Abstract：The aim of this study is to investigate the contamination of Campylobacter spp. from broiler slaughter and processing in chicken chain and its antimicrobial resistance, the drug resistant genes in Jiangsu Province, China. A total of 170 samples were collected from each section during chicken slaughtering and processing. CCDA plate was selected for separation and purification. PCR was performed to identify Campylobacter species including Campylobacter jejuni (C. jejuni) and Campylobacter coli (C.coli). Susceptibility of bacteria to 14 antibiotics belong to 6 categories was determined by the Kirby-Bauer (K-B) method as recommended by the CLSI. The class Ⅰ integron was detected through the int1 gene amplification by PCR. Tet(O) and aadE-sat4-aphA-3 were found by PCR. The Campylobacter isolates of the point mutation of gyrA gene at DNA sequence position 257 was examined by PCR and sequencing. Results showed that 124 Campylobacter isolates were obtained from 170 samples (72.95%). The resistant rates for Campylobacter isolates were 100% against Streptomycin, Tobramycin, Kanamycin, Ciprofloxacin, Ofloxacin, Nalidixic acid, Erythromycin and Tetracycline. The multi-drug resistance was common among Campylobacter isolates and the multiple resistant rate was 100%. The regnant drug-resistant spectrum is CTX-CRO-cDA-K-NOR-S-AZM-CIP-TE-NA-E-LEV-ENR-TOB. The class Ⅰ integron of Campylobcter isolates did not detected. The sequencing results showed that all of the Campylobacter isolates had the Thr-86-Ile point mutation in gyrA at quinolone resistant determining region. The detection rate of tet(O) and aadE-sat4-aphA-3 gene cluster were 100% and 90.32%, respectively. This study provided that the Campylobacter contamination occurred in the broilers slaughter and processing chain, and the antimicrobial resistance of Campylobacter isolates was relatively serious. There was significant correlation between antimicrobial phenotypes of Campylobacter isolates with their gene.
[1] Gillespie IA, O’Brien SJ, Frost JA, et al. A case-case comparison of Campylobacter coli and Campylobacter jejuni infection: a tool for generating hypotheses [J]. Emerg Infect Dis, 2002, 8(9): 937-942. DOI:10.3201/eid0809.010187 [2] Scallan E, Hoekstra RM, Angulo FJ, et al. Foodborne illness acquired in the United States--major pathogens [J]. Emerg Infect Dis, 2011, 17(1): 7-15. DOI: 10.3201/eid1701.091101p1 [3] 张小燕,周倩,唐梦君,等.江苏地区鸡源弯曲杆菌分离鉴定及耐药性研究[J].中国家禽,2017,69(18): 23-27. DOI:10.16372/j.issn.1004-6364.2017.18.005 [4] Clinical and Laboratory Standards Institute.Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fifth Informational Supplement. CLSI document M100-S25[S].Wayne, PA: Clinical and Laboratory Standards Institute, 2015,44-51. [5] 唐梦君, 周倩, 张小燕, 等.江苏省鸡肉产品中弯曲菌耐药特征及I型整合子分析[J].现代食品科技, 2018, 34(2):1-7. DOI:10.13982/j.mfst.1673-9078.2018.2.019 [6] Zirnstein G, Li Y, Swaminathan B, et al. Ciprofloxacin resistance in Campylobacter jejuni isolates: detection of gyrA resistance mutations by mismatch amplification mutation assay PCR and DNA sequence analysis [J]. J Clin Microbiol, 1999, 37(10):3276-3280. [7] Abdi-Hachesoo B, Khoshbakht R, Sharifiyazdi H,et al. Tetracycline resistance genes in Campylobacter jejuni and C. coli isolated from poultry carcasses [J]. Jundishapur J Microbiol, 2014, 7(9):12-129. DOI: 10.5812/jjm.12129 [8] 张艾煜,顾一心,梁昊,等. 我国607株不同宿主来源弯曲菌耐药基因簇aadE-sat4-aphA-3分布分析[J]. 疾病监测,2015,30(6):479-484. DOI: 10.3784/j.issn.1003-9961.2015.06.012 [9] Gripp E, Hlahla D, Didelot X, et al. Closely related Campylobacter jejuni strains from different sources reveal a generalist rather than a specialist lifestyle [J]. BMC Genomics, 2011, 12: 584. DOI: 10.1186/1471-2164-12-584 [10] Rasschaert G, Houf K, Van Hende J, et al. Campylobacter contamination during poultry slaughter in Belgium [J]. J Food Prot, 2006, 69(1): 27-33. [11] 焦新安,黄金林.世界禽病的研究进展[J].中国牧业通讯, 2005, (01): 76-79. [12] 张秀丽,炊慧霞,廖兴广,等.2011年河南省肉鸡养殖和屠宰加工过程中弯曲菌污染状况主动监测[J]. 中国卫生检验杂志, 2013,9(23): 2133-2135. [13] 焦扬,翟伟华,黄金林.鸡肉屠宰加工环节弯曲菌流行病学调查与分析[J]. 肉类工业, 2011,4:44-46. [14] Lim SK, Moon DC, Chae MH, et al. Macrolide resistance mechanisms and virulence factors in erythromycin-resistant Campylobacter species isolated from chicken and swine feces and carcasses [J]. J Vet Med Sci, 2017, 78(12): 1791-1795. DOI:10.1292/jvms.16-0307 [15] Nguyen TN, Hotzel H, Njeru J, et al. Antimicrobial resistance of Campylobacter isolates from small scale and backyard chicken in Kenya [J]. Gut Pathog,2016, 8(1): 39. DOI: 10.1186/s13099-016-0121-5 [16] Shobo CO, Bester LA, Baijnath S, et al. Antibiotic resistance profiles of Campylobacter species in the South Africa private health care sector [J]. J Infect Dev Ctries,2016, 10(11): 1214-1221. [17] 张艾煜. 弯曲菌抗生素敏感性及遗传特征分析[D]. 北京:中国疾病预防控制中心,2015. [18] Piccirillo A, Dotto G, Salata C, et al. Absence of class Ⅰ and class Ⅱ integrons among Campylobacter jejuni and Campylobacter coli isolated from poultry in Italy [J]. J Antimicrob Chemother, 2013, 20: 2683-2685. DOI: 10.1093/jac/dkt242 [19] 胡欣洁, 韩新锋, 朱冬梅,等. 肉鸡源多重耐药空肠、结肠弯曲菌的耐药分子特征[J]. 中国人兽共患病学报, 2015,31 (08): 694-699. DOI:10.3969/ j.issn.1002-2694.2015.08.002 [20] Wang Y, Huang WM, Taylor DE. Cloning and nucleotide sequence of the Campylobacter jejuni gryA gene and characterization of quinolone resistance mutantion [J]. Antimicrobian Agents Chemother, 1993, 37(3): 457-463. [21] Gibreel A, Tracz DM, Taylor DE, et al. Incidence of antibiotic resistance in Campylobacter jejuni isolated in AIberta, Canada, from 1999 to 2002, with special reference to tet(O)-mediated tetracycline resistance [J]. Antimicrob Agents Chemother, 2004, 48(9): 3442-3450. DOI: 10.1128/AAC.48.9.3442-3450.2004 [22] Qin SS,Wu CM,Wang Y,et a1.Antimicrobial resistance in Campylobacter coli isolated from pigs in two provinces of China [J].Int J Food Microbiol,2011,146(1):94-98. DOI: 10.1016/j.ijfoodmicro.2011.01.035
2018, Vol. 34 
