• Title/Summary/Keyword: Agar dilution

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Cytochrome b Gene-Based Assay for Monitoring the Resistance of Colletotrichum spp. to Pyraclostrobin

  • Dalha Abdulkadir, Isa;Heung Tae, Kim
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.616-628
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    • 2022
  • Resistance to pyraclostrobin due to a single nucleotide polymorphism at 143rd amino acid position on the cytochrome b gene has been a major source of concern in red pepper field infected by anthracnose in Korea. Therefore, this study investigated the response of 24 isolates of C. acutatum and C. gloeosporioides isolated from anthracnose infected red pepper fruits using agar dilution method and other molecular techniques such as cytochrome b gene sequencing, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and allele-specific polymerase chain reaction (PCR). The result showed that four isolates were resistant to pyraclostrobin on agar dilution method and possessed GCT (alanine) codon at 143rd amino acid position, whereas the sensitive isolates possessed GGT (glycine). Furthermore, this study illustrated the difference in the cytochrome b gene structure of C. acutatum and C. gloeosporioides. The use of cDNA in this study suggested that the primer Cacytb-P2 can amplify the cytochrome b gene of both C. acutatum and C. gloeosporioides despite the presence of various introns in the cytochrome b gene structure of C. gloeosporioides. The use of allele-specific PCR and PCR-RFLP provided clear difference between the resistant and sensitive isolates. The application of molecular technique in the evaluation of the resistance status of anthracnose pathogen in red pepper provided rapid, reliable, and accurate results that can be helpful in the early adoption of fungicide-resistant management strategies for the strobilurins in the field.

Antimicrobial Resistance of Seventy Lactic Acid Bacteria Isolated from Commercial Probiotics in Korea

  • Eunju Shin;Jennifer Jaemin Paek;Yeonhee Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.500-510
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    • 2023
  • In this study, lactic acid bacteria were isolated from 21 top-selling probiotic products on Korean market and their antimicrobial resistance were analyzed. A total 152 strains were claimed to be contained in these products and 70 isolates belonging to three genera (Bifidobacterium, Lactobacillus, and Lactococcus) were obtained from these products. RAPD-PCR showed diversity among isolates of the same species except for two isolates of Lacticaibacillus rhamnosus from two different products. The agar dilution method and the broth dilution method produced different MICs for several antimicrobials. With the agar dilution method, five isolates (three isolates of Bifidobacterium animalis subsp. lactis, one isolate of B. breve, one isolate of B. longum) were susceptible to all nine antimicrobials and 15 isolates were multi-drug resistant. With the broth microdilution method, only two isolates (one isolate of B. breve and one isolate of B. longum) were susceptible while 16 isolates were multi-drug resistant. In this study, only two AMR genes were detected: 1) lnu(A) in one isolate of clindamycin-susceptible and lincomycin-resistant Limosilactobacillus reuteri; and 2) tet(W) in one tetracycline-susceptible isolate of B. longum B1-1 and two tetracycline-susceptible isolates and three tetracycline resistant isolates of B. animalis subsp. lactis. Transfer of these two genes via conjugation with a filter mating technique was not observed. These results suggest a need to monitor antimicrobial resistance in newly registered probiotics as well as probiotics with a long history of use.

Antimutagenic and Antibacterial Activities of Korean and American Propolis (한국산과 미국산 프로폴리스의 항돌연변이 및 항균효과)

  • Jang, Il-Woong;Park, Jeong-Seob;Kwon, Hyoung-Cheol;Jung, Mun-Yhung;Choi, Dong-Seong
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.694-699
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    • 2009
  • The antimutagenic activities of ethanol extracts of Korean and American propolis were tested using Salmonella Typhimurium TA98 with two indirect mutagens of 3-amino-1,4-dimethyl-5H-pyrido [4,3-b]indole (Trp-P-1) and 2-aminoanthracene (2-AA) with S9 mix. Additionally, their antimicrobial activities against acne-related pathogenic strains of Propionibacterium acnes, Staphylococcus Epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa were evaluated using both paper disk method and agar dilution method. Ethanol extracts of Korean and American propolis showed strong inhibitory effects, in a dose dependant manner, against the mutagenicities induced by Trp-P-1 and 2-AA. The antimutagenic effect of ethanol extracts of Korean propolis showed significantly higher protective activity than that of American propolis against the Trp-P-1 induced mutagenicity of S. Typhimurium TA98 at the lower concentration ($1-10\;{\mu}g$), but significantly lower protective activity at the higher concentration ($50-200\;{\mu}g$). The antimutagenic effect of ethanol extract of Korean propolis showed significantly higher protective activity than that of American propolis against the 2-AA induced mutagenicity at the concentration of $1\;{\mu}g$, but significantly lower protective activity than that of the American at the higher concentration ($50-200\;{\mu}g$). Both extracts showed strong antimicrobial activities against all the acne-related pathogens tested, with minimal inhibitory concentration (MIC) values in the range $1,500-5,000\;{\mu}g/mL$.

