李轩;谢春;周藜;汪淑颖;向婧姝;黄靖宇;王翠桑;向红;
LI Xuan;XIE Chun;ZHOU Li;WANG Shu-ying;XIANG Jing-shu;HUANG Jing-yu;WANG Cui-sang;XIANG Hong;Guizhou Center for Disease Control and Prevention;The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education,School of Public Health, Guizhou Medical University;

• 1:贵州省疾病预防控制中心
• 2:贵州医科大学公共卫生与健康学院环境污染与疾病监控教育部重点实验室

摘要(Abstract)：

为了解鸡源弯曲菌分离株中毒力基因携带情况和耐药现状，探讨毒力基因的存在与耐药性的关系，本试验通过PCR方法对贵州省9个市(州)禽类养殖场分离的96株弯曲菌进行毒力基因检测，采用琼脂稀释法进行药敏试验，通过分类变量的关联性分析，计算关联系数(Cramer's V)后分析毒力基因与耐药性的相关性。结果显示，在弯曲菌分离株中携带率位于前3名的毒力基因是sodB、cheY、fur(99.0%、96.9%、96.9%),ciaB、virB11的携带率均为0;空肠弯曲菌优势毒力基因谱为flaA-cadF-cheY-cdtA-cdtB-cdtC-fur-katA-sodB,结肠弯曲菌优势毒力基因谱为flaA-cadF-cheY-fur-katA-sodB,结肠弯曲菌毒力基因谱型多于空肠弯曲菌。空肠弯曲菌和结肠弯曲菌分离株耐药率位于前3名的抗菌药均为萘啶酸(88.2%、93.5%)、四环素(85.2%、90.3%)和环丙沙星(73.5%、87.0%);结肠弯曲菌对红霉素、阿奇霉素、链霉素、氯霉素、泰利霉素和克林霉素的耐药水平高于空肠弯曲菌。红霉素与cdtC基因，阿奇霉素与cdtA、cdtB基因，环丙沙星与cdtB、cdtC和wlaN基因，链霉素与cdtA、cdtB、cdtC和wlaN基因，氯霉素与flaA基因，四环素与fur基因均存在弱相关性。结果表明，贵州省鸡源弯曲菌分离株毒力基因普遍存在，且菌株耐药情况严重，部分毒力基因与耐药性之间存在统计学关联。
In order to determine the presence of virulence genes and the current status of antibiotic resistance in Campylobacter chicken isolates, and to explore the relationship between the existence of virulence genes and drug resistance, this study utilized PCR for detecting virulence genes, agar dilution method for testing drug susceptibility, and correlation coefficient(Cramer's V) for assessing the correlation between virulence genes and antibiotic resistance on 96 Campylobacter isolates(34 Campylobacter jejuni isolates and 62 Campylobacter coli isolates) from poultry farms in 9 cities(prefectures) in Guizhou Province. The results showed that the top three virulence genes in Campylobacter isolates were sodB, cheY, and fur, with carrier rates of 99.0%, 96.9%, and 96.9%, respectively, while ciaB and virB11 were absent. The dominant virulence gene spectrum in Campylobacter jejuni(C. jejuni) was flaA-cadF-cheY-cdtA-cdtB-cdtC-fur-katA-sodB, while the dominant virulence gene spectrum in Campylobacter coli(C. coli) was flaA-cadF-cheY-fur-katA-sodB. The virulence gene spectrums of C. coli were more than those of C. jejuni. The top three resistance rates of C. jejuni and C. coli isolates were nalidixic acid(88.2% and 93.5%), tetracycline(85.2% and 90.3%) and ciprofloxacin(73.5% and 87.0%); C. coli was more resistant to erythromycin, azithromycin, streptomycin, chloramphenicol, telithromycin and clindamycin than C. jejuni. There was a weak correlation between erythromycin and cdtC gene, azithromycin and cdtA, cdtB genes, ciprofloxacin and cdtB, cdtC, wlaN genes, streptomycin and cdtA, cdtB, cdtC, wlaN genes, chloramphenicol and flaA gene, and tetracycline and fur gene. Thus, the virulence genes of Campylobacter chicken isolates in Guizhou Province are widespread, and Campylobacter strains show serious drug resistance; there is a statistical correlation between the existence of some virulence genes and drug resistance.

