• Title/Summary/Keyword: antibiotic resistance genes

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Characterization of Noble AmpC-Type $\beta$-Lactamases Among Clinical Isolates Using New Expression/Secretion Vector (발현ㆍ분비 벡터 및 임상 균주가 생성하는 신규 AmpC-type $\beta$-lactamase의 특성)

  • 정하일;성광훈;이정훈;장선주;이상희
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.104-110
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    • 2004
  • To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

Prevalence and Classification of Escherichia coli Isolated from bibimbap in Korea (비빔밥에서 분리한 대장균의 오염도 조사 및 특성 연구)

  • Lee, Da-Yeon;Lee, Joo-Young;Wang, Hae-Jin;Shin, Dong-Bin;Cho, Yong-Sun
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.126-131
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    • 2015
  • Pathogenic Escherichia coli is recognized as an important cause of diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome worldwide. This study was conducted to investigate the prevalence E. coli contamination in the Korean traditional food bibimbap. E. coli were isolated from 84 of 1142 (7.3%) bibimbap investigated from 2005 to 2011. Antibiotic resistance profiling demonstrated that 6 of the 84 isolates (7.2%) showed multiple drug resistance. Fifteen virulence genes specific for pathogenic E. coli such as Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli (EAEC) were examined by multiplex PCR for mixed bacterial cultures derived from bibimbap samples. The EPEC virulence gene (ent) was detected in 5 strains (5.9%), while ETEC, EAEC, and EIEC were not detected. STEC serotypes O103 (1.2%), O91 (1.2%), and O128 (6.0%) were found, but other serogroups such as O26, O157, O145, O111 and O121 were not detecded. Automated Repetitive-Sequence-Based PCR analysis showed different patterns.

Diversity of ampicillin resistant bacteria in domestic streams (국내 하천에 분포하는 ampicillin 내성균의 다양성)

  • Go, Eun Byeul;Chung, In-Young;Kim, Hyuk;Seok, Kwang-Seol;Kim, Byeori;Yoo, Yong-Jae;Jang, Yejin;Chae, Jong-Chan
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.440-443
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    • 2015
  • The widespread emergence of antibiotic resistant microorganisms in clinics and natural environments has attracted public concern. Especially, microorganisms inhabiting natural environment is considered as a source responsible for increasing the abundance of antibiotic resistant genes in ecosystem. In this study, the diversity of culturable bacteria resistant to ampicillin was investigated with water samples collected from seven locations in Korea. The genera belonging to Aeromonas and Acidovorax were dominant among the isolated 498 strains. The 66% of isolates showed multi-drug resistance against more than six antibiotics among tested fourteen ones and isolates resistant to seven antibiotics were the most prevalent with 19.7% abundance. Using the antibiotics susceptibility results, the intrinsic resistance profile was suggested for the most dominant genera, Aeromonas, Acidovorax, Pseudomonas, and Elizabethkingia.

Isolation and Identification of Ampicillin-resistant Bacteria in Changwon (창원근교에서의 ampicillin 내성세균의 분리 및 동정)

  • Bae, Young-Min
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1529-1535
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    • 2018
  • The number of antibiotic-resistant bacteria is increasing rapidly while the discovery rate of new antibiotics is in decline. A systematic study is therefore necessary to investigate which bacteria are resistant to medically important antibiotics and how high that resistance is. To that end, this study aimed to analyze which bacteria demonstrated resistance to ampicillin, one of the currently most-widely used medical antibiotics. Water samples were collected from the Changwon-Cheon that runs through Changwon City and from the pond in front of the dormitory building at Changwon University. Hundreds of ampicillin-resistant colonies were obtained and 22 morphologically distinct examples were chosen for further study. These bacteria were identified by amplifying their 16S rRNA genes and comparing those sequences with data in GenBank. The bacteria was identified as belonging to 10 families, 12 genera, and 17 species, and all were able to grow in the presence of $50{\mu}g/ml$ ampicillin while seven showed growth at ampicillin concentrations as high as 1.5 mg/ml.

