• Title/Summary/Keyword: minimal broth

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A Study of Antimicrobial Activity of Herbal Extracts on Clostridium difficile (Clostridium difficile에 대한 항균 한약재 탐색 연구)

  • Seong, Eunhak;Lim, Sookyoung;Lee, Myeongjong;Kim, Hojun
    • Journal of Korean Medicine Rehabilitation
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    • v.31 no.1
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    • pp.47-57
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    • 2021
  • Objectives This study was conducted to confirm the possibility of Clostridium difficile infection (CDI) treatment through natural herbal medicines. Methods After screening a total of 77 herbal medicines through the paper disc agar diffusion method, we selected the herbal medicines that showed a effectiveness compared to the positive control vancomycin. Afterwards, drugs that showed inhibitory effects compared to C. difficile without inhibition of Bifidobacterium bifidum and Lactobacillus plantarum, known as beneficial bacteria, were selected and minimal inhibitory concentration (MIC) was confirmed by applying the Broth microdilution method. Results The Coptidis Rhizoma, well known for its antimicrobial effect, was found to have antimicrobial effects on C. difficile, but also had inhibitory effects on the beneficial bacterium B. bifidum. 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus had antimicrobial effects on C. difficile without inhibiting the beneficial bacteria B. bifidum and L. plantarum. The MIC values of 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus were found to be 10 mg/mL, 20 mg/mL and 5 mg/mL, respectively. Conclusions Crataegi fructus, Corni fructus and Mume fructus were identified as candidate medicines for C. difficile. Further researchs will need to be done in vivo, and to find an optimal extraction method accompanied by economic evaluation.

In Vitro and In Vivo Anti-Clostridioides difficile Effect of a Probiotic Bacillus amyloliquefaciens Strain

  • Islam, Md Imtiazul;Seo, Hoonhee;Redwan, Asma;Kim, Sukyung;Lee, Saebim;Siddiquee, Mashuk;Song, Ho-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.46-55
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    • 2022
  • Clostridioides difficile infection (CDI) is a significant cause of hospital-acquired and antibiotic-mediated intestinal diseases and is a growing global public health concern. Overuse of antibiotics and their effect on normal intestinal flora has increased the incidence and severity of infections. Thus, the development of new, effective, and safe treatment options is a high priority. Here, we report a new probiotic strain, Bacillus amyloliquefaciens (BA PMC-80), and its in vitro/in vivo anti-C. difficile effect as a prospective novel candidate for replacing conventional antibiotics. BA PMC-80 showed a significant anti-C. difficile effect in coculture assay, and its cell-free supernatant (CFS) also exhibited a considerable anti-C. difficile effect with an 89.06 ㎍/ml 50% minimal inhibitory concentration (MIC) in broth microdilution assay. The CFS was stable and equally functional under different pHs, heat, and proteinase treatments. It also exhibited a high sensitivity against current antibiotics and no toxicity in subchronic toxicity testing in hamsters. Finally, BA PMC-80 showed a moderate effect in a hamster CDI model with reduced infection severity and delayed death. However, further studies are required to optimize the treatment condition of the hamster CDI model for better efficacy and identify the antimicrobial compound produced by BA PMC-80.

Anti-Oomycete Activity and Pepper Root Colonization of Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 against Phytophthora capsici

  • Elena, Volynchikova;Ki Deok, Kim
    • The Plant Pathology Journal
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    • v.39 no.1
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    • pp.123-135
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    • 2023
  • Previously, Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 from a sequential screening procedure were proven to effectively control Phytophthora blight caused by Phytophthora capsici. In this study, we further investigated the anti-oomycete activities of these strains against mycelial growth, zoospore germination, and germ tube elongation of P. capsici. We also investigated root colonization ability of the bacterial strains in square dishes, including cell motility (swimming and swarming motilities) and biofilm formation. Both strains significantly inhibited mycelial growth in liquid and solid V8 juice media and M9 minimal media, zoospore germination, and germ tube elongation compared with Bacillus vallismortis EXTN-1 (positive biocontrol strain), Sphingomonas aquatilis KU408 (negative biocontrol strain), and MgSO4 solution (untreated control). In diluted (nutrient-deficient) V8 juice broth, the tested strain populations were maintained at >108 cells/ml, simultaneously providing mycelial inhibitory activity. Additionally, these strains colonized pepper roots at a 106 cells/ml concentration for 7 days. The root colonization of the strains was supported by strong swimming and swarming activities, biofilm formation, and chemotactic activity towards exudate components (amino acids, organic acids, and sugars) of pepper roots. Collectively, these results suggest that strains YJR13 and YJR92 can effectively suppress Phytophthora blight of pepper through direct anti-oomycete activities against mycelial growth, zoospore germination and germ tube elongation. Bacterial colonization of pepper roots may be mediated by cell motility and biofilm formation together with chemotaxis to root exudates.

