, respectively under 1.0 vvm aeration and 400 rpm agitation. The spore yield was 88% based on the maximum viable cell number. As a result, it was confirmed that the production of high spore concentration could be obtained by a bench-scale culture using an industrial medium.

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Infection Structures on the Infected Leaves of Potato Pre-inoculated with Bacterial Strains and DL-3-amino Butyric Acid after Challenge Inoculation with Phytophthora infestans

  • Kim, Hyo-Jeong;Jeun, Yong-Chull
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.203-209
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    • 2007
  • Infection structures were observed using a fluorescence microscope at the penetration sites on the leaves of potato plants pre-inoculated with the bacterial strains Pseudomonas putida TRL2-3, Micrococcus luteus TRK2-2, and Flexibacteraceae bacterium MRL412, which mediated an induced systemic resistance on potato plants against late blight disease caused by Phytophthora infestans. In order to compare the infection structures on the leaves expressing systemic acquired resistance, the leaves of potato plants pre-treated with DL-3-amino butyric acid (BABA) were also observed after challenge inoculation with the same pathogen. The infection structures were investigated. The total number of germination and appressorium formation of P. infestans were counted. Furthermore, the frequencies of fluorescent epidermal cells at the penetration sites, which indicate a defense response of plant cell, were estimated. There were no differences on the germination rates of the fungal cysts among the untreated control, BABA pre-treated, and bacterial strains pre-inoculated plants. However, appressorium formation was slightly decreased on the leaves of BABA pre-treated plants compared to those of untreated as well as bacterial strains pre-inoculated plants. Furthermore, the frequencies of fluorescent cells of BABA pre-treated and bacterial strains pre-inoculated were higher than that of untreated plants, indicating an active defense reaction of the host cells against the fungal attack. On the other hand, the pre-treatment with BABA caused a stronger fluorescent of epidermal cells at the penetration sites compared to the pre-inoculation with the bacterial strains. Interestingly, the frequency of fluorescent cells by BABA, however, was lower than that by the bacterial strains. Based on the results it is suggested that the infection structures showing resistance reaction on the leaves of potato plants were different between by pre-inoculation with bacterial strains and by pre-treatment with BABA against the late blight pathogen.

Bacterial Community Dynamics during Composting of Food Wastes (음식물 쓰레기 퇴비화 과정에 따른 세균군집 구조의 변화)

  • Shin, Ji-Hye;Lee, Jin-Woo;Nam, Ji-Hyun;Park, Se-Yong;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.148-154
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    • 2009
  • Composting is a biological process converting solid organic waste into valuable materials such as fertilizer. The change of bacterial populations in a composting reactor of food waste was investigated for 2 months. Based on shifts in temperature profile, the composting process could be divided into the first phase ($2^{\circ}C\sim55^{\circ}C$), the second phase ($55^{\circ}C\sim97^{\circ}C$), and the third phase ($50^{\circ}C\sim89^{\circ}C$). The number of total bacteria was $1.66\times10^{11}$ cell/g, $0.29\times10^{11}$ cell/g, and $0.28\times10^{11}$ cell/g in the first, second, and third stages, respectively. The proportions of thermophiles increased from 33% to 89% in the second stage. T-RFLP analysis and nucleotide sequencing of 16S rRNA gene demonstrated that the change of bacterial community structure was coupled with shifts in composting stages. The structure of bacterial community in the ultra-thermophilic second stage reflected that of seeding starter. The major decomposers driving the ultra-thermophilic composting were identified as phylotypes related to Bacillus and Pseudomonas.

Analysis of the Changes in Metabolic Diversity of Microbial Community in pH-gradient Microcosm

  • Ahn, Young-Beom;Cho, Hong-Bum;Park, Yong-Keel
    • Journal of Microbiology
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    • v.37 no.1
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    • pp.1-9
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    • 1999
  • The Biolog redox technology was carried out for evaluation of acidification effect on microbial communities at each stage of pH gradient microcosm. While the number of heterotrophic bacterial population and activities of extracellular enzyme decreased as the pH decreased, the number of total bacteria in the microcosm was not affected. The average color development of sample at each pH-gradient showed a sigmoidal curve, and at higher pH, more overall color development appeared in Biolog plates. Average color development value in Biolog plates was stabilized at 50 hours as an optimum incubation time. The color production in the Biolog plates was caused by cell density at above pH 5.0, but by cell activity below pH 4.0. Principal component analysis of color responses revealed distinctive patterns among the pH-gradient microcosm samples.

