• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.5
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• pp.626-633
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• 2023

Vibrio spp. are aquatic bacteria that are ubiquitous in warm estuarine and marine environments. Especially, V. vulnificus and V. cholerae are currently known to cause potentially fatal infections in humans. This study investigated the distribution and antibiotic resistance of V. vulnificus and V. cholerae isolated from coastal areas of Gyeongsangnam-do in 2022. A total of 252 samples of water, shellfish and coastal sediment were collected from 7 locations along the coast, and 124 samples of fishery products were collected from markets. Among the 252 samples, forty-four V. vulnificus (11.7%) and fourteen V. cholerae non-O1/non-O139 (3.7%), none of which carried the ctx gene, were isolated. Out of the 124 samples, 6 (4.8%) tested positive for V. vulnificus and V. cholerae was not detected. The isolation rates of V. vulnificus and V. cholerae showed a significant correlation with environmental factors such as seawater temperature and salinity. In an antibiotic resistance test, V. vulnificus was susceptible to amikacin, gentamicin, imipenem trimethoprim/sulfamethoxazole, and ciprofloxacin, but resistant to cefoxitin (100.0%), followed by tetracycline (9.1%). Multidrug resistance was also observed. Continuous monitoring of Vibrio pathogens with water temperature and salinity is expected to help reduce the outbreaks, and rational use of antibiotic agents is needed to prevent the accession of antibiotic-resistant microorganisms in aquatic ecosystems.

• KSBB Journal
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• v.5 no.3
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• pp.247-253
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• 1990

These studies were carried out to obtain the transformant from carrot cells by using binary vector pGA472 with NPT II gene to confer kanamycin resistance in the plant cells. The binary vector pGA472 was mobilized from E. coli MC1000 into A. tumefaciens strains isolated in the Korea, C23-1. K29-1, and disarmed Ti-plasmid PC2760, and A28l using a tri-parental mating method with E. coli HB101/pRK2013. Transconjugants, C23-1/pGA472, K29-1/pGA472, PC2 760/pGA472 and A28l/pGA472 were obtaind on the minimum AB media containing tetracycline and kanamycin, were comfirmed to hold the Ti-plasmid and pGA472 binary vector on the 0.7% agarose gel. Transformed carrot calli were initiated on the MS media supplemented with l00$\mu\textrm{g}$/ml kanamycin and 250$\mu\textrm{g}$/ml carbenicillin after co-cultivation of carrot explant and transconjugant Agrobacteria. Selected callus was grown vigouousley for subculture on the medium containing 100$\mu\textrm{g}$/ml kanamycin, thus indication that the selected callus was transformed with NPT II gene.

• Korean Journal of Veterinary Research
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• v.51 no.4
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• pp.267-275
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• 2011

Methicillin-resistant Staphylococcus (S.) aureus (MRSA) is one of the most important nosocomial pathogens worldwide and the emergence of this strain has become a major clinical problem. In this study, we investigated the prevalence of MRSA and their genetic characteristics in 69 S. aureus isolated from humans and animals. In human isolates, higher antimicrobial resistance rates were observed against penicillin (80.6%), followed by erythromycin (11.9%) and tetracycline (9.0%). All of them were susceptible to clindamycin, enrofloxacin, novobiocin, pirlimycin, trimethoprim/sulfamethoxazole and vancomycin. The resistance patterns in animal isolates were similar to those of human isolates. Two (2.9%) MRSA strains were isolated from human (n = 1) and animal (n = 1), and these isolates were confirmed as carrying the mecA gene. One isolate originating from human was resistant to 7 drugs and the other isolate derived from animal was resistant to 11 drugs. Staphylococcal cassette chromosome mec (SCCmec) variant IIIB was identified in animal isolate but SCCmec type of an isolate from human was not exactly determined. Two MRSA isolates showed unrelated PFGE pattern between them. Our results indicated although the frequency of MRSA isolates from humans and animals was low, a continuous surveillance and monitoring should be called for to prevent the contamination and spread of MRSA in the community. To our knowledge, this is the first time that SCCmec type variant IIIB was detected from animals in Korea.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.3
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• pp.220-226
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• 2014

