大熊猫国家公园广元片区野生动物及其环境源大肠杆菌流行病学及耐药性调查

苏小艳; 杨梅; 燕霞; 侯蓉; 肖梅; 王劲松; 刘圣; 王路才; 张文平; 黄虹秀

doi:10.16829/j.slxb.150761

兽类学报 >

2023 , Vol. 43 >Issue 4: 430 - 442

DOI: https://doi.org/10.16829/j.slxb.150761

研究论文

大熊猫国家公园广元片区野生动物及其环境源大肠杆菌流行病学及耐药性调查

苏小艳 ,
杨梅 ,
燕霞 ,
侯蓉 ,
肖梅 ,
王劲松 ,
刘圣 ,
王路才 ,
张文平 ,
黄虹秀

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1 成都大熊猫繁育研究基地, 四川省濒危野生动物保护生物学重点实验室, 成都 610081;
2 大熊猫国家公园广元管理分局, 广元 628000;
3 四川省唐家河国家级自然保护区管理处, 广元 628109;
4 四川省毛寨自然保护区管理局, 广元 628401

苏小艳(1985-)，女，博士，副研究员，主要从事大熊猫等野生动物疾病防控研究.

收稿日期: 2022-11-24

修回日期: 2023-05-16

网络出版日期: 2023-08-01

基金资助

四川省自然科学基金(2022NSFSC0020)；成都大熊猫繁育研究基地自立课题(2021CPB-B15和2020CPB-B23)

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Epidemiology and antibiotics resistance of Escherichia coli from wild animals and their environment in Guangyuan area of Giant Panda National Park

SU Xiaoyan ,
YANG Mei ,
YAN Xia ,
HOU Rong ,
XIAO Mei ,
WANG Jinsong ,
LIU Sheng ,
WANG Lucai ,
ZHANG Wenping ,
HUANG Hongxiu

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1 Chengdu Research Base of Giant Panda Breeding, Sichuan Key Laboratory of Conservation Biology for Endangered Wildlife, Chengdu 610081, China;
2 Guangyuan Administrative Branch of Giant Panda National Park, Guangyuan 628000, China;
3 Sichuan Tangjiahe National Nature Reserve Management, Guangyuan 628109, China;
4 Sichuan Maozhai Nature Reserve Administration, Guangyuan 628401, China

Received date: 2022-11-24

Revised date: 2023-05-16

Online published: 2023-08-01

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摘要

为了解大熊猫国家公园广元片区野生动物及其生活环境中大肠杆菌流行病学特点，防止大肠杆菌在该区域造成流行性疫病，探明该区域野生动物对抗生素的耐药情况，本研究2022年8—9月采集大熊猫国家公园广元片区内野生动物粪便、水及土壤样品共计243份，采用16S rRNA、K-B法、高通量荧光定量PCR法和PCR技术对该区域的大肠杆菌进行分离鉴定、药敏实验、耐药基因和毒力基因检测。结果显示：243份样品中共分离鉴定出86株大肠杆菌，分离率为35.39%；该区域内分离到的大肠杆菌对阿莫西林有较高的耐药率(52.33%)，对链霉素、庆大霉素等抗生素的耐药率较低(1.16%~9.30%)，对头孢他啶、氯霉素、氟苯尼考、甲氧苄氨嘧啶均敏感；携带大量的耐药基因，其中多重耐药类(83.14%)、磺胺类(55.52%)、四环素类(48.52%)抗生素耐药基因携带率较高，其余耐药基因携带率为26.16%~35.72%；分离株携带1~6个毒力基因，irp2检出率(91.86%)最高，其次为ompA、ibeB和tsh (检出率分别为55.81%、69.77%和52.33%)，FyuA、fimH和PapA检出率较低(分别为17.44%、19.77%和1.16%)；86株大肠杆菌中均未检出5种致泻性大肠杆菌。本研究为大熊猫国家公园广元片区野生动物及环境源大肠杆菌的流行情况提供了科学数据参考，为人兽共患病的防控提供生态学依据。

关键词： 大熊猫国家公园广元片区; 大肠杆菌; 流行情况; 耐药性; 毒力基因

本文引用格式

苏小艳 , 杨梅 , 燕霞 , 侯蓉 , 肖梅 , 王劲松 , 刘圣 , 王路才 , 张文平 , 黄虹秀 . 大熊猫国家公园广元片区野生动物及其环境源大肠杆菌流行病学及耐药性调查[J]. 兽类学报, 2023 , 43(4) : 430 -442 . DOI: 10.16829/j.slxb.150761

Abstract

To prevent Escherichia coli from causing epidemic disease in the newly established Guangyuan area of the Giant Panda National Park (GAGPNP), located in Sichuan China, an investigation was conducted to explore the epidemiological characteristics of E. coli from wild animals and their environment as well as the antibiotic resistance of wild animals in the area. A total of 124 fecal samples from wild animals, 49 water and 70 soil samples were collected from the GAGPNP from August to September 2022. 16S rRNA, the K-B method, high-throughput fluorescent quantitative PCR and PCR technology were used to isolate and identify the E. coli, perform antibiotics sensitivity test, and for the detection of antibiotics resistance genes and virulence gene, respectively. The results showed that 86 strains of E. coli were identified, and the isolation rate was 35. 39%. The E. coli isolated from the GAGPNP had a high resistance rate to amoxicillin (52. 33%) and a low resistance rate to streptomycin, gentamicin and other antibiotics (1. 16% — 9. 30%), and were sensitive to ceftazidime, chloramphenicol, florfenicol and trimethoprim. A large number of antibiotics resistance genes were carried, including multiple antibiotics resistance genes (83. 14%), sulfonamides (55. 52%), and tetracyclines (48. 52%), resistace genes with a high carrying rate, while the other antibiotics resistance genes were 26. 16% — 35. 72%. E. coli isolates carried 1 — 6 virulence genes, and the detection rate of irp2 was the highest (91. 86%), followed by ompA, ibeB, and tsh, with the detection rates of 55. 81%, 69. 77%, and 52. 33% respectively. The detection rates of FyuA, fimH, and PapA were lower (17. 44%, 19. 77% and 1. 16% respectively). None of the 86 strains of E. coli detected 5 types of diarrhoeal E. coli. This study provides an scientific data reference for the prevalence of E. coli in wild animals in GAGPNP, and provides an ecological basis for the prevention and control of zoonosis.

Key words： Guangyuan area of Giant Panda National Park; Escherichia coli; Epidemic situation; Antibiotics resistance; Virulence genes

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