Mutation of rpsL Gene in Streptomycin-Resistant Pseudomonas syringae pv. actinidiae Biovar 3 Strains Isolated from Korea (국내에서 분리된 Streptomycin 저항성 Pseudomonas syringae pv. actinidiae Biovar 3 균주에서 rpsL 유전자의 돌연변이)

  • Lee, Young Sun;Kim, Gyoung Hee;Koh, Young Jin;Jung, Jae Sung
    • Research in Plant Disease
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    • v.28 no.1
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    • pp.26-31
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    • 2022
  • Pseudomonas syringae pv. actinidiae (Psa) is the causal agent responsible for the bacterial canker disease of kiwifruit plants. Psa strains are divided into five different biovars based on genetic and biochemical characteristics. Among them, biovar 2 and 3 strains of Psa were isolated and have been causing widespread damages in Korea. One of the most effective ways to control Psa is to use an antibiotic such as streptomycin. However, Psa strains resistant to this antibiotic were isolated in Korea, and an earlier study revealed that the resistance in the biovar 2 is associated with strA-strB genes. This study aimed to determine the molecular resistance mechanism of Psa biovar 3 strains to streptomycin. Sequencing the rpsL gene encoding ribosomal protein S12 from three streptomycin-resistant strains screened in the laboratory revealed that a spontaneous mutation occurred either at codon 43 or 88. Meanwhile, in four streptomycin-resistant strains of Psa biovar 3 isolated from two kiwifruit orchards, a single nucleotide in codon 43 of the rpsL, which is AAA in streptomycin-sensitive strain, was substituted for AGA causing an amino acid change from lysine to arginine. The resistant mechanism in all biovar 3 strains obtained in Korea was identified as a mutation of the rpsL gene.

The global regulator GacS of a biological bacterium Pseudomonas chlororaphis O6 regulates expression of the stationary-phase sigma factor rpoS and reduces survival in oxidative stress.

  • Kang, Beom-Ryong;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.100.2-101
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    • 2003
  • The global regulator, GacS (global antibiotic and cyanide sensor kinase), was required for the increased resistance to hydrogen peroxide occurring as cultures of the rhizobacterium, P. chlororaphis O6, matured. Specific stationary-phase peroxidase and catalase isozymes were absent in the GacS mutant, whereas a manganese-superoxide dismutase isozyme was expressed earlier and to a great extent than wild type. In the wild type cell, transcript accumulation of rpoS was higher in late logarithmic-phase cells than cells from mid logarithmic- or stationary-phase. Transcripts from rpoS in the GacS mutant were reduced in each of these growth phases compared to the wild type expression. The down stream sequence from rpoS lacked sequences encoding a small RNA, rsmZ, found in other pseudomonads and implicated in control of genes activated by the GacS system. These findings suggest that GacS-mediated regulation of RpoS plays role in control of oxidative stress in P. chlororaphis O6 by as yet an unknown mechanism.

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Food-Grade Expression and Secretion Systems in Lactococcus

  • Jeong, Do-Won;Hwang, Eun-Sun;Lee, Hyong-Joo
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.485-493
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    • 2006
  • Lactococcus species are noninvasive and nonpathogenic microorganisms that are widely used in industrial food fermentation and as well-known probiotics. They have been modified by traditional methods and genetic engineering to produce useful food-grade materials. The application of genetically modified lactococci in the food industry requires their genetic elements to be safe and stable from integration with endogenous food microorganisms. In addition, selection for antibiotic-resistance genes should be avoided. Several expression and secretion signals have been developed for the production and secretion of useful proteins in lactococci. Food-grade systems composed of genetic elements from lactic acid bacteria have been developed. Recent developments in this area have focused on food-grade selection markers, stabilization, and integration strategies, as well as approaches for controlled gene expression and secretion of foreign proteins. This paper reviews the expression and secretion signals available in lactococci and the development of food-grade markers, food-grade cloning vectors, and integrative food-grade systems.