Sixty-seven Vibrio parahaemolyticus isolates from surface seawater from the Wando area, on the southern coast of Korea, were analyzed for their susceptibility to 15 different antimicrobials and the presence of virulence genes. According to the disk diffusion susceptibility test, all of the strains studied were resistant to ampicillin and oxacillin, while decreasing percentages were resistant to vancomycin (64.2%), streptomycin (56.7%), amikacin (31.3%), kanamycin (22.3%), cephalothin (20.9%), erythromycin (10.4%), ciprofloxacin (4.5%), and tetracycline (3.0%). All of the strains were susceptible to five antimicrobials: chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. Fifty-nine isolates (88.1%) were resistant to three or more classes of antimicrobial and defined as multidrug resistant, and two strains were resistant to seven antimicrobial agents. The minimum inhibitory concentration (MIC) of the 67 V. parahaemolyticus isolates to ampicillin and oxacillin ranged from 512-2,048 and $64-512{\mu}g/mL$, respectively. All 67 isolates were also examined for the presence of the tdh and trh virulence genes using the polymerase chain reaction (PCR). However, no isolates possessed either tdh or trh. The VPA0477 (${\beta}$-lactamase) gene, present in all of the tested strains, was validated as a new specific marker gene in PCR assays for the accurate detection and identification of V. parahaemolyticus.

• Korean Journal of Veterinary Service
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• v.47 no.1
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• pp.27-33
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• 2024

At the present study, it was aimed to explore the states of antimicrobial resistant Staphylococcus aureus isolates from 1,360 chickens, pigs and cattle carcass (400 chickens, 480 pigs and 480 cattle) in Daegu province from January 2022 to December 2022. Among 1,360 samples, 81 of S. aureus were isolated cattle (1.4%), pigs (7.7%) and chickens (9.2%). In antimicrobial susceptibility test, all of the isolates were demonstrated susceptibility to rifampin. But the isolates were showed resistance other antibiotics in order of tetracycline (62.9%), ciprofloxacin (62.9%), tobramycin (58.0%), gentamicin (51.8%), amikacin (40.7%), penicillin (39.5%), clindamycin (35.8%), enrofloxacin (33.3%), trimethoprim/sulfamethoxazole (30.8%), oxacillin (30.8%), minocycline (29.6%), erythromycin (25.9%), quinupristin/dalfopristin (20.9%), chloramphenicol (12.3%), cefoxitin (9.8%). Among the 81 S. aureus isolates, 25 (30.8%) methicillin-resistant staphylococcus aureus (MRSA) were observed. Seven (28.0%) of 25 MRSA harbored mecA gene. About 96% of MRSA were multidrug resistance to at least 3 more drugs. A continuous monitoring and surveillance program to prevent antimicrobial resistance in livestock products is demanded.

• Journal of Food Hygiene and Safety
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• v.39 no.3
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• pp.231-238
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• 2024

Vibrio parahaemolyticus is a major seafood-borne pathogen commonly detected in marine environments. In Korea, V. parahaemolyticus-induced foodborne illnesses account for 7.5% of bacterial pathogen-related food poisonings. Moreover, the amount of antimicrobial agents used in aquatic cultures is continuously increasing. In this study, we isolated V. parahaemolyticus from seafood samples and performed antimicrobial susceptibility tests using the microbroth dilution method. Furthermore, using whole-genome sequencing, we identified antimicrobial resistance genes, virulence genes, and sequence types (STs). We could isolate V. parahaemolyticus from 47 (59.5%) of the 79 seafood samples we purchased from retail markets in Seoul and Chungcheong provinces. Antimicrobial susceptibility tests revealed that 2 and all of the 47 isolates were ampicillin-resistant (4.3%) and susceptible to all tested antimicrobial agents (100%), respectively. The genotype analysis revealed that all isolates carried beta-lactam-, tetracycline-, and chloramphenicol-associated antimicrobial resistance genes. However, we could detect fosfomycin resistance only in one isolate. Concerning the virulence genes, we detected T3SS1 and T3SS2-associated genes in all and one isolate, respectively. However, we could not detect the tdh and trh genes. Of the 47 isolates, 17 belonged to 15 different STs, including ST 658 with 3 isolates. The rest 30 isolates were identified as 25 new STs. The results of this study support the need for operating a continuous monitoring system to prevent foodborne illnesses and the spread of antimicrobial resistance genes in V. parahaemolyticus.

