• Title/Summary/Keyword: Disc diffusion method

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A monitoring survey on antimicrobial resistance of bacterial isolates from companion dogs in Incheon (인천지역 반려견 유래 세균의 항생제 내성 모니터링 조사)

  • Kim, Kyung-Mi;Cho, Min-Haneng;Lee, Seung-Hwan;Kim, Kyung-Ho;Lee, Jung-Gu;Lee, Sung-Mo
    • Korean Journal of Veterinary Service
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    • v.42 no.2
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    • pp.53-60
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    • 2019
  • This study was performed to investigate antimicrobial resistance in bacterial isolates obtained from companion dogs in veterinary hospitals and an animal shelter in Incheon. Drug resistance was examined respectively with the isolates of Escherichia coli, Enterococcus faecalis, and Staphylococcus pseudintermedius. The prevalence of drug resistance was calculated for each bacterial species towards 163 E. coli isolates, 156 E. faecalis isolates, and 86 S. pseudintermedius isolates by using selected antimicrobials. E. coli isolates were highly resistant to ampicillin, ciprofloxacin and tetracycline (47.9%, 28.2% and 28.2%, respectively). E. faecalis isolates were highly resistant to quinupristin-dalfopristin, tetracycline, kanamycin, rifampicin (69.8%, 66.0%, 53.8% and 51.9%, respectively). Higher levels of resistance were detected for ampicillin, penicillin, tetracycline, erythromycin, trimethoprim/sulfamethoxazole, telithromycin in S. pseudintermedius isolates (83.7%~52.6%, respectively). Occurrence of methicillin-resistant S. pseudintermedius (MRSP) was confirmed by oxacillin disc diffusion method, resulted in 23.3% occurrence among the S. pseudintermedius isolates (20/86 strains). The occurrence ratio of multidrug-resistance in the isolates of E. coli, E. faecalis, and S. pseudintermedius was 34.5%, 56.9%, and 67.9%, respectively. In this study, higher levels of antimicrobial drug resistance were observed in bacterial isolates obtained from dogs in Incheon. A regular monitoring and surveillance program should be implemented to prevent the emergence and spread of the drug-resistant bacteria carried in companion dogs.

Determination of Semen Quality and Antibacterial Susceptibility Pattern of Bacteria Isolated from Semen of Iraqi Subjects

  • Faisal, Anwer Jaber;Salman, Hamzah Abdulrahman
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.587-593
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    • 2021
  • Infertility is a key issue affecting mood and behavior in men. Microorganisms are one of the primary etiological agents that may be associated with infertility. The objective of the present study was to identify bacterial causative agents from the semen of infertile subjects and determine the effect of bacterial infection on sperm quality, as well as determine the susceptibility of these bacteria to drugs. Forty semen samples from 30 infertile patients and 10 fertile individuals were collected. The pH, volume, motility, and concentration of semen were analyzed. The samples were processed and identified by biochemical testing using API identification kits. The antibiotic susceptibility pattern was determined using the disc diffusion method. Abnormal sperm quality was observed. The mean age of the individual and their sperm morphology, concentration, progressive motility, pH level, and pus cell content were 31.9 years, 2.7%, 10.4 million/ml, 27.3%, 8.3, and 5.7, respectively. Among the tested samples, oligoasthenozoospermia was found to show the highest occurrence, at 27/30 samples, followed by teratozoospermia, at 25/30 samples, and asthenozoospermia, at 22/30 samples. Of the tested infertile patients' sperm, 19, 6, and 5 isolates were identified as Escherichia coli, Klebsiella pneumonia, and Staphylococcus epidermidis, respectively. The results also revealed multi-drug resistance in the bacteria. Compared to that shown by the other tested antibiotics, amikacin showed higher activity against all isolated bacteria. However, the bacteria exhibited maximum resistance against gentamicin, cefotaxime, levofloxacin, and ampicillin. In conclusion, leukocytospermia and bacterial infections are possibly responsible for sperm abnormalities. Multi-drug resistant bacteria were detected. Gentamicin, cefotaxime, levofloxacin and ampicillin were shown the highest resistance, while amikacin was the most effective antimicrobial agent against the isolated bacteria.

