两株鸡源印度不动杆菌的分离鉴定及系统进化分析  

Isolation,identification and phylogenetic analysis of two strains of chicken-derived Acinetobacter indicus

作　　者：邓宇 刘华园 王景景 王浩千 王新卫[1] DENG Yu;LIU Huayuan;WANG Jingjing;WANG Haoqian;WANG Xinwei(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;Nanyang City Wolong District Animal Health Supervision Office,Nanyang 473003,China)

机构地区：[1]河南农业大学动物医学院,郑州450046 [2]南阳市卧龙区动物卫生监督所,河南南阳473003

出　　处：《黑龙江畜牧兽医》2024年第9期71-76,119,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：河南科技攻关项目(212102110358)。

摘　　要：为了探究鸡源不动杆菌的生物学特性,试验对采自河南省信阳市固始县某养鸡场的鸡口咽拭子进行细菌的分离纯化,通过革兰氏染色、形态特征观察、生化试验,16S rRNA基因及rpoB基因zone1、zone2片段分析鉴定分离菌,并进行药敏试验、系统进化分析、核苷酸相似性分析和小鼠致病性试验。结果表明:从鸡口咽拭子样品中分离出2株细菌,分别命名为HNXY29R和HNXY36Q。2株分离菌在普通琼脂、麦康凯琼脂及血琼脂培养基上均能生长,经镜检确定为球杆状或短棒状的革兰氏阴性菌,生化特性与印度不动杆菌相符。2株分离菌的16S rRNA与GenBank中印度不动杆菌的核苷酸序列相似性在99%以上,其rpoB基因zone1、zone2可变区片段与GenBank中的印度不动杆菌的核苷酸相似性在98%以上,进一步确定这2株分离菌为印度不动杆菌。2株分离菌对青霉素、阿莫西林、新霉素、大观霉素、卡那霉素、红霉素、环丙沙星、诺氟沙星、利福平、多西环素、复方新诺明、氟苯尼考和替加环素耐药。用浓度为8.05×10^(8 )cfu/mL的菌液感染小鼠后,24 h内感染HNXY29R的小鼠有3只死亡,感染HNXY36Q的小鼠有2只死亡,直至第7天再无小鼠死亡。说明采集样本的河南省信阳市固始县某养鸡场的鸡群中存在印度不动杆菌,且该菌有一定的致病性。In order to investigate the biological characteristics of chicken-derived Acinetobacter indicus,in the experiment,chicken oropharyngeal swabs collected from chicken farms in Gushi County,Xinyang City,Henan Province,were carried out for the isolation and purification of Acinetobacter;the isolates were identified by Gram staining,morphological characterization,biochemical characterization,as well as analysis of the 16S rRNA gene and the zone1 and zone2 fragments of the rpoB gene;drug sensitivity test,phylogenetic analysis,nucleotide homology analysis,and pathogenicity test in mice were performed.The results showed that two bacterial isolates,named HNXY29R and HNXY36Q,were isolated from chicken oropharyngeal swab samples,which were able to grow on normal agar medium,MacConkey agar medium and blood plate,and were determined to be Gram-negative bacilli with bulbous rods or short rods by microscopic examination;and their biochemical characteristics were consistent with Acinetobacter indicus.The 16S rRNA of the two isolates had more than 99% nucleotide sequence homology with Acinetobacter indicus in GenBank;their rpoB gene zone1 and zone2 variable region fragments had more than 98% nucleotide homology with that of Acinetobacter indicus in GenBank,which further identified the two isolates as Acinetobacter indicus.Both isolates were resistant to penicillin,amoxicillin,neomycin,macromycin,kanamycin,erythromycin,ciprofloxacin,norfloxacin,rifampicin,doxycycline,cotrimoxazole,fosfenicol and tigecycline.After infecting mice with the bacterial solution at a concentration of 8.05×10~8 cfu/mL,three mice died within 24 h after infection with HNXY29R,and two mice died after infection with HNXY36Q,and no more mice died until the 7th day.The results suggested that Acinetobacter indicus was present in chickens in Gushi County,Xinyang City,Henan Province,and that the bacterium had a certain degree of pathogenicity.

关 键 词：印度不动杆菌 鸡 分离 鉴定 药敏试验 系统进化分析 

分 类 号：S852.61[农业科学—基础兽医学]