• Title/Summary/Keyword: Agar diffusion assay

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Estimation of Antibacterial Properties of Chlorophyta, Rhodophyta and Haptophyta Microalgae Species

  • Imran Bashir, Khawaja Muhammad;Lee, Jae-Hyeon;Petermann, Maike Julia;Shah, Abid Ali;Jeong, Su-Jin;Kim, Moo-Sang;Park, Nam-Gyu;Cho, Man-Gi
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.225-233
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    • 2018
  • In this exploratory study, eight types of microalgae from different phyla (Chlamydomonas reinhardtii, Chlorella species, Haematococcus pluvialis, Porphyridium purpureum, Porphyridium cruentum, Isochrysis species, Isochrysis galbana, and Pavlova lutheri) were tested for their antibacterial activities against eight target pathogenic bacterial strains. The agar well diffusion method and broth micro dilution assay were conducted to estimate the antibacterial activity. Microalgae cell-free supernatants, exopolysaccharides (EPS), water, and organic solvent extracts were used for inhibition analysis. EPS extracted from P. lutheri showed activity against Bacillus subtilis and Pseudomonas aeruginosa. Inhibition zone diameters of 14-20 mm were recorded on agar plates, while the minimum inhibitory concentrations in the broth micro dilution assay were $0.39-25mg\;ml^{-1}$. During this study, haptophyte microalgae, Isochrysis species, and P. lutheri extracts showed the highest activity against most of the tested pathogenic bacterial strains, while most of the extracts were active against the important foodborne pathogen P. aeruginosa. This study showed promising results regarding important microalgae phyla, which will further aid research related to extracts and exploitation of bioactive metabolic compounds in the food and pharmaceutical industries.

Biosurfactant Production from Novel Air Isolate NITT6L: Screening, Characterization and Optimization of Media

  • Vanavil, B.;Perumalsamy, M.;Rao, A. Seshagiri
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1229-1243
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    • 2013
  • In this paper, an air isolate (NITT6L) has been screened based on hemolytic activity, emulsification activity, drop collapsing test, and oil displacement test, as well as lipase activity. It was found that strain NITT6L was able to reduce the surface tension of the medium from 61.5 to 39.83 mN/m and could form stable emulsions with tested vegetable oils. Morphological, biochemical, 16S rRNA sequencing analyses, and fatty acid methyl ester analysis using gas chromatography confirmed that the air isolate under study was Pseudomonas aeruginosa. Characterization of the biosurfactant using agar double diffusion assay revealed that the biosurfactant was anionic in nature, and CTAB-methylene blue assay and Molisch test revealed its glycolipid nature. The FT-IR spectrum confirmed that the crude biosurfactant was a rhamnolipid. Using unoptimized medium containing sucrose as the carbon source, the isolate was found to produce 0.3 mg/ml of rhamnolipid in batch cultivation (shake flask) at $37^{\circ}C$ and pH 7. Optimization of the medium components was carried out using design of experiments and the yield of rhamnolipid has been enhanced to 4.6 mg/ml in 72 h of fermentation.

Comparison of the Antibacterial Activity of Domestic Cirsium japonicum Collected from Different Regions (지역별 국내 자생 엉겅퀴 추출물의 항균 활성)

  • Jang, Miran;Park, Hyejin;Hong, Eunyeong;Kim, Gun-Hee
    • Korean journal of food and cookery science
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    • v.30 no.3
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    • pp.278-283
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    • 2014
  • This study was investigated the antibacterial activities of Cirsium japonicum from extracts five regions(Chungnam, Gyeonggi, Gangwon, Jeju and Jeonnam) extract against six food-borne pathogenes(Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Listeria monocytogenes, Salmonella enterica and Vibrio vulnificus) using the broth dilution and agar diffusion method. At concentrations between 0 and $750{\mu}g/mL$ the extracts showed an antibacterial effect against all tested bacteria. The antibacterial activities of Cirsium japonicum from Jeju and Gangwon are stronger than others. The minimum inhibitory concentration(MIC) values against the six bacteria ranged from 93.75 to $750{\mu}g/mL$. In time killing assay(cell growth curves), the tested bacteria inactivated upon exposure to the extracts for 24h. At the 24h exposure to the extracts, all bacteria were inhibited to over 70% for growth. In particular, Bacillus subtilis, Salmonella enterica and Vibrio vulnificus conveyed an inhibition of growth to almost complete. It is anticipated that Cirsium japonicum extracts may have greater potential as natural food preservatives.

