• Korean Journal of Microbiology
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• v.38 no.4
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• pp.299-305
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• 2002

Clinically isolated Salmonella Enteritidis strain has a multi drug resistance plasmid, which confers ampicillin, chloramphe-nicol, sulfonamide, streptomycin and tetracycline, named pCAST2. We cloned a 7 kb Sacl fragment of pCAST2 which has sulfonamide, streptomycin and tetracycline resistance genes. The 7 kb SacI fragment showed the organization of sulII-strA-strB-tetR-tetA gene cluster which is different from the other clusters reported previously. In this study, we presented the method to detect this cluster by PCR analysis and showed that this cluster was found in Salmonella strains occurred sporadically at Kyungpook province in 2002.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.251-258
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• 1987

A total of 58 strains of Enterobacter species isolated from clinical specimens at Kyungpook National University Hospital in Taegu and Yonsei University Hospital in Seoul were tested for the molecular characterization to investigate the nosocomial infection through the study of R plasmids which might spread among Gram negative organisms regardless of their originated strains. All strains resistant to ampicillin, cefoxitin and cephalothin but susceptible to moxalactam were subjected to the further test for the determination of in detail MIC value against 23 drugs of common use including beta-lactam antibiotics and R plasmid profile analysis. The reistance frequency of strains against carbenicillin (53.4%) was similar to those against chloramphenicol, tobramycin, and sulfisomidine. Though the MIC values of resistance criteria against ceftazidime, aztreonam, imipenem, and norfloxacine in NCCLS manual were not available but MIC ranges of strains tested were very low. There were differences in patterns and frequencies of resistance between the strains isolated in Seoul and Taegu isolates. Seoul isolates showed a tendency of higher multiplicity of resistance than those of Taegu isolates. The resistances against cefoxitin, cephalothin, cefoperazone, cefotaxime, nalidixic acid, and rifampin were not conferred to the conjugally transferable R plasmid. The approximate molecular size of conjugally transferable R plasmids ranged 30 to 151 megadalton, and one or 2 to 3 R plasmids were identified in each transconjugants.

• 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 Journal of the Korean Society for Microbiology
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• v.22 no.1
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• pp.35-53
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• 1987

Multiply resistant Shigella strains isolated in Taegu area were subjected for the characterization of R plasmids. All strains isolated in 1984 and 1985 were susceptible to gentamicin, amikacin, and cephalothin, and most strains were susceptible to kanamycin (Km) and rifampin by agar dilution antimicrobial susceptibility test. The resistance frequency of S. flexneri against ampicillin (Ap) was higher than that of S. sonnei. The strains resistant to sulfisomidine (Su) and trimethoprim (Tp) were found at higher frequency in S. sonnei than in S. flexneri. The most prevalent resistance pattern of S. flexneri was chloramphenicol (Cm) tetracycline (Tc) streptomycin (Sm) Ap, followed by the pattern of CmTcSmSuApTp, CmTcSmSuApTp nalidixic acid, and CmTcSmSuAp in the decreasing order. The antibiogram of CmTcSmSuTp was found to be the most frequent pattern in S. sonnei. The ratio of conjugal transfer of S. flexneri was 47% and 75% of S. sonnei. The average number of plasmid harboring in Shigella was 4 and the size of plasmid ranged 1.3 to 134 megadalton (Mdal). Most S. flexneri carried plasmids of 2 to 3 Mdal and S. sonnei carried those of 3 to 4 Mdal size. The sizes of conjugative plasmids ranged 40-90 Mdal. The incompatibility group (Inc) F II plasmids (54-59 Mdal) were most frequent and rare Inc B plasmids (60 Mdal) of isolates in 1979 and 1980 and Inc FI (87 Mdal) of 1983 isolates were able to be classified by the colony test with standard reference plasmids. The R plasmids of known Inc group were tested for the restriction endonuclease analysis. The pattern of plasmids digested by EcoRl were apparently different by the Inc group but there was no significant difference between species or by the resistance patterns. Nonconjugative plasmids and their phenotypes were identified by transformation test. The transformants were resistant to less than two drugs. Colicin producing transformants carried the Col plasmid of 3.7 or 3.9 Mdal size. $Ap^r$ plasmids derived from S. sonnei were found to be mobilized by transfer factor RT641 to E. coli #CS100. $Ap^r$ plasm ids of same size shared by S. flexneri, S. sonnei, and E. coli were digested with Pstl. All of them showed two restriction fragments of 2.8 kilobase(kb) and 0.7kb. Other plasmids ($Sm^r\;Su^r$) derived from S. flexneri, S. boydii, and S. sonnei were digested with Pstl and they showed same restriction fragment patterns of 3.1kb and 2.9kb. The plasmid profiles of three strains of S. sonnei producing colicin and showing same resistance pattern of CmTcSmSuApTpKm appeared to be similar. Restriction patterns by EcoRl and the behavior of plasmids in conjugation or transformation process were also similar between those plasmids. The restriction patterns were significantly different between the plasmids of Inc FI group and those of unclassified Inc group.