Antimicrobial resistance in fecal Escherichia coli from different pig production systems

  • Mitchaothai, Jamlong;Srikijkasemwat, Kanokrat
    • Animal Bioscience
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    • v.35 no.1
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    • pp.138-146
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    • 2022
  • Objective: The objective of the current study was to investigate the influences of conventional (CO) and deep litter (DE) systems on antimicrobial resistance in fecal Escherichia coli (E. coli). Methods: A cross-sectional study was carried out to detect antimicrobial resistance to E. coli in swine fecal samples in CO and DE systems located in western and northeastern Thailand. Individual rectal swab samples were taken only from healthy pigs. A total of 215 individual and healthy pigs were randomly selected for isolation and antimicrobial susceptibility test of E. coli by the disc diffusion method. The test panel included amoxicillin (AMX), colistin, doxycycline (DOX), enrofloxacin, gentamicin (GEN), kanamycin, neomycin (NEO), and trimethoprim-sulfamethoxazole (SXT). Results: There were significant (p<0.05) lower resistance levels for GEN, NEO, and SXT in the DE farms compared to those in the CO farms. There was a lower number of antimicrobial resistance agents (p<0.001) in the DE farms compared to those in the CO farms. This result was consistent with those in western (p<0.01) and northeastern (p<0.01) Thailand. Overall, antibiograms of AMX-SXT and AMX-DOX-SXT were found in the CO (19.09% and 20.91%, respectively) and the DE (16.19% and 24.76%, respectively) farms. No antimicrobial resistance (5.71%) was found and AMX (13.33%) resistant pigs in the DE farms, whereas the pattern of AMX-GEN-SXT (6.36%) and AMX-DOX-GEN-SXT (11.82%) resistant pigs was found in the CO farms. Conclusion: The DE system for pig farming was superior to conventional pig farming by lowering the resistance level of fecal E. coli to GEN, NEO, and SXT, with decreasing the number of antimicrobial resistance agents and inducing a small proportion of pigs to be free from antimicrobial resistance.

Antibacterial activity of Bio-fermented Galla Rhios Extract (오배자 발효추출물의 항세균활성)

  • Doh, Eun Soo;Yoo, Ji Hyun
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.21-27
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    • 2014
  • Objectives : This experimental study was performed in order to investigate the antibacterial effect of bio-fermented Galla Rhois extract. Methods : The Galla Rhois extract was fermented by Streptococcus thermophilus, Saccharomyces cerevisiae and Lactobacillus delbrueckii, and their products was tested for antibacterial activity against six pathogenic microorganisms namely, Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Vibrio parahaemolyticus, Escherichia coli and Salmonella typhimurium by paper disc diffusion method. Results : The Galla Rhois fermented extract by Lactobacillus delbrueckii and Saccharomyces cerevisiae showed more effective antibacterial activity than not fermented extract against Bacillus subtilis and Vibrio parahaemolyticus. Antibacterial activity of fermented extract using especially Lactobacillus delbrueckii and Saccharomyces cerevisiae was proved that it was good with even 2 percents concentration. Antibacterial activity of Galla Rhois extract within pH 3 to pH 7 had been safe regardless of pH but low over pH 9. The growth of Bacillus cereus, Staphylococcus aureus, and Vibrio parahaemolyticus had a tendency to decrease depend on the increasing concentration of the extract. EtOEt, EtOAc and n-BuOH fractions of the Galla Rhois extract had a high level of antibacterial activity against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus and Vibrio parahaemolyticus, respectively. Surprisingly, EtOAc fractions of the Galla Rhois extract showed higher antibacterial activity against Vibrio parahaemolyticus alone. And antibacterial activity against six pathogenic microorganisms had a tendency to increase depend on the increasing concentration of the fractions of the Galla Rhois extract. Conclusions : Bio-fermented Galla Rhois extract, efficiently inhibited the growth of Bacillus cereus and Vibrio parahaemolyticus.

Antibacterial Activities of Eighteen Types of Essential Oils on S. mutans, P. gingvalis, and L. gasseri (에센셜 오일 18종의 S. mutans, P. gingivalis, L. gasseri에 대한 항균능 분석)

