• Title/Summary/Keyword: minimum inhibition concentration (MIC)

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Antibacterial Activity of Pharbitin, Isolated from the Seeds of Pharbitis nil, against Various Plant Pathogenic Bacteria

  • Nguyen, Hoa Thi;Yu, Nan Hee;Park, Ae Ran;Park, Hae Woong;Kim, In Seon;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1763-1772
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    • 2017
  • This study aimed to isolate and characterize antibacterial metabolites from Pharbitis nil seeds and investigate their antibacterial activity against various plant pathogenic bacteria. The methanol extract of P. nil seeds showed the strongest activity against Xanthomonas arboricola pv. pruni (Xap) with a minimum inhibition concentration (MIC) value of $250{\mu}g/ml$. Among the three solvent layers obtained from the methanol extract of P. nil seeds, only the butanol layer displayed the activity with an MIC value of $125{\mu}g/ml$ against Xap. An antibacterial fraction was obtained from P. nil seeds by repeated column chromatography and identified as pharbitin, a crude resin glycoside, by instrumental analysis. The antibacterial activity of pharbitin was tested in vitro against 14 phytopathogenic bacteria, and it was found to inhibit Ralstonia solanacearum and four Xanthomonas species. The minimum inhibitory concentration values against the five bacteria were $125-500{\mu}g/ml$ for the n-butanol layer and $31.25-125{\mu}g/ml$ for pharbitin. In a detached peach leaf assay, it effectively suppressed the development of bacterial leaf spot, with a control value of 87.5% at $500{\mu}g/ml$. In addition, pharbitin strongly reduced the development of bacterial wilt on tomato seedlings by 97.4% at $250{\mu}g/ml$, 7 days after inoculation. These findings suggest that the crude extract of P. nil seeds can be used as an alternative biopesticide for the control of plant diseases caused by R. solanacearum and Xanthomonas spp. This is the first report on the antibacterial activity of pharbitin against phytopathogenic bacteria.

Growth Inhibition of Helicobacter pylorio by Reynoutria elliptica Migo. (호장근에 의한 Helicobacter pylori의 생육 저해)

  • Lee, In-Seon;Im, Hyo-Gwon;Lee, Syng-Ook
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1182-1187
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    • 2003
  • This study was performed to evaluate the potentiality of Reynoutria elliptica Migo., being used as a folk remedy and a herb medicine for urethritis, cystitis, etc., on growth inhibition of Helicobacter pylori which is known as the ulcerogenic pathogen. The minimum inhibitory concentration (MIC) value of methanol extract from Reynoutria elliptica Migo, was determined to be 120 ppm for H. pylori and urease activity derived from H. pylori was inhibited over 80% by the extract at 2 mg/mL in urea broth. Among various solvent fraction of the methanol extract, the hexane fraction showed a significant inhibitory effect on the growth of H. pylori reducing both its growth and urease activity. Scanning and transmission electron micrographs of H. pylori treated with the methanol extract at 2 mg/mL for 3 hr showed that the cell walls and membranes were disrupted so that the cytoplasmic components were leaked from the body. These results suggest that Reynoutria elliptica Migo. possesses a therapeutic potential on the gastric disease caused by H. pylori.

Antioxidant, anti-inflammatory, and antibacterial activities of a 70% ethanol-Symphyocladia linearis extract