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.21-34
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• 1982

One hundred and twenty-one strains each of Escherichia coli isolated from stools of 60 patients who received various antimicrobial drugs in hospital for more than one week and apparently healthy 60 students who have no history of taking antimicrobial drugs during recent one month, were tested for their resistance to 13 antimicrobial drugs. The frequency of resistance strains was highest to tetracycline with 69.2%, and followed by streptomycin(Sm), sulfisomidine(Su), chloramphenicol(Cm), ampicillin(Ap), and carbenicillin(Cb) in the decreasing order, ranging from 61.2% to 39.3%. Strains resistant to kanamycin(Km), cephaloridine(Cr), and trimethoprim(Tp) occupied about one-fourth of strains, and only four strains were resistant either one or more of nalidixic acid, gentamicin and amikacin, and no strain was resistant to rifampicin. The frequency of resistant strains to Cm, Ap, Km, Cr, and Cb was much higher among patient isolates than student strains, but strains resistant to the other drugs showed almost the same frequencies between patient and student isolates. There was a marked difference in average minimum inhibitory concentrations of between resistant and susceptible strains, suggesting that the resistance to drugs is the plasmid origin. Seventy-six percent of strains were resistant to one to 10 drugs tested, and no much difference was observed between strains from patients and students. However, strains resistant to four or more drugs were much more frequently found among patient isolates than student strains, with the increasing tendency of multiply resistant strains among patient isolates following the increase in the number of resistant drugs. The transfer of drug resistance by conjugation was tested and 98 strains(67.5%) among 145 which were resistant to two or more drugs were found to transfer their drug resistance to E. coli. Among 74 strains resistant to 7 or more drugs, all except one transferred the resistance, and the number of strains with transferable resistance decreased, as the number of resistant drugs decrease. A R plasmid from randomly selected p13 strain was tested for the incompatibility group, and the plasmid was classified into Inc F II. R plasmM DNA bands were identified by polyacrylamide gel electrophoresis.

• Korean Journal of Veterinary Service
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• v.32 no.1
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• pp.61-76
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• 2009

At the present study, it was aimed to explore the molecular genetic characterization of multiple antimicrobial resistant Salmonella spp. isolates from pigs and cattle. A total of 138 Salmonella Typhimurium (S. Typhimurium) isolates were typed with phage, among them, 83.3% of S. Typhimurium tested could divide into a 10 phage types. Definitive type 193 (DT193) (25.4%) and DT195 (24.6%) were exhibited as the dominant types. DT104 and U302 were found from pigs and cattle. On the other hand, S. Enteritidis had 6 phage types, of them, phage type 21 (PT21) and PT11b were the popular types. In the plasmid profiles, 135 of S. Typhimurium isolates were exhibited 1 to 6 plasmid bands which molecular weight ranged from 90 to 2kb. 35 isolates (25.4%) harbored a 90kb plasmid which is thought to be the serotype specific virulence plasmid. Two of twenty five S. Enteritidis had common plasmids at 2 and 1.5kb. With multiplex polymerase chain reaction, virulence genes (invA and spvC) were detected from all Salmonella spp. from 167 of S. Typhimurium, S. Enteritidis and chloramphenicol resistant S. Schwarzengrund, but some drug resistant genes, such as PSE-1, cml/tetR and flo were not determined but other drug resistant genes, for example TEM and int were found. The detection rates of spvC, TEM and int gene was 35.3%, 29.3% and 72.5%, respectively. The TEM gene was highly popular in S. Typhimurium, which was detected from ampicillin and amoxicillin resistant strains as 95.9%. int gene was able to detect from all the isolates identified as multidrug resistsnt (MDR), particularly DT193 was thought as the most prevalent virulence and multidrug resistance isolate. The major plasmid profile and drug resistance pattern of DT193 were 90, 40, 10.5, 6.3, 3.0kb and ACCbDNaPSSuT, respectively. MDR was commonly found in other phage types, particularly DT104, U302 and DT203.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.295-300
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• 1987

Salmonella strains isolated from blood in the Dong-san hospital, Taegu during the period from 1971 to 1986 were studied for species distribution, drug resistance, and R plasmids. The number of strains was 2,527 and all of them were classified into S. typhi and S. paratyphi A. Approximately 300 strains were isolated in the period from 1974 to 1976 and 1978, 268 in 1982, and 204 in 1983, but the numbers isolated in the 1980's have a tendency to decrease as compared with those of the 1970's. S. typhi occupied 85% or more of strains isolated until 1976, but the isolation frequency decreased yearly with some variation, and S. paratyphi A increased gradually from 1974. Only 4 strains of S. paratyphi A were resistant to some drugs, and the resistance was not transferred to E. coli by conjugation. S. typhi resistant to drugs were 15 in 1971 through 1973, 24 in 1974, and 13 in 1975, but afterwards only few resistant strains were isolated. These strains were resistant to two or more drugs; chloramphenicol(Cm), tetracycline(Tc), streptomycin(Sm), sulfisomidine(Su), ampicillin(Ap), and kanamycin(Km) and no strain resistant to other drugs tested was found. Strains resistant to 3 or less drugs didn't transfer the resistance to E. coli by conjugation. There were 15 strains resistant to four or more drugs, and were isolated in years from 1972 to 1976. These strains transferred the resistance to E. coli, and the resistance was considered to be mediated by R plasmids. Transfer frequency was higher at $25^{\circ}C$ than at $37^{\circ}C$ and patterns of transferred resistance were Cm, Tc, Sm, Su; Ap, Km; Cm. R plasmids having markers of Cm, Tc, Sm and Su were classified into Inc H1.