关键词(KeyWords)： 弯曲菌;毒力基因;耐药性;相关性
Campylobacter;virulence gene;drug resistance;correlation

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技计划项目[黔科合基础(2017)1096];; 贵州省传染病预防与控制人才基地科研团队(RCJD2108)

作者(Authors): 李轩;谢春;周藜;汪淑颖;向婧姝;黄靖宇;王翠桑;向红;
LI Xuan;XIE Chun;ZHOU Li;WANG Shu-ying;XIANG Jing-shu;HUANG Jing-yu;WANG Cui-sang;XIANG Hong;Guizhou Center for Disease Control and Prevention;The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education,School of Public Health, Guizhou Medical University;

参考文献(References)：

• [1] Tacconelli E,Carrara E,Savoldi A,et al.Discovery,research,and development of new antibiotics:the WHO priority list of antibiotic-resistant bacteria and tuberculosis[J].Lancet Infect Dis,2018,18(3):318- 327.
• [2] Wei B,Cha S Y,Zhang J F,et al.Antimicrobial susceptibility and association with toxin determinants in clostridium perfringens isolates from chickens[J].Microorganisms,2020,8(11):1825.
• [3] Bessède E,Delcamp A,Sifré E,et al.New methods for detection of Campylobacters in stool samples in comparison to culture[J].J Clin Microbiol,2011,49(3):941- 944.
• [4] Koolman L,Whyte P,Burgess C,et al.Distribution of virulence-associated genes in a selection of Campylobacter isolates[J].Foodborne Pathog Dis,2015,12(5):424- 432.
• [5] 李健，时玉雯，刘辉.济南市鸡源空肠弯曲菌的流行特征及病原毒力、耐药性分析[J].现代预防医学，2021,48(22):4073- 4077.
• [6] Kahlmeter G,Brown D F,Goldstein F W,et al.European Committee on Antimicrobial Susceptibility Testing (EUCAST) technical notes on antimicrobial susceptibility testing[J].Clin Microbiol Infect,2006,12(6):501- 503.
• [7] Goualié B G,Akpa E E,Kakou-N'Gazoa S E,et al.Antimicrobial resistance and virulence associated genes in Campylobacter jejuni isolated from chicken in C?te d'Ivoire[J].J Infect Dev Ctries,2019,13(8):671- 677.
• [8] 唐梦君，周倩，张小燕，等.江苏部分地区鸡源与猪源弯曲菌耐药性分析与毒力基因检测[J].畜牧兽医学报，2020,51(9):2284- 2292.
• [9] Bardoň J,Pudová V,Kol■ková I,et al.Virulence and antibiotic resistance genes in Campylobacter spp.in the Czech Republic[J].Epidemiol Mikrobiol Imunol,2017,66(2):59- 66.
• [10] Sekizuka T,Murayama O,Moore J E,et al.Flagellin gene structure of flaA and flaB and adjacent gene loci in urease-positive thermophilic Campylobacter (UPTC)[J].J Basic Microbiol,2007,47(1):63- 73.
• [11] Nguyen T N,Hotzel H,El-Adawy H,et al.Genotyping and antibiotic resistance of thermophilic Campylobacter isolated from chicken and pig meat in Vietnam[J].Gut Pathog,2016,8:19.
• [12] Fraz?o M R,Medeiros M,Duque S,et al.Pathogenic potential and genotypic diversity of Campylobacter jejuni:a neglected food-borne pathogen in Brazil[J].J Med Microbiol,2017,66(3):350- 359.
• [13] Wieczorek K,Wo?kowicz T,Osek J.Antimicrobial resistance and virulence-associated traits of Campylobacter jejuni isolated from poultry food chain and humans with diarrhea[J].Front Microbiol,2018,9:1508.
• [14] 翟海华，王娟，盖文燕，等.胶东半岛地区鸡源空肠弯曲杆菌毒力基因携带状况调查[J].中国兽医杂志，2015,51(5):15- 18.
