• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.6
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• pp.850-857
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• 2022

Seventy-five Vibrio parahaemolyticus isolates from the surface seawater of the Geum River Estuary area, on the west coast of Korea, were analyzed for the presence of virulence genes and susceptibility to 17 different antimicrobials. All 75 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction; Only one of the isolates possessed the tdh or trh gene. According to the results of disk diffusion susceptibility tests, all of the strains were resistant to penicillin G, 92.0% were resistant to ampicillin, 82.7% were resistant to amoxicillin, 2.7% were resistant to ciprofloxacin, 2.7% were resistant to trimethoprim, 1.3% were resistant to cephalothin, and 1.3% were resistant to erythromycin. However, all of the strains were susceptible to amikacin, cefoxitin, chloramphenicol, gentamycin, kanamycin, nalidixic acid, nitrofurantoin, rifampin, streptomycin, and tetracycline. The average minimum inhibitory concentrations for ampicillin for V. parahaemolyticus was 557.6 ㎍/mL. These results not only provide novel insight into the necessity for seawater sanitation in Geum river estuary area, but they help reduce the risk of contamination of antimicrobial-resistant bacteria.

• International Journal of Industrial Entomology and Biomaterials
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• v.25 no.2
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• pp.195-203
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• 2012

In reared populations of Allomyrina dichotoma, commercial insects, the skin of last instar larvae was changed softer with opaque white, and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined their intestinal bacterial florae using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at $70^{\circ}C$ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on $MYPGP_{NAL}$ agar medium($12.5{\mu}g/ml$ of nalidixic acid) at $30^{\circ}C$. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn't be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

• Journal of Microbiology and Biotechnology
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• v.20 no.6
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• pp.1042-1052
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• 2010

Forty-seven Salmonella Typhimurium (33 zoonotic, 14 clinical) strains were tested for antimicrobial resistance using the standard disk diffusion method. The presence of relevant resistance genes and class 1 integrons were investigated by using PCR. Pulsed-field gel electrophoresis (PFGE) and plasmid profiling were carried out to determine the genomic diversity of Salmonella Typhimurium. Approximately 57.4% of the S. Typhimurium strains were multidrug resistant (MDR) and showed high resistance rates to tetracycline (70.2%), sulfonamides (57.4%), streptomycin (53.1%), ampicillin (29.7%), nalidixic acid (27.6%), kanamycin (23.4%), chloramphenicol (21.2%), and trimethoprim (19.1%). Resistance towards cephalosporins was noted for cephalothin (27.6%), cephradine (21.2%), amoxicillin clavulanic acid (17.0%), and cephalexin (17.0%). Resistance genes, $bla_{TEM}$, strA, aadA, sul1, sul2, tetA, tetB, and tetC, were detected among the drug-resistant strains. Thirtythree strains (70.2%) carried class 1 integrons, which were grouped in 9 different profiles. DNA sequencing identified sat, aadA, pse-1, and dfrA genes in variable regions on class 1 integrons. Thirty-five strains (74.4%) were subtyped to 22 different plasmid profiles, each with 1-6 plasmids (2.0 to 95 kb). PFGE subtyped the 47 strains into 39 profiles. In conclusion, high rates of multidrug resistance were found among the Malaysian Salmonella Typhimurium strains. The emergence of multidrug-resistant Salmonella Typhimurium to cephalosporin antibiotics was also observed. The strains were very diverse and no persistent clone was observed. The emergence of MDR Salmonella Typhimurium is a worldwide problem, and this report provides information for the better understanding of the prevalence and epidemiology of MDR S. Typhimurium in Malaysia.

• The Journal of the Korean Society for Microbiology
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• v.35 no.3
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• pp.239-250
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• 2000