• The Journal of the Korean Society for Microbiology
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• v.12 no.1
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• pp.11-18
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• 1977

A total of 665 strains of Escherichia coli isolated in Korea from stools of patients who were treated with antimicrobial drugs, doctors, and students were tested for the drug resistance and distribution of R plasmids. Approximately 25 to 41% of isolates were resistant to chloramphenicol, tetracycline, streptomycin, sulfisemidine and ampicillin(AP), and 9.5% were resistant to kanamycin. Nalidixic acid-resistant strains were only rarely encountered. The prevalence of resistant strains was significantly higher among patients than doctors and students. Strains multiply resistant to four or more drugs were significantly more prevalent among patient isolates than those from doctors and students, while no difference on the incidence of strains resistant to three or less drugs was noted among isolates from the three groups. The persons carrying strains resistant to four or more drugs were more frequently found among patients than doctors and students. Quite large proportions of drug-resistant strains transferred their resistance to drug-sensitive E. coli, with frequent transfer of whole resistance and AP resistance. Strains having higher multiplicity of resistance showed a tendency toward higher incidence of resistance transfer, irrespective of the origins of strains.

• Korean Journal of Veterinary Research
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• v.26 no.1
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• pp.79-85
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• 1986

This paper deals with the isolation of Salmonella on 3 herds during the period from May 1984 to May 1985. Isolated Salmonella were examined for serotypes, adhesive fimbriae, antibiotic susceptibility and detection of R plasmid. The results were as follows; Of total 1505 cattle, 24 Salmonella were isolated from 18 cattle (1.2%) and their serotypes were S. enteritidis (9 strains), S. derby(4 strains), S. infantis (1 strain), $C_1$ group(8 strains), $C_2$ group(1 strain) and untypable(1 strain). Twelve strains of Salmonella were isolated from 227 cattle with diarrhea and their serotypes were S. enteritidis(4 strains), S. derby(3 strains), S. infantis(1 strain), $C_1$ group(3 strains) and untypable(1 strain). The isolation rate of Salmonella in cattle varied from 1.6 to 0.9% in 3 herds, it was higher in summer and autumn and lower with the age. Of total 24 strains, 23 were adhesive type 1 fimbriae. Antibiotic susceptibility test of Salmonella isolated was performed by the agar dilution method, using 9 antibiotics as follows: ampicillin(Am), chloramphenicol(Cm), gentamicin(Gm), kanamycin(Km), nalidixic acid(Na), rifampicin(Rf), streptomycin(Sm), sulfadimethoxine(Su) and tetracycline(Tc). All the strains tested were susceptible to Am, Cm, Gm, Km, Na, Rf and Tc. Of total 24 strains, 23(95.8%) were resistant to Su and 14(58.3%) to Sm. Of the 23 resistant Salmonella strains, all the strains were found to carry R plasmid. Among them, two strains which had the R plasmid conferring SmSu resistance was transferable at $25^{\circ}C$ only.

• Korean Journal of Veterinary Research
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• v.26 no.1
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• pp.69-77
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• 1986

The purpose of this study was the examination for presence of K99 antigen (K99), enterotoxigenicity, 0-groups, colicin and antibiotic susceptibility among E. coil isolated from calves and cows. A total of 49(18.7%) among 262 strains, isolated from 30(26.5%) out of 113 calves and cows, possesed K99, and thirty three of 49 $K99^+$ strains produced ST. Of the strains of diarrheal calf origin which less than 15 days old, a high correlation was observed between enterotoxigenic ability and K99: 92.3% of the $K99^+$ strains produced heat stable enterotoxin(ST). In O group typing of 33 $ST^+$ strains, they belonged to O20(48.4%), O8(9.1%), O9(6.1%), O139(6.1%), O149(6.1%), O101(3.0%), O115(3.0%), except six which were untypable or autoagglutinable. Of 262 E. coil isolates, 30 strains(13.3%) produced colicin and K99 were detected in 6 strains. One hundred eighty eight strains(71.8%) of 262 E. coil isolates were resistance to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, tetracycline, alone or in combination thereof. One hundred and fourteen(60.6%) out of 188 drug resistance strains carried R factor($R^+$) which were transferable to the recipients by conjugation. Sixty five $R^+$ strains(57.0%) carried thermo-sensitive R plasmid.