  • Jeong Min Lee;Mi-Jin Yim;Hyun-Soo Kim;Seok-Chun Ko;Ji-Yul Kim;Gun-Woo Oh;Kyunghwa Baek;Dae-Sung Lee
    • Fisheries and Aquatic Sciences
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    • v.25 no.11
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    • pp.579-586
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    • 2022
  • Research on the potential biological activity of red alga Symphyocladia spp. has been limited to Symphyocladia latiuscula, which is widely used as a food ingredient in Korea. Here, we examined the biological activity of another species, Symphyocladia linearis, which is found in Korea and was reported as a new species in 2013. The aim of this study was to evaluate the antioxidant, anti-inflammatory, and antibacterial properties of a 70% ethanol extract of S. linearis. Antioxidant activity, which was evaluated using radical scavenging assays, revealed half maximal inhibitory concentration values for 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) of 34.57 and 11.70 ㎍/mL algal extract, respectively. Anti-inflammatory activity of the S. linearis ethanolic extract was evaluated using RAW 264.7 cells by measuring the inhibition of lipopolysaccharide-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production. The potential cytotoxicity of NO and PGE2 was first examined, confirming no toxicity at concentrations ranging from 10-100 ㎍/mL. NO production was inhibited 61.1% and 78.0% at 50 and 100 ㎍/mL S. linearis extract, respectively; and PGE2 production was inhibited 69.1%, 83.2%, and 94.8% at 25, 50, and 100 ㎍/mL S. linearis extract, respectively. Thus, the S. linearis extract showed very strong efficacy against PGE2 production. The cellular production of reactive oxygen species, measured using 2',7'-dichlorofluorescin diacetate fluorescence, was inhibited 48.8% by the addition of 100 ㎍/mL S. linearis extract. Antibacterial activity was evaluated using the disc diffusion method and minimum inhibitory concentration (MIC). S. linearis was effective only against gram-positive bacteria, exhibiting antibacterial activity against Staphylococcus aureus with a MIC of 256 ㎍/mL extract and against Bacillus cereus with a MIC of 1,024 ㎍/mL extract. Based on these results, we infer that a 70% ethanolic extract of S. linearis possesses strong anti-inflammatory properties, and therefore has the potential to be used in the prevention and treatment of inflammatory and immune diseases.

The Antimicrobial Effect of Pulsatilla Koreana Extracts to Oral Micro-Organism (백두옹 추출물의 치주 병인균에 대한 항균효과)

  • Chung, Sung-Wha;Chung, Chin-Hyung;Lim, Sung-Bin;Kim, Jung-Keun;So, Eun-Hee
    • Journal of Periodontal and Implant Science
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    • v.30 no.3
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    • pp.661-676
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    • 2000
  • Gingivitis and periodontitis are infectious diseases in that microorganisms are the primary extrinsic cause of the diseases. the occurrence of gingivitis has been associated clearly with the presence of microorganisms at the disease site, and the histologic nature of the tissue involved is indicative of an inflammatory response induced by microorganisms. additional evidence for the microbial etiology of periodontal disease is that numerous antimicrobial agents are effective in reducing plaque accumulation and periodontal diseases. the purpose of this article is to analyze the antimicrobial effects of Pulsatilla koreana. Well-dried Pulsatilla koreana purchased from herbs distributor was ground and extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH). we have then applied each solution to the bacteria samples(Bacteroides forsythus, Streptococcus mutans, Streptococcus sanguis, Porphylomonas gingivalis, Actinobacillus actinomycetemcomitans, Eikenella corrodens, Prevotella intermedia, Actinomyces viscosus, Prevotella nigrescens , Rothia dentocariosa, Fusobacterium nucleatum, Pseudomonas aeruginosa, Staphylococcus aureus) collected from several organizations. To conduct susceptibility test(Kirby-Bauer method), plate contained each periodontopathic bacteria is spread extracted into methanol(MeOH), ethylacetate(EtoAc), chlorform($CHCl_3$) and Butyl alcohol(BuOH) and to measure the minimum inhibition concentration(MIC) of the bacteria against the solutions to ultimately determine antimicrobial effects of the solutions, insert bacteria sample into $20{\mu\ell}/{m\ell}$, $10{\mu\ell}/{m\ell}$, $5{\mu\ell}/{m\ell}$, $2.5{\mu\ell}/{m\ell}$ of each solution and control group(not contained solution) 1. Solution extracted into methanol did not show clear zone against all bacteria samples. Only P.nigrescens, S. mutans and S. sanguis in solution extracted into ethylacetate, S. mutans and S. anguis in solutions extracted into chlorform and Butyl alcohol showed clear zone against all bacteria samples. Solution extracted into Butyl alcohol showed clear zone against 13 types of bacteria, excluding P. gingivalis. 2. In Solution extracted into methanol, the bacteria samples grew in the highest concentrated plate, showing minimal variation from control group. 3. In Solution extracted into Butyl alcohol, S. aureus, P. intermedia, E. corrodens, A. actinomycetemcomitans, B. forsythus, P. gingivalis et al. showed decreased growth in the highest concentrated plate. P. auruginosa, R. dentocariosa, A. viscosus, P. nigrescens, S. mutans et al. showed decreased growth at MIC $20{\mu\ell}/{m\ell}$ and S. sanguis showed decreased growth at MIC $10{\mu\ell}/{m\ell}$. 4. By analyzing the MIC level through considering the results from Kirby-Bauer method, Solution extracted into methanol did not reveal any antimicrobial effects and Solution extracted into Butyl alcohol showed the highest antimicrobial effects In conclusion, it can be used the extracts of Pulsatilla koreana as wide spectrum antimicrobial agent.

