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1 J. Exp. Biomed. Sci. 2012, 18(2): 152~159 pissn : Original Article Prevalence and Antimicrobial Susceptibility of Erythromycin-Resistant Campylobacter jejuni and Campylobacter coli Isolated from Swine Mi-Rai Choi 1, Shin-Moo Kim 2, Sang-Ha Kim 3, Wan-Soo Choi 4 and Young-Kwon Kim 5, 1 Department of Public Health and Welfare, Graduate school of Konyang University, Daejeon , Korea 2 Department of Clinical Laboratoty Scinence, Wonkwang Health Scinence University, Iksan , Korea 3 Department of Public Health and Welfare Graduate School of Konyang University, Daejeon , Korea 4 Department of Clinical Laboratoty Scinence Dong-eui Institute of Technology, Busan, , Korea 5 Department of Biomedical Laboratory Science, Konyang University, Daejeon , Korea Campylobacter species are known to the high optimum growth temperature (42 ) and the cause of enteritis in people. Erythromycin has a curative effect for enteritis caused by the bacteria. However, the rate of erythromycin-resistant bacteria was not well known until recently in Korea. Swine are one of sources of the infection with a Campylobacter species which cause the symptom of a high temperature. In this study, we cultured rectum fecal specimens of 100 pigs in an area of Buan-gun, Jeonbuk Province during July As a result, the detection rate of C. jejuni and C. coli and the rate of erythromycin-resistant bacteria for the separated Campylobacter species on the condition of high temperature were investigated. The possession or not of hipo and glya gene and ciprofloxacin-resistant gene gyra was also reviewed with biochemical characteristics and PCR. Key Words: Campylobacter species, C. jejuni, C. coli, Erythromycin resistant, hipo, Gene gyra 서 호열, 미산소성그람음성굽은막대균인 Campylobacter jejuni와 Campylobacter coli는각종야생동물과가축의장관내에널리분포되어있으며, 특히돼지를비롯한닭, 칠면조, 개, 소, 고양이등에보균률이높은것으로보고되고있다 (Smith et al., 1999; Friedman et al., 2000; Allos, 2001). Campylobacter 장염은사람에게서 Salmonella, Shigella, Yersinia, Escherichia coli 보다더높은장염을일으키는장염의중요한원인균으로알려져있다 (Gaunt et al., 1996; Altekruse et al., 1999). Campylobacter 감염에대한치료제로는 fluoroquinolones, erythromycin, ciprofloxacin이선택되어사용되어왔으나, * Received: 23 March, 2012 / Revised: 7 June, 2012 Accepted: 7 June, 2012 Corresponding author: Young-Kwon Kim. Korea Culture Collection of Medical Fungi (KCMF), Dean of Medical Science College, Konyang Univ. 685 Gasuwon-dong, Seo-gu Daejon , Korea. Tel: , Fax: ykkim3245@konyang.ac.kr C The Korean Society for Biomedical Laboratory Sciences. All rights reserved. 