• Title/Summary/Keyword: Antibiotic susceptibility test

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Real-Time PCR Detection of 16S rRNA Novel Mutations Associated with Helicobacter pylori Tetracycline Resistance in Iran

  • Dadashzadeh, Kianoosh;Milani, Morteza;Rahmati, Mohammad;Akbarzadeh, Abolfazl
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.8883-8886
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    • 2014
  • Background: Tetracycline is an antibiotic widely used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing due to increasing bacterial resistance. The aim of this study was to investigate the occurrence of 16S rRNA mutations associated with resistance or reduced susceptibility to tetracycline ofHelicobacter pylori by real-time PCR (RT-PCR) assays from culture. Materials and Methods: Tetracycline susceptibility and minimal inhibition concentration (MIC) was determined by the Epsilometer test (Etest) method. A LightCycler assay developed to detect these mutations was applied to DNA extracted from culture. The 16S rRNA of these isolates was sequenced and resistance-associated mutations were identified. From 104 isolates of H. pylori examined, 11 showed resistance to tetracycline. Results: LightCycler assay was applied to DNA extracted from 11 tetracycline-susceptible and 11 tetracycline resistance H. pylori isolates. In our study the sequencing of the H. pylori wild types in 16 s rRNA gene were AGA 926-928 with MIC (0.016 to $0.5{\mu}g/ml$), while the sequencing and MIC for resistant were GGA and AGC, (0.75 to $1.5{\mu}g/ml$), respectively. Also we found a novel mutation in 2 strains with $84^{\circ}C$ as their melting temperatures and exhibition of an A939C mutation. Conclusions: We conclude that real-time PCR is an excellent method for determination of H. pylori tetracycline resistance related mutations that could be used directly on biopsy specimens.

Distribution and antimicrobial susceptibility patterns of bacteria isolated from genital tract of riding mares (승용 씨암말의 생식기 유래 세균의 분포 및 항생제 감수성 양상)

  • Cho, Young-Jae;Lee, Yong-Duck;Jang, Jong-Duck;Shin, Kwang-Hyeu;Park, Yong-Soo;Yang, Jae-Hyuk;Kim, Sung-Joon;Cho, Gil-Jae
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.19-23
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    • 2015
  • This study was carried out to investigate the genital tract bacterial flora of riding mare in Jangsu stud farm during March to September, 2014. The specimens were collected from vaginal and uterus using a swab from 104 riding mares. Colonies were selected on blood and MacConkey agar plates, and identified as standard biochemical properties and Maldi-Tof MS. From this study, we isolated 148 strains including Escherichia (E.) coli (14.19%), Streptococcus (S.) equi subsp. zooepidemicus (2.7%), Streptococcus (S.) dysgalactiae subsp. equisimilis (2.03%), Klebsiella (K.) pneumonia (1.35%) and other strains from riding mares. In antimicrobial agents susceptibility test, it showed a high sensibility to the antibiotics of the most. E. coli and S. zooepidemicus were visible to have a high sensibility to almost antibiotics used in this study. However, K. pnemoniae showed a high antibiotic resistance patterns. These results may provide the basic information to establish strategies for the treatment and prevention of reproductive diseases in riding mares in Korea.

Distribution and Antimicrobial Susceptibility Patterns of Bacteria Isolated from Genital Tract in Thoroughbred Mares (더러브렛 씨암말의 생식기내 세균의 분포 및 항생제 감수성 양상)

