• Archives of Pharmacal Research
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• v.26 no.8
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• pp.638-643
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• 2003

Raw milk samples, and cow and chicken intestines were tested to isolate vancomycin-resistant, gram-positive bacteria. From these samples, we isolated seven vancomycin-resistant Streptococcus equinus, two vancomycin-resistant viridans Streptococcus and two vancomycin-resistant Enterococcus faecium. The MICs of several antibiotics, including vancomycin, against these strains were tested. Seven isolates of S. equinus showed high level resistance to vancomycin and teicoplanin (＞100 $\mu$ g/mL). The cell wall thickness of these strains was compared with that of the sensitive strain by TEM and no differences were obserbed between these strains. We compared the strains of vancomycin-resistant Streptococcus equinus using PCR with Microbial Uniprimer Kit. We concluded that it is necessary to combine other methods in order to cluster and identify all isolates of S. equinus.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.143-147
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• 2005

The non-pathogenic, non-glycopeptide-producing actinomycete Streptomyces coelicolor carries a cluster of seven genes (vanSRJKHAX) that confers inducible, high-level resistance to vancomycin. The van genes are organised into four transcription units, vanRS, vanJ, vanK and vanHAX, and these transcripts are induced by vancomycin in a vanR-dependent manner. vanHAX are orthologuous to genes found in vancomycin resistant enterococci that encode enzymes predicted to reprogramme peptidoglycan biosynthesis such that cell wall precursors terminate in D-Ala-D-Lac, rather than D-Ala-D-Ala. vanR and vanS encode a two-component signal transduction system that mediates transcriptional induction of the seven van genes. vanJ and vanK are novel genes that have no counterpart in previously characterised vancomycin-resistance clusters from pathogens. VanK is essential for vancomycin resistance in S. coelicolor and it is required for adding Gly branch to stem peptides terminating D-Ala-D-Lac. Because VanK can recognise D-Lac-containing precursors but the constitutively expressed femX enzyme, encoded elsewhere on the chromosome, cannot recognize D-Lac-containing precursors as a substrate, vancomycin-induced expression of VanHAX in a vanK mutant is lethal. Further, femX null mutants are viable in the presence of glycopeptide antibiotics but die in their absence. Bioassay using vanJp-neo fusion reporter system also showed that all identified inducers for van genes expression were glycopeptide antibiotics, but teicoplanin, a membrane-anchored glycopeptide, failed to act as an inducer.

• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.313-316
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• 2006

To obtain antibiotic resistant profiles of Bifidobacterium, minimum inhibitory concentrations (MIC) of 14 antibiotics for 93 Bifidobacterium isolates from Korean intestine origin were determined. All strains tested were sensitive to chloramphenicol, rifampicin, and amoxicillin, whereas resistant to aminoglycoside family, nalidixic acid, and vancomycin. Among vancomycin-resistant strains, 34% were resistant at more than $100\;{\mu}g/mL$, and showed variant resistances toward tetracycline, erythromycin, and penicillin. Their resistances against penicillin, cephalothin, and tetracycline were higher than ten years ago. MIC of ten isolates from commercial yoghurt products were very similar to those of strains from Korean intestine origin, and 20% strains showed resistance at higher than $100\;{\mu}g/mL$ vancomycin. These results indicated patterns of antibiotic resistance against Bifidobacterium from Korean intestine origin and commercial yoghurts were very similar,and prevalence of vancomycin resistance for Bifidobacterium was 20%. To develop new probiotic, antibiotic resistance of vancomycin and risks involved should be evaluated.

• Biomedical Science Letters
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• v.19 no.3
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• pp.280-283
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• 2013

Leuconostoc spp. is intrinsically resistant against vancomycin and rarely causes the infection in immunocompromised patients. In this report, we describe a fatal case of Leuconostoc lactis bacteremia in a patient with biliary tract stent insertion to resolve the biliary tract obstruction by multiple pseudocysts in the pancreatic head region. Leuconostic lactis isolated from the blood of the patients was confirmed by 16S rRNA sequencing and this isolate was susceptible against most antibiotics, including levofloxacin, penicillin, erythromycin and cefotaxime except vancomycin. The septic shock and multi-organ failure was abruptly progressed due to delayed use of adequate antibiotic. Using vancomycin as the empirical antibiotics in a bacteremic patient by Gram positive cocci, the treatment failures by the isolates with intrinsic resistance against vancomycin have to be considered. In addition, the prompt and accurate identification of Leuconostoc spp. are very important to select the adequate antibiotics.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.637-642
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• 2010

