• 미생물학회지
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• 제39권3호
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• pp.181-186
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• 2003

형광현미경을 이용한 기존의 직접생균수측정(DVC)법을 변형하여 산림토양의 각 층위에 분포하는 난배양성 토양세균의 정량적 평가를 시도하였다. 기존의 방법에서 사용된 DNA합성저해제(nalidixic acid; 100${\mu}g$/ml, pipemidic acid; 50 ${\mu}g$/ml, piromidic acid; 50 ${\mu}g$/ml)보다 5배 높은 농도를 사용한 결과 토양세균 군집의 세포분열이 배양24시간까지 분히 억제되어 기존의 DVC법보다 10배 이상 높은 계수치를 얻었다. 토양시료를 제균수로 원심 세척한 경우 DVC는 전균수(TDC)의 약 I% 미만을 나타내어 첨가된 기질의 이용능을 정확히 판정할 수 있었다. 형광현미경을 이용하여 DVC법에 의해 검출된 생균수는 전균수의 18-44%를 나타내었고, 평판법(PC)에 의해 계수된 생균수는 전균수의 1% 미만을 나타내어 DVC법이 평판법에 비해 100-2000배 이상 높은 계수치를 나타내었다. 이는 평판법으로는 배양할 수 없는 난배양성(VBNC)세균이 토양 내에 다수 존재해 있음으로 판단되었고, DVC법에 의해 난배양성 토양세균을 정량적으로 검출할 수 있는 가능성이 확인되었다. 영양기질로써 통상농도의 NB배지와 NB를 $10^{-2}$로 희석한 DNB배지를 사용하여 DVC법과 평판법에 의한 계수 결과를 비교한 결과 모든 시료에서 DNB배지에서의 생균수는 NB배지보다 5～10배 이상 높은 값을 나타내어 산림토양의 각 층위에 저영양세균(oligotrophic bacteria)이 다수 분포해 있음으로 추정되었다.

• Animal cells and systems
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• 제3권2호
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• pp.229-236
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• 1999

The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

• 대한수의학회지
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• 제27권2호
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• pp.227-233
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• 1987

This study was conducted to determine the epidemiological characteristics of Campylobacter enteritis. A total of 187 fecal specimens of Korean native goat were examined for the presence of C. jejuni and C. coli by direct plating. Fifty strains isolated were examined for biochemical and serological properties and susceptibility to 19 chemotherapeutic agents. A total of 29(15.5%) C. jejuni and 21 (11.2%) C. coli were isolated from the fecal specimen of 187 Korean native goats. Of the 50 isolates of C. jejuni and C. coli, 29 isolates of C. jejuni grouped as 7 biotypes (1,2,3,4,6,7 and 8) and biotypes 1(34.5%), 2(17.2%) and 3(20.7%) were encountered most frequently. Twenty-one C. coli strains were differentated into biotype I (61.9% of the isolates) and biotype II (38.1%). Of the 29 C. jejuni strains examined, 24(83.0%) were typable by the Lior serotyping scheme and five isolates were non typable. C. jejuni grouped as 8 serotypes, serotype 4(24.1%) and 26(20.7%) were encountered most frequently. In the case of 21 strains of C. coli grouped as 6 serotypes, the most frequent serotypes were 21(28.6%) and 25(23.8%). Total of 50 strains of isolated were all susceptible to amikacin, clindamycin and tobramycine. Overall 85% of isolates were sensitive to erythromycin, doxycycline, chloramphenicol, flume-quine, kanamycin, gentamicin, nalidixic acid, polymyxin B, colistin, tetracycline and ampicillin, but about 65% of isolates were resistant to cefamandole and ethyl hydrocuprein hydrochloride.

• 대한수의학회지
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• 제27권2호
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• pp.235-243
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• 1987

This paper deals with the distribution of Yersinia spp. isolated from the feces or the cecal contents of 1,755 pigs, 558 cows, 428 pigs slaughtered, 271 dogs slaughtered and 91 deer during the period of March 1985 to February 1986. Isolated Yersinia spp. were examined for serotype, biotype and antibiotic susceptibility of Y. enterocolitica. The results were as follows; One hundred and fourty-three stains of Yersinia spp. were isolated from 141(4.5%) out of 3,103 animals examined and their isolates were identified as Y. enterocolitica(138 strains), Y. kristensenii (3 strains), Y. intermedia(1 strain) and Y. pseudotuberculosis(1 strain). Yersinia spp. were isolated from 122(7.0%) of 1,755 pigs in piggeries, 15(3.5%) of 428 pigs slaughtered and 4(1.5%) of 271 dogs slaughtered, but no Yersinia spp. were isolated from cows and deer. The isolation rate of Yersinia spp. in pigs ranged from 5.9~8.0% in piggeries, it was higher in summer and autumn and highest in fattening pigs groups(10.4%), especially. One hundred and thirty-eight Y. enterocolitica isolates belonged to serotype 0 : 3(95 strains), 0 : 8(13 strains), 0 : 5(7 strains), 0 : 9(6 strains), 0 : 1, 2(1 strain) and untypable(16 strains), among them strains of serotype 0 : 3 biotype 3B(91 strains) were predominant. Antibiotic susceptibility test of 138 isolates of Yersinia spp. was performed by the agar dilution method, using 8 antibiotics as follows: ampicillin(Am), chloramphenicol, kanamycin, nalidixic acid(Na), rifampicin(Rf), streptomycin, sulfadimethoxine(Su) and tetracycline. All the strains tested were susceptible to Rf and Na, but resistant to Su, and 136 strains(98.6%) were also resistant to Am.