Enhancement of Erythrosine Photodynamic Therapy against Streptococcus mutans by Chlorhexidine (Streptococcus mutans에 대한 클로르헥시딘과 Erythrosine 광역동 치료의 상승효과)

  • Park, Jongcheol;Park, Howon;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.40 no.4
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    • pp.241-246
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    • 2013
  • The purpose of this study was to investigate the synergistic effects of erythrosine sensitized with a conventional halogen curing unit and sub-minimal bactericidal concentration(sub-MBC) of chorhexidine on bacterial viability of Streptococcus mutans in planktonic state. Sub-minimal bactericidal concentration of chlorhexidine was added into wells containing bacteria and erythrosine. The range of concentrations tested for chorhexidine was from 0.0000001% to 0.001%. The irradiation of the bacterial suspensions was performed for 15 sec with a conventional halogen curing unit light. In another set of experiment, the effects of 0.001% chlorhexidine were observed by adding chlorhexidine into wells containing the sub-minimal bactericidal concentration of erythrosine. At the concetration of 0.001% chlorhexidine, there were no antibacterial effects in the absence of erythrosine PDT(p < 0.05). At the concentraton of $1{\mu}M$ erythrosine, there was no photodynamic therapy effect in the absence of chlorhexidine(p < 0.05). But in the presence of sub-minimal bactericidal concentration of erythrosine with light exposure, the addition of 0.001% chlorhexidine increased the bactericidal rate(p < 0.05). A combination of erythrosine PDT with sub-MBC chlorhexidine resulted in a significant reduction in bacterial counts when compared to the case with the absence of chlorhexidine.

Multiplication of Escherichia coli DH5α::gfp on Strawberry Fruit Surface (딸기과실 표면에서 Eschercia coli DH5α::gfp 증식)

  • Yun, Hyejeong;Park, Kyeonghun;Ryu, Kyoung Yul;Yun, Jong-Chul;Kim, Byung Seok
    • Food Science and Preservation
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    • v.20 no.2
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    • pp.250-256
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    • 2013
  • To verify the multiplication of microorganisms on the surface of strawberries, the fate of E. coli $DH5{\alpha}::gfp$ at different temperatures, times and strawberry extract concentrations were measured. The population of E. coli $DH5{\alpha}::gfp$ rapidly increased by 7.36~7.78 log CFU/g at $25{\sim}30^{\circ}C$ for 24 hr and slowly increased by 6.49~8.49 log CFU/g at $10{\sim}20^{\circ}C$ for 48 hr. However, E. coli $DH5{\alpha}::gfp$ did not grow at $10{\sim}15^{\circ}C$ on the surface of the strawberries, regardless of the contact times with the bacterial suspension. E. coli $DH5{\alpha}::gfp$ reached 1.52~3.26 log CFU/g at $20^{\circ}C$ as the contact frequency increased from two to six times. The contact frequencies did not significantly differ. In the case of the six-time contact on the surface of the strawberry at 25 and $30^{\circ}C$, the E. coli $DH5{\alpha}::gfp$ increased by 5.17 and 5.01 log CFU/g. The effects of the strawberry extracts on the growth of E. coli $DH5{\alpha}::gfp$ showed that sterilization and non-sterilization do not affect the growth of microorganisms for 96 hr. In the minimal broth, the growth of E. coli $DH5{\alpha}::gfp$ increased by 1 log CFU/g for 96 hr. In less than 50 percent of the strawberry extracts, the growth rate of E. coli $DH5{\alpha}::gfp$ was higher than in the control and increased by 4 and 5 log CFU/g at 50 and 25 percent of strawberry extracts, respectively. Therefore, E. coli $DH5{\alpha}::gfp$ can multiply and survive on the surface of strawberries when it comes into contact with the fruit extract.

Plumbagin Inhibits Expression of Virulence Factors and Growth of Helicobacter pylori (Plumbagin에 의한 헬리코박터 파이로리균의 성장 및 병원성 인자 발현 억제효과)

  • Lee, Min Ho;Woo, Hyun Jun;Park, Min;Moon, Cheol;Eom, Yong-Bin;Kim, Sa-Hyun;Kim, Jong-Bae
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.218-226
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    • 2016
  • Helicobacter pylori primarily colonizes the human stomach. Infection by this bacterium is associated with various gastric diseases, including inflammation, peptic ulcer, and gastric cancer. Although there are antibiotic regimens for the eradication of H. pylori, the resistance of this species against antibiotics has been continuously increasing. The natural compound plumbagin has been reported as an antimicrobial and anticancer molecule. In this study, we analyzed the inhibitory effect of plumbagin on H. pylori strain ATCC 49503 as well as the expression of various molecules associated with H. pylori growth or virulence by immunoblotting and reverse transcription polymerase chain reaction (RT-PCR) analyses. We demonstrated the minimal inhibitory concentration of plumbagin on H. pylori through the agar dilution and broth dilution methods. Furthermore, we investigated the effect of plumbagin treatment on the expression of the RNA polymerase subunits and various virulence factors of H. pylori. Plumbagin treatment decreased the expression of RNA polymerase subunit alpha (rpoA), which is closely associated with bacterial survival. Moreover, the mRNA and protein levels of the major CagA and VacA toxins were decreased in plumbagintreated H. pylori cells. Likewise, the expression levels of urease subunit alpha (ureA) and an adhesin (alpA) were decreased by plumbagin treatment. Collectively, these results suggest that plumbagin may inhibit the growth, colonization, and pathogenesis of H. pylori by the mechanism demonstrated in this study.

