• Title/Summary/Keyword: tet gene

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Detection of Antibiotic Resistance and Resistance Genes in Enterococci Isolated from Sucuk, a Traditional Turkish Dry-Fermented Sausage

  • Demirgul, Furkan;Tuncer, Yasin
    • Food Science of Animal Resources
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    • v.37 no.5
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    • pp.670-681
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    • 2017
  • The aim of this study was to isolate enterococci in Sucuk, a traditional Turkish dry-fermented sausage and to analyze isolates for their biodiversity, antibiotic resistance patterns and the presence of some antibiotic resistance genes. A total of 60 enterococci strains were isolated from 20 sucuk samples manufactured without using a starter culture and they were identified as E. faecium (73.3%), E. faecalis (11.7%), E. hirae (8.3%), E. durans (3.3%), E. mundtii (1.7%) and E. thailandicus (1.7%). Most of the strains were found resistant to rifampin (51.67%) followed by ciprofloxacin (38.33%), nitrofurantoin (33.33%) and erythromycin (21.67%). All strains were found susceptible to ampicillin. Only E. faecium FYE4 and FYE60 strains displayed susceptibility to all antibiotics. Other strains showed different resistance patterns to antibiotics. E. faecalis was found more resistant to antibiotics than other species. Most of the strains (61.7%) displayed resistance from between two and eight antibiotics. The ermB, ermC, gyrA, tetM, tetL and vanA genes were detected in some strains. A lack of correlation between genotypic and phenotypic analysis for some strains was detected. The results of this study indicated that Sucuk manufactured without using a starter culture is a reservoir of multiple antibiotic resistant enterococci. Consequently, Sucuk is a potential reservoir for the transmission of antibiotic resistance genes from animals to humans.

Safety Assessment of Coagulase-Negative Staphylococci from Jeotgal, a Korean High-Salt-Fermented Seafood (젓갈 유래 Coagulase-Negative Staphylococci의 안전성 평가)

  • Jeong, Do-Won;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.84-90
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    • 2015
  • Seventeen ampicillin-sensitive coagulase-negative staphylococci (CNS) isolates identified in jeotgal were subjected to assessments for antibiotic susceptibility and safety hazards. Fifteen of the 17 CNS strains exhibited phenotypic resistances to at least one antibiotic, and their prevailing resistance was to penicillin G. The dfrA gene for trimethoprim and tetK for tetracycline were amplified by PCR from the two strains, respectively. α-Hemolytic activity was not detected from the 17 strains, while five strains presented δ-hemolytic activity. Among the five strains, two strains exhibited β-hemolytic activity. Biofilm was formed from twelve strains. All of the tested phenotypic characteristics were expressed in a strain-specific manner.

Examination of Improved Tetracycline Inducible Gene Expression System In Vitro (새로운 Tetracycline 유도적 유전자 발현 System의 In Vitro 검정)

  • Kwon, Mo Sun;Kim, Teoan;Koo, Bon Chul
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.109-115
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    • 2013
  • Until recently the most popular tetracycline-inducible gene expression system has been the one developed by Gossen and Bujard. In this study, we tested the latest version of same system and the results are summarized as follows: Compared with previous one, the difference of new system are minor changes of nucleotide sequences in transactivator and tetracycline response element (TRE) regions. Sensitivity to the doxycycline (a tetracycline derivative) was improved. Leakiness of GFP marker gene expression in non-inducible condition was significantly decreased. Higher expression of the marker gene was observed when the cells were fed with doxycycline-containing medium. Optimal insertion site of woodchuck posttranscriptional regulatory element (WPRE) sequence which was known to increase gene expression was different depending on the origin of cells. In chicken embryonic fibroblast, location of WPRE sequence at 3' end of TRE resulted in the highest GFP expression. In bovine embryonic fibroblasts, 3' end of transactivator was the best site for the GFP expression.

