• 한국동물위생학회지
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• 제47권1호
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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.

• 한국식품위생안전성학회지
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• 제31권2호
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• pp.94-98
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• 2016

HPLC-MS/MS를 이용하여 벌꿀의 동물용의약품 동시분석을 위한 분석법을 정립하고자 식품공전 중 벌꿀에 규격이 설정된 11종의 동물용의약품을 2개 그룹 Group 1(streptomycine, dihydrostreptomycine, neomycine)과 Group 2 (oxytetracycline, enrofloxacin, ciprofloxacin, cymiazole, chloramphenicol, amitraz, coumaphos, fluvalinate)으로 나눠서 동시에 분석 할 수 있는 방법을 연구 하였다. 두 그룹 모두 RT는 15분 이내였고, 검출한계는 $0.0056{\sim}0.0643{\mu}g/g$, 정량한계는 $0.0169{\sim}0.1948{\mu}g/g$으로 나타났으며 Group 1 ($0.05{\sim}1.0{\mu}g/g$의 농도범위)과 Group 2 ($0.01{\sim}1.0{\mu}g/mL$)의 검량선을 작성한 결과 각 동물용의약품의 직선상관계수($R^2$)는 0.9917~0.9987, 0.9923~1.000으로 매우 양호한 상태를 보였고, 최종 농도가 $0.25{\mu}g/g$, $1.0{\mu}g/g$에서의 회수율은 Group 1 65.1~80.6%, Group 2 64.2~90.3%를 나타났다, 또한 area 및 RT 에 대한 inter (n = 3), intra day (n = 6) RSD (%) 분석결과는 area는 10.92%이하, RT은 1.57% 이하로 나타나 양호한 수준을 보여 벌꿀 중 동물용의약품 동시분석 방법으로 적합하다고 판단된다.

• 생명과학회지
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• 제22권8호
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• pp.1114-1119
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• 2012

돼지 정액의 세균오염은 정자활력 감소나 수태율 저하 등을 유발하는데, 특히 Stenotrophomonas maltophilia는 자연환경에 널리 존재하여 비위생적으로 채취된 정액에 많이 오염될 뿐 아니라 사람에게도 병원성을 일으킨다. 본 연구에서는 S. maltophilia의 검사 시간 단축과 정확도를 높이고자 PCR 기법을 개발하였으며, 돼지 원정액에서 이들 세균의 오염율을 조사하고 유효 항생제를 선발하고자 하였다. 돼지 원정액에서 분리 빈도가 높은 18 균종을 대상으로 PCR을 적용한 결과, S. maltophilia에서만 445 bp의 특이 유전자 증폭산물이 확인되었다. 또한 순수 배양된 S. maltophilia는 $1.5{\times}10^3$ CFU/ml까지, 원정액에 혼합된 S. maltophilia에 대해서는 $1.5{\times}10^4$ CFU/ml까지 검출이 가능하였다. 2009년에 116건과 2011년 324건 등 총 440건의 원정액 중 26건(5.9%)에서 S. maltophilia가 분리되었으며, 여름과 가을철에 오염율이 상대적으로 높게 나타났다. 균분리법과 PCR간의 일치율은 98.9%, kappa value는 0.906이었으며, PCR의 민감도와 특이도는 각각 100%, 98.7%로 나타났다. 분리균주의 항생제 감수성 시험 결과, enrofloxacin (100%)과 florfenicol (92.3%)에 높은 감수성을 보였으나 amoxicillin/clavulanic acid, apramycin, ceftiofur, penicillin, spectinomycin에는 내성율이 높게 나타났다. 결론적으로 개발된 PCR기법은 민감도, 특이도, 일치율이 높아 균분리법을 보조하고 신속 정확하게 정액 내 오염세균을 확인할 수 있어 효율적인 정액관리가 가능할 것으로 기대된다.

• 한국동물위생학회지
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• 제35권4호
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• pp.295-305
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• 2012