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.241-247
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• 2000

The incidence of verotoxin-producing Escherichia coli(VTEC) was surveyed in domestic foods including hamburger, raw meats and vegetables from 1997 to 1999. The molecular biological characteristics of the isolates were analyzed. Three VTEC strain were isolated from 1,700 samples. Serotypes of those isolates were 0157 : H7, 026 H4, and 056 : Hl2, respectively. Serotype O26 : H4 produced VT I and VT II, and 055 Hl2 isolate produced VT I, however the 60 MDa plasmid DNA and eae gene were not found from both strains. One 0157 : H7 isolate produced VT II and harbour 60 MDa plasmid DNA, however eae gene was not found in the strain. Although they produced VT, it seemed that the virulence of two strains were relatively weak because of the lack of the eae gene. In addition, the serotype O157 : H7 isolate resistant to ampicillin and streptomycin, while isolates of serotype O26 : H4 and O55 : Hl2 were multi-resistant to antibiotics including ampicillin, carbenicillin , cephalothin, trimethoprim/sulfamethoxazole and tetracycline. Supernatants of cultures of all three isolates were showed cytotoxic effect to vero and HeLa cell

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.202-209
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• 2016

This study is aimed to determine the correlation between the toxin gene types and antibiotic resistance from MRSA (methicillin-resistant Staphylococcus aureus). Fifty-two strains of MRSA, between January 2014, and December 2014, were isolated from clinical specimens obtained from 2,664 cases in the intensive care unit of a hospital in Suncheon, Jeonnam, Korea. Genes encoding mecA, enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET), and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Toxin genes (seg and sei) were present in 40 strains (76.9%), followed by tst in 34 strains (65.4%). Other genes (eta, etb, sea, sed, see, seh, sej, and pvl) were not detected. Forty strains (76.9%) of MRSA had 2 or more toxin genes simultaneously; 5 coexistent toxin-genes (seb, sec, seg, sei, tst) were the most common in 28 strains (53.8%), and 6 strains (11.5%) had seg and sei genes. The coexistence of genes were 72.5~100%, showing a high correlation among genes (seb, sec, seg, sei and tst). As strains (seb, sec, tst) that had particular toxin genes (seb, sec, seg, sei, tst) in multiple showed 100% resistance to ciprofloxacin, clindamycin, erythromycin, we were able to find that seb, sec, and tst genes have a close relationship to the aforementioned antibiotics. It showed a higher resistance to ciprofloxacin, clindamycin, erythromycin, and tetracycline compared with strains that had toxin genes independent from multiple toxin genes.

• Journal of fish pathology
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• v.32 no.2
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• pp.97-104
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• 2019

The purpose of this study was to investigate the antimicrobial resistance and resistance transfer of Vibrio parahaemolyticus and Morganella morganii isolated from fish products purchased from fish markets in Yeosu April - December 2017. These bacteria were identified by biochemical test and PCR results, and the transfer of antimicrobial resistance was confirmed by the broth mating method. To isolate the transconjugants formed during conjugation, TSA medium containing 50 ㎍/ml of ampicillin (AMP), and 150 ㎍/ml of streptomycin (SM) or 30 ㎍/ml of oxytetracycline (OT) was used. M. morganii isolates showed low susceptibility to AMP, amoxicillin (AML), and colistin (CT), erythromycin, OT, and tetracycline, compared to V. parahaemolyticus resistance to AMP, AML, and CT. The conjugation of V. parahaemolyticus or M. morganii with Escherichia coli resulted in the separation of V. parahaemolyticus and M. morganii showing SM resistance as transconjugants. Meanwhile, Edwardsiella tarda transconjugants showing AMP and AML resistance were obtained from the broth mating of V. parahaemolyticus and E. tarda. But the transfer of the VPA0477 which is a β-lactamase gene of V. parahaemolyticus was not confirmed. These results suggest that resistance transfer between pathogenic bacteria is bidirectional and progresses in a wide variety of patterns.