三峡水库铜绿假单胞菌分离株的耐药性与毒理评价

黎毅; 张琪; 胡力凡; 刘凤燕; 肖国生; 戚文华

文章摘要

三峡水库铜绿假单胞菌分离株的耐药性与毒理评价

Drug Resistance and Toxicological Evaluation of Pseudomonas aeruginosa Isolates from Three Gorges Reservoir

DOI：doi:10.3969/j.issn.1005-7021.2024.04.009

英文关键词: Pseudomonas aeruginosa drug resistance drug resistance gene Caenorhabditis elegans toxicological evaluation

基金项目:国家自然科学基金项目(31702032)；重庆市科委自然科学基金项目(cstc2019jcyj-msxmX0410)

作者	单位
黎毅	1. 重庆三峡学院环境与化学工程学院，重庆 404100；2. 重庆市渝东北特色生物资源开发利用工程技术研究中心重庆三峡学院生物与食品工程学院，重庆 404100
张琪	重庆市渝东北特色生物资源开发利用工程技术研究中心重庆三峡学院生物与食品工程学院，重庆 404100
胡力凡	重庆市渝东北特色生物资源开发利用工程技术研究中心重庆三峡学院生物与食品工程学院，重庆 404100
刘凤燕	重庆市渝东北特色生物资源开发利用工程技术研究中心重庆三峡学院生物与食品工程学院，重庆 404100
肖国生	重庆市渝东北特色生物资源开发利用工程技术研究中心重庆三峡学院生物与食品工程学院，重庆 404100
戚文华	重庆市渝东北特色生物资源开发利用工程技术研究中心重庆三峡学院生物与食品工程学院，重庆 404100

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中文摘要:

铜绿假单胞菌（Pseudomonas aeruginosa）是一种重要的水源性和食源性致病菌，当机体免疫力下降时，易造成感染，了解铜绿假单胞菌的耐药性和致病力，对预防铜绿假单胞菌感染及其防治有重要意义。为探究三峡水库铜绿假单胞菌分离株的耐药性及其对秀丽线虫（Caenorhabditis elegans）的致病性，从三峡水库采集水样进行细菌分离，经形态学鉴定、生理生化鉴定、16S rDNA测序和oprI特异性基因鉴定筛选出铜绿假单胞菌，研究分离菌的耐药性、耐药基因和毒力基因，并利用模式生物秀丽线虫进行毒理评价。结果显示，从三峡水库分离鉴定出14株铜绿假单胞菌。分离株对青霉素G、氨苄青霉素、头孢唑林、卡那霉素、红霉素耐药，对阿米卡星、庆大霉素、诺氟沙星、环丙沙星、美罗培南、左氧氟沙星敏感。耐药基因bla TEM、aac(6′)-Ⅱ、qacEΔ1-sull、qnrS、mexR在所有分离菌株中呈阳性；耐药基因aac(3)-Ⅰ、tetA、tetB、tetD、tetE、cat、cmlA、qnrA、qnrD在所有分离菌株中呈阴性；此外，分离菌株携带多种毒力基因。分离菌PA7、PA8、PA12使秀丽线虫的平均寿命分别下降15%、14%、16%（P<0.05），身体弯曲频率分别下降18%、33%、23%（P<0.05），头部摆动频率分别下降10%、13%、11%（P<0.05），最长寿命缩短约3 d。三峡水库中铜绿假单胞菌分离株具有多重耐药性，部分菌株对秀丽线虫有致病性，可为水体铜绿假单胞菌的感染防治提供一定参考。

英文摘要:

Pseudomonas aeruginosa is an important water-borne and food-borne pathogen, which is easy to cause infection when the body′s immunity declines. Understanding the resistance and pathogenicity of P. aeruginosa is of great significance for the prevention and control of infection of P. aeruginosa. To investigate the resistance of P. aeruginosa isolated from Three Gorges Reservoir and its pathogenicity against Caenorhabditis elegans, bacteria were isolated from water samples collected from Three Gorges Reservoir. P. aeruginosa was screened out by morphological identification, physiological and biochemical identification, 16S rDNA sequencing and oprI specific gene identification. Drug resistance, resistance genes and virulence genes of the isolates were studied, and the model organism C. elegans was used for toxicological evaluation. The results showed that 14 strains of P. aeruginosa were isolated and identified from the Three Gorges Reservoir. The isolates were resistant to penicillin G, ampicillin, cefazolin, kanamycin and erythromycin, and sensitive to amikacin, gentamicin, norfloxacin, ciprofloxacin, meropenem and levofloxacin. The resistance genes bla TEM, aac (6′)-II, qacΔ1-sull, qnrS, mexR were positive in all isolates. Drug resistance genes aac(3)-Ⅰ, tetA, tetB, tetD, tetE, cat, cmlA, qnrA, qnrD were negative in all isolates. In addition, the isolates carried multiple virulence genes. After infection with isolates PA7, PA8 and PA12, the average lifespan of C. elegans decreased by 15%, 14%, and 16% respectively (P<0.05), and body bending frequency of decreased by 18%, 33% and 23% respectively (P<0.05), and head swing frequency of decreased by 10%, 13% and 11% respectively (P<0.05), and the longest lifespan was shortened by about 3 days. The results showed that the isolates of P. aeruginosa in the Three Gorges Reservoir had multiple drug resistance, and some of the isolates were pathogenic to C. elegans, which could provide some reference for the prevention and control of P. aeruginosa infection in water.

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