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Long-Term Starvation Induces the Viable-but-Nonculturable Condition in Lactobacillus crispatus KLB46

  • Lee, Seok-Yong;Kim, Ju-Hyeon;Jang, Jeong-Eun;Kim, Seung-Cheol;Yun, Hyeon-Sik;So, Jae-Seong
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.918-922
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    • 2001
  • In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

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Fermentation of a Potential Biocontrol Agent, Bacillus amyloliquefaciens SKU-78 Strain (풋마름병균의 길항세균 Bacillus amyloliquefaciens SKU-78의 대량 배양 조건 확립)

  • Kim, Shin-Duk;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.84-86
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    • 2014
  • Mass production of biocontrol agent is an essential step for its commercial use. Media composition and culture conditions for production of Bacillus amyloliquefaciens SKU-78, a potential biocontrol agent against bacterial wilts, were optimized by a flask culture. Low cost media combining nitrogen and carbon sources were tested. Maximum cell growth (> $2{\times}10^9$ CFU/ml) was obtained in a medium of 5% soy flour combined with 3% corn starch after 24 h cultivation. The optimum initial pH, temperature and shaking speed was 5.5, $30^{\circ}C$ and 150-250 rpm, respectively. Fermentation of SKU-78 was scaled up in 30 L fermenter and the profiles of cell density, pH, dissolved oxygen and spore formation were recorded. After 8 h lag phase, exponential growth occurred and reached at maximum viable cell number ($1.2{\times}10^{11}$ CFU/ml) after 20 h. The SKU-78 strain grown in a low cost medium exhibited the high suppression of bacterial wilts. The results indicate that SKU-78 strain can be produced in a low cost medium and provide a basis for scaling up to industrial level.

The Effects of Phellodendri Cortex Treatment on Hematological and Cyto-pathological Alterations in Non-Bacterial Prostatitis Rat Model (황백이 만성 비세균성 전립선염 모델에서 혈액학적 및 세포조직학적 변화에 미치는 영향)

  • Kim, Soon-Il;Ahn, Young-Min;Ahn, Se-Young;Doo, Ho-Kyung;Lee, Byung-Cheol
    • The Journal of Korean Medicine
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    • v.27 no.3 s.67
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    • pp.51-62
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    • 2006
  • Objective: Although chronic non-bacterial prostatitis is a common disease, it is very difficult to treat effectively. Lygodium japonicum has traditionally been used in treatment of urinary tract inflammation and voiding disturbance. In this study, we investigated the therapeutic effects and action mechanism of Lygodium japonicum in the rat model of non-bacterial prostatitis induced by castration and testosterone treatment. Methods: Five-month-old rats were treated with $17\beta-estradiol$ after castration for induction of experimental non-bacterial prostatitis, which is similar to human chronic prostatitis in histopathological profiles. Lygodium japonicum and testosterone were administered as an experimental specimen and a positive control, respectively. The prostates were evaluated by histopathological parameters including the epithelial score and epithelio-stromal ratio for glandular damage, proliferating cell nuclear antigen (PCNA) labeling index for cyto-proliferation and a TUNEL (deoxyuridine triphosphate biotin nick end-labeling) assay for cell apoptosis. Results: While prostates of control rats revealed severe acinar gland atrophy and stromal proliferation, the rats treated with Lygodium japonicum showed a lesser range of tissue damage. Epithelial score was improved in Lygodium japonicum than that of the control (P<0.05). The epithelio-stromal ratio was lower in Lygodium japonicum when compared to that of the control (P<0.05). Although there was no difference in PCNA and TUNEL positive cells of the glandular epithelia, we found an decreased number of PCNA positive cell and concurrent increase of TUNEL positive cells in the stroma of Lygodium japonicum treated rats (P<0.01). Conclusions: These findings suggest that Lygodium japonicum may protect the glandular epithelial cells and also inhibit stromal proliferation in association with suppression of cyto-proliferation and stimulation of apoptosis. We concluded that Lygodium japonicum may be a useful remedy agent for treating the chronic non-bacterial prostatitis.

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