• Title/Summary/Keyword: Disk diffusion method

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Specific Detection of Serratia marcescens Based on a PCR Assay and Antimicrobial Susceptibility of S. marcescens Isolated from Boar Semen (Serratia marcescens 검출을 위한 PCR 기법 개발 및 돼지정액 유래균주에 대한 항생제 감수성 양상)

  • Jung, Ji-A;Kim, Aeran;Seo, Byoung Joo;Jung, Suk Chan;Kim, In Cheul;Chung, Ki Hwa;Jung, Byeong Yeal
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1133-1139
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    • 2013
  • During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

Characterizations of Class 1 Integrons in Proteus mirabilis Isolated from Chickens at Chungcheong Province (충청지역의 닭으로부터 분리된 Proteus mirabilis 균주에 존재하는 Class 1 Integron의 유전형 분석)

  • Sung, Ji Youn;Byeon, Yong Gwan
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.2
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    • pp.65-70
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    • 2015
  • Antimicrobial agents or additives have commonly been used in domestic animals for the prevention and treatment of bacterial diseases. Unfortunately, this has resulted in the overgrowth of bacteria that is resistant to antimicrobial agents used by humans, and these might get disseminated to humans via the food. In this study, we investigated the prevalence of integrons, and characterized gene cassette arrays in Proteus mirabilis isolates obtained from chickens in Chungcheong province of Korea. Additionally, the correlation between gene cassette arrays and antimicrobial resistance rate was studied. A total of 26 Proteus mirabilis isolates were recovered from chickens in Chungcheong province in Korea. Antimicrobial susceptibility was determined by disk diffusion method. PCR and DNA sequencing were performed to characterize the gene cassette arrays. In addition, we employed repetitive extragenic palindromic sequence-based PCR (REP-PCR) method for clonality analysis of P. mirabilis strains. Of the 26 P. mirabilis isolates tested, 14 (53.8%) isolates carried class 1 integrons, while class 2 and class 3 integrons were not detected in our study. The class 1 integrons harbored genes encoding resistance to aminoglycosides (aacCA5, aadA2, aadA5 and aadA7), trimethoprim (dfrA17, and dfrA32), lincosamides (linF) and erythromycin (ereA). In particular, the presence of class 1 integron had a significant correlatation to a high resistance rate of aminoglycoside and trimethoprim. We confirmed that class 1 integrons are widely disseminated in P. mirabilis isolates from chickens, contributing to the resistance to diverse antimicrobial agents in Korea. To prevent further spreading of antimicrobial resistant genes among P. mirabilis isolates, constant monitoring and clinical policing will become necessary.

Improved Antibacterial Effect of Blending Essential Oils (블렌딩 에센션오일의 항균효과 증진)

  • Kwon, Pil Seung;Kim, Dae-Jung;Park, Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.3
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    • pp.256-262
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    • 2017
  • Essential oil from herb is known to exert pharmacological effects on the human body. In this study we investigated the antibacterial activity of 4 essential oils (teetree, rosemary, melisa, and lavender), as well as the blended mixture oil of teetree, rosemary, and melisa (TRM) on three bacteria, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Antibacterial analysis was performed using the standard disk diffusion method, and minimum inhibition concentration was determined by the broth microdilution method with different concentrations of essential oils (0.5, 1, 2 and 3 mg/mL). After incubation at $37^{\circ}C$ for 24 h, the antibacterial activity was assessed by measuring the zone of growth inhibition surrounding the disks. Herb oil with the inhibition zones showed varied values ranging from6 to 25 mm. However, the components of herb oil of TRM are as highly active as the teetree oil against pathogens, generating large inhibition zones for both gram negative and positive bacteria (13~22 mm and 8 mm inhibition zones). In the analysis for MIC, TRM showed growth-inhibitory effects at 0.0625% for S. aureus and E. coli, and 1.25% for P. aeruginosa. This result demonstrated that the anti-microbial activity of TRM was greater than a single herb oil, including oxacillin, rosemary, and teetrea. As a single herb oil, both rosemary and teetrea also had an anti-microbial effect by itself, and we can expect that the blended oil mixture may exert a synergistic effect against multidrug resistant bacteria, suggesting its future application in natural preservative agents for health food and cosmetics.

