圈养大熊猫肠道微生物分离、鉴定及细菌耐药性研究  

Isolation and Antimicrobial Resistance of Bacteria from Intestinal Tract of Giant Panda

作　　者：李蓓[1] 郭莉娟[1,2] 龙梅[1,2] 汤纯香[3] 周晓平[3] 罗燕[1] 邹立扣[1] 

机构地区：[1]四川农业大学都江堰校区微生物学实验室,森林资源保护与利用实验室,四川都江堰611830 [2]四川农业大学资源环境学院,成都611130 [3]中国保护大熊猫研究中心雅安碧峰峡基地,四川雅安625000

出　　处：《四川动物》2014年第2期161-166,共6页Sichuan Journal of Zoology

基　　金：四川省教育厅重点项目(10ZA055)资助

摘　　要：对中国卧龙自然保护区大熊猫研究中心-碧峰峡基地64只大熊猫分别采取粪样，用选择性培养基分离大肠杆菌、沙门氏菌，根据形态特征及16S rDNA序列对菌株进行鉴定；用PDA培养基分离真菌，真菌通用引物ITS4、ITS5进行扩增鉴定。采用K-B法（CLSI）药敏试验测试大肠杆菌和沙门氏菌对18种药敏纸片的耐药性。形态学与rDNA-ITS序列鉴定表明，共分离到真菌19株，鉴定为8个属。分离出大肠杆菌88株，沙门氏菌47株；大肠杆菌对各抗生素的耐药率为0%~35.22%，耐药率为TET（35.22%）、AML（12.50%）、S3（12.50%），共产生17种耐药谱，其中TET、AML-TET谱型占优势；沙门氏菌对各抗生素的耐药率为0%~42.55%，耐药率为TET（42.55%）、AML（40.43%）、S3（38.30%），也产生了17种耐药谱，AML和AML-TET谱型占优势。通过对细菌和真菌的分离鉴定以及细菌的耐药性试验，了解大熊猫肠道微生物与不同个体的关系，为研究大熊猫的营养及消化等奠定基础，对预防和诊断大熊猫肠道疾病，提高大熊猫的存活率提供依据。Feces samples of 64 giant pandas were collected from China＇s Panda Research Center Bifengxia base. Escherichia coli and Salmonella sp. were isolated using selective medium. Isolates were identified by morphological characteristics and confirmed by 16S rDNA sequence analysis. Fungi were isolated using PDA medium, and identified with universal primer （ITS4 and ITS5）. The susceptibility of isolates to eighteen antibiotics was performed according to the standard disk diffusion method recommended by the Clinical and Laboratory Standards Institute （CLSI）. The results showed that 19 fungi （8 genus） and 88 E. coli as well as 47 Salmonella sp. were obtained. The drug-resistant frequency of E. coil against all the tested antibiotics was 0%~35.55%. Of all the E. coli isolates, 35.22%, 12.50% and 12.50% were resistant to TET, AML and S3, respectively. Additionally, a total of 17 resistance profiles were observed, among which TET and AMP-TET were the dominant. In addition, 42.55%, 40.43% and 38.30% of Salmonella isolates were resistant to TET, AML and S3, respectively. Similarly, there were also 17 resistance profiles observed, and AML and AML-TET were the top two. Morphology and ITS sequencing showed that 19 species of fungi belonged to 8 different genus were isolated. The identification of the bacteria and fungi and the antimicrobial susceptibility testing could figure out the relationship between microorganism in intestinal tract and individuals of giant pandas. Moreover, this could facilitate the further study of the nutrition and digestion and providing basis for the enteric disease prevention, treatment and improvement of the livability of giant panda.

关 键 词：大熊猫 真菌 大肠杆菌 沙门氏菌 分离鉴定 耐药性 

分 类 号：S852.6[农业科学—基础兽医学]