• Title/Summary/Keyword: Disc assay

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Control Effect of Stenotrophomonas maltophilia BW-13 strain to the lettuce Bottom rot

  • Park, Jong-Young;Kim, Hyun-Ju;Bak, Joung-Woo;Lee, Kwang-Youll;Jun, Ok-Ju;Lee, Jin-Woo;Jung, Soon-Je;Moon, Byung-Ju
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.103.1-103
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    • 2003
  • An antagonistic bacteria, Stenotrophomonas maitophilia BW-13 strain which was effectively inhibited mycerial growth of Bottom rot pathogen, Rhizoctonia solani PY-1 strain was isolated from the rhizosphere of the lettuce in Uiryeong-Gun, Gyeongsangnam-Do from 2002 to 2003. For the biological control, the most suitable inoculum and its density of pathogen, PY-1 strain ware tested prior biological control test, For the pathogenicity test, A inoculum (wheat bran)sawdust+rice bran+PDB) showing disease incidence of 100% was selected as the most suitable inoculum, which showed more effective than B inoculum (sawdust+rice bran+DW) and mycelial disc. also, In selection of the amount of inoculum (40g, 50g, 60g, 70g, 80g), most suitable amount of inoculum of pathogen determined as 40g showing disease incidence of 80%. For the selection of effective microorganism to control bottom rot on lettuce, about 200 isolates were isolated from the diseased soil and lettuce leaves, and examined their antifungal activity to the pathogen on PDA. As the pots assay, BW-13 strain showed the highest control value as 90%, and followed by R-13 and R-26 strain as 80% and 60%, respectively. Selected BW-13 isolates identified as 5. maltophilia (GeneBank accession no. AJ293473.1, 99%) by 16S rRNA sequencing. This is the first report on the biological control using by S. maltophilia to the bottom rot pathogen, Rhizoctonia solani PY-1 strain.

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Anthocyanins from Clitoria ternatea Attenuate Food-Borne Penicillium expansum and its Potential Application as Food Biopreservative

  • Leong, Chean-Ring;Azizi, Muhammad Afif Kamarul;Taher, Md Abu;Wahidin, Suzana;Lee, Kok-Chang;Tan, Wen-Nee;Tong, Woei-Yenn
    • Natural Product Sciences
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    • v.23 no.2
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    • pp.125-131
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    • 2017
  • Clitoria ternatea or Commonly known blue pea, is a perennial climber crop native to Asian countries. The current study was aimed to evaluate the antimicrobial activity C. ternatea extract on food borne microorganisms and its antifungal effect on Penicillium expansum. The extract showed significant antimicrobial activity against 3 Gram positive bacteria, 2 Gram negative bacteria and 1 filamentous fungus on disc diffusion assay. The extract also showed good biocidal effect on all Gram positive bacteria tested and P. expansum. However, the kill curve analysis revealed that the fungicidal activity of the extract against P. expansum conidia was depend on the concentration of the extract and the time of exposure of the conidia to the extract. The scanning electron micrograph of the extract treated P. expansum culture showed alterations in the morphology of fungal hyphae. The germination of P. expansum conidia was completely inhibited and conidial development was totally suppressed by the extract, suggesting the possible mode of action of anthocyanin. Besides, the extract also exhibited 5.0-log suppression of microbial growth relative to control in the rice model. The results indicate the potential use of the C. ternatea anthocyanin as food biopreservative.

Emodin from Polygonum cuspidatum showed Angiogenesis Inhibiting Activity in vitro (호장근으로부터 분리된 emodin의 혈관신생 억제 활성)