론 1980년이후특히유럽지역을중심으로 fluoroquinolone 내성이증가하고있으며 (Smith et al., 1999), 벨기에의 Vanhoof 등은사람에서분리된 C. jejuni 중 8.4% 가 erythromycin에내성임을보고하였고 (Vanhoof et al., 1978), 기타여러나라에서보고된 C. jejuni와 C. coli의 erythromycin 내성률은 Table 1과같이보고하였다 (Gilbreel and Taylor, 2006). Erythromycin 내성 C. coli는돼지에서높은분리율을나타내고, C. jejuni는닭에서높은분리율을나타내고있어 (Arestrup et al., 2001; Chuma T et al., 2001; Van Looveren M et al., 2001), 사람에게서분리되는 Campylobacter species 감염에대한치료과정중 macroride계항균제에대한내성이나타날수있기때문에, 이들항균제에대한내성패턴의연구와감시가요구되고있다. 또한, Campylobacter species에대한배양과항균제감수성검사가매우까다롭기때문에우리나라의임상미생물검사실에서는 Campylobacter species에대한일상적인검사가이루어지고있지않고, 경험적항균제치료에의존하고있어, Campylobacter species에대한정확한항균제치료의가이드라인이절실하게요구되고있다. 따라서본연구에서는사람에게중요한 Campylobacter species의감염

2 Table 1. Sequence of oligonucleotides Primer name Target gene Sequence (5' to 3') Amplicon size (bp) CJF C. jejuni ACTTCTTTATTGCTTGCTGC CJR hipo a GCCACAACAAGTAAAGAAGC 323 CCF C. coli GTAAAACCAAAGCTTATCGTG CCR glya a TCCAGCAATGTGTGCAATG 126 CampyMAMAgryA1F TTTTTAGCAAAGATTCTGAT gyra b CampyMAMAgryA5R CAAAGCATCATAAACTGCAA 265 a Wang et al., 2002, b Zirnstein et al., 원인돼지로부터호열성 (42 ) Campylobacter species를분리하여검출률을조사하였으며, 항균제감수성검사를실시하여 Campylobacter species의치료에사용하는항균제들에대한내성률을조사하여항균제치료에대한참고자료를제시하고자하였다. 연구대상및방법돼지에서 Campylobacter jejuni와 C. coli의분리 Campylobacter jejuni와 C. coli의생화학적성상. 본연구에사용한시료는 2009년 7월 2일부터 7일사이에전북부안지역일개농장의돼지 100마리를대상으로직장변검체를면봉으로채취하여 Stuart 수송배지 (BBL, Copan, Italy) 에넣어보존수송하였다. Campylobacter의분리를위해선택배지인 Preston agar (Oxoid, Basingstoke, UK) 에면봉으로도말한후백금이로획선접종하였다. 10.0% CO 2 배양기에 42, 48시간배양한후, 용혈이없고원형의회백색집락을재차분리하였다. 순수분리된집락중그람음성, 굽은막대균, oxidase 양성, hippurate hydrolysis 양성, 30 μg의 nalidixic acid 디스크에내성, 30 μg의 cephalothin 디스크에내성인성상을 C. jejuni로예비동정하였고, 위성상중 hippurate hydrolysis 음성인균주를 C. coli로예비동정하였다. 분리된균주는 10.0% CO 2 조건에서 37, 48시간 blood agar plate (BAP) 에배양하였고, 배양된균체는 15.0% glycerol이포함된 BHI broth 1 ml에현탁하여다음실험시까지 -70 에냉동보관하였다. Campylobacter jejuni와 C. coli의 hipo, glya 유전자의검출. C. jejuni의확인동정을위해 hipo 유전자를, C. coli는 glya 유전자를대상으로 PCR을수행하였다. BAP 배지에 C. jejuni와 C. coli를각각접종하여 42, 10.0% CO 2 배양기에서 48시간배양한후집락을증류수에현탁시켰다. 균현탁액을 100 에서 10분간끓인다음 10,000 rpm 에서 5분간원심분리한후상층액을 PCR을위한주형 DNA로사용하였다. 본실험에사용된 primer의염기서열은 Table 1과같으며 Genotech Co. (Daejeon, Korea) 에의뢰하여합성하였다. PCR을위한반응액조성은 primer 각 1 μl, PCR mastermix (dntp, taq polymerase, MgCl 2, Daejeon, Korea) 4 μl, 8-MOP 12 μl, 주형 DNA 2 μl를넣어최종 20 μl로하였다. PCR 반응은 thermal cycler (GeneAmp system, PerkinCetus, Norwalk. CT, USA) 를사용하였다. 유전자증폭을위한반응조건은 predenaturation 95 에서 5분, denaturation 95 에서 30초, annealing 50 에서 30초, extention 72 에서 30초간 30회반복하였고, 72 에서 5 분간 last extention 시킨후 4 에서보관하였다. 증폭된 PCR 반응산물은 2.0% agarose gel (Cambrex bion science inc. Rockland, USA) 상에서전기영동하여확인하였다. 항균제감수성시험 Erythromycin 과 quinolone 내성균의분리. 내성균의분리를위해표준디스크확산법 (Clinical and Laboratory Standards Institute, 2012) 을수행하였다. 배지는 5.0% 면양혈액을넣은 Mueller-Hinton agar (MHA, Difco Laboratories, Sparks, MD, USA) 를사용하였다. 순수배양된집락을 Tryptic soy broth (TSB, Difco) 2 ml에현탁하여 1 McFarland 탁도로맞추고, 면봉으로배지의표면에고루바른후디스크를놓고 37, 10.0% CO 2 배양기에서 48시간배양후결과를판독하였다. 디스크는 amikacin 30 μg, ampicillin 10 μg, cephalothin 30 μg, ciprofloxacin 5 μg, erythromycin 15 μg, imipenem 10 μg, nalidixic acid 30 μg, tetracycline 30 μg (Becton-Dickinson, Cockeysville, MD, USA) 를사용하였다. 우무희석법 (Agar dilution method) 을위한배지는 CLSI 표준방법에따라 5.0% 혈액을첨가한 MHA 평판배지를사용하였다. 항생제는 ampicillin, cephalothin, clindamycin,