Honokiol as an Effective Antimicrobial Compound against Causative Agent of American foulbrood, Paenibacillus larvae

  • Song, Hyunchan;Kim, Ki-Young
    • Journal of Apiculture
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    • v.34 no.2
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    • pp.131-136
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    • 2019
  • Recently, number of honeybees (Apis mellifera) has visibly decreased because they are vulnerable to some diseases like American foulbrood disease. American foulbrood disease, which is caused by Paenibacillus larvae, is emerged as great cause of decrease in number of honeybees. After antibiotic-resistant strain emerged, it is now more difficult to treat those pathogens successfully. Researches on finding alternative antibacterial compound are ongoing. In this study, we examined the antibacterial effect of honokiol on P. larvae. Honokiol showed great antibacterial effect with minimum inhibitory concentration of 12.5 ㎍/mL and minimum bactericidal concentration of 50 ㎍/mL. An agar diffusion test also confirmed the anti-Paenibacillus larvae activity of honokiol with an inhibitory zone of 9±0.5 mm. Since honokiol is known to interact membrane of some bacteria, we measured 260 nm absorbing particles, which could be induced by leakage of cells, and confirmed that the leakage of P. larvae occurred in dose-dependent manners. However, result of crystal violet assay suggested that honokiol has only mild anti-biofilm formation effect on P. larvae, which means honokiol controls the bacteria by inducing the bursting of membrane. Finally, an additive effect of honokiol with tetracycline and terramycin was found using a checkerboard assay with a fractional inhibitory concentration index value of 0.5.

A study on Enterotoxigenic Escherichia coli (대장균의 장내 독소 생성 균주에 관한 연구)

  • 이영남
    • Korean Journal of Microbiology
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    • v.16 no.4
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    • pp.161-169
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    • 1978
  • Escherichiae-like organisms were isolated from rectal specimens of 56 children who were either in preschool age or in elementary school. The isolated strains were subjected to tests to screen enteropathogens producing heat-labile enterotoxin and susceptibility test to various antibiotics by disc diffusion method on agar plates. Production of heat-labile enterotoxin by the strains was assyed in the sensitive and reproducible cultured adrenal tumor cell system. The assay was sterodogenesis of the cell in the presence of heat-labile enterotoxin. Among 56 strains, gave positive reaction in the test of toxin production. This meant that about 10% of the children population objected to the study harbored the toxigenic strain of enteropathogenes. Some of these toxigenic strains were resistant to the antibiotics employed in the test. This study suggested that considerable population in Korea may harbor entertoxigenic E. coli as a part of intestinal normal flora. The toxigenic strains which are resistant to antibiotics may bring issue of public health in the future.

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Grape Seed Extract Protects Mice against Disseminated Candidiasis

  • Han, Yong-Moon
    • Natural Product Sciences
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    • v.9 no.2
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    • pp.60-63
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    • 2003
  • Effect of grape seed extract (GSE) against Candida albicans was examined under in-vitro and in-vivo conditions. The GSE was extracted in ethanol. In-vitro results from an agar diffusion susceptibility assay showed the GSE inhibited C. albicans growth. This anticandidal effect was at dose-dependency. In experiments with animals, mice that received the GSE (0.5 mg per mice), intravenously (i.v.), before i.v.-infection wish viable C. albicans yeast cells survived longer than diluent (buffer)-received control mice. In contrast, when GSE was given to mice after the mice were infected with the yeast cells, these mice showed a similar survival rate as compared to control mice that received no treatment with the GSE. Taken together, these data indicate that GSE has prophylactic effect but not therapeutic effect against disseminated candidiasis.

