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患白内障病黑斑蛙米尔伊丽莎白菌的分离鉴定与PNGase基因克隆 被引量:1

Isolation,identification of Elizabethkingia miricola of Pelophylax nigromaculatus with cataract and cloning of PNGase gene
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摘要 为给安徽舒城地区患白内障病黑斑蛙(Pelophylax nigromaculatus)提供治疗方案,对病蛙进行了细菌分离、鉴定和药敏检测。从典型患病黑斑蛙的肝脏、眼睛、脑和脊柱中分离出4株优势菌株,经16S rRNA和gyrB基因序列分析,结合生理生化鉴定,结果表明,4株菌株均为米尔伊丽莎白菌(Elizabethkingia miricola)。对分离菌的N-糖苷酶(PNGase)基因进行了克隆和分析,结果显示,其包含PNGase F和PNGase F-Ⅱ两种类型,进化分析结果同样显示,其与米尔伊丽莎白菌聚为一支,表明PNGase基因序列可作为伊丽莎白菌属内部菌分类的参考依据。药敏试验结果显示,4株分离株仅对酰胺醇类中的氟苯尼考敏感,对香豆素类中的新生霉素、大环内酯类中的红霉素和利福霉素类中的利福平等3种抗生素中介,而对测试的其他14种抗生素耐药。实验结果为伊丽莎白菌属细菌的分类鉴定提供了参考方法,且为黑斑蛙细菌病防治提供参考依据。 In order to provide treatment protocols for the cataract disease of black-spotted pond frog(Pelophylax nigromaculatus)in Shucheng area,Anhui Province,four dominant bacterial strains were isolated from the liver,eyes,spine and brain,respectively.The 16S rRNA gene and gyrB gene sequences of the isolated strains were analyzed,and the results showed that the four strains were Elizabethkingia miricola,which was confirmed by physiological and biochemical identification.N-glycosidase(PNGase)genes of the isolated strain was cloned and analysed,and the results showed it contained two types,PNGase F and PNGase F-Ⅱ.The phylogenetic analysis of PNGase F and PNGase F-Ⅱboth showed that the isolated strain was clustered with Elizabethkingia miricola in Elizabethkingia genus,which indicated that they can be used as reference genes for the classification of Elizabetha genus.Antimicrobial susceptibility test showed that the four isolates were only sensitive to florfenicol in amidols,intermediary to neosporin in coumarin,erythromycin in macrolide and rifampin in rifamycin,and resistant to other 14 antibiotics.The experiment results will provide a reference for the prevention and treatment of the bacterial diseases of black-spotted pond frog.
作者 龚保荣 吴红军 李本镇 徐大洋 邹文腾 曲君艺 鲍传和 朱若林 GONG Baorong;WU Hongjun;LI Benzhen;XU Dayang;ZOU Wenteng;QU Junyi;BAO Chuanhe;ZHU Ruolin(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China)
出处 《浙江农业学报》 CSCD 北大核心 2023年第6期1297-1306,共10页 Acta Agriculturae Zhejiangensis
基金 安徽省高校自然科学基金(KJ2019A0164) 安徽省大学生创新创业训练计划项目(201910364056)。
关键词 黑斑蛙 米尔伊丽莎白菌 16S rRNA GYRB N-糖苷酶 药敏试验 Pelophylax nigromaculatus Elizabethkingia miricola 16S rRNA gyrB N-glycosidase antimicrobial susceptibility test
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