冀南地区滑液支原体分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2023.10.041

摘要/Abstract

摘要： 【目的】了解中国冀南地区滑液支原体(Mycoplasma synoviae,MS)的流行和耐药情况,为该地区防控MS感染提供科学依据。【方法】于2019—2022年采集冀南地区疑似MS感染的7个规模化鸡场样品(发病鸡的气囊、腭裂或爪垫渗出液)进行病原分离培养及PCR鉴定,对分离株进行vlhA基因分型序列分析,采用微量肉汤稀释法测定分离株最小抑菌浓度(MIC)。【结果】所采集样品中分离出9株疑似MS菌株,均可使液体培养基颜色变为橘黄,在固体培养基中菌落形态为典型的"煎荷包蛋"样;PCR鉴定及测序结果显示,9株分离株与MS HN01株相似性为100%,表明分离株均为MS;vlhA基因分型序列结果显示,9株MS富含脯氨酸重复序列(PRR)区均为35个氨基酸,属于L型,核苷酸序列相似性为99.7%~100%,与中国其他地区L型菌株亲缘关系较近,与泰国L型菌株亲缘关系较远;MIC测定结果显示,9株MS对泰乐菌素、替米考星、螺旋霉素、林可霉素、沃尼妙林、泰妙菌素、多西环素、土霉素、氟苯尼考、恩诺沙星的MIC_50/90值分别为0.25/0.25、0.5/0.5、0.5/0.5、1/1、≤0.016/≤0.016、0.062/0.125、1/1、2/2、4/4和32/32 μg/mL。【结论】本研究于冀南地区成功分离到9株L型MS,与国内其他地区MS差异较小、遗传进化相对稳定,治疗当地MS感染可选用泰妙菌素、沃尼妙林、泰乐菌素、替米考星、螺旋霉素,其次为林可霉素、多西环素、土霉素。本研究结果可为当地防控MS感染提供科学依据。

关键词: 滑液支原体; 分离鉴定; vlhA基因; 最小抑菌浓度

Abstract: 【Objective】 The prevalence and antibiotic resistances of Mycoplasma synoviae (MS) isolates were investigated in the Southern region of Hebei province of China,so as to provide scientific basis for the control of MS infection in this region.【Method】 Samples (the exudate from the airbag,cleft palate,or paw pad of diseased chickens) were collected from 7 large-scale chicken farms suspected of MS infection in the Southern region of Hebei province from 2019 to 2022.Strains were isolated from the samples and identified by PCR.DNA sequence analysis of the vlhA gene was conducted for genotyping of the isolated strains.Minimal inhibitory concentration (MIC) values of the strains were determined by the microbroth dilution method.【Result】 Nine suspected MS strains were isolated from the collected samples.All the strains could turn the liquid culture medium orange and showed typical "fried eggs" colonies on solid culture medium.PCR identification and sequencing results showed that the 9 isolated strains had 100% similarity with the typical HN01 strain,indicating that all of which were MS strains.The vlhA gene typing sequence analysis showed that 9 strains all had 35 amino acis in the proline-rich repeats (PRR),which hence belonged to the L-type,with a nucleotide sequence similarity of 99.7% to 100%.The phylogenetic relationships of isolated strains were close with those L-type strains isolated from other regions of China,whereas far from the L-type strains reported in Thailand.MIC_50/90 values of the 9 MS strains for tylosin,tilmicosin,spiramycin,lincomycin,valnemulin,tiamulin,doxycycline,oxytetracycline,florfenicol,and enrofloxacin were 0.25/0.25,0.5/0.5,0.5/0.5,1/1,≤0.016/≤0.016,0.062/0.125,1/1,2/2,4/4 and 32/32 μg/mL,respectively.【Conclusion】 Nine L-type MS strains were isolated in the Southern region of Hebei province,which showed tiny differences from strains isolated in other regions of China and were relatively stable in genetic evolution.The tiamulin,valnemulin,tylosin,tilmicosin,spiramycin could be recommended for the therapy of MS infections in the region followed by lincomycin,doxycycline,oxytetracycline.The presented data might serve as a guide for the prevention and control of MS infection in the region.

Key words: Mycoplasma synoviae; isolation and identification; vlhA gene; minimum inhibitory concentration

中图分类号:

S852.62

刘子卿, 熊威振, 任豆豆, 周守长, 刘冠慧, 许金朋. 冀南地区滑液支原体分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2023, 50(10): 4270-4278.

LIU Ziqing, XIONG Weizhen, REN Doudou, ZHOU Shouchang, LIU Guanhui, XU Jinpeng. Isolation,Identification and Drug Resistance Analysis of Mycoplasma synoviae in the Southern Region of Hebei Province[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4270-4278.

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