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Competitive Enzyme-Linked Immunosorbent Assay for Detection of Gentamicin Residues in Edible Animal products (축산식품 중에 잔류하는 Gentamicin 검사를 위한 ELISA 개발에 관한 연구)

  • Kim, Jae-Myung;Lee, Mun-Han;Lee, Hang;Ryu, Pan-Dong;Cho, Myung-Haing;Park, Jong-Myung
    • Journal of Food Hygiene and Safety
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    • v.9 no.3
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    • pp.123-131
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    • 1994
  • An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

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Multidrug-Resistant Tuberculosis Presenting as Miliary Tuberculosis without Immune Suppression: A Case Diagnosed Rapidly with the Genotypic Line Probe Assay Method

  • Ko, Yousang;Lee, Ho Young;Lee, Young Seok;Song, Junwhi;Kim, Mi-Yeong;Lee, Hyun-Kyung;Shin, Jeong Hwan;Choi, Seok Jin;Lee, Young-Min
    • Tuberculosis and Respiratory Diseases
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    • v.76 no.5
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    • pp.245-248
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    • 2014
  • Miliary tuberculosis (TB) is a rare extrapulmonary form of TB, and there have been only two reports of miliary TB associated with infection with multidrug-resistant (MDR)-TB pathogen in an immunocompetent host. A 32-year-old woman was referred to our hospital because of abnormal findings on chest X-ray. The patient was diagnosed with MDR-TB by a line probe assay and was administered proper antituberculous drugs. After eight weeks, a solid-media drug sensitivity test revealed that the pathogen was resistant to ethambutol and streptomycin in addition to isoniazid and rifampicin. The patient was then treated with effective antituberculous drugs without delay after diagnosis of MDR-TB. To the best of our knowledge, this is the first case of miliary TB caused by MDR-TB pathogen in Korea.

Nuclear Progression through In Vitro Meiotic Maturation of Bovine Oocytes (우 난포란의 체외성숙시 핵의 발달과정)

  • 주영국;공일근;정미경;강대진;박충생
    • Journal of Embryo Transfer
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    • v.7 no.2
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    • pp.125-131
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    • 1992
  • The nudear changes of bovine oocytes during 24 hrs. of culture for mejotic maturation were examined. Bovine oocytes were collected from small(<2 mm), medium(2~6 mm) and large(>6mm) follicles and classified into three grades by their morphological characteristics. A total of 242 oocytes collected were obtained:from 184 small, 157 medium and 1 large follicles, respectively and were classified into 95 grade I, 155 grade H and 92 grade III oocytes. All the bovine oocytes collected and graded were washed with a basal medium and incubated in groups of 10 for 24 hrs in 5% $CO_2$ and 39$^{\circ}C$. The basal medium used was composed of TCM-199 supplemented with sodium bicarbonate, sodium pyruvate, streptomycin, penicillin G and 10% FCS. The oocytes were cultured in drops of 50,$\mu$l basal medium supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH and 1$\mu$g /ml estradiol-17$\beta$. The oocytes were fixed and examined on their chromosomal status by 1% acetorcein staining in the interval of 3 hrs. Most of the grade I oocytes developed to germinal vesicule stage at 0 to 3 hrs., germinal vesicle breakdown at 6 hrs., metaphase I at 9 to 15 hrs., anaphase I and telophase I at 18 hrs., and metaphase II and the first polar body at 24 hrs. after culture for meiotic maturation. However, it was found that compared to grade I oocytes, grade H and W oocytes reached earlier to germinal vesicle breakdown and most of them developed earlier to M II stage at 21 hrs. after culture.

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Effects of Sugar Type on Viability of Frozen-Thawed Canine Spermatozoa

  • Lim, Y.H.;Son, J.M.;Shin, Y.J.;Kim, Y.S.;Lee, D.S.;Yoon, K.Y.;Shin, S.T.;Cho, J.K.
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.239-243
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    • 2008
  • This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at $4^{\circ}C$ for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into $37^{\circ}C$ water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.

Antimicrobial Resistance of Enterococcus Species Isolated from Chicken in Turkey

  • Sanlibaba, Pinar;Tezel, Basar Uymaz;Senturk, Esra
    • Food Science of Animal Resources
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    • v.38 no.2
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    • pp.391-402
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    • 2018
  • The aim of the present work was to provide information about Enterococcus strains isolated from pre-packaged chicken samples in Ankara (Turkey), focusing on their prevalence, phenotypic and genotypic characteristics, and antibiotic resistance. We report the first study on the occurrence of antibiotic resistant enterococci in pre-packaged chicken samples in Ankara. A total of 97 suspicious enterococcal isolates were identified from 122 chicken samples. All isolates were identified to species level by phenotypic and molecular methods. In the 16S rDNA sequence analysis, Enterococcus faecium (61.85%) and Enterococcus faecalis (38.15%) were found to be the most frequently detected Enterococcus spp. Of the 97 isolates tested for hemolytic activity, 12.37% enterococcal strains were ${\beta}$-hemolytic. ${\beta}$-Hemolysin was most prevalent among E. faecium (58.33%) compared to E. faecalis (41.66%). Disk diffusion method was used for determining of antibiotic resistance. The analysis of the antimicrobial resistance of the 97 Enterococcus isolates revealed that the resistance to kanamycin (98.96%), rifampicin (80.41%) and ampicillin (60.82%) was most frequent. Furthermore, resistance to erythromycin (38.14%) and ciprofloxacin (34.02%) was also observed. The frequencies of resistance to tetracycline (9.27%), penicillin G (8.24%), and chloramphenicol (3.09%), gentamicin (2.06%) and streptomycin (1.03%) were low. None of the isolates was resistant to vancomycin. Multi-drug resistance was found in 97.93% of Enterococcus strains. E. faecium strains showed a more resistant phenotype than E. faecalis strains according to the antibiotic resistance levels. The results of this study indicated that chicken meat is a potential reservoir for the transmission of antibiotic resistance from animals to humans.