3 Table 2. Antimicrobials usceptibility patterns of C. jejuni isolated from swine MIC (µg/ml) Percentage of Organisms (No. tested) Antimicrobial agent Range 50% 90% resistant (%) Erythromycin-resistant (26) Ampicillin 2~ Cephalothin 64~ Ciprofloxacin 4~ Clindamycin 8~ Erythromycin Imipenem Nalidixic acid 32~ Tetracycline 4~ Erythromycin-susceptible (7) Ampicillin 4~ Cephalothin Ciprofloxacin 8~ Clindamycin 0.5~ Erythromycin 2~ Imipenem Nalidixic acid 128~ Tetracycline 8~ erythromycin, gentamicin, nalidixic acid, ciprofloxacin, tetracycline (Sigma Chemical, St. Louis, MO, USA) 및 imipenem (Merk Sharp & Dohme, Rahway, NJ, USA) 을사용하였다. 순수배양된집락을 TSB 2 ml에현탁하여 1 McFarland 탁도로맞추고, replicator 로접종하여 37, 10.0% CO 2 배양기에서 48시간배양후증식을억제시킨항균제의최소농도를최소발육억제농도 (minimum inhibitory concentration, MIC) 로측정하였다. 정도관리를위해 Escherichia coli ATCC 25922를동시에시험하였다. Ciprofloxacin 내성 gyra 유전자의검출. Ciprofloxacin 내성 Campylobacter의확인을위해서 gyra 유전자를 PCR 법으로확인하였다. PCR 반응조건은 hipo와 glya 유전자에사용된방법과동일하며, 사용한 primer 의염기서열은 Table 1과같다. 결과돼지에서 C. jejuni와 C. coli의분리 2009년 7월 2일부터 7일사이에부안지역일개농장에서돼지 100마리의직장변검체에서분리된 Campylobacter species의검출률은 55주 (55.0%) 이었으며, 이중 C. jejuni는 33주 (33.0%), C. coli는 22주 (22.0%) 가분리되었다. C. jejuni와 C. coli의생화학적성상. 돼지에서분리한 C. jejuni 33주와 C. coli 22주의생화학적성상은 Catalase 양성, Urease 음성, Oxidase는양성을보였으며, hippurate hydrolysis 시험에서는 C. jejuni는진한자주색을보여양성이었고, 무색인 C. coli는음성을보였다. Inodoxyl acetate hydrolysis는두균주모두양성을보였으며, TSI의 H 2 S 성상은모두음성이었다. Nalidixic acid와 Cephalothin 감수성시험은두균주모두내성으로동정되었다. 항균제감수성시험 Erythromycin 과 quinolone 내성균의분리. Campylobacter 균종감염의장염치료제로사용되고있는 erythromycin 을포함한 8종의항균제에대한감수성시험에서 erythromycin 내성률, MIC 범위, MIC 50 및 MIC 90 은 Table 2, 3과같다. 본시험에사용된 C. jejuni에대한 erythromycin의 MIC 범위는 128 μg/ml, MIC 90 은 128 μg/ml이었고, erythromycin 내성균주에대해가장항균력이높은것은 imipenem이었으며, 그 MIC 범위는 μg/ml, MIC 90 은 μg/ml이었고, ampicillin의 MIC 범위는 4~ 8 μg/

4 Table 3. Antimicrobial susceptibility patterns of C. coli isolates from swine MIC (µg/ml) Percentage of Organisms (No. tested) Antimicrobial agent Range 50% 90% resistant (%) Erythromycin-resistant (14) Ampicillin 2~ Cephalothin 64~ Ciprofloxacin 8~ Clindamycin 0.5~ Erythromycin Imipenem 0.06~ Nalidixic acid 128~ Tetracycline 0.5~ Erythromycin-susceptible (8) Ampicillin 4~ Cephalothin Ciprofloxacin 4~ Clindamycin 16~ Erythromycin 0.5~ Imipenem 0.06~ Nalidixic acid 64~ Tetracycline 2~ ml, MIC 90 은 8 μg/ml이었으며, tetracycline의 MIC 범위는 4~128 μg/ml, MIC 90 은 128 μg/ml, clindamycin 의 MIC 범위는 8~64 μg/ml, MIC 90 은 64 μg/ml이었다. 