唾液乳杆菌无细胞上清液对金黄色葡萄球菌溶血活性的抑制作用研究  

Inhibition of CFCS of Lactobacillus salivarius on Hemolytic Activity of Staphylococcus aureus

作　　者：邢国锋 党乙凡 赵耀 苏帆帆 吴自豪 李涛[1] XING Guofeng;DANG Yifan;ZHAO Yao;SU Fanfan;WU Zihao;LI Tao(Key Laboratory of Livestock and Forage Resources Utilization around Tarim,Ministry of Agriculture and Rural Affairs,Engineering Laboratory for Tarim Animal Diseases Diagnosis and Control of Xinjiang Production&Construction Corps,College of Animal Sciences and Technology,Tarim University,Alar 843300,China;College of Life Sciences and Technology,Tarim University,Alar 843300,China)

机构地区：[1]塔里木大学动物科学与技术学院,新疆生产建设兵团塔里木动物疫病诊断与防控工程实验室,农业农村部环塔里木畜草资源利用重点实验室,阿拉尔843300 [2]塔里木大学生命科学与技术学院,阿拉尔843300

出　　处：《中国畜牧兽医》2025年第4期1884-1894,共11页China Animal Husbandry & Veterinary Medicine

基　　金：兵团指导性科技计划项目(2022ZD111、TDZKKY202203);国家级大学生创新创业计划项目(202410757003)。

摘　　要：【目的】探究唾液乳杆菌无细胞上清液(CFCS)对金黄色葡萄球菌溶血活性的影响,并比较不同培养条件对唾液乳杆菌CFCS抑菌效果的影响。【方法】通过肉汤稀释法测定唾液乳杆菌CFCS对金黄色葡萄球菌的最小抑菌浓度(MIC)与亚抑菌浓度(SIC);利用溶血活性半定量法检测唾液乳杆菌CFCS对金黄色葡萄球菌溶血活性的影响,利用实时荧光定量PCR检测金黄色葡萄球菌α-溶血素(hla)和AraC/XylS家族转录调控因子rbf基因相对表达量;通过小鼠致病性试验,检测唾液乳杆菌CFCS对小鼠死亡率、脏器载菌量和组织病理变化的影响;使用牛津杯双层琼脂扩散法分别测定不同培养基和培养条件对唾液乳杆菌CFCS抑菌活性的影响。【结果】唾液乳杆菌CFCS对金黄色葡萄球菌的MIC为31.25 mg/mL,SIC为3.906 mg/mL。经MIC唾液乳杆菌CFCS处理后,与阳性对照组相比,金黄色葡萄球菌的溶血活性极显著下降(P<0.01),hla基因相对表达量显著降低(P<0.05),rbf基因相对表达量极显著升高(P<0.01)。小鼠致病性试验结果显示,经MIC唾液乳杆菌CFCS处理后,金黄色葡萄球菌感染小鼠肝脏、肾脏和脾脏载菌量极显著降低(P<0.01),肝脏、肾脏和脾脏脏器组织病变明显减轻。MRS培养基中发酵的唾液乳杆菌CFCS对金黄色葡萄球菌的抑菌效果最好,抑菌圈直径为17.64 mm, TPY培养基次之(12.83 mm)。以MRS培养基为基础,在初始葡萄糖含量为4%、初始氮源含量为1.1%、初始pH 4.0、发酵时间48 h时,唾液乳杆菌CFCS对金黄色葡萄球菌的抑菌效果最好。【结论】唾液乳杆菌CFCS可抑制金黄色葡萄球菌的溶血活性,降低其致病性。培养条件的改变可对唾液乳杆菌CFCS抑菌活性产生显著影响。【Objective】The objective of this experiment was to investigate the effect of cell-free culture supernatant(CFCS)of Lactobacillus salivarius(L.salivarius)on the hemolytic activity of Staphylococcus aureus(S.aureus)and to compare the effect of different culture conditions on the bacteriostatic effect of CFCS of L.salivarius.【Method】The minimum inhibitory concentration(MIC)and subinhibitory concentration(SIC)of CFCS of L.salivarius on S.aureus were determined by broth dilution method.The influence of CFCS of L.salivarius on the hemolytic activity of S.aureus was detected by semi-quantitative method.The relative expression levels ofαhemolysin(hla)and AraC/XylS family transcriptional regulators rbf gene in S.aureus were determined by Real-time quantitative PCR.The effects of CFCS of L.salivarius on mortality,organ bacteria load and histopathological changes of mice were detected by pathogenicity test.The effects of different medium and culture conditions on the antibacterial activity of CFCS of L.salivarius were determined by Oxford Cup double-layer agar diffusion method,respectively.【Result】The MIC and SIC of CFCS of L.salivarius against S.aureus were 31.25 and 3.906 mg/mL,respectively.After treatment with CFCS of L.salivarius with MIC,compared with positive control group,the hemolytic activity of S.aureus was extremely significantly decreased(P<0.01),the relative expression level of hla gene was significantly decreased(P<0.05),and the relative expression level of rbf gene was extremely significantly increased(P<0.01).The results of pathogenicity test in mice showed that after treatment with MIC of CFCS of L.salivarius,the bacteria load in liver,kidney and spleen of mice infected with S.aureus was extremely significantly decreased(P<0.01),and the lesions of liver,kidney and spleen tissues were significantly reduced.The fermentation of CFCS of L.salivarius in MRS medium had the best inhibitory effect on S.aureus,and the inhibitory zone diameter was 17.64 mm,followed by TPY medium(12.83 mm).Based on MRS me

关 键 词：唾液乳杆菌 无细胞上清液(CFCS) 金黄色葡萄球菌 溶血活性 致病性 

分 类 号：S831.5[农业科学—畜牧学]