Antibiotic Resistance of Enterococcus Isolated from the Processed Grain Foods, Saengsik and Sunsik

  • Kim, Soo-Hwan;Kim, Jong-Shin;Park, Jong-Hyun
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.470-476
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    • 2007
  • To evaluate the vancomycin resistance of Enterococcus spp. (VRE) from Saengsik and Sunsik, Enterococcus were isolated and identified from 25 Saengsik and 35 Sunsik samples, and resistance of Enterococcus to other antibiotics was also assessed. Thirty nine Enterococcus, 16 strains from Saengsik, and 23 strains from Sunsik, were ultimately isolated. The most frequently collected Enterococcus isolates in Saengsik were E. casseliflavus and E. hirae, and were E. casseliflavus and E. faecium in Sunsik. However, E. faecalis was not detected in those foods. Minimum inhibitory concentrations of vancomycin against the isolates were below $4\;{\mu}g/mL$ and no strains evidenced profound levels of resistance. The isolates were found to be susceptible to vancomycin with the exception of eight E. casseliflavus and three E. gallinarum. All Enterococcus isolates proved resistant to streptomycin and chloramphenicol. 23% of the isolates were resistant to penicillin; however, all of the isolates were sensitive to tetracycline. Six and 48%, respectively, of the strains from the Saengsik and Sunsik proved resistant to erythromycin. All of E. mundtii and E. hirae isolates from Saengsik, and 20% of E. gallinarum and E. casseliflavus isolates from Sunsik were found to be ampicillin-resistant. All of E. gallinarum, E. casseliflavus, and E. faecium were rifampin-resistant. The antibiotic resistances of Enterococcus were relatively low, and this low vancomycin resistance was similar to that evidenced by Enterococcus isolates obtained from the other foods. However, there may be a need for some review of the accepted antibiotics criteria for Enterococcus and VRE in ready-to-eat foods.

Antibacterial Activity In Vitro and Primary Dermal Irritation Test in Rabbits of Surfactin Produced Bacillus subtilus Complex BC2121 (Bacillus subtilus Compulex BC1212에서 생산된 Surfactin의 시험관내 항균활성 및 토끼에서 일차피부자극시험)

  • Hwang Mi-Hyun;Yun Hyo-In;Lim Jong-Hwan;Kim Kil-Soo;Rhee Man-hee;Kim Nam-Woo;Kim Jong-Choon;Park Seung-Chun
    • Toxicological Research
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    • v.21 no.1
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    • pp.39-43
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    • 2005
  • The present study was carried out to investigate the antibacterial activity against Staphylococcus aureus with antibiotic-resistance in vitro and the skin irritation in rabbits with surfactin produced by Bacillus subtilus Complex BC1212. The antibacterial activities of selected antimicrobial agents (surfactin, amoxacillin, colistin, norfloxacin and streptomycin) were evaluated by using the broth microdilution method. As the results, the minimum inhibitory concentration (MIC) of the surfactin was less than 15.6 ㎍/ml. In the skin irritation test, two out of 4 rabbits showed very slight edema at 24 h after the administration of surfactin, and then recovered at 72 h. The change of body weight was normal during the skin irritation test. The primary irritation index in accordance with the Draize evaluation of topical reaction was calculated to be '0.125', which meant not irritating. Based on these results, it could be concluded that the test agent, surfactin, was a non-irritant. We could also think that the surfactin may be useful for the treatment of S. aureus infections such as bovine mastitis.

Antimicrobial Susceptibility of Gram-Negative Bacteria from Dogs and Cats (개와 고양이에서 분리된 그람음성균의 항생제 감수성 양상)

  • Kim, Dae-Keun;Shin, Dong-Ho;Kim, Ha-Young;Byun, Jae-Won;Lee, Kyeong-Hyun;Lee, O-Soo;Jung, Byeong-Yeal
    • Journal of Veterinary Clinics
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    • v.28 no.4
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    • pp.348-351
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    • 2011
  • The purpose of this study was to determine the distribution of gram-negative bacteria isolated from companion animals with sepsis, and to investigate the antimicrobial susceptibility patterns of the isolates. Bacterial pathogens were isolated from specimens of dogs and cats submitted to National Veterinary Research and Quarantine Service between 2008 and 2009. A total of 44 gram-negative pathogens were isolated from necropsied organs. The most common isolates were E. coli (n = 33), K. pneumoniae (n = 4) and B. bronchiseptica (n = 4). Most of gram-negative isolates were susceptible to ceftiofur (68.2%), colistin (84.1%), florfenicol (84.1%) and spectinomycin (61.4%). Most of those were resistant to ampicillin (77.3%), erythromycin (86.4%), flumequine (65.9%), lincomycin (97.7%), oxytetracycline (61.4%), penicillin (100%), streptomycin (63.6%), spiramycin (97.7%), sulfamethoxazole (90.9%), tylosin (97.7%) and tiamulin (100%). In conclusion, colistin and florfenicol could be useful against sepsis due to gram-negative bacteria.

