广西三黄鸡肠道中抗鸡白痢乳酸菌的分离鉴定  

Isolation and Identification of Lactobacillus Resistant to Salmonella Pullorum from Intestinal Tract of Guangxi Three-yellow Chicken

作　　者：农庭镔 黄颖 马美红 张雪悦 杨梦雅 李宜海 何秀苗 NONG Tingbin;HUANG Ying;MA Meihong;ZHANG Xueyue;YANG Mengya;LI Yihai;HE Xiumiao(Guangxi Key Laboratory of Polysaccharide Materials and Modification,College of Marine Sciences and Biotechnology,Guangxi Minzu University,Nanning,Guangxi 530008)

机构地区：[1]广西民族大学海洋与生物技术学院,广西多糖材料与改性重点实验室,广西南宁530008

出　　处：《中国家禽》2024年第9期103-111,共9页China Poultry

基　　金：广西自然科学基金项目(2022GXNSFAA035520);国家自然科学基金项目(32160824);广西民族大学2017年度校级引进人才科研启动项目(2017KJQD005)。

摘　　要：为开发鸡源益生菌资源,筛选能抑制鸡白痢沙门氏菌(SP)的乳酸菌,试验以散养三黄鸡为研究对象,以融钙圈法从鸡肠道中分离出乳酸菌,以双层琼脂平板扩散法筛选出对SP具有明显抑制效果的菌株,并结合形态学观察、生理生化试验和16S rRNA基因序列进行初步鉴定。在分析目的菌株的抑菌谱、耐酸耐胆盐特性以及对抗生素的敏感性后,以乳酸菌发酵液连续灌胃1日龄雏鸡14 d,研究其对雏鸡的安全性。结果显示:从鸡肠道中筛选出2株具有益生潜力的乳酸菌A1和K41,其无细胞上清液对SP的抑菌圈直径约为17 mm,其中A1为植物乳杆菌,K41为唾液乳杆菌;A1和K41的无细胞上清液对鼠伤寒沙门氏菌、大肠杆菌、铜绿假单胞菌、藤黄微球菌和金黄色葡萄球菌均有明显的抑菌效果;A1和K41在pH 3.0条件下的存活率分别为97.24%和89.03%,在0.3%胆盐浓度下的存活率分别为89.81%和87.20%;A1和K41对β-内酰胺类抗生素阿莫西林、氨苄西林和头孢曲松等敏感,对卡那霉素、磺胺异噁唑和环丙沙星等耐药;口服A1和K41的雏鸡生长良好,生产性能与对照组相比无显著差异(P>0.05)。综上,植物乳杆菌A1和唾液乳杆菌K41对SP和多种病原菌有明显的抑制效果,且耐酸耐胆盐能力良好,对雏鸡安全无害。In this study,lactic acid bacteria that could inhibit Salmonella Pullorum(SP)were screened for exploiting poultryderived probiotic resources.LAB was isolated from the chicken's intestines by the calcium-dissolving circle method.Strains with significant bacteriostatic activity against SP were chosen through the double-layer agar plate diffusion method.Target strains were preliminarily characterized by morphological observation,physiological and biochemical experiments,and 16S rRNA gene sequenc⁃ing.The target strains were subsequently analyzed for the antibacterial spectrum,the tolerance towards acid and bile salt,and the an⁃tibiotic susceptibility.The culture of the target strains was continuously fed to the 1-day-age chicks for 14 days to evaluate their bios⁃ecurity.The results showed that two potential probiotics,A1 and K41,were isolated from the intestinal tract of the chickens,and the diameter of the inhibition zone of the cell-free supernatants of the two strains on SP was about 17 mm.A1 and K41 were preliminarily identified as Lactobacillus plantarum and Lactobacillus salivarius,respectively.The in vitro antibacterial test results showed that the cell-free supernatants of A1 and K41 had significant inhibition effects against Salmonella Typhimurium(ST),Escherichia coli,Pseu⁃domonas aeruginosa,Micrococcus luteus and Staphylococcus aureus.The survival rates of A1 and K41 at pH 3.0 were 97.24%and 89.03%,respectively,and the survival rates of A1 and K41 at 0.3%bile salt concentration were 89.81%and 87.20%,respectively.A1 and K41 were sensitive toβ-lactam antibiotics such as amoxicillin,ampicillin and ceftriaxone,and resistant to kanamycin,sulfa⁃furazole and ciprofloxacin,etc.The chicks fed with the culture of A1 and K41 grew well,and there was no significant difference on growth performance compared with the control group(P>0.05).In conclusion,Lactobacillus plantarum A1 and Lactobacillus salivari⁃us K41 had significant bacteriostatic effects on SP and multiple pathogenic bacteria,and good acid and bi

关 键 词：鸡白痢沙门氏菌 植物乳杆菌 唾液乳杆菌 益生菌 

分 类 号：S831.5[农业科学—畜牧学]