罗非鱼无乳链球菌疾病微生态防治研究——拮抗益生菌的分离及鉴定.docVIP

罗非鱼无乳链球菌疾病微生态防治研究 ——拮抗益生菌的分离及鉴定 大学生：吴川良 杨慧 熊文丽 指导教师：周永灿 王世锋 （海南大学，海南海口，570228） 摘要：本文利用径向扩散法，以具有高致病性的罗非鱼无乳链球菌为指示菌，从海南不同的水体中分离对无乳链球菌具有较强拮抗作用的益生菌，并利用16S rDNA进行初步鉴定，和利用毒性耐受性试验测定其安全性。结果表明，不同水体中能够分离拮抗益生菌的难易程度不同，在有机质含量多、不用或少用抗生素的水体中较易分离到益生菌，而在有机物含量少，或者是有机物含量高，但是抗生素使用量较大的水体中，较难获得拮抗益生菌；从采集的9个地点共分离到1904株细菌，从中分离获得59株对对无乳链球菌具有较强拮抗作用的菌株，抑菌圈直径平均大小为22.61mm，经16S rDNA序列测定表明，所分离的拮抗益生菌主要为枯草杆菌，以及部分苏云金芽孢杆菌、表皮葡萄球菌、蜡样芽孢杆菌等。59株益生菌经腹腔注射法测定其对罗非鱼的毒性，其中22株拮抗菌对罗非鱼无毒性，为罗非鱼无乳链球菌疾病的微生态防治奠定了良好基础。 关键词：罗非鱼；无乳链球菌；拮抗益生菌 The?Ecological?Therapy?of?the?Tilapia?Disease?Caused?by? Streptococcus?agalactiae 桾the?separation?and?identification?of?antergic?probiotics Chuanliang Wu, Hui Yang, Wenli Xiong, Yongcan Zhou*, Shifeng Wang* (Hainan University, Haikou, Hainan China, 570228) Abstract：This study was conducted to separate and identify the antergic probiotics. We separated the probiotics from different water bodies of Hainan which had strong antagonism effect for Streptococcus agalactiae. First, we used the radial diffusion method with highly pathogenic tilapia Streptococcus agalactiae as the indicator bacteria, and then these probiotics were identified by 16 S rDNA technology. The security of these selected probiotics were detected by toxicity tolerance tests. It turned out that the amount of probiotics that we could separate were different in different water bodies. There were more probiotics in the water that contained more organic matter content or less antibiotics. It was difficult to separate probiotics from the water that contained more inorganic qualitative content and antibiotics. In this study, we acquired 59 strains which had strong antagonism effect for Streptococcus agalactiae from the total 1904 strains of bacteria achieved from 9 sampling sites. It indicated that most probiotics were Bacillus subtilis, Bacillus thuringiensis, Staphylococcus epidermidis and Bacillus cereus. Through the toxicity tolerance tests, we found 22 strains which were nontoxic to the tilapia. The resu