We investigated the prevalence and the molecular characteristics of vancomycin-intermediate Staphylococcus aureus (VISA) among methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from clinical samples at tertiary or general hospitals participating in a nationwide surveillance program for VISA and vancomycin-resistant Staphylococcus aureus (VRSA) in Korea during an 8-week period in each year from 2001 to 2006. Of 41,639 MRSAs isolated, 37,856 were screened and 169 grew on brain heart infusion agar supplemented with 4 ${\mu}g/ml$ vancomycin. A vancomycin MIC of 4 ${\mu}g/ml$ was confirmed for 33 VISA isolates of the 169 isolates. Eighteen of the 33 isolates were classified as hetero-VISA (hVISA) by the population analysis profile (PAP) method. All VISA isolates were susceptible to linezolid, tigecycline, and quinupristin-dalfopristin. Most VISA isolates (MIC 4 ${\mu}g/ml$) showed a PFGE C pattern with sec, seg, and sei enterotoxin genes, including ST5-SCCmec type II, or a PFGE A pattern with sea, including ST239-SCCmec type III.

• Biomedical Science Letters
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• v.26 no.3
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• pp.149-156
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• 2020

We developed a simple and rapid method for detecting vancomycin resistance genes, such as vanA and vanB, using loop-mediated isothermal amplification (LAMP). To identify not only vancomycin resistance genes, but also the genus Enterococcus, primers were designed for vanA, vanB, and 16S rRNA. Screening for vancomycin susceptibility in Enterococcus was performed using Etest (bioMérieux Inc). The results of the LAMP assay were compared to those of real-time RT-PCR. The optimal conditions for the LAMP assay were 65℃ for 60 min. The detection limits of the LAMP assay for vanA, and vanB were 2 × 10² copies/reaction. Compared to RT-PCR, the sensitivities and specificities of LAMP for 16S rRNA, vanA, and vanB were 100/100%, 100/100%, and 100/100%, respectively. The vanA genotype-vanB phenotype accounted for 57.5% (46/80) of the vancomycin-resistant Enterococci samples collected from 2016 to 2019. In conclusion, the LAMP assay developed in this study showed high sensitivity and specificity for vancomycin-resistant genes. Moreover, due to the simplicity and rapidity of the LAMP assay, its use can be very useful in clinical microbiology laboratories.

• Food Science and Biotechnology
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• v.15 no.4
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• pp.511-515
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• 2006

Antibiotic resistance of foodborne pathogens adapted to garlic (Allium sativum Linn.) was determined in order to understand the relationship between antibiotic resistance and garlic. The Gram (-) strains of Escherichia coli and Salmonella typhimurium and the Gram (+) strains of Bacillus cereus and Staphylococcus aureus were subcultured consecutively in a garlic broth, and the surviving colonies on the agar were selected as the adapted strains. Minimal inhibitory concentrations (MIC) for 15 antibiotics on the adapted strains were determined on Muller-Hinton Infusion agar. Adaptation to 1.3%(v/v) garlic juice increased MIC for vancomycin, aminoglycoside, and erythromycin on B. cereus, and for ampicillin and erythromycin on E. coli O157:H7. MIC of aminoglycosides, chloramphenicol, and vancomycin on the adapted S. aureus increased. The adapted S. typhimurium was more resistant to penicillin and vancomycin than the non-treated strain. The adapted S. typhimurium and S. aureus lost their antibiotic resistance in non-garlic stress conditions. However, the adapted B. cereus was still resistant to erythromycin and vancomycin, and the adapted E. coli was also resistant to erythromycin. Antibacterial garlic might increase the antibiotic resistance of E. coli, B. cereus, S. aureus, and S. typhimurium and this resistance can continue even without the stress of garlic. Therefore, garlic as a food seasoning could influence the resistance of such pathogens to these antibiotics temporarily or permanently.