Sixty-eight clinical isolates of Stenotrophomonas maltophilia from inpatients of 2 university hospitals in Taegu were epidemiologically analyzed by using the minimum inhibitory concentrations of 25 antimicrobial drugs, biochemical reaction, pulsed-field gel elctropgoresis (PFGE), and PCR with enterobacterial repetitive intergenic consensus sequences as primer (ERIC-PCR). 1. All the strains were susceptible to minocycline. More than 57% were susceptible to sulfisomidine (Su), ciprofloxacin (Ci), Ofloploxacin (Of), nalidixic acid (Na), and chloramphenicol (Cm), and $19{\sim}35%$ to ceftazidime (Cd), trimethoprim (Tp), Ticacillin-clavulanic acid, and cefoperazone-sulbactam. Most isolates were resistant to ${\beta}$-lactam antibiotics such as ampicillin (Ap), carbenicillin (Cb), cefotaxim (Ct), cefoxitin (Cx), and aminoglycosides including gentamicin (Gm), tobramycin (Tb), amikacin (Ak). 2. All the isolates were multiply resistant of 5 to 17 drugs and showed 40 different resistance pattern types. 3. All the strains showed very similar biochemical reactions except ${\beta}$-galactosidase and nitrate reduction test. Fourteen strains selected randomly were classified 10 different pattern type by PFGE and ERIC-PCR. These two methods showed identical result. Four strains isolated from wound in 1994 showed similar MIC pattern and identical API 20NE profile, PFGE, and ERIC-PCR pattern indicating episodes of cross-infection among patients. These results indicate that PFGE or ERIC-PCR profile has comparable discriminatory power for epidemiological typing of S. maltophilia.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.5
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• pp.564-572
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• 2016

In this study, we isolated and characterized Escherichia coli from mussels and inland pollution sources in or in proximity to the Changseon area on the southern coast of Korea in 2014. A total of 147 strains of E. coli were isolated from 54 mussels and 32 pollution-source samples. The susceptibility of the isolates to 24 antimicrobial agents was analyzed. The resistance of E. coli isolates to rifampin was highest at 100%, followed by cephalothin (98.6%), tetracycline (91.8%), amikacin (81.0%), ampicillin (79.6%), cefazolin (79.6%), streptomycin (73.5%), piperacillin (70.7%), gentamicin (37.4%), cefoxitin (35.4%), cefamandole (34.7%), tobramycin (29.9%), amoxicillin/clavulanic acid (24.5%), nalidixic acid (21.8%), trimethoprim (19.0%), chloramphenicol (17.7%), cefotaxime (12.9%), trimethoprim (10.9%), ceftazidime (10.2%), aztreonam (7.5%), imipenem (2.7%), cefepime (2.0%), and cefotetan (0.0%). In addition, the antimicrobial resistance of E. coli isolates from inland pollution sources was slightly greater than or similar to that of isolates from mussels.

• Korean Journal of Veterinary Service
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• v.37 no.1
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• pp.19-27
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• 2014

This study was conducted to investigate the antimicrobial resistance, presence of ${\beta}$-lactamase genes and integrons in 83 ESBL-producing Escherichia coli isolated from Nakdong river and Geumho river in Daegu. Among the ${\beta}$-lactam antimicrobials, all isolates were resistant to ampicillin, cephalothin, cefamandole and cefotaxime, followed by piperacillin (98.8%), ampicillin/sulbactam (86.7%), aztreonam (60.2%) and cefepime (59.0%), whereas resistance to piperacillin/tazobacram, ticarcillin/clavulanic acid and cefoxitin was less than 30%. Many of the ESBL-producing Escherichia coli were also resistant to non-${\beta}$-lactams antimicrobials such as nalidixic acid (83.1%), sulfonamides (72.3%), ciprofloxacin (62.7%) and gentamicin (38.6%). All isolates showed resistance to seven or more antimicrobial agents. The most frequently detected gene was $bla_{TEM+CTX-M}$ (49.4%), followed by $bla_{CTX-M}$ (27.7%), $bla_{TEM}$ (6.0%) and $bla_{OXA}$ (1.2%). But $bla_{SHV}$ was not found. Class 1 integrons were found in 61.4% (51 isolates) of isolates, however, class 2 and 3 integrons were not detected. The results showed water from Nakdong river and Geumho river is contaminated with ESBL-producing E. coli isolates. These results suggest the need for further investigation of antibiotic resistant bacteria to prevent public health impacts in the water environment.

• Korean Journal of Veterinary Research
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• v.21 no.1
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• pp.25-31
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• 1981