Isolation of Salmonella from the layer chickens reacting in pullorum-typhoid agglutination test (추백리 혈청검사 양성 산란계로부터 Salmonella속균 분리)

  • 류재윤;전무형;장경수;손현수;곽학구;박경재;우용구
    • Korean Journal of Veterinary Service
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    • v.22 no.3
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    • pp.221-237
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    • 1999
  • To investigate the specificity of rapid slide agglutination test for pullorum-gallinarum diseases and to obtain a basic data for avian salmonellosis control, salmonella isolation was peformed for the layer chickens positively reacted in pullonlm-typhoid agglutination test. The biochemical, serological and antimicrobial properties of the isolates were examined. The results obtained through this study were summarized as follows; 1. Of 2,384 chickens tested by the agglutination test, 606 chickens (25.4%) were positive reactors. 154 of 606 reactors and 49 of the non-reacting chickens were investigated for salmonella isolation, resulting in isolation of 68 strains of salmonellae from 27 chickens. 2. By organs, the isolation frequency from liver, cecum, spleen, ovary and gall bladder showed 8.9% (18 strains), 8.9% (18 strains), 7.4% (15 strains), 4.4% (9 strains) and 3.9% (8 strains), respectively. 3. By culture medium the combination of selenite broth and MacConkey agar revealed the highest isolation rate and the enrichment culture by delayed secondary enrichment culture method was found the most effective for salmonella isolation. 4. The serotypes of 68 salmonella isolates were identified as 3 strains of S pullorum, 24 strains of S gallinarum, 15 strains of S typhimurium, 8 strains of S enteritidis, 7 strains of S paratyphi A, 5 strains of S typhimurium and 6 strains of the other salmonellae. 5. The serotypes of 8 salmonella strains isolated from 49 chickens non-reacting in pullorum-typhoid agglutination test were identified as 3 strains of S typhimurium and 5 strains of S infantis. 6. When 24 chickens of which 68 strains of salmonellae isolated were examined by microplate agglutination test, the average antibody titer for pullorum antigen was $2^{5.25}$. The chickens at antibody titer between $2^3$ and $2^5$ showed the higher frequency of isolation as compared with the chickens at the other titers. 7. When salmonella isolates were tested the antimicrobial drug sensitivity by disk diffusion method, S paratyphi A were highly sensitive by 100% to ATM and GM, S typhimurium, by 88% to AM, CIP, IMP and TN, S infantis, by 100% to AM, CRO, ENR and PIP, S enteritidis,by 100% to IMP and PIP, S pullorum, by 100% to ATM, CRO, ENR and PIP and S gallinarum, by 92% to CRO, CIP and PIP.

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Antimicrobial Activities of Corn Silk Extract of Klebsiella pneumoniae (옥수수수염 추출액의 Klebsiella pneumoniae에 대한 항균활성)

  • Kang, Hyun-Kyung;Bae, Il Kwon
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1399-1407
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    • 2015
  • Klebsiella pneumoniae is found in the normal flora of the skin, mouth, respiratory tract, urinary tract, and intestines in human. However, the stain is opportunistic pathogen, which is the causative agent of community acquired pneumonia. Corn silk has been known to be effective for antimicrobial activity against pathogenic bacteria, including K. pneumoniae, Staphylococcus aureus, Bacillus subtilis, Shigella spp., Salmonella spp., Escherichia coli, Pseudomonas aeruginosa, et al. In this study we focused on the antimicrobial properties of con silk water extract of K. penumoniae. K. pneumoniae isolates K. pneumoniae ATCC 13883 and broad-spectrum β-lactamase (BSBL), exteded-spectrum β-lactamase (ESBL), carbapenemase-producers. Antimicrobial susceptibilities were determined by the disk diffusion method. Searches for bla genes were performed by PCR amplication and direct sequencing. MacConkey agar plate medium was prepared using the corn silk extracts (50% or 100%) instead of distilled water for antimicrobial activity test. The microbial growth inhibitory potential of K. pneumoniae was determined by using the MacConkey agar plate spreading method, and the plate was incubated 18 hr at 37℃. Genes encoding β-lactamases including SHV-1 (n=8), SHV-2a (n=8), SHV-5 (n=2), SHV-11 (n=2), SHV-12 (n=18), TEM-1 (n=10), CTX-M-3 (n=2), CTX-M-14 (n=2), CTX-M-15 (n=1), GES-5 (n=5), KPC-2 (n=6), KPC-3 (n=4), and NDM-1 (n=2) were detected. The corn silk extract showed significantly antimicrobial activity against K. pneumoniae ATCC 13883, but BSBLs, ESBLs, and carbapenemase producers were not. Therefore, corn silk extract is thought to be able to assist in the prevention and rapid recovery of infectious disease caused by K. pneumoniae.