Antifungal Activity of Borneolum (Borneo-Camphor) from Dryobalanops aromatica against Malassezia furfur (Dryobalanops aromatica 유래 용뇌의 Malassezia furfur에 대한 항균 활성)

  • Kim Young-ju;Hwang Guen-bae;Seu Young-Bae
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.236-239
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    • 2005
  • Ethyl acetate extracts of traditional medicinal herbs were screened for their antifungal activity against Malassezia furfur through the liquid growth inhibition method with 96 well plate and the paper disk agar diffusion assay. Among the 91 kinds of herbal solvent extracts, borneolum (borneo-camphor, No. 56) was finally selected as the best antifungal effective medicinal herb. The MIC value of No. 56 against M. furfur was 0.3 mg/ml ($8.2{\times}10^6$ CFU/ml) and it was three times more efficient than midazole which is the known antifungal agent in clinical medicines.

In Vitro Effects of Essential Oils from Ostericum koreanum against Antibiotic-Resistant Salmonella spp

  • Shin, Seung-Won
    • Archives of Pharmacal Research
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    • v.28 no.7
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    • pp.765-769
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    • 2005
  • The essential oil fraction of Ostericum koreanum was analyzed by GC-MS. Inhibiting activities of this oil and its main components were tested by the broth dilution assay and disk diffusion test against one antibiotic-susceptible and two resistant strains of Salmonella enteritidis and S. typhimurium, respectively. The GC-MS analysis revealed thirty-four compounds; the main components were $\alpha$-pinene (41.12%), $\rho$-cresol (17.99%) and 4-methylacetophenone (7.90%). The essential oil of O. koreanum and its main components were significantly effective against the tested antibiotic-susceptible strains as well as against the resistant strains of the two Salmonella species, with MICs (minimum inhibitory concentrations) ranging from 2 mg/mL to 16 mg/mL. The anti-Salmonella effects of the oils were dose-dependent on $M\"{u}ller-Hinton$ agar plates in this experiment. Additionally, checkerboard titer test results demonstrated significant combined effects of streptomycin and O. koreanum oil or cresol, one of the main components of this oil, against the two streptomycin resistant strains of S. typhimurium, with FICIs ranging from 0.12 to 0.37.

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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Antibacterial Activities of Honeys on the Staphylococcus aureus (Staphylococcus aureus에 대한 벌꿀의 항균 활성)

  • 백승화;정동현
    • The Korean Journal of Food And Nutrition
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    • v.15 no.2
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    • pp.158-164
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    • 2002
  • This study examined for the investigation the effect of honey on antibacterial activity. The experimental honey were used the domestics, or chestnut honey, multiflower honey, acassia honey, native honey and the foreign, or manuka honey, clover honey, canola honey, and the artificial honey, made with the diluted solution of each 12.5%, 25.0%, 50.0%. The result of compared the occasion of added-catalase with not added-catalase about the honey's antibacterial activity on Staphylococcus aureus by agar well diffusion assay were as follows. When the catalase was not added, manuka honey antibacterial activity was superior to chestnut honey's in the diluted honey of 12.5% and on the occasion of the diluted honey of 25.0%, it was approved in the order of manuka honey > chestnut honey > multiflower honey 〉 native honey > clover honey > acassia honey and the occasion of the diluted honey of 50.0%, it was approved in the order of manuka honey > chestnut honey > canola honey > native honey > multiflower honey > clover honey > acassia honey(p > 0.01). The clear zone representing inhibition of growth in diluted honey of 12.5, 25.0, 50.0 % with non-treat catalase ranged from 5.85 to 6.60, 4.26 to 8.27, 5.24 to 11.49 mm, respectively. When the catalase was added, antibacterial activity only showed in the manuka honey of 12.5% and on the occasion of the diluted honey of 25.0%, manuka honey's antibacterial activity was superior to chestnut honey (p > 0.01). On the occasion of the diluted honey of 50.0%, antibacterial activity was high in the order of manuka honey > chestnut honey > clover honey > canola honey > native honey(p > 0.01). The correlation was approved significantly among the manuka honey, chestnut honey, clover honey, canola honey and native honey.