鸡源益生菌的分离鉴定及其益生特性分析  

Isolation and Identification of Chicken Probiotics and Analysis of Their Probiotic Properties

作　　者：郭晴 王宏莉 冯雨轩 张雨萌 马琪茵 程佳 刘明超 GUO Qing;WANG Hongli;FENG Yuxuan;ZHANG Yumeng;MA Qiyin;CHENG Jia;LIU Mingchao(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,保定071001

出　　处：《中国畜牧兽医》2025年第4期1852-1861,共10页China Animal Husbandry & Veterinary Medicine

基　　金：河北农业大学大学生创新创业训练计划项目(2024059);国家自然科学基金(31902328)。

摘　　要：【目的】研究鸡源唾液乳杆菌(Lactobacillus salivarius)对大肠杆菌标准株(CVCC1450)的抑菌作用,为防治鸡大肠杆菌病提供益生菌菌株。【方法】本研究采用选择性培养基结合分子生物学方法从健康白羽蛋鸡泄殖腔中分离益生菌株。通过形态学观察和16S rDNA测序进行鉴定,对分离株的抗菌活性和抑菌物质进行分析,建立分离株生长曲线和产酸曲线,探究其在消化道中的耐受性,通过药敏试验和溶血试验评判其安全性。【结果】分离得到2株益生菌,均是唾液乳杆菌,对大肠杆菌抑菌圈直径均达20 mm以上,均有良好的抑菌作用。将分离菌上清液pH调整至7.0后,抑菌活性显著下降,说明2株唾液乳杆菌的主要抑菌物质为有机酸。2株唾液乳杆菌生长特性和产酸能力良好,培养10 h后到达其平台期,培养22 h后pH降低至3.75;对胆盐溶液和人工胃肠液的耐受性较强,在0.3%胆盐溶液中存活率均达27%以上,在人工胃肠液中存活率均达55%以上。2株唾液乳杆菌具有安全性,对链霉素、氨苄西林、头孢他啶、氧氟沙星、青霉素、头孢唑林、多西环素等多数抗菌药敏感;对庆大霉素、卡那霉素、呋喃唑酮3种抗菌药耐受,无溶血性。【结论】本研究分离得到2株唾液乳杆菌,对大肠杆菌有较好的抑菌活性,为防治大肠杆菌导致的鸡等禽类腹泻提供安全高效、无残留污染的绿色微生态制剂奠定基础。【Objective】The purpose of this experiment was to study the bacteriostatic effect of chicken derived Lactobacillus salivarius on Escherichia coli(CVCC1450)standard strain,and provide probiotic strains for the prevention and treatment of chicken colibacillosis in the future.【Method】In this study,selective medium combined with molecular biology method was used to isolate probiotic strains from cloaca of healthy White laying hens.Morphological observation and 16S rDNA sequencing were used to identify the isolates,and the antibacterial activity and antibacterial substances of the isolates were analyzed.The growth curve and acid production curve of the isolates were established to explore their tolerance in the digestive tract,and their safety was evaluated by drug sensitivity test and hemolysis test.【Result】Two strains of probiotics were isolated,all of which were Lactobacillus salivarius,and had good inhibitory effect on Escherichia coli with a diameter of more than 20 mm.After pH of the supernatant was adjusted to 7.0,the antibacterial activity of the isolates decreased significantly,indicating that the main antibacterial substance of the two strains of Lactobacillus salivarius was organic acid.Two strains of Lactobacillus salivarius had good growth characteristics and acid production ability.After 10 h of cultivation,they reached their plateau phase,and after 22 h of cultivation,the pH decreased to 3.75.The tolerance to bile salt solution and artificial gastrointestinal fluid of the two strains was strong,with a survival rate of over 27%in 0.3%bile salt solution and over 55%in artificial gastrointestinal fluid.Two strains of Lactobacillus salivarius were safe and sensitive to most antibiotics such as streptomycin,ampicillin,cefotaxime,ofloxacin,penicillin,cefazolin and doxycycline,and resistant to three antibiotics gentamicin,kanamycin and furazolidone,without hemolytic activity.【Conclusion】In this study,two strains of Lactobacillus salivarus were isolated,which had good antibacterial activity again

关 键 词：唾液乳杆菌 生物学特性 大肠杆菌 抑菌作用 

分 类 号：S852.6[农业科学—基础兽医学] R378.992[农业科学—兽医学]