본시험에사용된 Erythromycin 내성 C. coli에대한 erythromycin의 MIC 범위는 128 μg/ml, MIC 90 은 128 μg/ml이었으며, erythromycin 내성균주에대해항균력이높은것은 ampicillin, clindamycin, imipenem이었으며, imipenem의 MIC 범위는 0.06~0.125 μg/ml, MIC 90 은 μg/ml이었고, ampicillin 의 MIC 범위는 2~ 8 μg/ml, MIC 90 은 8 μg/ml이었으며, tetracycline의 MIC 범위는 0.5~128 μg/ml, MIC 90 은 128 μg/ ml, clindamycin의 MIC 범위는 0.5~4 μg/ml, MIC 90 은 4 μg/ ml이었다. C. jejuni의 erythromycin 감수성균주에대해항균력이높은것은 imipenem이었으며, imipenem의 MIC 범위는 μg/ml, MIC 90 은 μg/ml이었고, erythromycin 의 MIC 범위는 2~4 μg/ml, MIC 90 은 4 μg/ml이었으며, ampicillin clindamycin의 MIC 범위는각각 4~64 μg/ml와 0.5~1 μg/ml, MIC 90 은 64 μg/ml와 1 μg/ml이었고, tetracycline 의 MIC 범위는 8~128 μg/ml, MIC 90 은 128 μg/ml이었다. Imipenem은 erythromycin 내성과감수성 Campylobacter 균종에서가장낮은 MIC 90 를보였으며, erythromycin 내성 Campylobacter 균종의장염치료제는 ampicillin이추천되었다. 또한 erythromycin 감수성 Campylobacter 균종에 대한 clindamycin은낮은 MIC 90 을보였다. 모든 Campylobacter 균종은 ciprofloxacin에내성이었으며, erythromycin 내성과감수성 C. jejuni는 tetracycline에각각 38.5% 와 71.4% 의내성률을, erythromycin 내성과감수성 C. coli는 tetracycline에각각 64.3% 와 62.5% 의내성률을보였다. C. jejuni와 C. coli의 hipo, glya와 gyra 유전자의검출. 생화학적동정시험에서 C. jejunie로동정된 33균주는모두 C. jejunie 특이유전자인 hipo를확인할수있었으며, C. coli로동정된 22균주는모두 C. coli 특이유전자인 glya를확인할수있었으며, nalidixic acid와 ciprofloxacin에내성을나타내는 C. jejuni와 C. coli의모든균주에서변이된유전자인 gyra가 266 bp에서관찰되어, ciprofloxacinresistant Campylobacter species임을알수있었다 (Table 4, Fig. 1-3). 고찰 Campylobacter는닭, 칠면조, 돼지, 개, 소, 고양이등에서보균률이높으며, 각종야생동물및가축의장관내에널리분포한다. 특히 C. jejuni는닭과소에서주로보균하고있으며, C. coli는돼지에서많이보균하고있어이

5 Table 4. Detection of hipo, glya and grya gene for C. jejuni and C. coli isolated from swine Source of isolation Detection of indicated genes by PCR hipo glya gyra CJ-01 * CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CJ CC CC CJ CC CJ CJ CJ CC CJ CJ CC CJ CC CC CC CJ CJ CC CC CJ CC CC CC CJ Table 4. Detection of hipo, glya and grya gene for C. jejuni and C. coli isolated from swine (contineued) Source of isolation Detection of indicated genes by PCR hipo glya gyra CC CC CC CC CJ CC CJ CJ CC CC CJ CJ CC CC Total * NO. of isolates tested. CJ, Campylobacter jejuni; CC. Campylobacter coli. 러한동물들로부터사람에감염되어 Campylobacter 장염을일으키는것으로알려져있다 (Guévremont et al., 2006; Shin et al., 2010). 이세균들은공기중의노출이나건조, 낮은 ph, 가열에대한저항성이낮으며, 식품중에는증식하기어려우나, 적은세균수의감염으로도장염을일으킬수있어, 여러나라에서세균성식중독의빈번한원인균으로보고되어왔다 (Pedersen, et al., 2003). Campylobacter 균종의감별을위한동정실험은생화학적성상으로쉽지않은것으로알려져왔으며, C. jejuni와 C. coli의분리동정법으로 hippurate hydrolysis 시험, nalidixic acid와 cephalothin 감수성시험이사용되어왔으나, nalidixic acid와 cephalothin에대한내성균주의출현으로인하여그이용가치가점차적어지고있다. Dassanayake 등은 C. jejuni의 13.0% 는 hippurate hydrolysis 음성이라고보고하여기존의생화학적동정실험만으로는정확한동정을기대할수없게되었다. Wang 등은 C. jejuni만이보유하고있는특이유전자를 hipo 유전자로확인이가능하며, C. coli만이보유하고있는특이유전자인 glya로유전자로확인이가능하다고보고한이래 hipo 유전자와 glya 유전자의검출은 C. jejuni와 C. coli 를동정하는데중요한지표로사용되고있다 (Wang et