Bacterial Community of Galchi-Baechu Kimchi Based on Culture-Dependent and - Independent Investigation and Selection of Starter Candidates

  • Kim, Tao;Heo, Sojeong;Na, Hong-Eun;Lee, Gawon;Kim, Jong-Hoon;Kwak, Mi-Sun;Sung, Moon-Hee;Jeong, Do-Won
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.341-347
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    • 2022
  • In this study, the bacterial community of galchi-baechu kimchi was determined using culture-based and culture-independent techniques (next generation sequencing:NGS), and showed discrepancies between results. Weissella koreensis and Pediococcus inopinatus were the dominant species according to the NGS results, while Bacillus species and P. inopinatus were dominant in the culture-dependent analysis. To identify safe starter candidates, sixty-five Bacillus strains isolated from galchi-baechu kimchi using culture-dependent methods were evaluated for their antibiotic resistance, presence of toxin genes, and hemolytic activity. Strains were then assessed for salt tolerance and protease and lipase activity. As a result, four strains-B. safensis GN5_10, B. subtilis GN5_19, B. velezensis GN5_25, and B. velezensis GT8-were selected as safe starter candidates for use in fermented foods.

Effects of Lactobacillus reuteri MG5346 on Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL)-Induced Osteoclastogenesis and Ligature-Induced Experimental Periodontitis Rats

  • Yu-Jin Jeong;Jae-In Jung;YongGyeong Kim;Chang-Ho Kang;Jee-Young Imm
    • Food Science of Animal Resources
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    • v.43 no.1
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    • pp.157-169
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    • 2023
  • Effects of culture supernatants of Lactobacillus reuteri MG5346 (CS-MG5346) on receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclastogenesis were examined. CS-MG5346 treatment up to 400 ㎍/mL significantly reduced tartrate-resistant acid-phosphatase (TRAP) activity, the phenotype biomarker of osteoclast, without affecting cell viability. CS-MG5346 inhibited the expression of osteoclast specific transcriptional factors (c-fos and nuclear factor-activated T cells c1) and their target genes (TRAP, cathepsin, and matrix metallo-proteinase-9) in a dose-dependent manner (p<0.05). The administration of L. reuteri MG5346 (2×108 CFU/day) for 8 wks significantly improved furcation involvement, but no difference was observed in alveolar bone loss in ligature-induced experimental periodontitis rats. The elevated RANKL/osteoprotegerin ratio, the biomarker of periodontitis, was significantly lowered in the gingival tissue by administration of L. reuteri MG5346 (p<0.05). L. reuteri MG5346 showed excellent stability in simulated stomach and intestinal fluids and did not have antibiotic resistance. Based on the results, L. reuteri MG5346 has the potential to be a promising probiotic strain for oral health.

Emodin Attenuates Inflammasome Activation Induced by Helicobacter pylori Infection through Inhibition of VacA Translocation

  • Thach Phan Van;Anh Duy Do
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.507-516
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    • 2023
  • Eradication of Helicobacter pylori infection is an essential strategy to decrease the risk of developing gastric cancer. However, the standard triple therapy has negative aspects associated with side effects and the emergence of antibiotic resistance. Therefore, alternative therapies are required to enhance the management of H. pylori infection effectively. In this study we examined the effect of emodin on the amelioration of inflammatory response due to H. pylori infection. Our results indicated that emodin treatment effectively decreased the expression of virulence genes, including sabA, vacA, cagL, cagA, sabA, and suppressed the adhesion ability of H. pylori to AGS cells. Emodin has been shown inhibitory effects on the inflammasome pathway through reductions in VacA translocation, lowering ROS stress, cleaved Caspase-1, NLRP3, and cleaved Gasdermin D levels, thereby lowered pyroptosis in infected cells. In summary, our study demonstrated that emodin has the ability to attenuate inflammation caused by H. pylori by modulating virulence gene expression and decreasing VacA translocation. Further study is required to evaluate the therapeutic efficacy of emodin in treating H. pylori infection and better understand the underlying mechanisms.