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In vitro selection of fungicides for control of leaf blight of sweet persimmon tree caused by Pestalotiopsis theae (In vitro에서 단감나무 둥근갈색무늬병 방제를 위한 살균제 선발)

  • Chang, T.H.;Lim, T.H.
    • The Korean Journal of Pesticide Science
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    • v.5 no.4
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    • pp.50-56
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    • 2001
  • In vitro experiments, several fungicides including prochloraz, tebuconazole, fluazinam, fludioxonil, and iminoctadine-triacetate showed more than 85% inhibition of mycelial growth of Pestalotiopsis theae (SP-3). Dichlofluanid and chlorothalonil inhibited mycelial growth at the rate of 10 and 33%, however benomyl did not inhibit mycelial growth of the fungus. Minimum inhibitory concentration(MICs) of iminictadine-triacetate on the mycelial growth of SP-3 isolate was $10{\mu}g/m{\ell}$, but that of P. theae $\underline{MAFF}$ 752002 and P. longiseta $\underline{MAFF}$ 752001 was $1{\mu}g/m{\ell}$. MIC of benomyl, chlorothalonil, dichlorofluanid was $1,000{\mu}g/m{\ell}$, and that of fludioxonil, fluazinam, tebuconazole was $10{\mu}g/m{\ell}$. Conidial germination was inhibited more than 80% in tile medium which contained $1{\mu}g/m{\ell}$ of prochloraz, tebuconazole, fluazinam, fludioxonil, and minoctadine-triacetate. Control values of benomyl, chlorothalonil and dichlofluanid were 40, 60 and 30%, respectively. The controlling effect of iminoctadine-triacetate ($10{\mu}g/m{\ell}$) aganist P. theae (SP-3) in leaf disc test was more than 93%, but benomyl and dichlofluanid could not control leaf blight disease caused by P. theae (SP-3). Tebuconazole, fluazinam, fludioxonil, and iminoctadine-triacetate showed more than 94% of protective and curative effect of leaf blight of sweet persimmon, while, benomyl and dichlofluanid did not show any control effect of the disease.

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Inhibition of Microbial Quorum Sensing Mediated Virulence Factors by Pestalotiopsis sydowiana

  • Parasuraman, Paramanantham;Devadatha, B;Sarma, V. Venkateswara;Ranganathan, Sampathkumar;Ampasala, Dinakara Rao;Reddy, Dhanasekhar;Kumavath, Ranjith;Kim, In-Won;Patel, Sanjay K.S.;Kalia, Vipin Chandra;Lee, Jung-Kul;Siddhardha, Busi
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.571-582
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    • 2020
  • Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.

Antimicrobial Activity of Extracts and Fractions of Ginkgo biloba Leaves, Seed and Outer Seedcoat (은행 잎, 종실 및 외종피 추출물의 항균활성)

  • Park, Saet-Byoul;Cho, Gyu-Seong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.7-13
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    • 2011
  • This study was conducted to investigate the antimicrobial activity of Ginkgo biloba L. leaves, seed and outer seedcoat against bacteria. Antimicrobial effects of Ginkgo biloba L. leaves (GBL), seed (GBS) and outer seedcoat (GBO) were examined by paper disc method and optical density method to determine minimum inhibition concentration (MIC), and observed by scanning electron microscope (SEM) to figure out the morphological change on the surface when Ginkgo biloba leaves extract was treated. The extracts of GBL, GBS and GBO were extracted by solvents such as methanol, ethanol, water. The methanol extract of GBL and GBO showed the highest antimicrobial activity against Bacillus cereus, Bacillus subtilis, Klebsiella pneumoniae, Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, Yersinia enterocolitica except Escherichia coli and thus was further fractionated. The MICs of the chloroform fraction of GBL methanol extract were $125{\mu}g$/mL against B. subtilis, and L. monocytogenes; GBO methanol extract were $62.5{\mu}g$/mL against B. cereus and $125{\mu}g$/mL against B. subtilis, and L. monocytogenes. The microorganisms were treated with chloroform extracts ($2000{\mu}g$/mL) of GBL and GBO methanol extracts. It was observed by scanning electron microscope (SEM). The cells were expanded and a part of cell wall was completely destructed by GBL and GBO. Thus Ginkgo biloba L. leaves and outer seedcoat could be further developed into a natural antimicrobial agent.