Antimicrobial Resistance and Implicated Genes of E. coli Isolated from Commercial and Cooked Foods in Seoul (서울시내 유통식품에서 분리한 대장균의 항생제 내성 및 내성유전자)

  • Yoo, Young-Ah;Kim, Moo-Sang;Kim, Kyong-Sik;Park, Sun-Hee;Jung, Sung-Kuk
    • Journal of Food Hygiene and Safety
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    • v.25 no.3
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    • pp.220-225
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    • 2010
  • Distribution of foodborne E. coli strains, antimicrobial resistant genes and antimicrobial susceptibility have been carried out on E. coli isolated from commercial and cooked foods distributed food in Seoul. Of total 1,313 samples, fifty samples(3.8%) were found E. coli that included one of the ETEC and EPEC, respectively. The serotype of ETEC in seasoning raw meat was E. coli O26 and produced Verotoxin 2. Fifty percentage of total isolates were susceptible to all antimicrobial agents. Specially, there were ampicillin(36%), amoxicillin/clavulanic acid(32%) and tetracycline(22%) etc. Resistant gene (tetB) were found in four tetracycline resistant E. coli strains, and TEM gene was found in one ampicillin resistant E. coli isolate.

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.

Antimicrobial Resistance Patterns and Resistance genes assay of Shigella sonnei Isolated in Korea for Five Years (최근 5년 동안 국내에서 분리된 Shigella sonnei의 항균제 내성 유형과 내성유전자형 분석)

  • Huh, Wan;Lee, Sang-Jo;Kwon, Gi-Seok;Jang, Jong-Ok;Lee, Jung-Bok
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.31-39
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    • 2007
  • This study has been carried out for investigating the relatedness of representative 135 Shigella sonnei strains isolated from 2000 to 2004 by using biotyping and antimicrobial resistance. All strains showed typical biochemical characterisics of Shigella strain. Among 135 strains,79 (58.5%) strains were biotype "g",54 (40.0%) strains were biotype "a" and 2 (1.5%) strains were biotype "e". The results of susceptibility test against 16 antimicrobial agents were like this. Most of strains were susceptible to AN, CIP, C and GM. 129 (95.6%) strains were resistant to SXT, 126 (93.3%) strains were resistant to TE and 122 (90.4%) strains were resistant to SM. One hundred thirty two (97.8%) strains were resistant to more than two antimicrobial agents. R28 type (antimicrobial resistance patterns 28: resistant to AM, SAM, TE, TIC, SXT, K, SM and AmC) were 42 strains (31.1%). The other strains were showed 33 kinds of R patterns. The results of $bla_{TEM}$, sulII, tetA and strA gene detection were coincided with phenotype of antimicrobial resistance by disk diffusion method. But some strains which had sulII and strA genes were not showed the resistance against SXT and SM.

Co-occurrence Analyses of Antibiotic Resistance Genes and Microbial Community in Human and Livestock Animal Feces (사람 및 가축 유래 분변 미생물 군집과 항생제 내성 유전자 간 상관 관계에 대한 연구)

  • Jiwon Jeong;Aprajita Bhandari;Tatsuya Unno
    • Korean Journal of Environmental Agriculture
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    • v.41 no.4
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    • pp.335-343
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    • 2022
  • BACKGROUND: Antibiotics used in animal husbandry for disease prevention and treatment have resulted in the rapid progression of antibiotic resistant bacteria which can be introduced into the environment through livestock feces/manure, disseminating antibiotic resistant genes (ARGs). In this study, fecal samples were collected from the livestock farms located in Jeju Island to investigate the relationship between microbial communities and ARGs. METHODS AND RESULTS: Illumina MiSeq sequencing was applied to characterize microbial communities within each fecal sample. Using quantitative PCR (qPCR), ten ARGs encoding tetracycline resistance (tetB, tetM), sulfonamide resistance (sul1, sul2), fluoroquinolone resistance (qnrD, qnrS), fluoroquinolone and aminoglycoside resistance (aac(6')-Ib), beta-lactam resistance (blaTEM, blaCTX-M), macrolide resistance (ermC), a class 1 integronsintegrase gene (intI1), and a class 2 integrons-integrase gene (intI2) were quantified. The results showed that Firmicutes and Bacteroidetes were dominant in human, cow, horse, and pig groups, while Firmicutes and Actinobacteria were dominant in chicken group. Among ARGs, tetM was detected with the highest number of copies, followed by sul1 and sul2. Most of the genera belonging to Firmicutes showed positive correlations with ARGs and integron genes. There were 97, 34, 31, 25, and 22 genera in chicken, cow, pig, human, and horse respectively which showed positive correlations with ARGs and integron genes. In network analysis, we identified diversity of microbial communities which correlated with ARGs and integron genes. CONCLUSION(S): In this study, antibiotic resistance patterns in human and livestock fecal samples were identified. The abundance of ARGs and integron genes detected in the samples were associated with the amount of antibiotics commonly used for human and livestocks. We found diverse microbial communities associated with antibiotics resistance genes in different hosts, suggesting that antibiotics resistance can disseminate across environments through various routes. Identifying the routes of ARG dissemination in the environment would be the first step to overcome the challenge of antibiotic resistance in the future.

Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

  • Maeng, Chang-Jae;Kim, Hyung-Kwoun;Park, Sun-Yang;Koo, Bon-Tag;Oh, Tae-Kwang;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.85-91
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    • 2001
  • To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The $P_{C27}$ promoter cloned from the clostridial chromosmal DNA showed a 5-fold higher promoter strength than the $P_{SP02}$ promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, $P_{C27}$ exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3'end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of $P_{C27}$ was distinctively repressed in the presence of glucose. Using $P_{C27}$ as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolyticus KCTC 8957P was expressed in B. subtilis. When compared with the lipase expression by the T7 promoter induced by IPTG in E. coli, the $P_{C27}$ promoter showed about a 1.5-fold higher expression level in B. subtilis than that without induction.

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Construction of New P4-Derived Vector Plasmid Containing Tetracyclin Resistance Marker for the Bacteriophage P2-P4 System (박테리오파아지 P2-P4 시스템을 위한 tetracyclin resistance marker 함유 P4 유도체 벡터 플라스미드 조성)

  • Kim, Kyoung-Jin
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.118-122
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    • 2003
  • To develop vector plasmid for the bacteriophage P2-P4 system which is a useful experimental tool for the study of viral capsid assembly, we constructed a new P4-derived vector plasmid starting from P4 ash8 sid71 With recombinant DNA technology, a portion of P4 genome was deleted and tetracyclin resistance gene (terR) was introduced into P4 genome to give P4 selectivity. Resulting P4 ash8(sid71) terR was 12.09 kb long and could be converted to a viable bacteriophage with P2 infection. The burst size of induced bacteriophage form of P4 ash8(sid71) terR was determined. The CsCl buoyant equilibrium density gradient experiment of new P4 derivative suggested the upper limit of packaging capacity in P2-size head.

Molecular subtyping and antimicrobial susceptibility of Streptococcus dysgalactiae subspecies equisimilis isolates from clinically diseased pigs

  • Oh, Sang-Ik;Kim, Jong Wan;Kim, Jongho;So, Byungjae;Kim, Bumseok;Kim, Ha-Young
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.57.1-57.11
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    • 2020
  • Background: Streptococcus dysgalactiae subspecies equisimilis (SDSE) acts as an etiological agent for lameness, neurological signs, and high mortality in pigs. Despite its importance in pig industries and zoonotic potential, little is known about the effects of this pathogen. Objectives: This study aimed to determine the molecular characteristics and antimicrobial resistance of SDSE strains isolated from diseased pigs. Methods: A total 11 SDSE isolates were obtained from diseased pigs. Bacterial identification, PCR for virulence genes, emm typing, and antimicrobial resistance genes, multilocus sequence typing, and antimicrobial susceptibility test were performed. Results: Nine isolates were from piglets, and 8 showed lameness, sudden death, or neurological signs. The isolates were PCR-positive for sla (100%), sagA (100%), and scpA (45.5%), and only 1 isolate amplified the emm gene (stL2764). Eight different sequence types were detected, categorized into 2 clonal complexes and 4 singletons. All the isolates in this study were included in a small cluster, which also contained other strains derived from humans and horses. The minimum inhibitory concentrations for the tested beta-lactams were low, while those for macrolides, tetracyclines, and fluoroquinolones were relatively high. PCR analysis of the macrolide and tetracycline resistance genes demonstrated that the isolates carried erm(B) (18.2%, n = 2), mef(A/E) (9.1%, n = 1), tet(M) (18.2%, n = 2), and tet(O) (90.2%, n = 10). Two isolates presented a mutation in parC, which is associated with fluoroquinolone resistance. Conclusion: This study provided insight into swine-derived SDSE, as it is related to veterinary medicine, and elucidated its zoonotic potential, in the context of molecular epidemiology and antimicrobial resistance in public health.