• Journal of Korean Society of Forest Science
• /
• v.87 no.2
• /
• pp.164-172
• /
• 1998

This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra${\times}$P. maximowiczii was MS medium supplemented with $0.1mg/{\ell}$ NAA, $0.5mg/{\ell}$ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was $100mg/{\ell}$ kanamycin or $60mg/{\ell}$ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7%. The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

• Korean Journal of Veterinary Service
• /
• v.20 no.1
• /
• pp.103-125
• /
• 1997

An epidemiologic study on pleuropneumonia in the slaughter pigs(Chonan and Asan area, Chungnam province, Korea) during the period of January 1994 through December 1995 was conducted. Isolation of A pleuropneumoniae was attempted in 425 pigs with pneumonic lesions. Biochemical properties, antimicrobial susceptibility, serotypes and pathogenicity of isolated A pleuropneumoniae were investigated. In addition, outer membrane protein(OMP) of the Isolates were extracted to determine its properties and immunogenicity in both mice and piglets The results obtained through this study were summarized as followed ; 1. Of 3, 395 slaughter pigs, pleuropneumonia was observed in 425 pigs(10.6%). A pleuropneumoniae was isolated from 22 pigs(5.2%) out of 425 pigs with pneumonic lesions. The biochemical properties of all isolates were same as those of reference A pleuropneumoniae strain. Among 22 isolates, 9, 1 and 12 isolates were serovar 2, 3 and 5, respectively. 2. The results of antimicrobial susceptibility test revealed that the isolates showed high susceptibility to ciprofloxacin and cephalothin, moderate susceptibility to amikacin, gentamicin, kanamycin and streptomycin, and low susceptibility to erythromycin, tylosin and sulfadimethoxin. 3. The isolates were varied in pathogenicity to mice. Median lethal dose of LE9402(serovar 2) and LE9511(serovar 5) were $9.2{\times}10^7$ CFU and $2.8{\times}10^7$%CFU, respectively. Specific pneumonic lesions were observed from the infected mice with clinical signs. Bacteria recovery rate was high in the lung, but low In heart blood and tracheas. 4. Serovar 2 was found to be more pathogenic than serovar 5 in guinea pig. Mortality on guinea pigs inoculated with serovar 2($5.4{\times}10^8-5.4{\times}10^6$CFU) and serovar 5($2.8{\times}10^8-2.8{\times}10^6$ CFU) was 20~40% and 40~80%, respectively. A severe hemorrhagic lesions and focal pneumonic lesions were observed from dead guinea pigs. Bacteria recovery rate was relatively higher in the lung than that of other organs. 5. In the SDS-PAGE analysis, OMP-enriched fractions of both isolates and reference strains contain common peptide bands equivalent to molecular weight of 17, 27, 42, 52 and 95Kd. In addition to common peptide bands, the bands which are specific to each isolate were also observed. The profiles of Sephadex G25 fractions showed 3 major peaks. The common peptide bands which were observed by SDS-PAGE of the crude OMPs were found in the peaks 1 and 2. 6. The OMPs extracted from serovar 2(LE9402) and serovar 5(LE9511) provided high level of protection in mice(70~80%) and pigs(100%). All animals inoculated with OMPs were seroconverted, showing micro-agglutination titer of 640 to 1280.

• Korean Journal of Veterinary Research
• /
• v.46 no.4
• /
• pp.337-345
• /
• 2006

The shigellae are common etiological agents of bacillary dysentery in humans and primates. During four years from 2002 to 2005, 22 strains of Shigella spp. were isolated from the diarrheic patients in Seoul region. All of them were identified as S. flexneri by biochemical tests and serotyping. The prevalence of serotypes were variable by year, but the major serotypes were 2a and 3a. In an antimicrobial susceptibility test, all of the isolates were resistant to streptomycin and tetracycline, and susceptible to amikacin, kanamycin, cefoxitin, and gentamicin. All of the isolates showed the multi-resistant patterns over 3 drugs. By analysis of the plasmid profile the isolates were classified into 7 groups (P1~P7). Serotypes 2a and 2b were distributed to P1, P2, P3, and P4. Serotype 3a was differentiated to P5 and serotype 3b, to P6 and serotype 4a, to P7. PCR results showed that all isolates were positive for two virulence genes, ipaH and ial, but none of the strains had stx gene. The set1A and set1B genes were detected from 12 isolates (54.5%) that belonged to serotype 2a and 2b. The sen gene was detected from 19 isolates (86.4%). The 22 isolates showed 12 to 17 DNA fragments in the sizes ranging from 20.5 kb to 1135 kb, resulting in 13 patterns by the PFGE with Not I digestion. The PFGE patterns of the isolates showed the close relation with the serotypes, but no relations with year of isolation and antimicrobial resistance.

