珲春敬信湿地候鸟白额雁恶臭假单胞菌的分离鉴定及药物敏感性分析

doi:10.16431/j.cnki.1671-7236.2025.01.042

摘要/Abstract

摘要： 【目的】了解中、俄、朝边境地区珲春敬信湿地候鸟携带病原菌情况,为候鸟疫病防控提供科学依据。【方法】本研究在珲春敬信湿地采集155份候鸟白额雁新鲜粪便样本,用PBS进行稀释,接种到LB固体培养基和血液琼脂培养基上培养,挑取单个菌落进行分离纯化后,采用革兰染色镜检、生化鉴定、16S rRNA基因PCR扩增及测序等方法进行细菌分离鉴定,并对分离菌进行耐药性分析。【结果】分离菌为革兰阴性菌,呈紫红色短杆状。生化鉴定结果显示,分离菌尿素、赖氨酸脱羧酶、鸟氨酸脱羧酶、半固体等呈阳性;硫化氢、苯丙氨酸、葡萄糖酸盐等呈阴性。PCR扩增出大小为1 500 bp的单一条带,扩增序列与NCBI中恶臭假单胞菌MZ836867序列相似性最高,为99.6%。系统进化树显示,分离菌与恶臭假单胞菌MZ836867属于同一个分支,说明分离菌为恶臭假单胞菌。药敏试验结果显示,分离菌对氯霉素、复方新诺明、诺氟沙星等13种药物敏感;对多黏菌素B、克林霉素、亚胺培南等9种药物中介;对万古霉素、氟苯尼考、林可霉素、苯唑西林等8种药物耐药,其中对林可霉素、苯唑西林2种药物耐药性最强。【结论】本研究从珲春敬信湿地候鸟白额雁粪便中成功分离出具有耐药性的恶臭假单胞菌,为珲春敬信湿地候鸟白额雁肠道菌群的深入研究提供了参考。

关键词: 珲春; 白额雁; 恶臭假单胞菌; 分离鉴定; 耐药性

Abstract: 【Objective】 This study was aimed to understand the China-Russia-North Korea border area prevalence of pathogenic bacteria carried by migratory birds in Hunchun Jingxin wetland,so as to provide scientific basis for migratory bird disease prevention and control. 【Method】 This study collected 155 fresh fecal samples from the migratory bird species Anser albifrons located at the Hunchun Jingxin wetland.The collected Anser albifrons fecal samples were diluted in PBS and cultured on LB agar and blood agar media.Individual colonies were isolated and purified,and the bacteria were identified using Gram staining,biochemical tests,PCR amplification and sequencing of the 16S rRNA gene,and other methods. The drug resistance of the isolated bacteria was analyzed. 【Result】 The isolated bacteria were Gram negative bacteria,appearing as purple red short rod-shaped bacteria.Biochemical tests of the isolate strain revealed positive results for urease,lysine decarboxylase,ornithine decarboxylase,semisolid,etc.,and hydrogen sulfide,phenylalanine,gluconate,etc.were negative.A single band with a size of 1 500 bp was amplified by PCR,and the amplified sequence had the highest similarity with the sequence of Pseudomonas aeruginosa MZ836867 published in NCBI,which was 99.6%.Phylogenetic analysis revealed that the isolated strain belonged to the same branch with strain MZ836867.This indicated that the isolated strain was Pseudomonas putidis.The results of drug sensitivity test showed that the isolate was sensitive to 13 kinds of drugs,such as chloramphenicol,cotrimoxazole and norfloxacin,intermediated to 9 kinds of drugs such as polycolistin B,clindamycin and imipenem,and it was resistant to 8 drugs including vancomycin,flufenicol, lincomycin and oxacillin, etc.,among which lincomycin and oxacillin were the most resistant. 【Conclusion】 This study successfully isolated multidrug-resistant Pseudomonas putida from the fecal samples of Anser albifrons in Hunchun Jingxin wetland,providing a reference for further research on the gut microbiota of migratory birds in this wetland.

Key words: Hunchun; Anser albifrons; Pseudomonas putida; isolation and identification; drug resistance

中图分类号:

S852.61

马振, 王隆胜, 唐泽宇, 闵鹏飞, 赵健豪, 孟凡奇, 薛书江, 贾立军. 珲春敬信湿地候鸟白额雁恶臭假单胞菌的分离鉴定及药物敏感性分析[J]. 中国畜牧兽医, 2025, 52(1): 461-469.

MA Zhen, WANG Longsheng, TANG Zeyu, MIN Pengfei, ZHAO Jianhao, MENG Fanqi, XUE Shujiang, JIA Lijun. Isolation,Identification and Drug Sensitivity Analysis of Pseudomonas putidis from Anser albifrons in Hunchun Jingxin Wetland[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 461-469.

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