• 대한수의학회지
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• 제54권2호
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• pp.91-99
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• 2014

An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1605-1612
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• 2016

Quinolone-resistant Salmonella strains were isolated from patient samples, and several quinolone-sensitive strains were used to analyze mutations in the quinolone resistance-determining region (QRDR) of gyrA, gyrB, parC, and parE and to screen for plasmid-mediated quinolone resistance. Among the 21 strains that showed resistance to nalidixic acid and ciprofloxacin (MIC 0.125-2.0 μg/ml), 17 strains had a mutation in QRDR codon 87 of gyrA, and 3 strains had a single mutation (Ser83 → Phe). Another cause of resistance, efflux pump regulation, was studied by examining the expression of acrB, ramA, marA, and soxS. Five strains, including Sal-KH1 and Sal-KH2, showed no increase in relative expression in an analysis using the qRT-PCR method (p < 0.05). In order to determine the genes involved in the resistance, the Sal-9 isolate that showed decreased susceptibility and did not contain a mutation in the gyrA QRDR was used to make the STM (MIC 8 μg/ml) and STH (MIC 16 μg/ml) ciprofloxacin-resistant mutants. The gyrA QRDR Asp87 → Gly mutation was identified in both the STM and STH mutants by mutation analysis. qRT-PCR analysis of the efflux transporter acrB of the AcrAB-TolC efflux system showed increased expression levels in both the STM (1.79-fold) and STH (2.0-fold) mutants. In addition, the expression of the transcriptional regulator marA was increased in both the STM (6.35-fold) and STH (21.73-fold) mutants. Moreover, the expression of soxS was increased in the STM (3.41-fold) and STH (10.05-fold) mutants (p < 0.05). Therefore, these results indicate that AcrAB-TolC efflux pump activity and the target site mutation in gyrA are involved in quinolone resistance.

• 한국수산과학회지
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• 제47권3호
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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.

• 한국동물위생학회지
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• 제32권4호
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• pp.325-334
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• 2009

Salmonella spp. are the important pathogens both economically and clinically in animals as well as human. Some of them have highly zoonotic potentials even though they are asymptomatic in animals. Therefore, the prevalence of Salmonella spp. in animals is highly concerned for human health. The present study was carried out to investigate the prevalence, antimicrobial resistance and PFGE patterns of Salmonella spp. isolated from chickens at slaughterhouse in Incheon area. The overall isolation rate of Salmonella spp. from cloaca and cecum specimens was 7.3 % (37/510). Thirty seven isolates of Salmonella spp. were identified to 5 serotypes; S. Enteritidis, S. Newport, S. Typhimurium, S. Gallinarum, and S. Derby with prevalence of 46.0%, 40.5%, 8.1%, 2.7%, and 2.7%, respectively. Resistance to nalidixic acid was found in 97.3% of Salmonella spp. isolated, followed by streptomycin (16.2%), tetracycline (16.2%), ampicillin (5.4%). Only 6 isolates (16.2%) showed resistance to more than two antimicrobials. In PFGE analysis of chicken and human isolates with Xba I, S. Enteritidis isolates from chicken showed very high similarity over 82.8% and also the similarity was very high in the comparison with human isolates. However, the higher similarity (100%) was observed among chicken isolates of S. Typhimurium. These results suggest the close genetic relatedness of Salmonella spp. isolated from chickens with human.

• 대한수의학회지
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• 제26권1호
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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.

• 대한수의학회지
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• 제48권3호
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• pp.275-285
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• 2008

Antimicrobial drugs are widely used in poultry industry as growth promoters or to control infectious diseases. However, this practice is reported to have caused high resistance to antimicrobial drugs in normal chicken flora and pathogens. Antimicrobial resistance to Escherichia coli (E. coli) from chicken has been mainly reported in normal flora, but rare in pathogenic organism in Korea, recently. Therefore, this study was conducted to investigate prevalence of antimicrobials resistance, transfer of R plasmid, and association between antimicrobial drug resistance and O serotype of 203 pathogenic E. coli from poultry in Korea during the period from April 2003 to December 2005. These isolates showed a high resistance to tetracycline (Tc, 93.6%), nalidixic acid (Na, 92.6%), streptomycin (Sm, 81.8%), ampicillin (Ap, 77.3%), ciprofloxacin (Ci, 70.9%), sulfisoxazole (Su, 66.5%), and trimethoprim (Tp, 58.1%). Two hundred-one (99.0%) of the isolates were resistant to one or more drugs. They showed 57 different resistant patterns, and the most prevalent resistant pattern among them was Tc, Sin, Su, Ap, Tp, Ci, Na. Sixty-eight (33.8%) of the isolates transferred all or a part of their antimicrobial resistant pattern to the recipient strain by R plasmid. The most common antimicrobial resistant pattern was Tc, Sm, Su, Ap, Tp, Ci, Na in serotype O78, O88 and O15, respectively. These results exhibit high individual and multiple resistance to antimicrobials of pathogenic E. coli from poultry in Korea. They also suggest the needs for surveillance to monitor antimicrobial resistance in pathogenic bacteria that can be potentially transmitted to humans from food animals and to regulate the abuse of antimicrobials on food-producing animals in Korea.