Effect of hydrothermal extract from Codonopsis lanceolata Benth. et Hook on the growth of lactic acid bacteria and harmful bacteria

  • Kim, Se-young;Lee, Jinsil;Geong, Ga-Jin;Yi, Seh-Yoon
    • Proceedings of the Korean Society of Food and Cookery Science Conference
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    • 2003.10a
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    • pp.75-75
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    • 2003
  • The objective of this study is to investigate the effect of Codonopsis lanceolata Bentk et Hook on the growth of lactic acid bacteria and some harmful bacteria. Extract of C. lanceolata was obtained by hydrothermal extraction method and used intactly without concentration. To investigate the inhibitory effect on the growth of harmful bacteria, we used agar dilution method. On agar plate containing more than 25% extract, they showed decreased number of colonies. (omitted)

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Changes in the Oral Microflora in Patients with Acute Myeloid Leukemia during the Period of Induction Therapy (항암 화학요법중인 급성 골수성 백혈병 환자의 구강내 세균변화에 관한 연구)

  • Byul-Hee Lee;Chong-Youl Kim
    • Journal of Oral Medicine and Pain
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    • v.18 no.1
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    • pp.73-82
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    • 1993
  • To investigate the changes in aerobic and facultative anaerobic oral microflora during remission-induction chemotherapy in patients with acute myeloid leukemia, 10 consecutive patients were studied during a period of 28 days. One day before, during and after the induction therapy, patients were given 10% Betadine solution for mouthrinses after breakfast and kept from eating and drinking. After 3 hours, paraffin-stimulated whole saliva was obtained for 2 minutes and transported to the laboratory. The samples were dispersed and homogenized by use of vortex mixer for 20 seconds. From these samples 10-fold serial dilutions (from 10-1 through 10-3) were prepared. Each dilution of 0.1 ml was plated on duplicate set of one nonselective medium (Blood agar) and four selective media (Sabourauds dextrose agar, Mannitol salt agar, Mac-Conkey agar, SF medium ) using applicator woods. All agar plate were incubated at 37$^{\circ}C$ for 48 hours. The total number of microorganisms was calculated and the percentage distribution of the various microorganisms from each specimen was drawn. 1. The salivary flow rate decreased by 66%, going from 5.38 ml/2min to 1.81 ml/2min over two days during the chemotherapy. 2. The total number of microorganisms in saliva increased by 22%, going from 4.88$\times$105/ml to 6.00$\times$105/ml over two days during the chemotherapy. 3. The salivary flow rate and the total number of microorganisms in saliva were recovered within 28 days after the chemotherapy. 4. The quantitative alteration in oral Enterobacteria, Enterococci, Staphylococci, Cndida during the chemotherapy had no statistical significance. 5. In saliva of the patients with acute myeloid leukemia who ahd intraoral ulcer, Enterobacteria was quantitatively predominent. Our study suggests that chemotherapy-induced transient xerostomia may induce acute oral infection. Consequently, the use of saliva substitute, the removal of intraoral infection source and the consistent oral hygiene care seem to be required to avoid the transmission of potential pathogenes in this group of patients.

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Continuous Production of Agarooligosaccharides Using Packed-Bed Reactor (Packed-Bed 반응기를 이용한 한천올리고당의 연속생산)

  • 임동중;김종덕;강양순;공재열
    • KSBB Journal
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    • v.16 no.4
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    • pp.398-402
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    • 2001
  • Enzymatic hydrolysis of agar was carried out continuously to produce agarooligosaccharides by immobilized agarase in Packed-Bed Reactor. The reactor was constructed using a acryl tube with an internal diameter of 10 mm and a useful height of 140 mm. The Packed-Bed Reactor was 11 mL reactor volume as its length : diameter ratio was 14 : 1. The operation condition of reaction was performed with an 1 g/L agar concentration at 40$^{\circ}C$, 10 mM MOPS buffer(pH 7.0) and with the flow rate 3 mL∼48 mL/h at a dilution rate of 1.09∼5.45 h$\^$-1/. The hydrolysis products was identified DP6, DP4 and DP2 by HPLC. The conversion rate of agar was about 80% and amount of total agarooligosaccharide was 0.88 mg/mL at Packed-Bed Reactor.