The aim of this study was to isolate microorganisms from half milk samples of dairy goats by California mastitis test (CMT) during the lactation period and to further investigate the susceptibility of isolated organisms to antimicrobial drugs. From a total of 235 half milk samples with CMT scores of 2 or above from 366 dairy goats distributed throughout Jeonnam province, microorganisms were isolated from 198 (83.5%) samples either singly (99.0%) or in combination (1.0%). The most prevalent microorganism was the coagulase-negative Staphylococcus spp., (44.4%, n=88) followed by Staphylococcus aureus (24.2%, n=48), Escherichia coli (11.1%, n=22) and Streptococcus spp. (7.6%, n=15). Isolated bacteria also included Bacillus spp. (2.5%, n=5), Pseudomonas spp. (2.5%, n=5), Micrococcus spp. (1.5%, n=3), Corynebacterium spp. (1.5%, n=3), Enterococcus facium (1.0%, n=2), Morganella morganii (0.5%, n=1) and Streptococcus agalactiae (0.5%, n=1). During the summer season, a high prevalence of all microorganisms were observed in which Staphylococcus spp. (30.8%), Escherichia coli (8.6%), and Streptococcus spp. (5.6%) were among the most prevalent bacteria isolated. Staphylococcus spp. was also shown to be high in the winter (21.7%). In most samples, the presence of bacterial pathogens in goat milk led to the increase in the total somatic cell count (SCC). Most of the half milk samples of dairy goats with bacterial contamination showed SCC of ${\geq}1{\times}10^6cells/ml$ (90.4%). Minor pathogens (11.4%) were more detected from milk samples with SCC of < $1{\times}10^6cells/ml$ than major pathogens (4.1%), while the major pathogens tended to be higher from samples with SCC of ${\geq}3{\times}10^6cells/ml$. Susceptibility of these bacteria to 12 antimicrobial agents was tested by the Kirby-Bauer disc diffusion method. Results indicated that more than 90% of bacteria isolated from CMT 2+ dairy goat half milk samples were susceptible to trimethoprim/sulfamethoxazole, amoxicillin/clavulanic, enrofloxacin and cephalothin while they were resistant to tetracycline (44.7%).

• 한국동물위생학회지
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• 제23권1호
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• pp.45-59
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• 2000

The present study was conducted to investigate the epidemiological properties of salmonellosis of poultry farms in Kyongbuk province during the relied from November 1998 to November 1999 And antibiotic susceptibility and biochemical characteristics of 120 Salmonella cultures isolated from chicken samples were also investigated. The results obtained through this study were summarized as follows, 1. Among 667,200 chickens of 31 flocks in 17 layer farms and 9 broiler farms, 61,350 chickens of 31 flocks were died with salmonellosis. The death rate of 25 farms varied from 0.1% to 75.0%, and the average death rate was 9.2%. 2. According to etiological agents, fowl typhoid was shown the most predominant outbreak among the salmonellosis during a year, which accounted for 8R.0%(22/25) of the total case 3. The serotypes of 120 Salmonella isolates were identified as 7 strains(5.8%) of S pullorum, 10 strains(8.4%) of S typhimurium and 103 strains(85.8%) of S gallinarum. 4. Most outbreak of fowl typhoid were prevalent on the layer chicken farms(77.1%), and the summer season(45.5%) also appeared the most hazardous season during the year. 5. It seemed that the Hyline breed(70.6%) was the most susceptible among the layer chicken breeds, and followed by Isabrown(23.5%), Tetra(5.9%) in order. 6. In layers, 76.4% of fowl typhoid occurred commonly from 14 to 40 weeks including the early laying peroid, but in broiler farms, all cases was outbreak within first second weeks. 7 All the strains of S pullorum were resistant to lincomycin(Lm), penicillin(Pm), and steptomycin(Sm), but sensitive to amikacin(Ak), ampicillin(Am), cephalothin(Ce), ciprofloxacin (CiP), chloramphenicol(Cm), colistin(Co), enrofloxacin(Enr), furazolidone(Fu), gentamicin(Gm), kanamycin(Km), neomycin(Nm), polymyxin(Po), and teracycline(Tc). All the strains of S typhimurium were resistant to Lm(100%), Pm(100%), Po(90%), and Sm(90%), but were sensitive to Ak, Am, Ce, CiP, Cm, Co, Enr, Fu, Gm, Km, Nm, and Tc. 8. Minimum inhibitory concentration(MIC) of 103 strains of S gallinarum were also evaluated and their patterns were much more variable than others. All the strains of S gallinarum were sensitive to Ak, Am, Ce, Cip, Cm, Enr, Fu, Km, and Nm, but resistant to Lm(100%), and Sm(100%), 99(96.1%) to Co, 83(80.6%) to Pm, and 83(80.6%) to Po, 55(53.4%) to Gm, and 33(32.0%) to Tc. 9. The multiple drug resistance patterns of 120 Salmonella strains were CoLmPmPo Sm pattern(34.2%), CoGmLmPmPoSmTc(20.8%), CoGmLmPmPoSm(13.3%), CoGmLmSm(7.5%), LmPmSm(7.5%), LmPm(6.7%), CoLmSmTc(3.3%),, CoGmLmSmTc(1.7%), GmLmSmTc(1.7%), CoGmLmPoSm(0.1%), LmPmPo(0.1%), CoLm Sm(0.1%), and LmSm(0.1%), in order.

