羊源沙门菌分离鉴定、耐药性及中草药抑菌效果研究

doi:10.16431/j.cnki.1671-7236.2025.08.044

摘要/Abstract

摘要： 【目的】对羊源沙门菌进行分离鉴定，检测其对常见抗菌药的耐药性以及中草药的抑菌效果，旨在为沙门菌病的防治提供参考依据。【方法】从湛江市某山羊场采集患有腹泻流涕症状的雷州山羊肛拭子样本33份和鼻拭子样本17份并进行细菌分离纯化，采用革兰染色、PCR扩增测序对病原菌进行鉴定。通过微量肉汤稀释法测定15种常用抗菌药对分离株的最小抑菌浓度(MIC)，利用试管二倍稀释法测定分离株对12种中草药的敏感性，并进一步探究不同中草药联合应用对分离株的体外抑菌作用。采用结晶紫染色法测定分离株的生物被膜形成量。【结果】分离菌在麦康凯琼脂培养基上呈圆形、边缘整齐、轻微隆起、无色透明单菌落；在营养琼脂可见白色、圆形、边缘整齐的单菌落。革兰染色可观察到两端钝圆、无芽孢、无荚膜的革兰阴性杆菌。16S rRNA基因PCR扩增结果显示，获得片段大小约1 500 bp条带。BLAST比对结果显示，分离菌株与肠炎沙门菌(登录号：AE016841)相似性＞97%。从采集的50份样本中共分离获得15株羊源沙门菌，分离率为30%。药敏试验结果显示，分离株对青霉素、头孢他啶的耐药率分别为90%和70%，对阿奇霉素和磺胺嘧啶钠的耐药率均为40%，对头孢唑林钠、土霉素、链霉素和乙酰甲喹的耐药率均为10%；分离株对阿米卡星、多西环素、卡那霉素、磺胺间甲氧嘧啶、哌拉西林、小诺霉素、恩诺沙星表现敏感。中草药抑菌试验结果显示，乌梅和五味子对分离株的MIC为15.62 mg/mL，黄连对分离株的MIC为62.5 mg/mL。中草药联合用药试验结果显示，以黄连＋苍术、五味子＋诃子以及五味子＋广藿香为组合的中草药复方的MIC分别为15.62、7.81和7.81 mg/mL。生长曲线与生物被膜检测结果显示，2MIC黄连、乌梅和五味子提取物抑菌效果较好，其中五味子抑制沙门菌生物被膜形成效果最佳。【结论】本研究分离获得15株羊源沙门菌，中草药提取物对其具有显著的体外抑菌效果，且部分中草药组合使用时能够增强其抑菌活性。研究结果对于中草药在防治沙门菌感染中有着重要指导意义，也为后续进一步探索沙门菌病的综合防治策略提供了的数据支撑。

关键词: 山羊; 沙门菌; 耐药性; 中草药; 抑菌效果

Abstract: 【Objective】 The purpose of this experiment was to isolate and identify Salmonella from goat,detect its resistance to common antibacterial drugs and the bacteriostati effect of Chinese herbal medicines,aiming to provide a reference basis for the prevention and treatment of Salmonella-related diseases. 【Method】 A total of 33 anal swab samples and 17 nasal swab samples of Leizhou goats with symptoms of diarrhea and rhinorrhea were collected from a goat farm in Zhanjiang city,and bacterial isolation and purification were carried out.The pathogenic bacteria were identified by Gram staining,PCR amplification and sequencing.The minimum inhibitory concentration (MIC) of 15 commonly used antibacterial drugs against the isolated strains was determined by microbroth dilution method.The sensitivity of the isolates to 12 kinds of Chinese herbal medicines was determined by the double dilution method in test tubes.Subsequently,the in vitro bacteriostati effect of the combined application of different Chinese herbal medicines on the isolates was further explored.The biofilm formation amount of the isolates was determined by crystal violet staining. 【Result】 The isolated bacteria presented as round,neatly edged,slightly raised,colorless and transparent single colonies on Maclonkey agar medium.White,round and neatly edged single colonies appeared in the nutrient agar.Gram staining showed Gram-negative bacilli with blunt and round ends,no spores and no capsules.The PCR amplification results of 16S rRNA gene showed that a band with a fragment size of 1 500 bp.BLAST comparison results showed that the similarity between the isolated strains and Salmonella Enteritis (accession number:AE016841) was more than 97%.A total of 15 goat-derived Salmonella were isolated from 50 collected samples,with an isolation rate of 30%.The results of the drug susceptibility test showed that the resistance rates of the isolates to penicillin and ceftazidime were 90% and 70% respectively,the resistance rates to azithromycin and sulfadiazine sodium were both 40%,and the resistance rates to cefazolin sodium,oxytetracycline,streptomycin and acetoquinine were all 10%.The isolated strains were sensitive to amikacin,doxycycline,kanamycin,sulfamonomethoxil,piperacillin,minocythromycin and enrofloxacin.The results of the antibacterial test of Chinese herbal medicines showed that the MIC of Prunus mume and Schisandra chinensis against the isolates was 15.62 mg/mL,and the MIC of Coptis chinensis against the isolates was 62.5 mg/mL.The results of the combined medication test of Chinese herbal medicines showed that the MIC of the Chinese herbal medicine compound prescriptions composed of Coptis chinensis＋Atractylodes lancea,Schisandra chinensis＋Terminalia chebula,and Schisandra chinensis＋Pogosteom cablin was 15.62,7.81 and 7.81 mg/mL respectively.The results of growth curve and biofilm detection showed that the 2MIC Coptis chinensis,Prunus mume and Schisandra chinensis had the good antibacterial effect,among which Schisandra chinensis had the best effect in inhibiting the formation of Salmonella biofilms. 【Conclusion】 In this study,15 goat-derived Salmonella were isolated and obtained.The extracts of Chinese herbal medicines had a significant in vitro antibacterial effect,and when some Chinese herbal medicines were used in combination,the antibacterial activity could be enhanced.This results had significant guiding significance for Chinese herbal medicine in the prevention and treatment of Salmonella infections,and also provided data support for further exploration of comprehensive prevention and treatment strategies for Salmonella diseases in the future.

Key words: goat; Salmonella; drug resistance; Chinese herbal medicine; bacteriostati effect

中图分类号:

S852.61

唐其东, 瓦仁乐乐, 梁铭俊, 罗晓虹, 杜树浩, 丁月霞. 羊源沙门菌分离鉴定、耐药性及中草药抑菌效果研究[J]. 中国畜牧兽医, 2025, 52(8): 3965-3975.

TANG Qidong, WA Renlele, LIANG Mingjun, LUO Xiaohong, DU Shuhao, DING Yuexia. Isolation,Identification and Drug Resistance of Salmonella from Goat and the Bacteriostatic Effects of Chinese Herbal Medicines[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3965-3975.

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