  • Choi, Seong-Kyoon;Lee, Soo-Gil;Yang, Jae-Hyek;Cho, Gil-Jae
    • Journal of Veterinary Clinics
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    • v.24 no.1
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    • pp.19-25
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    • 2007
  • This study carried out to investigate the genital tract bacterial flora of Thoroughbred mare in Jeju province during March and July, 2006. The specimens were collected from vaginal ucosa and clitorial fossa using a culture swab (BBL, USA) from 100 Thoroughbred mares. Colonies were selected blood and MacConkey agar plate, and identified as standard biochemical properties using Biolog system (Thermo, USA). In this study, 470 gram-negative strains were isolated more frequently than 249 gram-positive strains. We were Isolated Escherichia coli (19.8%), Proteus mirabillis (14.9%), Enterobacter nimipressuralis (7.4%), Enterobacter mobilis (4.7%), Aeromonas encheleia (4.3%), Pseudomonas aeroginosa (3.0%), Staphylococcus aureus (14.9%), Staphylococcus epidermidis (11.2%), Coagulasenegative Staphylococcus spp. (10.0%), Enterococcus faecalis (9.2%), Enterococcus faecium (8.0%), Actinomyces viscosus (7.2%), Micoroccus diversus(6.8%), Streptococcus dysgalactiae subsp. equisimilis(5.2%), Streptococcus equi subsp. zooepidemicus (3.2%), Other non-beta hemolytic Streptococcus spp. (2.0%) and many others from vaginal mucosa and clitorial fossa in Thoroughbred mares. No significant bacteria (Taylorella equigenitalis and Klebsiella pneumonia) were isolated from the mare genital tract. In antimicrobial agents susceptibility test, it shows a high sensibility in the antibiotics of the most which excepts the streptomycin and neomycinm, kanamycin, spectinomycin, compound sulfonamides. Especially, Staphylococcus epidermidis, Enterococcus spp. and Streptococcus spp. were visible a high sensibility in the all antibiotics. However, Staphylococcus aureus, Pseudomonas aeruginosa, Proteus spp. and E. coli were showed a high antibiotic resistance patterns. These results may provide the basic information to establish strategies for the treatment and prevention of reproductive disease in Thoroughbred mares in Korea.

A study on the characteristics and pathogenicity of Aeromonas veronii isolated from infected goldfish (Carassius auratus) (피부 궤양이 발생한 금붕어(Carassius auratus)에서 분리한 Aeromonas veronii의 특성 및 병원성 분석)

  • Hyeon Ki Jung;Min Su Kim;Sok Ho Kim;Min Soon Choi
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.79-88
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    • 2024
  • Aeromonas spp. infections have been reported to cause significant economic losses not only in the ornamental fish industry but also in aquaculture. In December 2022-January 2023, an Aeromonas infection occurred on a goldfish in korea, A gram-negative bacterium was isolated from the skin and internal organs of infected goldfish (Carassius auratus). The results showed that the isolate was identified as Aeromonas veronii using 16S rDNA targeted oilgpnucleotide primers, furthermore characteristics of A. veronii was confirmed by enterotoxin gene, infectious experiment, antibiotic resistance. In-vivo pathogenicity of isolates to goldfsh resulted in 100% mortality in challenged host within one week of post experiment injection. As a result of PCR analysis targeting three enterotoxin-encoding genes, cytotoxic enterotoxin (act) was identified in A. veronii isolate in this study. Antimicrobial susceptibility pattern of isolate showed it was to susceptible to most antimicrobial agents tested but resistant to ampicillin, imipenem, meropenem and clindamycin.

Improved Eradication Rate of Standard Triple Therapy by Adding Bismuth and Probiotic Supplement for Helicobacter pylori Treatment in Thailand

  • Srinarong, Chanagune;Siramolpiwat, Sith;Wongcha-um, Arti;Mahachai, Varocha;Vilaichone, Ratha-Korn
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9909-9913
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    • 2014
  • Background: Helicobacter pylori (H. pylori) remains an important cause of gastric cancer and peptic ulcer disease worldwide. Treatment of H. pylori infection is one of the effective ways to prevent gastric cancer. However, standard triple therapy for H. pylori eradication is no longer effective in many countries, including Thailand. This study was designed to evaluate the efficacy of adding bismuth and probiotic to standard triple therapy for H. pylori eradication. Materials and Methods: In this prospective single center study, H. pylori infected gastritis patients were randomized to receive 7- or 14-day standard triple therapy plus bismuth with probiotic or placebo. Treatment regimen consisted of 30 mg lansoprazole twice daily, 1 g amoxicillin twice daily, 1 g clarithromycin MR once daily and 1,048mg bismuth subsalicylate twice daily. Probiotic bacteria composed of Bifidobacterium lactis, Lactobacillus acidophilus and Lactobacillus paracasei. Placebo was conventional drinking yogurt without probiotic. CYP2C19 genotyping and antibiotic susceptibility tests were also done. H pylori eradication was defined as a negative $^{13}C$-urea breath test at least 2 weeks after completion of treatment. Results: One hundred subjects were enrolled (25 each to 7- and 14-day regimens with probiotic or placebo). Antibiotic susceptibility tests showed 36.7% metronidazole and 1.1% clarithromycin resistance. CYP2C19 genotyping revealed 40.8%, 49% and 10.2% were rapid, intermediate and poor metabolizers, respectively. The eradication rates of 7- or 14 regimens with probiotics were 100%. Regarding adverse events, the incidence of bitter taste was significantly lower in the 7- day regimen with the probiotic group compared with 7- day regimen with placebo (40% vs. 64%; p=0.04). Conclusions: The 7-day standard triple therapy plus bismuth and probiotic can provide an excellent cure rate of H. pylori (100%) in areas with low clarithromycin resistance such as Thailand, regardless of CYP2C19 genotype. Adding a probiotic also reduced treatment-related adverse events.