  • Lee, Tae-Kyoo;Kim, Jong-Hwa;So, June-No
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.50-54
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    • 2003
  • Polygonum cuspidatum has been used as a fork medicine for a long time. Emodin was purified from the root of P. cuspidatum by thin layer chromatography (TLC) and preparative high perfomance liquid chromatography (HPLC). The effects of emodin on the migration of endothelial cells and in vitro angiogenesis stimulated with vascular endothelial cell growth factor (VEGF) were examined, using human umbilical vein endothelial cells (HUVECs) and porcine pulmonary arterial endothelial cells (PPAECs). Emodin potently inhibited the VEGF-induced migration of (HUVECs) at relatively low cocentrations $(0.1-10\;{\mu}g/ml)$; the inhibition of endothelial cells by emodin was 75.4% at $0.1\;{\mu}g/ml$ and about 90% at $1\;{\mu}g/ml$. Emodin also inhibited VEGF-induced sprout formation in vitro at concentrations of $0.1-10\;{\mu}g/ml$. Emodin was also evaluated for the inhibitory potential on in vivo angiogenesis in a growing chick embryo chorioallantoic membranes (CAM). At a concentration of $1.0\;{\mu}g/ml$ Per disc, emodin was able to induce avacular zone in the CAMs. These findings suggest that emodin is a potent angiogenesis inhibitor and P. cuspidatum is a useful herb in the development of therapeutics for angiogenesis dependent diseases.

Effect of Scutellariae Radix as a Novel Antibacterial Herb on the ppk(Polyphosphate Kinase) Mutant of Salmonella typhimurium

  • Hahm, Dae-Hyun;Yeom, Mi-Jung;H.Lee, Eun-Joo;Shim, In-Sop;Lee, Hye-Jung;Kim, Hong-Yeoul
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1061-1065
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    • 2001
  • The antibacterial effects of water extracts of Scutellariate Radix (a dried root of Scutellaria baicalensis GEORGI) and its major flavonoid components, Baicalin and Baicalein, on Salmonella typhimurium, a representative enteric pathogen, were studied. Through a Kriby-Bauer disc analysis, the growth-inhibition activity of Scutellariae Radix against. S. typhimurium was found to be compatible with commercial antibiotics, such as ampicillin, chloramphenicol, and streptomycin. In contrast, the growth of a nonpathogenic E. coli strain was unaffercted by Scutellariae Radix. To examine the effect of polyphosphate kinase (ppk), a putative virulence factor, on the antibacterial activity of Scutellariae Radix, the growth profile of a ppk mutant of S. typhimurium was investigated in a tryptic soy broth containing different concentrations of water extracts of Scutellariae Radix. The ppk mutant was able to grow in 6 mg/ml of water extracts of Scutellariae Radix, whereas in 6 mg/ml of water extracts of Scutellariae Radix, whereas the wild-type could not, implying that the inactivation of ppk made S. typhimurium more resistant to the antibacterial activity of Scutellariae Radix. No enhanced resistance was observed in a ppk mutant of S. typhimurium complemented with a ppk expression vector. The attenuation of the virulence by ppk inactivation was also observed in a virulence assay using BLAB/c mice. Neither Baicalin nor Baicalein exhibited any growth-inhibition activity against S. typhimurium. The water extracts of Scutellariae Radix stimulated the transcription of ppk, especially in the early growth-stage of S. typhimurium.

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Synergistic Antimicrobial Effect of Lonicera japonica and Magnolia obovata Extracts and Potential as a Plant-Derived Natural Preservative

  • Lee, Ye Seul;Lee, Yun Ju;Park, Soo Nam
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1814-1822
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    • 2018
  • Most people use cosmetics to protect their skin. Preservatives are often used to prevent their contamination upon use. There has been a great demand for natural preservatives due to recent reports on the side effects of parabens. Therefore, we evaluated the antimicrobial activities of Lonicera japonica and Magnolia obovata extracts and determined their potential as natural preservatives. We found that the 50% ethanol extract from L. japonica had antibacterial activity only against S. aureus and P. aeruginosa, while the ethyl acetate fraction showed antimicrobial activity against all six microbial strains tested. On the other hand, the 70% ethanol extract and the ethyl acetate fraction from M. obovata showed antimicrobial activity against all six strains. A synergistic effect against S. aureus, B. subtilis, and C. albicans was confirmed when two ethyl acetate fractions having antimicrobial activity against all six strains were used in combination. Synergistic activity against B. subtilis was also confirmed through kill-time analysis. High-performance liquid chromatography was performed to identify the components of each extract. Based on the minimum inhibitory concentration and the results of a disc diffusion assay, we confirmed that caffeic acid and luteolin influenced the antimicrobial activity of L. japonica and that the antimicrobial activity of M. obovata was influenced by the interaction of magnolol and honokiol with other components. Therefore, this study suggests that the combination of L. japonica and M. obovata extracts may be used as a plant-derived natural preservative.