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Antimicrobial Effects of Lonicera japonica against Gram Positive and Gram Negative Anaerobic Bacteria

  • Rhee, Ki-Hyeong;Lee, Keyong-Ho
    • Natural Product Sciences
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    • v.17 no.1
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    • pp.23-25
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    • 2011
  • It has been shown that the butanol extract of Lonicera japonica has antimicrobial and other potentially useful biological activities. The purpose of this study was to determine the in vitro activity of Lonicera japonica compared to other antimicrobial agents against anaerobic bacteria. Specifically, the in vitro activity of the butanol extract was investigated against 104 clinical isolates of anaerobic bacteria using an agar dilution method and the results were compared to erythromycin, cefoxitin, imipenem, clindamycin, and metronidazole. It was found that Lonicera japonica and imipenem were the most active antimicrobial agents tested.

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.

Comparative Study Using MIC Test for Cefazedone and other First Generation Cephalosporins (1세대 세파계 항생제인 Cefazedone의 MIC Test를 통한 비교 연구)

  • Bok, Hae-Sook;Kim, Myung-Min;Choi, Kyung-Eob
    • Korean Journal of Clinical Pharmacy
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    • v.5 no.2
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    • pp.13-16
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    • 1995
  • The Minimum Inhibitory Concentration (MIC) of a first-generation cephalosporin derivative, Cefazedone (CZD; $PAZERON^R$ inj.) was determined by the two-fold serial agar dilution method. The in-vitro antibacterial activity of CZD against a wide variety of clinical isolates was compared with those of other first generation cephalosporins such as Methylol Cephalexin (CEX), Cefazolin (CEZ), Cefadroxil (CDX), Cephradine (CED), Ceftezol (CTZ) and one of second generation cephalsporin antibiotics, Cefotaxime (CTX). CZD had the most potent inhibitory effect against Gram-positive strains, when compared to the first-generation cephalosporin antibiotics tested in this study and CTX. The geometric MIC mean of CZD for Gram-positive strains was calculated as 0.386 kg/m{\ell}$, and those of CEX, CEZ, CDX, CTZ, CED, and CTX were 6.073, 0.894, 3.399, 0.748, 7.884 and 1.502 $kg/m{\ell}$, respectively. In addition, the geometric mean of CZD for staphylococclJs aureus strains was obtained as 0.340 $kg/m{\ell}$ and those of CEX, CEZ, CDX, CTZ, CED, and CTX 6.145, 0.534, 4.126, 0.442, 10.51, and 2.500 $kg/m{\ell}$, respectively. Against Gram-negative strains, CZD showed better antibacterial activity than CEZ, CDX, CTZ, and CED.

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Drug susceptibility of bacteria and M pachydermatis isolated from canine external ear canals (개 외이도에서 분리한 세균 및 M pachydermatis의 약제감수성에 대하여)

  • Kim, Ki-hyang;Choi, Won-phil;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.560-565
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    • 1999
  • The present work was conducted to investigate the drug susceptibility of microorganisms isolated from canine external ear canals. Antifungal susceptibility test of M pachydermatis (17 strains) was perfomed by agar dilution method, using 11 antifungal drugs including amphotericin B(A), nystatin(N), pimaricin(P), griseofulvin(G), bifonazole(B), clotrimazole(C), miconazole(M), econazole(E), ketoconazole(K), tolnaftate(T), 5-fluorocytosine(F). All isolates were highly sensitive to K, M, T(geometric mean MIC ; GM $MIC{\leq}0.16{\mu}g/ml$) but they weren't sensitive to P, F and G(GM $MIC{\geq}92.37{\mu}g/ml{\sim}{\geq}128{\mu}g/ml$). Antibacterial susceptibility test against 119 isolates of bacteria was performed by agar dilution method, using 9 antibacterial drugs including erythromycin(ET), chloramphenicol(CP), gentamycin(G), vancomycin(V), ampicillin(AP), amoxacillin(AX), chlortetracycline(CT), ciprofloxacin(CF), enrofloxacin(EF). All isolates of Staphylococcus spp(101 strains) were highly sensitive to EF, CF, G(GM MIC $0.33{\sim}1.47{\mu}g/ml$). In other gram positive cocci(4 strains), they were highly sensitive to EF, CF, V(GM MIC $1{\sim}4.76{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). In gram positive rods(13 strains), they were highly sensitive to EF, CF, G(GM $MIC{\leq}0.19{\sim}1{\mu}g/ml$). In Pseudomonas aeruginosa(1 strain), it was highly sensitive to AX, EF, ET, CF(GM MIC $0.06{\sim}1{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). All isolates weren't sensitive to AP(GM MIC 16~>$32{\mu}g/ml$).

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