A total of 250 enteric bacteria (148 Escherichia coli, 41 Klebsiella pneumoniae, 46 Enterobacter spp. and 15 Proteus spp.) isolated from bovine udder infections in 1979 through 1980 were examined for drug resistance and prevalence of R. plasmids. The drug tested were streptomycin (SM), kanamycin (KM), ampicillin (AP), chloramphenicol (CP), tetracycline (TC), gentamicin (GM), oxolinic acid (OA) and nalidixic acid (NA). The detection of R. plasmids was performed with Escherichia coli ML 1410 NAr as the recipient. Of the 148 Escherichia coli isolated, 68(45.9%) were found to be resistant to one or more drugs tested, and about 50% of the resistant strains were multiply resistant. of the 68 drugresistant strains, 13(19.1%) were found to carry R. plasmids which were capable of performing a conjugal transfer. CP resistance was transfered together with the other resistance. Of 41 strains of Klebsiella pneumoniae isolated, 90.2% were resistant to the drugs, alone or in combination thereof. Strains resistant to AP and TC were 63.4%, and 48.8%, respectively. R. plasmids were detected in 78.4% of the drug-resistant strains, and these strains transfered all or a part of their drug resistance pattern. AP and CP resistance were transfered in 100% of AP and CP-resistant strains. Eleven (37.9%) of 29 R. plasmids showed a thermosensitive transfer. Of the 46 strains of Enterobacter spp. isolated, 37(80.4%) were resistant to the drugs tested. A high percentage of resistance was noted for AP(65.2%). All strains resistant to four or more drugs transferred their resistances to Escherichia coli ML 1410, but strains resistant to three or fewer drugs did not transfer the resistances. All of the 15 Proteus strains isolated were resistant to more than two drugs. of them, 6 were quadruple resistance to SM, KM, CP and TC, and 9 were double one to AP and TC. Three (20.0%) of the drug-resistant isolates had R.plasmids conferring AP and TC resistance. GM, OA and NA of the drugs tested were very active to all of 250 Gram-negative enteric bacteria isolated from bovine udder infections.

• Journal of fish pathology
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• v.10 no.1
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• pp.1-14
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• 1997

Major object of this study was to investigate the causative organism of th e diseased cultured fingerling of sea bass, L japonicus. The experimental results are summarized as follows ; Staphylococcus epidermidis, isolated from the liver, kidney, spleen and brain, was considered to be the causative organism. External symptoms of this disease were congestion and hemorrhages in eyes. Anatomical symptoms were hemorrhage of brain, congestion of liver, and slight swelling of kidney and spleen. Growth of the isolates was good on BHIA, HIA and Staphylococcus No. 110. The growth occurred at a range(optimum) of $10\sim45^{\circ}C(35\sim40^{\circ}C)$, 0~9% (1~3%) of NaCl concentration and pH 4~10(8). DNase and coagulase production of all isolated strains were nagative, but was positive in hemolysis. Urease was positive reaction, and novobiocin resistance was nagative. Acid was produced anaerobically from glucose and maltose. Acid was produced aerobically from glucose, galactose, sucrose, maltose and dextrine. But gas was not produced from any carbohydrates. When the isolated strain was injected intramuscularly on fingerling of sea bass, L japonicus, it had virulence at $1.7{\times}10^{10}$ viable cells/$m\ell$ for all fish examined but no virulence at $1.7{\times}10^4$ viable cells/$m\ell$. Bacitracin, Erythromycin and Nofloxacin were observed as bacteriostatic agents to the strain, but Colistin, Gentamicin and Nalidixic acid were not. There were remarkable congestion of the brain, regressive necrosis of the liver, and showed necrosis of the epithelial cells of renal tubules in kidney tissues.

• 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.

• The Korean Journal of Food And Nutrition
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• v.14 no.6
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• pp.548-561
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• 2001

Listeria spp. from sea water, fishes and shellfishes have been troubled in many countries. So we exam ined its distribution rates, biochemical characteristics of a separated strain, growth curve of pH at set times to 4 species of standard strain, and yes or no of growth inhibition for precautionary measure of food poisoning by L. monocytogenes, garlic, mustard, wasabi, and green tea extracts including sensitivity of antibiotics 10 species. As its results, check numbers of its positivity to Listeria spp. were 32 species in total examination body 200 species, and its isolation rates were 16%, L. innocua was 14.0%, L. monocytogenes 1.0%, and L. seeligeri 1.0% by the strain species. All the standard strain of 4 species showed growth inhibition bellow pH 3.0, its pH conditions of the optimum growth at 7.0∼8.0, and its growth was more active in alkali co]tuition than in acid condition. Its growth inhibition examination by garlic extracts had an the worst effects with O.D values of 0.078∼0.210. But the case of mustard and wasabi had weakened effect, and the case of green tea had some effect as the time went by. The results of sensitivity examination of antibiotics 10 species were as fellows. L. innocua of the 16 cases showed sensitivity of 100% in all 5 species, Ampicillin, etc, and Ciprofloxacin showed sensitivity of 43.7% and gentamicin, 93.7%. But tetracycline showed tolerance of 31.3% , cefotaxine. 75%, nalidixic acid, 100%. L. monocytogenes of the 6 cases showed sensitivity of 100% in all 6 species, ciprofloxacin, etc.