贵州中蜂蜂房哈夫尼菌的分离鉴定及药敏试验

doi:10.16431/j.cnki.1671-7236.2018.04.027

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (4): 1050-1058.doi: 10.16431/j.cnki.1671-7236.2018.04.027

贵州中蜂蜂房哈夫尼菌的分离鉴定及药敏试验

邱吉宇¹, 主性¹, 钟纯燕¹, 罗亚琰¹, 侯萍², 樊莹², 范文穗³, 邓位喜⁴, 韩建伦⁵

1. 贵州大学动物科学学院, 贵阳 550025;
2. 贵州省农业委员会, 贵阳 550000;
3. 麻江县农业局, 麻江 557600;
4. 遵义职业技术学院, 遵义 563000;
5. 道真县珍珍园中蜂养殖场, 道真 563500

收稿日期:2017-12-07 出版日期:2018-04-20 发布日期:2018-04-25
通讯作者: 主性(1972-),男,贵州毕节人,副教授,硕士生导师,研究方向:临床兽医,E-mail:815657446@qq.com E-mail:815657446@qq.com
作者简介:邱吉宇(1992-),男,贵州开阳人,硕士,研究方向:中西结合兽医与中草药应用,E-mail:327507456@qq.com
基金资助:
遵义中蜂品种选育与开发利用（黔科合NY[2015]3029号）；中华蜜蜂主要疫病绿色防治关键技术研究（遵市科合（2016）52号）

Isolation, Identification and Drug Sensitive Test for Hafnia alvei from Apis cerana in Guizhou

QIU Jiyu¹, ZHU Xing¹, ZHONG Chunyan¹, LUO Yayan¹, HOU Ping², FAN Ying², FAN Wensui³, DENG Weixi⁴, HAN Jianlun⁵

1. College of Animal Science, Guizhou University, Guiyang 550025, China;
2. Agricultural Committee of Guizhou Province, Guiyang 550000, China;
3. Agricultural Bureau of Majiang County, Majiang 557600, China;
4. Zunyi Vocational & Technical College, Zunyi 563000, China;
5. Zhenzhen Garden Chinese Bee Farms, Daozhen 563500, China

Received:2017-12-07 Online:2018-04-20 Published:2018-04-25

摘要/Abstract

摘要：

为准确诊断贵州某蜂场发生的细菌性中蜂疾病，并提供防制方案，本试验进行了细菌分离纯化、形态学观察、生化试验，应用细菌16S rRNA基因通用引物进行基因扩增测序，测序结果在NCBI上进行比对分析，选取同源性较高的5个属的30株细菌的16S rRNA序列进行同源性分析并构建系统进化树，并对该分离菌进行动物回归试验和药敏试验。结果显示，分离菌为革兰氏阴性短杆菌，生化试验初步鉴定为蜂房哈夫尼菌；16S rRNA序列与哈夫尼杆菌属同源性最高，在97.0%~99.6%之间；遗传进化树表明，分离菌在哈夫尼菌菌属这一分支；动物回归试验显示，分离菌对中蜂有一定致病性，表明该蜂场受到蜂房哈夫尼菌感染，造成了蜜蜂的副伤寒；药敏试验结果表明，分离菌对青霉素、红霉素、克林霉素等耐药，对阿米卡星、复方新诺明、氧氟沙星等敏感。本试验结果为中蜂感染蜂房哈夫尼菌病的研究提供了一定参考，并能对蜂场提供合理用药意见。

关键词: 中蜂; 蜂房哈夫尼菌; 16S rRNA; 分离鉴定

Abstract:

For the diagnosis,prevention and treatment of Apis cerana's bacterial disease in Guizhou,a pathogen was identified in this paper by isolation and purification,morphological observation and biochemical test.The bacterial 16S rRNA gene universal primers were used for amplification and sequencing.The results of the sequencing were analyzed on NCBI.16S rRNA sequences of 30 bacteria of 5 genera with high homology were selected to analyze the nucleotide homology and construct the phylogenetic tree,and animal regression test and drug sensitivity test of the isolate were carried out.The results showed that the isolate was short gram-negative bacilli,and the biochemical tests were initially identified as Hafnia alvei.16S rRNA sequences shared the highest homology with Hafnia,ranging from 97.0% to 99.6%.The phylogenetic tree indicated that the isolate belonged to the branch of Hafnia.The animal regression test showed that the isolate had some pathogenicity to Apis cerana,indicating that the bees were infected by Hafnia alvei.Antibiotic sensitive test showed that the isolate was resistant to penicillin,erythromycin and clindamycin,and was sensitive to amikacin,sulfamethoxazole and ofloxacin.The results of this study provided a reference for the study of honeybee Hafnia infection in Apis cerana and provided reasonable drug advice for the bee farms.

Key words: Apis cerana; Hafnia alvei; 16S rRNA; isolation and identification

中图分类号:

S891

邱吉宇, 主性, 钟纯燕, 罗亚琰, 侯萍, 樊莹, 范文穗, 邓位喜, 韩建伦. 贵州中蜂蜂房哈夫尼菌的分离鉴定及药敏试验[J]. 《中国畜牧兽医》, 2018, 45(4): 1050-1058.

QIU Jiyu, ZHU Xing, ZHONG Chunyan, LUO Yayan, HOU Ping, FAN Ying, FAN Wensui, DENG Weixi, HAN Jianlun. Isolation, Identification and Drug Sensitive Test for Hafnia alvei from Apis cerana in Guizhou[J]. , 2018, 45(4): 1050-1058.

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贵州中蜂蜂房哈夫尼菌的分离鉴定及药敏试验

Isolation, Identification and Drug Sensitive Test for Hafnia alvei from Apis cerana in Guizhou

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