Antimicrobial Effects of Ethanol Extract of Yongdamgosam-hwan against Streptococcus mutans (용담고삼환(龍膽苦參丸) 에탄올 추출물의 Streptococcus mutans에 대한 항균활성에 관한 연구)

  • Yun, Yong-Il;Lee, Hae-Soo;Jung, Min-Ji;You, Seong-Il;Song, Yung-Sun;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.55-61
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    • 2015
  • Objectives : Yongdamgosam-hwan(YGH) has been used as a traditional medicine from old times for antiinflammatory effects. Streptococcus mutans(S. mutans) is known as a prime bacteria responsible for causing caries by forming a biofilm referred to as dental plaque on the tooth surface. But antimicrobial activity of YGH with dental disease is not sufficiently understood. This study was designed to investigate the effects of YGH ethanol extract on antimicrobial effect against Streptococcus mutans.Methods : The antimicrobial effect of YGH ethanol extract was assessed by the paper disk diffusion method and optical density method to determine minimum inhibition concentration(MIC), also observed by fractional inhibitory concentration index(FICI) and time-kill assay to figure out the synergic effect on the combination of YGH ethanol extract with antibiotics.Results : The YGH ethanol extract 500 μg was 7.5-8.5 mm diameter of clear zone of inhibition against Streptococcus mutans in a concentration-dependent manner and MIC was 250 μg/mL. The administration of the ethanol extract in combination with gentamicin and streptomycin induced a reduction of ≥4-8-fold in all tested bacteria. Furthermore, time-kill study was found that a combination of YGH ethanol extract with oxacillin and streptomycin produced a more rapid decrease in the concentration of bacteria CFU/mL than the YGH ethanol extract or antibiotics alone.Conclusions : As a result, the YGH ethanol extract has good antimicrobial effects. And the results suggest that YGH could be employed as a natural antibacterial agent in dental care products.

A Study of Antibiotic Resistance of Enterococci Isolated from Recycling Water of Floor Fountains in Gwangju Area (광주지역내 바닥분수 재이용수에서 분리한 장구균의 항생제 내성 연구)

  • Kim, Yeon-Hee;Song, Hyeong-myeong;Choi, Yeong-Seop;Lee, Yoon-Kook;Park, Jin-Hwan;Kim, Sun-Jung;Kim, Ha-Ram;Kang, Yu-Mi;Bae, Seok-Jin;Cho, Yong-Gwan
    • Journal of Environmental Health Sciences
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    • v.45 no.3
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    • pp.222-230
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    • 2019
  • Objectives: This study aims to evaluate water quality in terms of microorganisms and identify the antibiotic resistance of Enterococci isolated from the recycling water in floor fountains at three parks and one reservoir in the Gwangju area. Methods: Water samples were analyzed for Enterococci using membrane Enterococcus indoxyl ${\beta}$ d glucoside agar (mEI) as described in USEPA Method 1600. The vancomycin-resistant Enterococci with VanA and VanB were identified by PCR. An examination of the antibiotic resistance of isolates against 14 antibiotics was performed by the disk diffusion method. Results: The drinking water quality criterion was exceeded for total colony counts in 68% of all recycling water samples. The average concentration of total califorms and fecal coliforms was 139,325 and 413 CFU/100 mL, respectively. VanA and VanB were not detected from the isolates. We found the antibiotic resistant Enterococci strains to be E. faecalis, E. faecium, E. durans, E. mundtii, E. hirae, and E. thailandicus. The isolates were resistant to Rifampin (50%), Erythromycin (25.8%), Tetracycline (10.2%), Nitrofurantoin (8.1%), Minocycline (3.1%), Erythromycin (1.2%), Penicillin (0.7%), Norfloxacin (0.5%), and Teicoplanin (0.5%) among the 14 antibiotics tested. Antibiotic resistance tests for Enterococci from the recycling water of floor fountains resulted in 30.2% showing resistance to two or more antibiotics. Conclusions: These results showed that the multi-antibiotic resistance of Enterococci, E. coli, and others should be investigated continuously in each environment field.