Fig. 1. Agarose gel electrophoresis of PCR products by using primer pairs hipo gene for identification of C. jejuni. Lane 1 to 12, C. jejuni; M, size marker; lane 1, strain no.

CJ-9; lane 10, strain no. CJ-10; lane 11, strain no. CJ-11; lane 12, strain no. CJ-12. Fig. 2. Agarose gel electrophoresis of PCR generated by using primer pairs for glya gene for identification of C.

CC-30; lane 7, strain no. CC-31; lane 8, strain no. CC-32; lane 9, strain no. CC-35; lane 10, strain no. CC-36; lane 11, strain no. CC-38; lane 12, strain no. CC-39; lane 13, strain no.

6 Fig. 1. Agarose gel electrophoresis of PCR products by using primer pairs hipo gene for identification of C. jejuni. Lane 1 to 12, C. jejuni; M, size marker; lane 1, strain no. CJ-1; lane 2, strain no. CJ-2; lane 3, strain no. CJ-3; lane 4, strain no. CJ-4; lane 5, strain no. CJ-5; lane 6, strain no. CJ-6; lane 7, strain no. CJ-7; lane 8, strain no. CJ-8; lane 9, strain no. CJ-9; lane 10, strain no. CJ-10; lane 11, strain no. CJ-11; lane 12, strain no. CJ-12. Fig. 2. Agarose gel electrophoresis of PCR generated by using primer pairs for glya gene for identification of C. coli. Lane M, size marker; lane 1, strain no. CC-18; lane 2, strain no. CC-19; lane 3, strain no. CC-21; lane 4, strain no. CC-25; lane 5, strain no. CC-28; lane 6, strain no. CC-30; lane 7, strain no. CC-31; lane 8, strain no. CC-32; lane 9, strain no. CC-35; lane 10, strain no. CC-36; lane 11, strain no. CC-38; lane 12, strain no. CC-39; lane 13, strain no. CC-40; lane 14, strain no. CC-42; lane 15, strain no. CC-43; lane 16, strain no. C. coli ATCC Fig. 3. Agarose gel electrophoresis of PCR products by using primer pairs grya gene for detection of ciprofloxacin-resistant Campylobacter species Lane 1 to 12, Campylobacter species; M, size marker; lane 1, strain no. CJ-1; lane 2, strain no. CJ-2; lane 3, strain no. CJ-3; lane 4, strain no. CJ-4; lane 5, strain no. CJ-5; lane 6, strain no. CJ-6; lane 7, strain no. CJ-7; lane 8, strain no. CJ-8; lane 9, strain no. CJ-9; lane 10, strain no. CJ-10; lane 11, strain no. CJ-11; lane 12, strain no. CJ-12. al., 2002). 1994년 Sánchez 등은 Campylobacter 감염치료에널리사용되는 ciprofloxacin에대한내성조사보고에의하면스페인에서분리한균주중에는 ciprofloxacin 내성균이분리되지않았지만, 1992년에서는 49균주가내성임을보고하였으며, ciprofloxacin에대한내성은변이된 gyra 유전자에기인하는것으로보고하였다. 따라서 Campylobacter 균종의동정과 ciprofloxacin 내성연구에서 여러가지유전자검출방법들이보고되어왔으나, 본연구에서는 Wang 등이보고한 hipo와 glya 유전자중 hipo 양성은 C. jejuni로, glya 양성은 C. coli로동정하였으나 (Wang et al., 2002), Zirnstein 등은 ciprofloxacin 내성을나타내는균주에서변이된 gyra 유전자를검출하여 C. jejuni와 C. coli의 ciprofloxacin 내성세균의지표유전자로확인하였다 (Zirnstein et al., 1999)