  • Yoon, Hyun-Seo;Park, Chung-Mu
    • Journal of The Korean Society of Integrative Medicine
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    • v.10 no.4
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    • pp.103-111
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    • 2022
  • Purpose : In recent years, essential oils have been produced using natural extracts for various uses. Their functionality is currently being tested not only for cosmetics and perfumes but also for other categories of products. Therefore, this study verified their antibacterial effects on S. mutans, P. gingivalis, and L. gasseri which are the representative strains that cause oral diseases. Methods : Eighteen types of natural essential oils were made at a concentration of 50 % (v/v) using Tween 20, and their antibacterial effects were verified by applying S. mutans, P. gingivalis, and L. gasseri. The antibacterial effects were measured with the disc diffusion method. All the experiments were repeated three times, and the mean value of three measurement values for each variable was used for data analysis. A one-way analysis of variance was conducted using these mean values. Results : Of the eighteen types of essential oils tested, sixteen types showed antibacterial effects on S. mutans, and sixteen and fifteen types exhibited antibacterial effects on P. gingivalis and L. gasseri respectively. The types of essential oils with high-level antibacterial activities were geranium, may chang, and bergamot for S. mutans, lemongrass, bergamot, and eucalyptus for P. gingivalis, and lemongrass, machan, and geranium for L. gasseri in order of antibacterial effect. This result was statistically significant (p<.001). In addition, in the case of mandarin oil, it was found that there was no antibacterial activity in all three strains. Conclusion : This study proved the antibacterial activities of essential oils, which are used for various purposes in daily life, against dental caries and periodontal diseases. The study results will likely be applied to different prevention programs for oral health and broadly used to develop products such as oral care items and dentifrices.

Production and Identification of Secondary Metabolite Gliotoxin-Like Substance Using Clinical Isolates of Candida spp.

  • Noorulhuda Ojaimi Mahdi, Al-Dahlaki;Safaa Al-Deen Ahmed Shanter, Al-Qaysi
    • Microbiology and Biotechnology Letters
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    • v.50 no.4
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    • pp.488-500
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    • 2022
  • Most fungal infections by opportunistic yeast pathogens such as Candida spp. are the major causes of morbidity and mortality in patients with lowered immune. Previous studies have reported that some strains of Candida secret secondary metabolites play an important role in the decreasing of immunity in the infected patient. In this study, 110 Candida spp. were isolated from different clinical specimens from Baghdad hospitals. Candida isolates were identified by conventional methods, they were processed for Candida speciation on CHROMagar. The results of identification were confirmed by internal transcribed spacer (ITS) sequencing. Phylogenetic trees were analyzed with reference strains deposited in GenBank. Antifungal susceptibility testing was evaluated by the disc diffusion method and performed as recommended by the Clinical and Laboratory Standard Institute (CLSI) M44-A document. Candida isolates investigated produce secondary metabolites gliotoxin with HPLC technique and quantification. Out of 110 Candida isolates, C. albicans (66.36%) was the most frequent isolate, followed by the isolates of C. tropicalis (10.9%) and C. glabrata (6.36%) respectively. Concerning the antifungal susceptibility test, Candida isolates showed a high level of susceptibility to Miconazole (70.9%), Itraconazole (68.2%), and Nystatine (64.5%). The ability of obtained isolates of Candida spp. to produce gliotoxin on RPMI medium was investigated, only 28 isolates had the ability to secret this toxin in culture filtrates. The highest concentrations were detected in C. albicans (1.048 ㎍/ml). Gliotoxin productivity of other Candida species was significantly lower. The retention time for gliotoxin was approximately 5.08 min.

Virulence gene profiles and antimicrobial susceptibility of Salmonella Brancaster from chicken

  • Evie Khoo ;Roseliza Roslee ;Zunita Zakaria;Nur Indah Ahmad
    • Journal of Veterinary Science
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    • v.24 no.6
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    • pp.82.1-82.12
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    • 2023
  • Background: The current conventional serotyping based on antigen-antisera agglutination could not provide a better understanding of the potential pathogenicity of Salmonella enterica subsp. enterica serovar Brancaster. Surveillance data from Malaysian poultry farms indicated an increase in its presence over the years. Objective: This study aims to investigate the virulence determinants and antimicrobial resistance in S. Brancaster isolated from chickens in Malaysia. Methods: One hundred strains of archived S. Brancaster isolated from chicken cloacal swabs and raw chicken meat from 2017 to 2022 were studied. Two sets of multiplex polymerase chain reaction (PCR) were conducted to identify eight virulence genes associated with pathogenicity in Salmonella (invasion protein gene [invA], Salmonella invasion protein gene [sipB], Salmonella-induced filament gene [sifA], cytolethal-distending toxin B gene [cdtB], Salmonella iron transporter gene [sitC], Salmonella pathogenicity islands gene [spiA], Salmonella plasmid virulence gene [spvB], and inositol phosphate phosphatase gene [sopB]). Antimicrobial susceptibility assessment was conducted by disc diffusion method on nine selected antibiotics for the S. Brancaster isolates. S. Brancaster, with the phenotypic ACSSuT-resistance pattern (ampicillin, chloramphenicol, streptomycin, sulphonamides, and tetracycline), was subjected to PCR to detect the corresponding resistance gene(s). Results: Virulence genes detected in S. Brancaster in this study were invA, sitC, spiA, sipB, sopB, sifA, cdtB, and spvB. A total of 36 antibiogram patterns of S. Brancaster with a high level of multidrug resistance were observed, with ampicillin exhibiting the highest resistance. Over a third of the isolates displayed ACSSuT-resistance, and seven resistance genes (β-lactamase temoneira [blaTEM], florfenicol/chloramphenicol resistance gene [floR], streptomycin resistance gene [strA], aminoglycoside nucleotidyltransferase gene [ant(3")-Ia], sulfonamides resistance gene [sul-1, sul-2], and tetracycline resistance gene [tetA]) were detected. Conclusion: Multidrug-resistant S. Brancaster from chickens harbored an array of virulence-associated genes similar to other clinically significant and invasive non-typhoidal Salmonella serovars, placing it as another significant foodborne zoonosis.

Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Antimicrobial Synergistic Effects of Gallnut Extract and Natural Product Mixture against Human Skin Pathogens (피부 병원성균에 대한 오배자 천연 복합물의 시너지 항균 효과)

  • Kim, Ju Hee;Choi, Yun Sun;Kim, Wang Bae;Park, Jin Oh;Im, Dong Joong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.2
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    • pp.155-161
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    • 2021
  • This study was attempted to investigate natural materials with antimicrobial activity and to apply as natural preservatives in cosmetics. The disc diffusion method was used to search for nine species of natural antibacterial material for three species of skin pathogenic bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa) and Candida albicans. As a result of measuring the size of inhibition zone, Rhus Semialata gall (Gallnut) extract, Oak vinegar, and ε-polylysine were showed strongest antibacterial activities (> 10 mm). The Minimum Bactericidal Concentration (MBC) of gallnut and oak vinegar ranged from 10 to 20 mg/mL and from 20 to 40 mg/mL against five human skin pathogens. The MBC of ε-polylysine ranged from 0.5 to 2 mg/mL in fungus. The synergic effect of gallnut extract/oak vinegar mixture and gallnut extract/ε-polylysine mixture were evaluated by checkerboard test. Compared to when used alone, the MBC of gallnut extract/oak vinegar mixture were at 4 times lower concentration against E. coli, C. albicans, and A. brasiliensis. Also Furthermore, the MBC of gallnut extract/ε-polylysine mixture were at 4 times lower concentration against C. albicans and A. brasiliensis. It was confirmed that the combination of gallnut extract with oak vinegar or ε-polylysine resulted in synergistic antibacterial effect against three human skin pathogens. Thus, it is expected that gallnut extract and natural product mixture can not only demonstrate antibacterial synergies, but also be applied in cosmetics as a natural preservative system with a wide antibacterial spectrum.

Effects of Artemisia capillaris Extracts on Intestinal Microflora In vitro and In vivo (In vitro 및 In vivo에서 인진쑥 추출물이 장내미생물에 미치는 영향)

  • Oh, Mi-Hyun;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1587-1594
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    • 2010
  • This study was designed to investigate the effect of Artemisia capillaris extracts on the intestinal microflora. In agar diffusion method, the solvent fractions of Artemisia capillaris showed growth inhibition against the intestinal microflora. In particular, the chloroform fraction of Artemisia capillaris had strong antibacterial activity against Clostridium perfringens, Clostridium difficile, Eubacterium limosum, and Bacteroides fragilis, but did not show antibacterial activity against Bifidobacterium bifidum and Lactobacillus acidophilus. Most chloroform fraction of Artemisia capillaris inhibitory activities were not reduced by heat treatment or pH variation against C. perfringens, C. difficile, E. limosum, and B. fragilis. MICs of the chloroform fraction were 1.25 mg/mL against C. perfringens, E. limosum and B. fragilis and 2.5 mg/mL against C. difficile. MBCs of chloroform fraction were 5 mg/mL against C. perfringens, E. limosum and 2.5 mg/mL against C. difficile, B. fragilis. The ethyl acetate fraction of Artemisia capillaris showed $3.08{\pm}0.03$ mg/10 mg total polyphenol and $1.91{\pm}0.03$ mg/10 mg total flavonoid contents. In vivo tests were performed to investigate the influence of Artemisia capillaris extract on the intestinal microflora in rats. The results showed the possibilities of utilizing Artemisia capillaris extracts as a functional food component to control intestinal microflora.