Adhesion control of Campylobacter jejuni in chicken skin using emulsifiers (유화제를 이용한 계육 표면에서 Campylobacter jejuni의 부착 제어)

  • Oh, Do Geon;Kim, Kwang Yup
    • Korean Journal of Food Science and Technology
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    • v.52 no.6
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    • pp.670-677
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    • 2020
  • To prevent contamination by Campylobacter jejuni during chicken carcass processing, the effect of emulsifiers on C. jejuni inoculated on chicken skin was investigated using confocal laser scanning microscopy. Among the 8 emulsifiers (SWA-10D, L-7D, M-7D, S-1670, L-1695, P-1670, polysorbate 20, polysorbate 80) tested for antimicrobial activity by the paper disk method, 4 emulsifiers (L-7D, L-1695, polysorbate 20, polysorbate 80) were screened further. Emulsifier L-1695 showed the largest clear zone at a concentration of 200 mg/mL. The 4 emulsifiers subjected to primary screening were screened for heat and pH stability. In the contact surface test, emulsifier L-1695 showed the lowest log CFU/㎠ value on both stainless steel and ceramic surfaces. When emulsifier L-1695 was applied via general and electrostatic spray methods, the number of C. jejuni entrapped inside chicken skin follicles was significantly reduced in both methods. In conclusion, the emulsifier L-1695 could be employed as a microbial detachment agent in the chicken carcass processing industry.

Anti-microbial Activity of Grapefruit Seed Extract and Processed Sulfur Solution against Human Skin Pathogens (피부질환 원인균에 대한 자몽종자추출물과 법제유황수의 항균 효과)

  • Ha, Yu-Mi;Lee, Bo-Bae;Bae, Hee-Jung;Je, Kyoung-Mo;Kim, Soon-Rae;Choi, Jae-Suk;Choi, In-Soon
    • Journal of Life Science
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    • v.19 no.1
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    • pp.94-100
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    • 2009
  • This study was carried out to examine the antimicrobial effects of grapefruit seed extract (GSE) and processed sulfur solution (PSS) against human skin pathogens: Malassezia furfur, M. restricta, Propionibacterium arnes, Trichophyton mentagrophytes and T. rubrum. The antimicrobial effects of GSE and PSS were tested by agar diffusion method and micro broth dilution method. As the results, the MIC values of GSE against M. furfur, M. restricta, P. acnes, T. mentagrophytes and T. rubrum were 3.91, 3.91, 0.004, 0.024, and $0.012{\mu}l/ml$, respectively. The MIC values of PSS were 0.03, 0.03, 0.156, 0.003, and $0.012{\mu}l/ml$, respectively. Antimicrobial activity of skin care emulsion products containing 0.5% GSE and 0.5% PSS against human skin pathogens were 5.2, 4.3, 8.0, 9.5 and 12.8 mm, respectively. Refractive index, pH, viscosity and color value of skin care emulsions containing GSE and PSS were measured. According to these results, it was concluded that the GSE and PSS were the promising sources of antibacterial agent which could be useful for skin and hair care products as well as for the alternative medicine development in treatment of certain types of skin ailments.

Screening of Antifungal Activities of Medicinal Plants for the Control of Turfgrass Fungal Disease (잔디 병해 방제를 위한 약용식물의 항균작용 탐색)

  • Kang, Jae Young;Kim, Dae Ho;Lee, Dong Gu;Kim, In Seob;Jeon, Min Goo;Lee, Jae Deuk;Kim, Ik Hwi;Lee, Sanghyun
    • Weed & Turfgrass Science
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    • v.2 no.1
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    • pp.70-75
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    • 2013
  • Seven medicinal plant extracts were tested for antifungal activities against six species of the major turfgrass pathogenic fungi (Colletotrichum graminicola, Pythium spp., Rhizoctonia cerealis, Rhizoctonia solani AG1-1, Rhizoctonia solani AG2-2, and Sclerotinia homoeocarpa) using paper disk diffusion method. Three medicinal plant extracts, including Pinus densiflora showed antifungal activities. In suppression of mycelium growth test, on medium adding P. densiflora extract showed that inhibition rate of mycelium growth were above 80% in 10 mg/10 ml concentration of the extract. The inhibition rate of Pythium spp. was 100% and C. graminicola was 84.3% in 10 mg/10 ml concentrations of P. densiflora extract, respectively. In particularly, the inhibition rate of Pythium spp. was 89.5% in 2 mg/10 ml concentrations of P. densiflora extract. As a result, P. densiflora extract showed high antifungal activity to Pythium spp. and C. graminicola of the turfgrass pathogen in in vitro test.