Antimicrobial efficacy of endophytic Penicillium purpurogenum ED76 against clinical pathogens and its possible mode of action

  • Yenn, Tong Woei;Ibrahim, Darah;Chang, Lee Kok;Ab Rashid, Syarifah;Ring, Leong Chean;Nee, Tan Wen;Noor, Muhamad Izham bin Muhamad
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.193-199
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    • 2017
  • This study was aimed to evaluate the antimicrobial activity of Penicillium purpurogenum ED76 on several clinically important microorganisms. The endophytic fungus P. purpurogenum ED76 was previously isolated from Swietenia macrophylla leaf. The antimicrobial efficacy of P. purpurogenum ED76 dichloromethane extract was determined via disc diffusion and broth microdilution assay. A kill curve study was conducted and the morphology of extract treated bacterial cells were viewed under scanning electron microscope. The dichloromethane extract showed significant inhibitory activity on 4 test bacteria and 2 test yeasts. The minimal inhibitory concentration of the extract ranged from 125 to $1,000{\mu}g/ml$, which indicates the different susceptibility levels of the test microorganisms to the fungal extract. The kill curve study has revealed a concentration-dependent inhibition for all test microorganisms. With the increase of the extract concentration, the microbial growth was significantly reduced. The scanning electron micrograph of dichloromethane extract-treated Staphylococcus aureus cells showed the total damage of the cells. The cell wall invagination of the bacterial cells also indicates the loss of cellular materials and metabolic activity. The gas chromatography mass spectrometry analysis of the extract also showed that the major compound was stigmasterol, which constitutes 45.30% of the total area. The dichloromethane extract of P. purpurogenum ED76 exhibited significant inhibitory activity on several clinically important bacteria and yeasts. The study proposed a possible mode of action that the extract cause significant damage to the morphology of S. aureus cells.

Chitosan Based Silver Nanocomposites (CAgNCs) Display Antibacterial Effects against Vibrio ichthyoenteri

  • Beom, Seo Seung;Shin, Sang Yeop;Dananjaya, S.H.S.;De Silva, A.B.K.H.;Nikapitiya, Chamilani;Cho, Jongki;Park, Gun-Hoo;Oh, Chulhong;Kang, Do-Hyung;De Zoysa, Mahanama
    • Journal of Veterinary Clinics
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    • v.34 no.4
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    • pp.261-267
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    • 2017
  • The aim of this study was to investigate the antibacterial properties of chitosan silver nanocomposites (CAgNCs) using pathogenic Vibrio ichthyoenteri as a bacterial model. Results of agar disc diffusion and turbidimetric assays showed that CAgNCs could inhibit the growth of V. ichthyoenteri in concentration dependent manner. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CAgNCs were 75 and $125{\mu}g/mL$, respectively. Furthermore, CAgNCs treatment induced the reactive oxygen species (ROS) level in V. ichthyoenteri cells in concentration and time dependent manner, suggesting that it generates oxidative stress, leading to bacterial cell death. The field emission scanning electron microscope (FE-SEM) images of CAgNCs treated V. ichthyoenteri exhibited strong cell membrane damage than un-treated control bacteria. MTT assay results showed the highest cell viability (22%) at $75{\mu}g/mL$ of CAgNCs treated bacteria samples. The results from this study suggest that CAgNCs is a potential antibacterial agent to control fish pathogenic bacteria.