Inhibition of Foodborne Pathogens and Spoilage Bacteria and Their Structural Changes by Ethanol Extract of Schizandra chinensis Baillon (오미자 에탄올 추출물에 의한 식품위해성 세균의 증식 억제 및 세포구조 변화)

  • Kim, Se-Ryoung;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.1
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    • pp.109-119
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    • 2012
  • This study analyzed the antibacterial activity of the ethanol extract of Schizandra chinensis Baillon against food pathogenic microorganisms to determine its capabilities as a natural antimicrobial agent. A paper disc diffusion test, minimum inhibitory concentration (MIC) determination, and time-kill assay showed that the ethanol extract strongly inhibits the growth of Listeria monocytogenes, Bacillus cereus, Escherichia coli O157:H7, and Pseudomonas aeruginosa. Release of cytoplasmic ${\beta}$-galactosidase was detected in E. coli, E. coli O157:H7, S. aureus, and P. aeruginosa treated with the ethanol extract. An increase of outer membrane permeability caused by the ethanol extract was also observed. An outward flow of cell constituents was detected in the Gram negative strains treated with the ethanol extract. These results imply that the inner and outer membranes of cells were partially destroyed and cell constituents were released by the treatment of the S. chinensis Baillon ethanol extract. The results of this study indicate that ethanol extract of S. chinensis Baillon evidences a fairly good antibacterial effect.

Antibacterial activity of Tonghyeonipal-dan against Methicillin-resistant Staphylococcus aureus (통현이팔단 에탄올 추출물의 Methicillin Resistant Staphylococcus aureus에 대한 항균활성)

  • KIM, In-Won;KANG, Ok-Hwa;KONG, Ryong;KWON, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.30 no.5
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    • pp.15-21
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    • 2015
  • Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

Antimicrobial Activities of Eight Compounds Purified from the Roots of Polygala tenuifolia Willdenow and the Aerial Bulbils of Dioscorea batatas Decene (원지와 영여자로부터 분리한 8종 화합물의 항균효과)

  • Chin, Hwi-Seung;Son, Rak-Ho;Lee, Yong-Hwa;Ham, Ah-Rom;Mar, Woong-Chon;Kim, Won-Ki;Nam, Kung-Woo
    • YAKHAK HOEJI
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    • v.54 no.2
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    • pp.106-111
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    • 2010
  • The chemical structures of eight compounds purified from two plants (Polygala tenuifolia Willdenow and Dioscorea batatas Decene) were determined and their anti-microbial activity against three microbial strains (Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans) was tested. The three micro organisms were cultured in 96-well plates or Petri dishes without (control) or with the eight compounds added at concentrations of 100 to 0.01 ${\mu}M$ (wt/vol). The growth of the microorganisms in the medium was examined after a 24-h incubation. The inhibitory effect of each compound on the growth of the microorganisms was calculated from the optical density measured at 595 nm, turbidity, and size of the inhibition zone around the treated paper disc. The minimum inhiitory concentration (MIC) of compounds 4 to 7 against S. aureus was 0.08, 0.05, 1.3 and 0.02 ${\mu}M$, respectively, and 0.09, 0.1, 0.2 and 100 ${\mu}M$ against C. albicans. The $IC_{50}$ (50% inhibition) values of compounds 5 and 6 were 3.1 and 6.4 ${\mu}M$ against S. aureus, respectively, and 10 and 2.4 ${\mu}M$ against C. albicans. Therefore, compounds 4 to 6 were the most potent anti-microbial agents among the eight compounds tested.