Isolation of vibrio species from cultured flounders (paralichthys olivaceus) with uicers and ascites in the southern coast of Korea during the winter season (동절기 한국 남해안의 궤양증 및 복수증 양식 넙치로부터 vibrio종의 분리)

  • Lee, Hun-Ku;Kim, Hee-Je;Kim, Il
    • Korean Journal of Microbiology
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    • v.29 no.5
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    • pp.319-328
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    • 1991
  • this study was performed for the purpose of surveying the distribution and characteristics of the Vibrio species in the diseased farm flounders (Paralichthys olivaceus) during the winter season in Korea. Samples were collected along the southern coast in the Sinam, Keoje Island, and Teosu areas from November '90 to Feburary '91. Nine species of Vibrio were identified as V. tubiashii (85 strains), V. damsela (78 strains), V. anguillarum (21 strains), V. compbelli (9 strains), V. fluvialis (9 strain), V. costicola (1 strain), V. alginolyticus (1 strain), V. gazogenes (1 strain), V. marinus (1 strain), and unidentified 3 groups of bacteria (154 strains) which may have related with V. tubiashii were isolated. Eleven kinds of discs (BBL Co.) were used for antibiotic susceptibility test and 116 strains were selected for the experiment. Most bacteria were powerfully inhibited their growth by 4 antibiotics such as cephalothin, chloramphenicol, colistin, and tetracycline, but they were poorly inhibited by 4 antibiotics such as carbenicillin, kanamycin, penicillin G, and streptomycin.

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Isolation of Streptococcus thermophilus and Lactobacillus delbrueckii as Starter Culture Candidate Originated from Indonesian Cow's Milk

  • Andrian, Danish;Rizkinata, Denny;Susanto, Tan Steven Ryan;Lucy, Jap;Jan, Tan Tjie
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.201-209
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    • 2018
  • Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus fermentum and Lactobacillus casei were successfully isolated from indigenous Indonesian fresh milk based on the general morphological and biochemical classification as described in Bergey's manual. Verification was conducted by sequencing of 16S rRNA after selection using the classification method mentioned in the manual. All isolates exhibited antimicrobial activity against Escherichia coli and Staphylococcus aureus in the well diffusion test. The susceptibilities of the isolated S. thermophilus 24/S1 and L. delbrueckii 94/L4 against 22 different antibiotics were determined by the disc diffusion method and variable susceptibility patterns were observed. Both isolates were susceptible to amoxicillin, the most commonly prescribed antibiotic, and resistant to sulfonamide. The presence of a plasmid was not detected after extraction. S. thermophilus 24/S1 and L. delbrueckii 94/L4 starter cultures were prepared for yogurt production after 9.5 h of incubation and the yogurt was evaluated for its flavor and quality by 30 volunteers. A score of $4.93{\pm}0.45$ out of 7 was obtained as compared to the yogurt prepared using commercial starter cultures which yielded a score of $4.76{\pm}0.30$ out of 7.