The isolation and antimicrobiol susceptibility of Escherichia coli O157:H7 on bovine feces and carcass (소 분변과 도체에서 E coli O157:H7의 분리와 항생제 감수성)

  • Chae Hee-Sun;Kim Jong-Hwa;Kim Gyu-Hyeon;Choi Tae-Seok;Shin Bang-Woo;Lee Duck-Joo;Lee Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.28 no.1
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    • pp.71-79
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    • 2005
  • In this study, a total of 2,119 samples was taken from bovine feces and carcass from March 2002 to December 2003. And those were examined for the presence of enterohemorrhagic E coli O157:H7. The properties of the isolates were characterized for biochemical features, serotypes, virulence genes and antimicrobial susceptibility. Forty five strains($3.7\%$) of E coli O157:H7 were isolated from 1,208 fecal samples and were not detected in carcass using immunomagnetic separation technique and selective media. In multiplex PCR using stx1, stx2, eaeA and hlyA primers, the amplified bands at 180 bp, 255bp, 384bp and 534bp were observed, respectively. In antimicrobial susceptibility test, all isolates were susceptible to amoxicillin/clavulanic acid and cefazolin. The isolates were most resistant to sulfisoxazole($24.4\%$), followed by streptomycin($22.2\%$), tetracycline($20.0\%$). Eight strains($17.8\%$) of 45 isolates showed the multi-resistant patterns with over 3 drugs.

The in vitro Drug Resistance of Escherichia coli Isolated from Piglets, Calve, Lamb and Goats with Diarrhoea (대장균 설사중에 이환된 소, 돼지, 양에서 분리한 대장균의 약제감수성)

  • Kim, Bong Hwan;Rhee, Jai Chin;Kim, Dong Sung
    • Korean Journal of Veterinary Research
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    • v.19 no.2
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    • pp.121-126
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    • 1979
  • 대장균 설사중에 걸린 어린 돼지, 송아지, 어린 양에서 분리한 대장균 126주의 항생제와 화학요법제(15종)에 대한 감수성을 disc diffusion technique로 조사한 성적은 다음과 같다. 1. 어린 돼지에서 분리한 62주의 대장균은 gentamicin(GM)에 100%, colistin(CL)에 96.8%, kanamycin(KM)에 93.5%, neomycin(NM)에는 91.9%가 감수성을 가지고 있었으나 ampicillin(AM), erythromycin(EM), lincomycin (LM), novobiocin (NB), penicillin (PC), streptomycin(SM), tetracycline(TC), sulfaisodimidin(SU)에는 내성을 가지고 있었다. chloramphenicol(CP), sulfamethoxazole-trimethoprim(SXT), cephalosporin(CE)에는 각각 75.8%, 64.5%, 50%가 감수성이 있었다. 2. 송아지에서 분리한 32주의 대장균은 GM에 100%, CL에는 87.5%, SXT에는 66.7%가 감수성이 있었으나 CP와 KM에는 각각 40.6%, SU에는 56.2% NM에는 62.5%, SM에는 87.5%가 내성을 가지고 있었다. EM, LM, NB, PC에는 전혀 감수성이 없었으며 AM, SM, TC에도 고도의 내성을 가지고 있었다. 3. 어린 양에서 분리한 32주의 대장균은 GM과CL에 100%, CP에 96.9%, KM과 NM에 90.6%가 감수성이 있었다. SM과 SU에도 71.9%나 감수성이 있었으나 CM, EM, LM, PC, TC에는 대부분 내성을 가지고 있었다. AM에는 21.9%가 감수성이 있었다. 4. 2종류 이상의 약제에 내성을 가진 대장균의 AM, CE, CP, CL, GM, KM, NM. SM, TC, SU등 10종의 약제에 대한 multiple drug resistance pattern(MDRP)을 조사한 바 돼지 유래 약제내성 대장균의 MDRP는 18가지였으며 이중 가장 빈도가 높은 것은 AM, CE, SM, TC, SU 내성형으로 전체의 24.2%나 되었다. 송아지 유래 약제내성 대장균의 MDRP는 17가지였으며, AM, CE, CP, KM, NM, SM, TC, SU 내성형이 28.1%로 가장 빈도가 높았다. 반면에 어린 양에서 분리한 대장균의 MDRP는 9가지였으며 AM, CE, SU의 3약제 내성형이 40.6%로 가장 많았다.

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