7 Fluoroquinolone이개발된초기에는여러균종에대한항균력이높아 (Segreti et al., 1989) Campylobacter 장염치료에유용하게사용되었으며 (Goodman et al., 1990), fluoroquinolone이널리사용됨에따라서 fluoroquinolone 내성균주가증가하고있음이보고되었다 (Zirnstein et al., 1999). Payot 등 (2004) 은프랑스돼지에서분리한 C. coli 균주중 fluoroquinolone 내성 Campylobacter 균주가 65.8% 임을보고하였고, Kim 등 (2008) 은우리나라익산지역의닭과환자에서분리한 C. jejuni가모두 nalidixic acid와 ciprofloxacin 내성임을보고하였다. 본연구에서돼지를대상으로분리된 C. jejuni와 C. coli에서는 nalidixic acid와 ciprofloxacin에모두내성을나타내었으며, nalidixic acid의 MIC 50 와 MIC 90 은각각 128 μg/ml이었다. Ciprofloxacin 내성유전자인 gyra 유전자가본연구에서분리한 C. jejuni와 C. coli 모두에서검출되어, Campylobacter 감염치료에 fluoroquinolone을사용할수없을뿐만아니라, nalidixic acid 디스크시험은 Campylobacter species 감별에도이용할수없으며, Campylobacter 감염치료에 fluoroquinolone을사용할때에는항균제감수성시험이반드시필요할것으로사료된다. 본실험에사용된 Campylobacter 균종 55주는 cephalothin, ciprofloxacin, erythromycin, nalidixic acid에내성이었고, erythromycin 내성 C. jejuni는 ampicillin에는 3.8%, clindamycin에 88.4%, imipenem에 0%, tetracycline에 38.5% 가내성이었으며, 가장낮은 MIC 90 를보인항균제는 ampicillin이었다. Erythromycin 감수성 C. jejuni는 ampicillin에 14.2%, clindamycin 과 imipenem에 0%, tetracycline에는 71.4% 가내성이었으며, 가장낮은 MIC 90 를보인항균제는 clindamycin이었다. 한편 erythromycin 내성 C. coli는 ampicillin, clindamycin, imipenem은 0%, tetracycline은 64.3% 가내성이었고, 가장낮은 MIC 90 를보인항균제는 clindamycin이었으며, erythromycin 감수성 C. coli는 ampicillin과 imipenem은 0%, clindamycin은 92.8%, tetracycline은 62.5% 가내성이었으며, 가장낮은 MIC 90 를보인항균제는 erythromycin 으로나타났다. 따라서본연구에사용된 Campylobacter species 55주모두에서퀴롤론계항균제에대해서 100.0% 내성을나타내앞으로 Campylobacter species 감염치료에사용할수없다는결론을얻었으며, erythromycin과 tetracycline에대해서도내성균이점차증가하고있어 Campylobacter species 감염에대한치료시에는반드시항균제감수성시험이나관련유전자의검출시험을한후에치료가필요할것으로사료된다. REFERENCES Allos BM. Campylobacter jejuni Infections: update on emerging issues and trends. Clin Infect Dis : Altekruse SF, Stern NJ, Fields PI, Swerdlow DL. Campylobacter jejuni an emerging foodborne pathogen. Emerg Infect Dis : Arestrup FM, Engberg J. Antimicrobial resistance of thermophilic Campylobacter. Vet Res : Chuma T, Ikeda T, Maeda T, Niwa H, Okamoto K. Antimicrobial susceptibilities of Campylobacter strains isolated from broilers in the southern part of Japan from 1995 to J Vet Med Sci : Clinical and Laboratory Standards Institute: Methods for antimicrobial dilution and disk susceptibility testing of infrequently isolated or fastidious bacteria; approved guideline. CLSI Document M45-A, Dassanayake RP, Zhou Y, Hinkley S, Stryker CJ, Plauche G, Borda JT, Sestak K, Duhamel GE. Characterization of cytolethal distending toxin of Campylobacter species isolated from captive macaque monkeys. J Clin Microbiol : Gaunt PN, Piddock LJ. ciprofloxacin resistant Campylobacter species in humans: an epidemiological and laboratory study. J Antimicrob Chemother : Goodman LJ, Trenholme