Screening of Antibiotics that Selectively Inhibit a Bacterial Species Associated with a Recurrent Aphthous Stomatitis Risk

  • Lee, Ahreum;Kim, Yunji;Choi, Youngnim
    • International Journal of Oral Biology
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    • v.42 no.3
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    • pp.123-128
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    • 2017
  • Recurrent aphthous stomatitis (RAS) is a common oral mucosal disorder for which no curative treatment is available. We previously reported that decreased Streptococcus salivarius and increased Acinetobacter johnsonii on the oral mucosa are associated with RAS risk. The purpose of this study was to identify antibiotics that selectively inhibit A. johnsonii but minimally inhibit oral mucosal commensals. S. salivarius KCTC 5512, S. salivarius KCTC 3960, A. johnsonii KCTC 12405, Rothia mucilaginosa KCTC 19862, and Veillonella dispar KCOM 1864 were subjected to antibiotic susceptibility test using amoxicillin, cefotaxime, gentamicin, clindamycin, and metronidazole in liquid culture. The minimal inhibitory concentration (MIC) was defined as the concentration that inhibits 90% of growth. Only gentamicin presented a higher MIC for A. johnsonii than MICs for S. salivarius and several oral mucosal commensals. Interestingly, the growth of S. salivarius increased 10~200% in the presence of sub-MIC concentrations of gentamicin, which was independent of development of resistance to gentamicin. In conclusion, gentamicin may be useful to restore RAS-associated imbalance in oral microbiota by selectively inhibiting the growth of A. johnsonii but enhancing the growth of S. salivarius.

Rapid Detection of Vancomycin-resistant Enterococci (VRE) in Clinical Samples from University Hospital

  • Yang, Byoung-Seon;Park, Jung-Yeon;Choi, Seung-Gu
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.1
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    • pp.16-20
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    • 2013
  • Outbreaks of vancomycin-resistant enterococci (VRE) are being reported more frequently in many countries. While seven glycopeptide resistance genotypes have been described in Enterococci, vanA and vanB are the most common resistance genotypes. The aim of this study was to detect antibiotic susceptibilities of 23 Enterococcus faecium strains, which caused an outbreak in a University hospital by a disk diffusion test to investigate the presence of the species specific gene, and the resistant genotypes, vanA and vanB by duplex PCR. PCR for vanA and vanB was performed on 23 enterococci. Twenty three were identified as E. faecium and were tested positive for the vanA genotype. This study will report on the validation of a simple and accurate VRE detection method that can be easily incorporated into the daily routine of a clinical laboratory. Early detection of VRE strains, including those with susceptibility to vancomycin, is of paramount clinical importance as it allows rapid initiation of strict infection control practices, as well as the therapeutic guidance for confirmed infections. The PCR method developed in the present study is simple and reliable for the rapid characterization of VRE.

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Microbiologic Pollution of Indoor Air in Industrial Work-Places (산업체 작업환경의 실내 공기에서 미생물 오염도)

  • 강경희;장명웅
    • Journal of Life Science
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    • v.9 no.3
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    • pp.314-327
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    • 1999
  • This study was investigated to isolate identify the total bacteria and fungi from the indoor air of work-place of the shoes, paint, stainless steel, and plastic industries. The number of bacterial colonies on the nutrient agar plates were calculated by the open petridish method for 30 minutes in indoor air of work-places at the autumn and winter. The isolated bacteria were identified by Gram stain and biochemical test using API Staph and API 20E kits. The isolated fungal colonies were identified by gross appearance of the giant colonies and microscopic examination of their spore and hyphal characteristics on the slide culture method. The minimum inhibitory concentration (MIC) of several antibiotics against isolated bacteria was determined by the microdilution method with Mueller-Hinton broth. The 70-400 colonies in autumn and 54-236 colonies in winter were isolated from the indoor air of work-places of several industry. The isolation rates of Gram positive cocci, Gram positive bacilli, Gram negative bacilli, and Gram negative cocci were 46.3%, 19.8%, 17.3%, and 16.1%, respectively. In Gram positive cocci, the most strains were identified as Aerococcus spp, Micrococcus spp, and Staphylococcus spp. In Gram positive and negative bacilli, and Gram negative cocci were identified as Bacillus spp, Pseudomonas spp, and Neisseria spp, respectively. The frequently isolated fungi were Aspergillus spp, Penicillium spp and Rhizopus spp, respectively. The frequently isolated Aerococcus spp, Micrococcus spp, and Staphylococus spp were highly resistance against ampicillin, erythromycin, methicillin, and tetracycline. These results arouse our attention to microbiologic pollution in the indoor air of work-places of industries.

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