• 한국동물위생학회지
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• 제42권4호
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• pp.289-296
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• 2019

A novel rapid procedure with liquid chromatography tandem mass spectrometry (LC-MS/MS) detection has been developed by changing various conditions including sample preparation such as QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) methodology. This work has been involved the optimization and validation of detection method for fluoroquinolones which are widespread used in livestock especially in the chicken. Five grams of homogenized chicken muscle were extracted with QuEChERS EN and acetonitrile containing 5% formic acid and cleaned with anhydrous magnesium sulfate and C₁₈ sorbent. The separation was performed on Acquity UPLC HSS T3 (2.1 mm×100 mm, 1.8 ㎛) column. The mobile phase A and B were composed of water containing 0.1% formic acid and acetonitrile containing 0.1% formic acid, respectively. Flow rate was 0.25 mL/min and column temperate was 40℃. LC-MS/MS with multiple reaction monitoring has been optimized for ten fluoroquinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, marbofloxacin, norfloxacin, ofloxacin, orbifloxacin, pefloxacin and sarafloxacin). The method developed in this study has been presented good linearity with correlation coefficient (R²) of 0.9971~0.9998. LOD and LOQ values ranged from 0.09 to 0.76 ppb and from 0.26 to 2.29 ppb, respectively. The average recoveries were from 77.46 to 111.83% at spiked levels of 10.0 and 20.0 ㎍/kg. Relative standard deviation (%) ranged 1.28~11.90% on intra-day and 3.10~8.38 % on inter-day, respectively. This analysis method was applicable to the livestock residue laboratories and was expected to be satisfactory for the residue surveillance system.

• 한국어병학회지
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• 제7권1호
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• pp.37-46
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• 1994

서해안 일원의 가온 양만장 사육조로부터 분리한 E. tarda균에 대한 약제내성검사 및 내성균의 내성인자 전달 시험을 실시하였다. 공시한 항생제에 대한 내성률은 LM, PM, SD, SF 등에는 100%, AP, CH, SM, OT 및 CP 등에는 89% 이상, OA 및 KM에는 70% 정도 그리고 AK, GM 및 EF 등에는 30% 이하의 내성률을 보였으며, 이와같은 내성률은 각 양만장에서 분리한 균주간에 차이를 보이지 않았다. 내성유형은 총 34유형으로 모든 분리균주가 8~14제의 다제내성을 보였으며, 그 중 LM PM SD SF CP CH SM OT AP OA KM(22주, 21.4%) 및 LM PM SD SF CP CH SM OT AP OA(12주, 11.7%) 유형등이 고빈도로 출현하였다. 내성균의 내성인자 전달유형은 총 55형으로 단제에서 13제까지의 다양한 내성인자가 전달되었는데, 그 중 LM PM SD SF CP CH SM OT AP OA(10주. 10%), LM PM SD SF CP CH SM OT AP OA KM(7주, 7%) 및 LM PM SD SF CP CH AP OA(7주, 7%) 등의 다제내성유형이 고빈도의 전달률을 보였으며, 약재별로는 GM, EF 및 AK 등에는 저빈도(0~20%), OT, SM, KM 및 OA 등에는 중등도(49~62%), AP, PM, LM, SF, CP, CH 및 SD 등에는 고빈도(71~94%)로 내성인자를 전달하였다.

• 한국동물위생학회지
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• 제37권2호
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• pp.85-96
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• 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.

• Journal of Microbiology
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• 제43권5호
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• pp.391-397
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• 2005

Escherichia coli is a common inhabitant of the intestinal tracts of animals and humans. The intestines of animals also represent an ideal environment for the selection and transfer of antimicrobial resistance genes. The aim of this study was to investigate the resistance of E. coli isolated from chicken fecal samples to fluoroquinolones and to analyze the characterization of mutations in its gyrA and parC gene related resistance. One hundred and twenty-eight E. coil isolates showed a high resistance to ciprofloxacin (CIP; $60.2\%$), enrofloxacin (ENO; $73.4\%$) and norfloxacin (NOR; $60.2\%$). Missense mutation in gyrA was only found in the amino acid codons of Ser-83 or Asp-87. A high percentage of isolates ($60.2\%$) showed mutations at both amino acid codons. Missense mutation in parC was found in the amino acid codon of Ser-80 or Glu-84, and seven isolates showed mutations at both amino acid codons. Isolates with a single mutation in gyrA showed minimal inhibitory concentrations (MIC) for CIP (${\le}0.5\;to\;0.75{\mu}g/ml$), ENO (1 to $4{\mu}g/ml$) and NOR (0.75 to $4{\mu}g/ml$). These MIC were level compared to isolates with two mutations, one in gyrA and one in parC, and three mutations, one in gyrA and two in parC (CIP, ${\le}0.5\;to\;3{\mu}g/ml;\;ENO,\;2\;to\;32<{\mu}g/ml;\;NOR,\;1.5\;to\;6\;{\mu}g/ml$). However, the isolates with two mutation in gyrA regardless of whether there was a mutation in parC showed high MIC for the three fluoroquinolones (CIP, 0.75 to $32{\le}{\mu}g/ml;\;ENO,\;3\;to\;32{\le}{\mu}g/ml;\;NOR,\;3\;to\;32{\le}{\mu}g/ml$). Interestingly, although the E. coil used in this study was isolated from normal flora of chicken, not clinical specimens, a high percentage of isolates showed resistance to fluoroquinolones and possessed mutations at gyrA and parC associated with fluoroquinolone resistance.