GM, Kaplan RL, Segreti J, Hines D, Petrak R, Nelson JA, Mayer KW, Landau W, Parkhurst GW, Stuart Levin. Empiric antimicrobial therapy of domestically acquired acute diarrhea in urban adults. Arch lntern Med : Guévremont E, Nadeau E, Sirois M, Quessy S. Antimicrobial susceptibilities of thermophilic Campylobacter from humans, swine, and chicken broilers. Can J Vet Res : Kim SM, Kim EC, Choi MR, So HA, Shim ES, Kim ES, Park SC, Seong CN, Chong Y. Cytolethal Distending Toxin Production, Genotypes and Atimicrobial Susceptibility of Campylobacter jejuni Isolates from Diarrhea Patients and Chickens. Journal of Bacteriology and Virology : Payot S, Avrain L, Magras C, Praud K, Cloeckaert A, Chaslus- Dancla E. Relative contribution of target gene mutation and efflux to fluoroquinolone and erythromycin resistance, in French poultry and pig isolates of Campylobacter coli. Int J Antimicrob Agents :

8 Pedersen K, Wedderkopp A. Resistance to quinolones in Campylobacter jejuni and Campylobacter coli from Danish broilers at farm level. J Appl Microbiol : Sánchez R, Fernández-Baca V, Díaz MD, Muñoz P, Rodríguez- Créixems M, Bouza E. Evolution of susceptibilities of Campylobacter species to quinolones and macrolides. Antimicrob Agents Chemother : Segreti J, Nelson JA, Goodman LJ, Kaplan RL, Trenholme GM. In vitro activities of lomefloxacin and temafloxacin against pathogens causing diarrhea. Antimicrob Agents Chemother : Shin E, Lee Y. Characterization of erythromycin-resistant porcine isolates of Campylobacter coli. Microb Drug Resist : Smith KE, Besser JM, Hedberg CW, Leano FT, Bender JB, Wicklund JH, Johnson BP, Moore KA, Osterholm MT. Quinolone-resistant Campylobacter jejuni infections in Minnesota, Investigation Team. N Engl J Med ; 340: Van Looveren M, Daube G, De Zutter L, Dumont JM, Lammens C, Wijdooghe M, Vandamme P, Jouret M, Cornelis M, Goossens H. Antimicrobial susceptibilities of Campylobacter strains isolated from food animals in Belgium. J Antimicrob Chemother : Wang G, Clark CG, Taylor TM, Punknell C, Barton C, Price L, Woodward DL, Rodgers FG. Colony multiplex PCR assay for identification and differentiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis and C. fetus subsp. J Clin Microbiol : Zirnstein G, Li Y, Swaminathan B, Angulo F. Ciprofloxacin resistance in Campylobacter jejuni isolates: detection of gyra resistance mutations by mismatch amplification mutation assay PCR and DNA sequence analysis. J Clin Microbiol :

Microsoft PowerPoint - PDFCoTemp.PPT

Microsoft PowerPoint - PDFCoTemp.PPT 주요유방염세균의검출및 항생제감수성검사 임숙경 1. 검사시료 1. 시료채취 유방염우유 2. 체세포수측정 3. 대상세균 - Staphylococcus aureus - Coagulase negative staphylococci -Escherichia coli 3. 균분리방법 E. coli CHEC Agar 직접도말 (37 18-24h) 보라색집락 EMB Agar