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.343-352
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• 1999

Vancomycin resistant enterococci (VRE) have emerged as an important nosocomial pathogen. Since 1989 the Center for Disease Control, United States, has reported a rapid increase in the incidence of enterococcal bacteremia and endocarditis infection by VRE. It was suggested that the use of avoparcin was associated with the appearance of VRE in animal husbandry. To date, several detection methods have been used based on conventional methods of culture and gene detection. However, these methods have some limitations such as time-consuming, laborious and additional differential needs. Therefore, In this study a multiplex PCR method was established to detect and differentiate resistance types of enterococci which specifically amplify the four van genes encoding vancomycin resistance elements. Using the method, we investigated the incidence rates and types of VRE from farms using or not using avoparcin. A total of 1091 animal fecal samples were collected from 70 pig and 32 poultry farms. A total of 425 of enterococci were isolated from samples. Of the 425 isolates, 11 of the them showed a pattern of high-level vancomycin resistance (MIC : $64{\sim}256{\mu}g/ml$) which was associated with the presence of the vanA or vanB gene. Fifty-seven isolates showed a pattern of low-level vancomycin resistance (MIC : $3{\sim}8{\mu}g/ml$) associated with the vanC-1 or vanC-2 gene. Interestingly, all isolates with high-level vancomycin resistance were from farms that have never used avoparcin. Moreover, the high-level VRE isolation rate in Korea (2.58%) was much lower than that of other countries (50% in England, 7% in Belgium) where avoparcin have been used. In conclusion, the multiplex PCR method established in this study could be applied for detection of VRE.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.5
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• pp.714-720
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• 2012

To evaluate the antibiotic risk of $Enterococcus$ in fermented soy paste, $Enterococcus$ spp. were isolated and identified from 31 $Cheongkukjang$ and 17 $Doenjang$, samples. Exactly 123 $Enterococcus$ spp., 119 from $Cheongkukjang$ and four from $Doenjang$, were ultimately isolated. The most frequently collected $Enterococcus$ isolates in $Cheongkukjang$ were 69 strains of $E.$ $faecium$ and 20 strains of $E.$ $faecalis$. All four $Enterococcus$ spp. from $Deonjang$ were identified as $E.$ $faecium$. All isolates were sensitive to ampicillin, chloramphenicol, penicillin, and tetracyclin $E.$ However, they showed broad spectra from sensitivity to resistance to erythromycin, ripampin, and streptomycin. Vancomycin minimum inhibition concentration (MIC) of $Enterococcus$ spp. from $Cheongkukjang$ ranged from 0.25 to 8 ${\mu}g/mL$. Almost all strains were sensitive to vancomycin, but eight strains showed intermediate resistance to vancomycin. Seventeen strains showing the highest MIC of 8 ${\mu}g/mL$ among all isolates were evenly distributed among $E.$ $faecalis$, $E.$ $faecium$, $E.$ $gallinarum$, and $E.$ $casselifalvus$, in which the strong resistant genes of $van$A and $van$B for vancomycin were not detected. Overall antibiotic resistance of $Enterococcus$ isolates was relatively low and particularly low vancomycin resistance was similar to those of $Enterococcus$ isolates obtained from other foods. Therefore, the antibiotics resistance of $Enterococcus$ and especially vancomycin-resistant $Enterococcus$ spp. from $Cheongkukjang$ and $Doenjang$, is not hazardous.

• Biomedical Science Letters
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• v.6 no.3
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• pp.201-208
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• 2000

The vancomycin, one of the family of glycopeptide antibiotics, inhibits the synthesis of bacterial cell wall peptidoglycan and has been widely used against gram-positive bacterial infections, especially for a treatment of methicillin resistant S. aureus infection. However, clinical isolate which was intermediately resistant to vancomycin (Mu50: MIC 8 $\mu\textrm{g}$/ml) was isolated in recent years. In this study we performed vancomycin susceptibility test with the increment method and population analysis with clinical isolates S. aureus. Also we did several kinds of tests with three selected isolates (s129: MIC 7 $\mu\textrm{g}$/ml, s134: MIC 7 $\mu\textrm{g}$/ml, s135: MIC 8 $\mu\textrm{g}$/ml) to find out possible mechanism of vancomycin resistance. As a result, the prevalence of vancomycin resistant S. aureus isolates among S. aureus strains resistant to methicillin was 23.3% (25/107). The vancomycin resistances of isolated strains of S. aureus were between those of Mu5O and Mu3 strains. By PCR analysis, none of the isolates with decreased vancomycin susceptibility contained known vancomycin resistant genes such as vanA, vanB, vanC1, vanC2, and vanH. Major bands of 81 kDa, 58 kDa, 33 kDa, 28 kDa were demonstrable in whole cell lysates by SDS-PAGE from all three isolates as well as reference strains. And especially,45 kDa protein was overproduced in Mu50 strains. Among them increased production of NAD$^{+}$-linked-$_{D}$-lactate dehydrogenase (dnLDH) were detected from one clinical strain (s135) and Mu5O strain. From these data, we suggest that the mechanism of vancomycin resistance in these isolates are distinct from that in enterococci.