Physiological Studies on the Formation of Hairy Root by the Agrobacterium rhizogenes ; IV. Culture of Hairy Root and Survey of the Culture Condition. (Agrobacterium rhizogenes 에 의한 hairy root 형성에 대한 생리학적 연구 ; IV. Hairy root 배양 및 배양 조건에 관한 조사)

  • Hwang, Baik;An, Jun-Cheul;lee, Jae-Hyuk
    • KSBB Journal
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    • v.4 no.3
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    • pp.246-253
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    • 1989
  • Hairy roots of carrot were induced by Agrobacterium rhizogenes $A_4$ strain within about 2-4 weeks after inoculated from root disc. Early axonic culture is established in RCM agar medium and following is in MS rigid medium. After 15 days culture, the hairy roots were vigrous growth in about 10 times of initial inoculum. Anthocyanin contents of hairy roots were more than of ordinary roots. 2, 4-D ($10^{-4}mg/ l$), sucrose (5%), nitrogen source (0.03M) contained medium was optimized to growth of hairy root and contents of anthocyanin. Phenotypic alterations of leaves are observed in transformed plants and determined the transformation of hairy roots and the transformed plants by opine assay.

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Antimicrobial Activity of Myagropsis yendoi Extract (애기외톨개모자반 (Myagropsis yendoi) 추출물의 항균활성)

  • Park, Na-Bi;Song, Eu-Jin;Kim, Koth-Bong-Woo-Ri;Lee, Chung-Jo;Jung, Ji-Yeon;Kwak, Ji-Hee;Choi, Moon-Kyoung;Kim, Min-Ji;Nam, Ki-Wan;Ahn, Dong-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.6
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    • pp.642-647
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    • 2010
  • This study examined the antimicrobial activity of Myagropsis yendoi (MY). The antimicrobial activity of ethanol and aqueous MY extracts were measured using a paper disc assay. The MY ethanol extract had the best antimicrobial activity. It inhibited the growth of Candida tropicalis and Gram-positive bacteria such as Bacillus subtilis, Listeria innocua, L. monocytogenes, and Staphylococcus aureus at 4 mg/mL. The minimum inhibitory concentration (MIC) of the MY ethanol extract ranged from 0.1 to 0.5%. The MY ethanol extract inhibited the growth of B. subtilis and L. innocuaat concentrations of 0.15% and 0.1%, respectively. The antimicrobial activity of the MY ethanol extract was not affected by heating at $121^{\circ}C$ for 15 min or pH 2.10. Therefore, the main substances responsible for the antimicrobial activity of the MY ethanol extract are believed to be stable with changing heat and pH.

Antimicrobial activities of Ramaria botrytis (Fr.) against oral bacteria (싸리버섯 추출물의 구강세균에 대한 항균활성)

  • Kim, Ki-Hwa;Han, So-Ra;Kim, Byeol-Lee;Jung, Sang-Hee;Oh, Tae-Jin
    • Journal of Korean society of Dental Hygiene
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    • v.17 no.3
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    • pp.493-504
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    • 2017
  • Objectives: This study aimed to find out the antimicrobial activities of Ramaria botrytis (Fr.) extracts against oral pathogens. Methods: The antimicrobial activities of Ramaria botrytis (Fr.) extracts were evaluated against oral pathogens by the disc diffusion assay, and the minimum inhibitory concentrations (MICs) of ethyl acetate extracts were determined by broth dilution method. The strains used in this study were Staphylococcus aureus, Streptococcus mutans, Streptococcus sanguinis, Streptococcus sobrinus, Streptococcus anginosus, Streptococcus ratti, Streptococcus criceti, Actinomyces israelii, Actinomyces viscosus and Aggregatibacter actinomycetemcomitans. Results: The ethyl acetate extract of Ramaria botrytis (Fr.) effectively inhibited the growth of oral bacteria compared with acetone or ethanol extract. The ethyl acetate extract exhibited MIC values ranging from 3.75 to 15.00 mg/ml, and it showed antimicrobial activity against both Gram-positive and negative oral bacteria. Conclusions: The ethyl acetate extracts from Ramaria botrytis (Fr.) showed the antimicrobial activities against ten oral bacteria. Thus, the extract of Ramaria botrytis (Fr.) may be considered as an effective natural antimicrobial agent for the prevention of oral pathogens.