野生动物源乳酸菌的分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2025.12.040

摘要/Abstract

摘要： 【目的】乳酸菌是对人体、动物和植物有益的一类菌群，在食品工业、医药健康及农业领域中广泛应用，野生动物因其生存环境与畜禽和人类存在较大差异，其肠道内菌群也存在明显多样性。本试验旨在从野生动物粪便中分离乳酸菌，并分析其生物学特性及抑菌活性，以期获得微生物制剂的候选株。【方法】收集江苏某野生动物园内不同动物的570份粪便样本，用MRS选择培养基进行初筛，通过菌落形态观察、革兰染色镜检、生化鉴定等进一步进行乳酸菌鉴定，从中筛选出不溶血的分离株，用16S rRNA基因序列比对确定其种属，对分离株进行耐酸、耐胆盐、抑菌活性、药敏试验、自凝性和疏水性、动物安全性等一系列体外试验。【结果】从样本中分离得到40株疑似乳酸菌，其中3株有溶血现象，其余37株经16S rRNA基因鉴定后发现其可分属于包括融合魏斯氏菌、罗伊氏乳杆菌、唾液联合乳杆菌等在内的11个种属。经多项体外试验筛选后发现，分离株发酵乳杆菌ZX2性能突出，与其他分离株相比高度耐酸，可以直接在pH 3.0的MRS培养基中生长，且可耐受0.3%胆盐；对金黄色葡萄球菌和绿脓杆菌有一定的体外抑菌能力，对D型产气荚膜梭菌G07和超级耐药大肠杆菌DJ2无抑菌能力；自凝性在5 h内可达16%，疏水性可达95%以上；对氯霉素、四环素、青霉素、氨苄西林、头孢唑林、氧氟沙星均存在一定敏感性；对小鼠无致病性。【结论】本研究在野生动物粪便中分离出37株乳酸菌，其中发酵乳杆菌ZX2具有良好的益生特性，肠道定植能力优良，可作为潜力益生菌菌株应用于开发微生态制剂。

关键词: 野生动物; 乳酸菌; 分离与鉴定; 益生特性

Abstract: 【Objective】 Lactic acid bacteria (LAB) are a group of beneficial microorganisms for humans,animals and plants,which are widely applied in food industry,pharmaceutical health,and agricultural fields.Due to the significant differences in living environments between wild animals and livestock/poultry/humans,their intestinal microbiota exhibited distinct diversity.This study aimed to investigate the presence,biological characteristics,and antibacterial activity of LAB in fecal samples of different wild animals,with the goal of identifying candidate strains for microbial preparations. 【Method】 A total of 570 fecal samples were collected from various animals in a wildlife park in Jiangsu province.The samples were first subjected to primary screening using MRS selective medium.Isolates were preliminarily identified as LAB through colony morphology observation,Gram staining microscopy,and biochemical tests.Non-hemolytic isolates were further screened,and their species/genus were confirmed by 16S rRNA gene sequence alignment.A series of in vitro tests were conducted,including acid tolerance,bile salt tolerance,antibacterial activity,drug sensitivity,auto-aggregation ability,hydrophobicity,and animal safety evaluation. 【Result】 Forty suspected LAB strains were isolated,among which three exhibited hemolytic activity.The remaining thirty-seven strains were identified by 16S rRNA gene analysis as belonging to eleven species,including Weissella confusa,Lactobacillus reuteri and Lactobacillus salivarius,etc.After screening through a series of in vitro tests,it was found that the isolate Lactobacillus fermentum ZX2 exhibited outstanding performance.Compared with other isolates,it had high acid tolerance,being capable of growing directly in MRS medium at pH 3.0,and could tolerate 0.3% bile salt.It showed a certain degree of in vitro antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa,but no antibacterial activity against Clostridium perfringens type D-G07 and super drug-resistant Escherichia coli DJ2.Its auto-aggregation rate could reach 16% within 5 h,and its hydrophobicity could exceed 95%.Additionally,it showed certain sensitivity to chloramphenicol,tetracycline,penicillin,ampicillin,cefazolin and ofloxacin,and was non-pathogenic to mice. 【Conclusion】 In this study,thirty-seven strains of lactic acid bacteria were isolated from the feces of wild animals.Among them,Lactobacillus fermentum ZX2 exhibited excellent probiotic properties and intestinal colonization ability,could be used as a potential probiotic strain for the development of microecological preparations.

Key words: wild animals; lactic acid bacteria; isolation and identification; probiotic characteristics

中图分类号:

S816.3

徐昀眺, 叶思明, 李自盼, 卞泓凯, 潘新语, 丁香, 王海燕, 刘文博. 野生动物源乳酸菌的分离鉴定及生物学特性研究[J]. 中国畜牧兽医, 2025, 52(12): 5967-5978.

XU Yuntiao, YE Siming, LI Zipan, BIAN Hongkai, PAN Xinyu, DING Xiang, WANG Haiyan, LIU Wenbo. Isolation,Identification and Biological Characteristics of Lactic Acid Bacteria from Wild Animals[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(12): 5967-5978.

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