牛源致病性大肠杆菌的敏感药物筛选及腐殖酸钠的抑菌试验

doi:10.16431/j.cnki.1671-7236.2020.10.036

摘要/Abstract

摘要： 为了解黑龙江地区部分规模化牛场犊牛腹泻性大肠杆菌毒力基因和耐药基因分布情况，筛选敏感药物以及分析腐殖酸钠对牛源致病性大肠杆菌的防治作用，本研究收集了40份病料，通过分离纯化、染色镜检、生化鉴定以及分子生物学的方法对分离菌进行鉴定；采用PCR技术进行毒力基因和耐药基因的检测；采用K-B法进行药敏检测；利用稀释平板计数法研究不同浓度腐殖酸钠对大肠杆菌的抑制效果；腐殖酸钠灌胃小鼠研究其对攻毒小鼠的保护作用。结果显示，共分离出牛源大肠杆菌30株；其中25株检出毒力基因，在检测的10种毒力基因中，有8种毒力基因被检出，检出率达83.3%，同时还存在多种毒力基因组合；在检测的7种耐药基因中bla_TEM基因检出率最高，为100%，bla_SHV基因最低，为13.3%，其他耐药基因检出率也较高；药敏试验结果显示，30株分离菌对美罗培南最为敏感，其次为阿米卡星；稀释平板计数结果显示，5%腐殖酸钠的抑菌效果优于其他浓度；攻毒试验结果显示，腐殖酸钠能明显提高攻毒小鼠的存活率。本研究对该地区抗菌药物的合理使用以及腐殖酸钠的临床应用提供了依据。

关键词: 大肠杆菌; 毒力基因; 耐药基因; 药敏试验; 腐殖酸钠

Abstract: In order to get more information about the distribution of virulence genes and drug resistance genes of Escherichia coli(E.coli) isolated from calf diarrhea in some large-scale cattle farms in Heilongjiang,sensitive drugs were screened to analyze the preventive and therapeutic effect of sodium humate on E.coli.In this study,40 samples were collected and identified by separation and purification,staining microscopy,biochemical identification and molecular biology,and the virulence and drug resistance genes were detected by PCR and the drug sensitivity was detected by K-B test.The inhibitory effect of different concentrations of sodium humate on E.coli was measured by serial dilution and plate counts.Moreover,the protective effect of sodium humate on mice were studied through gastric administration.The results showed that 30 strains of E.coli were isolated,of which 25 strains had virulence genes and 8 virulence genes were detected out of 10,with detection rate of 83.3%.There were many virulence gene combinations.The highest detection rates for bla_TEM and bla_SHV genes were 100% and 13.3% respectively,and the detection rate of other resistance genes was also high.The results of drug sensitive test showed that 30 isolated strains were most sensitive to meropenem,followed by amikacin.The results of serial dilution and plate counts showed that 5% sodium humate was the most bacteriostatically effective,and the results of toxicity test showed that sodium humate could improve the survival rate of mice.This study provided a basis for the rational use of antibiotics and the clinical application of sodium humate in this area.

Key words: Escherichia coli; virulence genes; drug resistance genes; antimicrobial susceptibility test; sodium humate

中图分类号:

S853

王栋, 金圣子, 杜垣逸, 关桐旭, 李婉莹, 刘云. 牛源致病性大肠杆菌的敏感药物筛选及腐殖酸钠的抑菌试验[J]. 中国畜牧兽医, 2020, 47(10): 3361-3370.

WANG Dong, JIN Shengzi, DU Yuanyi, GUAN Tongxu, LI Wanying, LIU Yun. Screening of Sensitive Drugs for Bovine Pathogenic Escherichia coli and Bacteriostatic Test of Sodium Humate[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3361-3370.

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