Strain Improvement of Leuconostoc paramesenteroides as a Acid-Resistant Mutant and Effect on Kimchi Fermentation as a Starter. (Leuconostoc paramesenteroides의 내산성 변이주로의 개량과 starter로의 첨가효과)

  • 김영찬;정은영;김은해;정대현;이옥숙;권태종;강상모
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.151-160
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    • 1998
  • The Leuconostoc paramesenteroides dominated at refrigeration temperature range was isolated from kimchi, and improved its growth properties by mutation for competitive growth against Lactobacillus plantarum at lower pH. It was found that the minimal pH for the wild type Leuconostoc paramesenteroides Pw growth was pH 4.5 adjusted with HCI and pH 5.0 adjusted with organic-mixture (lactic acid:acetic acid=1:2), respectively. The mutant P-100 could grow in pH 4.0, 4.5, respectively, in MRS broth. Two strains Pw and P-100 were added into kimchi as starter and compared the quality characteristics of kimchi. The total acceptability of Pw and P-100 inoculated kimchi were evaluated better than that of control kimchi (no starter added) by sensory test and extended the optimal pH range of kimchi up to about 2.2, 2.5 times, respectively. In kimchi added P-100, the succinic acid was more abundant than others and the total number of Lactobacillus plantarum was down about 2.5 times in contrast to control kimchi.

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Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Mycelial Culture Conditions of Lepista nuda and Extracellular Enzyme Activity (민자주방망이버섯(Lepista nuda) 균사체 배양조건 및 효소활성)

  • Kim Sang-Dae;Kim Ji-Hye;Kim Jong-Bong;Han Yeong-Hwang
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.164-167
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    • 2005
  • The culture condition and medium composition for the enhanced mycelial growth of Lepista nuda DGUM 26501 were investigated. The optimal temperature and pH for the mycelial growth were $24^{\circ}C$ and $7.0\~8.0$, respectively. The partial pressure of oxygen for the enhanced mycelial growth was more than $10\%\;O_2$. When Czapek-Dox medium was used as a minimal medium, manitol and xylitol were very good carbon sources. Organic nitrogen sources were better than inorganic ones for mycelial growth. As the nitrogen source tested, com steep liquor, soytone and protease peptone were the best as a source of organic nitrogen sources. When ammonium phosphate as phosphorus sources was used, the enhanced mycelial growth was shown. Nicotinic acid was proved to be the most appropriate source of vitamin. After the mycelia of L. nuda DGUM 26501 was cultivated at $24^{\circ}C$ for 10 days in LNM broth (pH 7.0), the activities of extracellular enzyme were determined. The specific activity of $\alpha-amylase$ was much higher than those of other enzymes. However, little or no enzyme activities of $\beta-glucosidase$, CMCase, laccase and lipase were found.

ANTIBACTERIAL EFFECT OF POLYPHOSPHATES ON MUTANS STREPTOCOCCI (Mutans streptococci에 대한 polyphosphate의 항균효과)

  • Kang, Kye-Sook;Choi, Yeong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.80-91
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    • 2003
  • Mutans streptococci, especially S. mutans and S. sobrinus strongly implicated in pathogenesis of dental caries, the major cause of tooth loss in children. Use of an antibacterial agent controlling dental caries has been rationalized. The present study was performed to observe the antibacterial effect of inorganic polyphosphates (polyP) on S. mutans and S. sobrinus. S. mutans GS5 and S. sobrinus 6715 were grown in brain-heart infusion broth with or without polyP. Minimal inhibitory concentration (MIC) of polyP for S. mutans GS5 was determined to be 0.08% and that for S. sobrius 6715 was 0.17%. PolyP 15 added to the growing culture of S. mutans GS5 and S. sobrinus 6715 at their exponential phase was as effective in inhibiting the growth of S. mutans GS5 and S. sobrinus 6715 as polyP added at the very beginning of the culture. More than 85% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. mutans GS5 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. And more than 99.9% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. sobrinus 6715 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. Intracellular nucleotide release from S. mutans CS5 and S. sobrinus 6715 was increased in the presence of polyP 15 for 5h but was not really reversed by the addition of divalent cations like $Ca^{++}\;and\;Mg^{++}$. The majority of the cells appeared to be atypical in their shape, demonstrating accumulation of highly electron-dense granules and ghost cells. The overall results suggest that polyP have a strong bactericidal activity against S. mutans and S. sobrinus in which lysis in relation to chelation may not play the major role but unknown mechanism that possibly affects the viability of the bacterium may be involved. PolyP may be used as an agent for prevention of dental caries.

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