• Korean Journal of Veterinary Service
• /
• v.37 no.2
• /
• pp.85-96
• /
• 2014

Total 99 strains of Campylobacter spp. were isolated from 117 cases of duck's fecal samples. Among 99 strains of Campylobacter spp. isolates, 93 strains (93.9%) were C. jejuni and 6 strains (6.1%) were C. coli. Prevalence of virulence and GBS associated genes of 72 C. jejuni isolates was determined by m-PCR. Among the 10 kinds of virulence associated genes, cadF, dnaJ, flaA and ceuE genes were detected in all of C. jejuni isolates from ducks, racR, pldA, iamA, ciaB, virB11 and docC genes were 87.5%, 84.7%, 77.8%, 48.6%, 13.9% and 11.1%, respectively. Antimicrobial susceptibility test was performed on 72 C. jejuni isolates. The rate of resistance were 62.5% for oxytetracycline, 55.6% for kanamycin, 54.2% for enrofloxacin, 50% for ciprofloxacin, 37.5% for tetracycline and nalidixic acid, 18.1% for ampicillin, 15.3% for streptomycin, and 6.9% for ofloxacin. All isolates were susceptible to erythromycin. The adherence (intracellular and extracellular bacteria) abilities of the 20 isolates to INT-407 cells were between $4.21{\pm}1.27{\times}10^4$ CFU/well and $1.053{\pm}0.451{\times}10^6$ CFU/well from the isolates of cj-55 and cj-52, respectively, and that can be expressed as 0.1033% to 5.2655% to the infecting inoculum. The invasion (intracellular bacteria) abilities of the 20 isolates to INT-407 were between $1.00{\pm}1.73{\times}10^3$ CFU/well and $8.47{\pm}5.16{\times}10^4$ CFU/well from the isolates of cj-13 and cj-47, respectively, and that can be expressed as 0.0050% to 0.4235% to the infecting inoculums. The average CFU/well of 20 campylobacters isolated from ducks for adherence to and invasion were $2.646{\pm}2.886{\times}10^5$ and $3.03{\pm}2.7{\times}10^4$ respectively, and that was $1.3230{\pm}1.2139%$ and $0.1516{\pm}0.1343%$ of the starting viable inoculum. There was considerable correlation ($R^2$=0.627) between the adherence and invasion ability of C. jejuni isolates for INT-407 cell.

• Korean Journal of Veterinary Service
• /
• v.44 no.4
• /
• pp.271-281
• /
• 2021

Pathogenic Escherichia coli (E. coli) is one among the most important agents of diarrhea in calves. From January to December 2021, 108 isolates from feces of calves with diarrhea were investigated for enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), shiga toxin-producing E. coli (STEC), enteroaggregative E. coli (EAEC), and enteroinvasive E. coli (EIEC) using real-time PCR. In addition, the genes for F5, F17 and F41 fimbriae were detected by PCR. The most frequently isolated pathotypes were EPEC/STEC (29 isolates), and ETEC/EPEC/STEC (29 isolates). ETEC/EPEC, and ETEC/STEC were also found in 10 isolates. EPEC, STEC, and ETEC were detected in 13, 11, and 6 respectively. EAEC, and EIEC was not detected. Antimicrobial resistance test was carried out by agar disc diffusion method with 14 antimicrobials. Among 108 pathogenic E. coli isolates, 107 isolates were resistant to at least one of 14 antibiotics used in this study, 99 (91.7%) were resistant to two or more antimicrobials, and a single remarkable isolate was resistant to 14 antimicrobials. The isolates were primarily resistant to penicillins, streptomycin, tetracycline, ceftiofur, Trimethoprim/sulfamethoxazole, Kanamycin, and Ciprofloxacin. The high rate of resistance in pathogenic E. coli, sometimes to multiple drugs, may complicate future options for treating human infections. These results may bu used for diagnosis and therpeitic purposes in calves with diarrhea.

• Journal of environmental and Sanitary engineering
• /
• v.3 no.2 s.5
• /
• pp.23-41
• /
• 1988

Two hundred and eighty-six strains of Salmonella species were isolated from the twelve provincial institutes of health and 19 general hospitals of urban and rural areas in Korea from January to December in 1986. The antimicrobial susceptibility test of these cultures was done by the method of agar diluton. The resistance frequency of Salmonella cultures was $29.7\%$. Among these resistant cultures, the most provalent resistance pattern of Salmonella was ampicillin, carbenicillin, chloramphenicol, tetracycline, streptomycin, and its resistance frequency was $15\%$. In plasmid profile of resistance strains, average number of plasmid harboring in Salmonella was 1-4 and molecular weight of plasmid ranged 1.6 to 70 megadalton (Md.). Plasmid pattern of strains isolated from Seoul and Kang-won showed the same or similar profiles. Plasmid pattern was identical in the same resistance pattern.