• [15] Guirado P,Paytubi S,Miró E,et al.Differential distribution of the wlaN and cgtB genes,associated with Guillain-Barré syndrome,in Campylobacter jejuni isolates from humans,broiler chickens,and wild birds[J].Microorganisms,2020,8(3):325.
• [16] Wysok B,Wojtacka J,Kivist? R.Pathogenicity of Campylobacter strains of poultry and human origin from Poland[J].Int J Food Microbiol,2020,334:108830.
• [17] Jelena J,Ivana B L,Tatjana B,et al.The prevalence of four virulence genes in strains of Campylobacter jejuni isolated from broilers in Serbia[J].Veterinarski Arhiv,2020,90(1):39- 45.
• [18] Du Y,Wang C,Ye Y,et al.Molecular identification of multidrug-resistant Campylobacter species from diarrheal patients and poultry meat in Shanghai,China[J].Front Microbiol,2018,9:1642.
• [19] Shobo C O,Bester L A,Baijnath S,et al.Antibiotic resistance profiles of Campylobacter species in the South Africa privatehealth care sector[J].J Infect Dev Ctries,2016,10(11):1214- 1221.
• [20] Li B,Ma L,Li Y,et al.Antimicrobial resistance of Campylobacter species isolated from broilers in live bird markets in Shanghai,China[J].Foodborne Pathog Dis,2017,14(2):96- 102.
• [21] 余峰玲，许艳平，童晶，等.徐州市禽源弯曲菌的分离鉴定及耐药性分析[J].食品安全质量检测学报，2020,11(18):6628- 6632.
• [22] Gharbi M,Béjaoui A,Ben H C,et al.Prevalence and antibiotic resistance patterns of Campylobacter spp.isolated from broiler chickens in the north of Tunisia[J].Biomed Res Int,2018,2018:7943786.
• [23] García-Fernández A,Dionisi A M,Arena S,et al.Human campylobacteriosis in Italy:emergence of multi-drug resistance to ciprofloxacin,tetracycline,and erythromycin[J].Front Microbiol,2018,9:1906.
• [24] Wang Y,Dong Y,Deng F,et al.Species shift and multidrug resistance of Campylobacter from chicken and swine,China,2008—14[J].J Antimicrob Chemother,2016,71(3):666- 669.
• [25] Abraham S,Sahibzada S,Hewson K,et al.Emergence of fluoroquinolone-resistant Campylobacter jejuni and Campylobacter coli among Australian chickens in the absence of fluoroquinolone use[J].Appl Environ Microbiol,2020,86(8):e02765-19.
• [26] Ghunaim H,Behnke J M,Aigha I,et al.Analysis of resistance to antimicrobials and presence of virulence/stress responsegenes in Campylobacter isolates from patients with severe diarrhoea[J].PLoS One,2015,10(3):e119268.
• [27] Almofti Y A,Dai M,Sun Y,et al.Impact of erythromycin resistance on the virulence properties and fitness of Campylobacter jejuni[J].Microb Pathog,2011,50(6):336- 342.
• [28] Raeisi M,Khoshbakht R,Ghaemi E A,et al.Antimicrobial resistance and virulence-associated genes of Campylobacter spp.isolated from raw milk,fish,poultry,and red meat[J].Microb Drug Resist,2017,23(7):925- 933.
• [29] Ghunaim H,Behnke J M,Aigha I,et al.Analysis of resistance to antimicrobials and presence of virulence/stress responsegenes in Campylobacter isolates from patients with severe diarrhoea[J].PLoS One,2015,10(3):e119268.