• Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.24-28
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• 2009

A tetracycline resistant Vibrio parahaemolyticus, capable of growing on TCBS medium containing tetracycline, was isolated from cultivated fishes. A gene responsible for the tetracycline resistance was cloned from chromosomal DNA of the V. parahaemolyticus strain using Escherichia coli KAM3, which lacks major multi-drug efflux pumps (${\Delta}acrB$) as host cells. The nucleotide sequence and homology analysis revealed an open reading frame (ORF) for tetracycline resistance protein (TetB). In order to characterize the antibiotic resistance of TetB originated from the V. parahaemolyticus strain, the gene was sub cloned into plasmid pSTV28. The resulting plasmid was designated as pSTVTetB and transformated into E. coli KAM3. E. coli KAM3 cells harboring the recombinant plasmid pSTVTetB are able to grow on plates containing tetracycline and oxytetracycline but not doxycycline, indicating that the tetB gene confers the tetracycline- and oxytetracycline-resistance to the host cell.

• 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 [bla_TEM], 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.

• Fisheries and Aquatic Sciences
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• v.9 no.2
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• pp.97-100
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• 2006

To determine whether the tetracycline resistance genes tet (34), tet (M), and tet (S) can be transferred among bacteria, we used a filter mating experiment allowing intimate cell-cell contact between donor and recipient. The tet(34) gene, conveyed on a chromosome of Vibrio species (No. 6 and SW-42) was not transferred to Escherichia coli JM109, suggesting that it is not transferred among bacterial species. The tet (M) gene was transferred from three Vibrio strains (4-E, SW-18, and SW-38) to E. coli at frequencies of $8.5{\times}10^{-5}\;to\;2.1{\times}10^{-6}$. The tet(S) gene was transferred from Lactococcus garvieae KHS98032 to E. coli at a frequency of $1.8{\times}10^{-6}$. Transconjugated recipients showed increased minimum inhibitory concentrations against oxytetracycline. Although the donors possess the Tn916-Tn1545 transposons, they were not detected in transformed recipients, suggesting that the transfer of tet(M) and tet(S) is mediated by elements or mechanisms. Two ribosomal protect protein genes were also transmissible from marine bacteria to E. coli, suggesting gene hopping among marine, terrestrial, and human environments.

• 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.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.863-870
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• 2005

One hundred and twenty three strains of Mycoplasma pneumoniae were isolated from patients with respiratory diseases from February 2002 to April 2005 in Busan, Korea. The MICs of tetracycline and erythromycin up to $90\%$ of the 123 M. pneumoniae isolates tested were $0.5\~1.0$, and $0.5\~512{\mu}/ml$, respectively. Plasmid DNA was not isolated from all of the M. pneumoniae isolates. Out of 323 strains of M. pneumoniae, 57 ($46.3\%$) stains contain tetM gene on their chromosomal DNA, and 60 ($48.8\%$) strains were mutated in domain V of 23S rRNA for erythromycin resistance. Out of 63 strains of M. pneumoniae which were not mutated in domain V of 235 rRNA for erythromycin resistance, 36 ($57.1\%$) strains contained tetM gene, and out of 60 strains of M. pneumoniae which were mutated in domain V of 23S rRNA for erythromycin resistance, 21 ($35.0\%$) strains contained tetM gene. These results suggest that the isolation rate of erythromycin and tetracycline resistant M. pneumoniae is higher than those of other countries, and erythromycin and tetracycline are not first choice drug for M. pneumoniae infection in Korea, and it need confirm by a nationwide surveilance of antimicrobial resistance.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.4
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• pp.388-400
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• 2023

Aeromonas spp. and Pseudomonas spp. are opportunistic pathogens widely distributed in the aquatic environment. To test the antibiotic susceptibility, the MIC of the 18 antibiotics mainly used in aquaculture were measured. Aeromonas spp. and Pseudomonas spp. straoms had different resistance patterns against most antibiotics. The MIC of tetracycline for four Aeromonas spp. strains (10.5%) was < 0.25 ㎍/mL. However, 0.5-4 ㎍/mL tetracycline inhibited most Pseudomonas spp. strains. The tet resistance performance of 14 genes including tet(B), tet(E), and tet(M) were investigated. Investigating, the tetracycline resistance gene of 38 Aeromonas spp. strains detected tet(A) in 21 strains (55.3%). Two Pseudomonas spp. strains showed high MIC values and no inhibition zone. tet gene analysis detected tet(D) in only one strain (5%).