• Korean Journal of Veterinary Research
• /
• v.44 no.2
• /
• pp.217-224
• /
• 2004

This study was carried out to investigate the antibiotic resistance pattern of Listeria spp. and Staphylococcus aureus. A total of 17 (14.8%) L. monocytogenes, 13 (11.3%) L. innocua, 7 (7%) L. welshimeri, and 83 (72.2%) S. aureus were isolated from commercial poultry carcasses in Seoul and Kyonggi province during the period between 2001 and 2003. Antibiotic susceptibility test of all Listeria strains isolated was performed by the disk agar diffusion method. Antibiotics used in the study were as follows; Amikacin (An), Ampicillin (Am), Cephalothin (Cf), Chloramphenicol (C), Ciprofloxacin (Cip), Erythromycin (E), Gentamicin (Gm), Imipenem (Ipm), Kanamycin (K), Minocycline (Mi), Neomycin (N), Norfloxacin (Nor), Ofloxacin (Ofx), Penicillin (P), Streptomycin (S), Tetracycline (Te), Tobramycin (Nn), Trimethoprim (Tmp), Trimethoprim/Sulfamethoxazloe (Sxt), and Vancomycin (Va). The antibiotic resistance pattern of S. aureus isolates was performed by the disk agar diffusion method. For the latter program, antibiotics used to the study were as follows; Cf, C, Cip, Clindamycin (Cc), E, Gm, Ipm, Nafcillin (Nf), Oxacillin (Ox), P, Te, Sxt, and Va. Of the 17 L. monocytogenes isolates, 94.1% were resistant to Te, 88.2% to Mi, 11.8% to Nor, 11.8% to S, 5.9% to Cip, and 5.9% to C. Of 13 L. innocua, 53.8% were resistant to Te, 23.1% to Mi, 23.1% to S, 7.7% to Cip, and 7.7% to Nor. Of 7 L. welshimeri, 57.1% were resistant to Te, and 14.3% to Am. Of 83 S. aureus, 100% were resistant to Te, 86.7% to Gm, 34.9% to P, 15.7% to Cip, 12% to Cc, 9.6% to E. The multiple antibiotic resistance patterns of L. monocytogenes isolates were observed in Te Mi Cip (5.9%), Te Mi Nor (5.9%), Te Mi (76.5%), and Te Nor (5.9%). Multiple antibiotic resistance was also found in L. innocua isolates. Resistant to Te Mi S Cip Nor was 7.7%, Te Mi S (7.7%), Te Mi (7.7%), and was 7.7% to Te S. Antibiotic resistance patterns for S. aureus isolats were demonstrated to Te Gm P Cip Cc E (6.0%), Te Gm Cip Cc E (3.6%), Te Gm P Cc (1.2%), Te Gm P (15.6%), Te Gm Cip (2.4%), Te P Cip (2.4%), Te Gm Cc (1.2%), Te Gm (56.6%), Te P (9.6%), and to Te Cip (1.2%). The results of this study suggest a high incidence of Lsteria spp. and S. aureus on poultry carcasses. The contaminated poultry carcasses may be a potential vehicle for foodborne infections due to multiple antimicrobial resistant organisms.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.30 no.5
• /
• pp.796-801
• /
• 2001

A total of 277 mineral spring water samples in Kangwon province from 1999 to 2000 were analyzed for the presence of Yersinia spp. by the conventional Food and Drug Administration protocol, and presumptive strains were identified by morphological, cultural and biochemical tests according to Bergey’s manual. Also, the biotypes, serotypes, and susceptibility to 12 antibiotics were tested. Among the total 277 mineral spring water samples, 40 samples (14.4%) were found to be contaminated with Yersinia species. Among the 40 strains of Yersinia spp. isolates, 33 strains (82.5%) for Yersinia enterocolitica, 4 strains (10%) for Yersinia frederiksenii, 2 strains (5%) for Yersinia intermedia, and 1 strain (2.5%) for Yersinia sakazaki were identified, respectively. Of 40 Yersinia spp. isolates, Yersinia enterocolitica (82.5%) was the most predominant species in the mineral spring water samples compared to other Yersinia species. Compared to direct culture method after KOH treatment and KOH treatment method after cold enrichment for better isolation ratio of according to comparision of Yersinia species, the detection ration (18.5%) of KOH treatment method after cold enrichment was about 3 times better than that (6.1%) of direct culture method after KOH treatment. According to serotypes of Y. enterocolitica isolates, O : 5 (12.9%) was the most predominant and followed by O : 3 (9.7%), O : 8 (6.5%), and O : 9 (3.2%), and others. For biotypes of Y. enterocolitica isolates, 1A (71.0%) was the most predominantly abundant and followed by 3A (12.9%), 3B (9.7%), 1B (3.2%) and 5 (3.2%). Also, an antibiotic susceptibility test showed that Yersinia spp. isolates were very susceptible to the antibiotics tested, but they were very strongly resistant to ampicillin, cephalothin and carbenicillin.