• Korean Journal of Veterinary Service
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• v.32 no.3
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• pp.227-239
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• 2009

At the present study, it was aimed to detect virulence genes and antimicrobial resistance genes among 102 strains of 12 Salmonella serotypes isolated from pigs and cattle. In polymerase chain reaction (PCR), invA was detected from all strains of Salmonella spp., spvC was detected from Salmonella enterica serotype Enteritidis (S. Enteritidis) (100%), S. Bradenburg (75%), and S. Typhimurium (20.4%). Drug resistance related genes of 12 types were detected from all strains. TEM ($bla_{TEM}$) gene was detected from 51 (92.7%) of 55 $\beta$-lactams (54 ampicillin or 1 amoxicillin) resistance strains. 55 (100%) of 55 chloramphenicol resistance strains, 3 (100%) of 3 gentamicin resistance strains and 5 (100%) of 5 kanamycin resistance strains did contain cml, aadB, and aphA1-Iab, respectively. strB (89.9%), strA (88.4%), aadA2 (84.1%) and aadA1 (72.5%) were detected from 69 streptomycin resistance strains. sulII and dhfrXII were detected from 49 (100%) of 49 sulfamethoxazole/trimethoprim resistance strains, but sulI was not detected. tetA (97.9%) and tetB (21.6%) were detected from 97 tetracycline resistance strains. int gene was detected from 58 (56.9%) of 102 strains. 54 S. Typhimurium of 102 Salmonella spp. were attempted to detect drug resistance genes. TEM was detected from 44 (95.7%) of 46 $\beta$-lactams (45 ampicillin or 1 amoxicillin) resistance strains. cmlA was detected from 51 (100%) of 51 chloramphenicol resistance strains. aadA2 (100%), strA (100%), strB (100%), and aadA1 (79.6%) were detected from 54 streptomycin resistance strains. sulII (100%) and dhfrXII (100%) were detected from 49 sulfamethoxazole/trimethoprim resistance strains. tetA was detected from 54 (100%) of 54 tetracycline resistance strains. int gene was detected from 54 (100%) of 54 strains. The major drug resistance pattern and resistance gene profile were ampicillin, chloramphenicol, streptomycin, sulfamethoxazole/trimethoprim and tetracycline (ACSSuT) and TEM, cmlA, aadA1, aadA2, strA, strB, sulII, dhfrXII, tetA and int, respectively.

• Korean Journal of Veterinary Service
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• v.36 no.2
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• pp.73-78
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• 2013

This study was conducted to investigate antibiotic resistance among Salmonella Typhimurium isolates from diseased pigs in Gyeongbuk province during the period 1998~2011. One hundred forty one isolates were tested for antibiotic resistance using the standard disk diffusion method and were examined for presence of resistance gene by PCR method. S. Typhimurium showed high drug resistance rates to tetracycline (95.7%), streptomycin (93.6%), ampicillin (86.5%), cephalothin (80.1%), gentamicin (79.4%), and trimethoprim/sulfamethoxazole (72.3%). Resistance gene, blaTEM, blaPSE1, tetA, tetB, tetG, sul1, sul2, aadA, strA, grm, and temA were detected among the antibiotic resistance isolates and temB, tetC, aadB gene were not detected. One hundred twenty one (89.6%) tetA, two (1.5%) tetB and one (0.7%) tetG gene were detected in the 135 tetracycline resistant isolates. Two (1.6%) temA gene were detected in one hundred twenty two ampicillin resistance isolates and temB was not detected.

• Korean Journal of Microbiology
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• v.46 no.4
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• pp.326-333
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• 2010

A total of 105 nontyphoid Salmonella isolated from infants in Seoul from 2003 to 2009 was investigated for their serotype, antimicrobial resistance, characterization of integron, and the patterns of Pulsed-field gel electrophoresis (PFGE). Eighteen serotypes were detected in 105 isolates, and the two most common serotypes were S. Enteritidis (47.6%) and Montevideo (15.2%). Among the Salmonella serovars, a high level of antimicrobial resistance was found to ampicilin (60%), tetracycline (46.7%), streptomycin (35.2%) and nalidixic acid (28.6%). In the multi-drug resistance patterns, the predominant patterns were only nalidixic acid (15.7%), ampicillin-ampicillin/sulbactam-tetracycline (14.5%), and ampicillin-streptomycin-chloramphenicol-tetracycline (10.8%). PCR and DNA sequencing analysis revealed the presence of class 1 integron in 20 isolates (19%). Of the class 1 integron positive isolates 20% harboured the integron-associated gene cassettes : aadA2, blaP1, dfr17-aadA5, dfrA12-aadA2, and aadA7. PFGE was carried out to examine the genetic relatedness among S. Enteritidis isolates. Except for three strains, fifty strains were divided by three pulsotypes.

• Journal of Microbiology
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• v.45 no.5
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• pp.379-387
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• 2007

A hundred and seventeen antibiotic-resistant Escherichia coli strains were isolated from public tap and spring waters which were polluted by fecal coliforms. There were no significant differences between two water sources as to the coliform pollution level (p> 0.05). All E. coli isolates were detected to be resistant to one or more antibiotics tested. Nearly 42% of the isolates showed multiresistant phenotype. Three (2.5%) of these isolates contained class 1 integron. Sequencing analysis of variable regions of the class 1 integrons showed two gene cassette arrays, dfr1-aadA1 and dhfrA17-aadA5. Resistance to ampicillin, tetracycline or trimethoprim-sulfamethoxazole was transferable according to the results of conjugation experiments. The rate of tetracycline resistance was 15%. tet(A)-mediated tetracycline resistance was widespread among tetracycline-resistant E. coli isolates. Genotyping by BOX-polymerase chain reaction (BOX-PCR) showed that some of the strains were epidemiologically related. This is the first report on the prevalence and characterization of class 1 integron-containing E. coli isolates of environmental origin in Turkey.