解淀粉芽胞杆菌对乳鼠感染轮状病毒防治作用研究

发布时间：2018-05-03 08:04

本文选题：解淀粉芽胞杆菌 + 分离鉴定　；参考：《吉林农业大学》2015年硕士论文

【摘要】：轮状病毒病是由轮状病毒感染引起的初生动物消化机能紊乱的一种肠道传染病,临床上以腹泻、脱水为主要特征,给我国的畜牧业生产造成重大损失。目前对于轮状病毒病的治疗没有特效药物,一般以预防为主。因此,寻找安全、有效的预防和治疗轮状病毒病的制剂显得尤为重要。研究表明,芽胞杆菌中的个别种类可以产生抗病毒物质。所以本试验旨在获得既具有抗菌又具有抗病毒活性的益生芽胞杆菌,为拓展微生态制剂的功能奠定一定基础。试验采集20份不同地点的土壤样品,分别以金黄色葡萄球菌和大肠杆菌为指示菌,采用平板涂布法,筛选出具有抗菌作用的芽胞杆菌4株,分别命名为YI-1、YI-3、YI-7、YI-8。经过小鼠毒性试验及耐人工胃酸、人工肠液试验对这4株菌进行复筛,结果发现菌株YI-1具有安全、可抗人工胃酸、人工肠液的特点。经分子生物学和柠檬酸盐试验鉴定该菌为解淀粉芽胞杆菌。用MA-104细胞培养轮状病毒,测定病毒滴度。试验乳鼠随机分为5组,分别为阴性对照组、阳性对照组、治疗组、预防组、细菌与病毒体外作用组,6日龄灌服病毒,每日观察乳鼠精神状态、排便情况,并在接种病毒后的48h、96h、168h,每组随机取2只脱颈处死后,剖检观察消化道病变。同时分别取空肠、回肠组织,用甲醛固定,做石蜡切片,用HE染色后测定各组的空肠、回肠绒毛高度。结果显示在接种病毒后的96h,5组乳鼠的腹泻率分别为0%、100%、66.7%、28.5%和0%;剖检可见发病鼠空肠、回肠内充满气体,肠壁变薄,肠黏膜脱落;制备的病理切片经HE染色可见发病乳鼠空肠、回肠部分肠壁水肿,肠上皮细胞间连接被破坏;肠绒毛肿胀、变短;微绒毛排列紊乱。用SPSS 17.0统计软件对测得的空肠、回肠绒毛高度进行分析,结果显示各组间肠绒毛高度差异显著,具有统计学意义。
[Abstract]:Rotavirus disease is a kind of intestinal infectious disease caused by rotavirus infection in primary animals. It is characterized by diarrhea and dehydration in clinic and causes great loss to animal husbandry production in China. At present, the treatment of rotavirus disease has no special drugs, generally to prevent. Therefore, the search for safe, effective prevention and treatment of rotavirus disease preparation is particularly important. Studies have shown that individual species of Bacillus can produce antiviral substances. The purpose of this study was to obtain probiotic Bacillus with both antibacterial and antiviral activities, and to lay a foundation for the development of the function of microecological agents. Twenty soil samples from different sites were collected. Staphylococcus aureus and Escherichia coli were used as indicator bacteria, and 4 strains of Bacillus spp. The four strains were screened again by mice toxicity test, artificial stomach acid tolerance test and artificial intestinal fluid test. The results showed that the strain YI-1 had the characteristics of safety, resistance to artificial gastric acid and artificial intestinal fluid. The strain was identified as Bacillus amylolyticus by molecular biology and citrate test. Rotavirus was cultured in MA-104 cells and the titer of rotavirus was measured. The experimental neonatal rats were randomly divided into five groups: negative control group, positive control group, treatment group, prevention group, bacteria and virus in vitro group. At 48h, 96h and 168h after inoculation of the virus, 2 rats in each group were killed at random, and the pathological changes of digestive tract were observed. At the same time, jejunum and ileum tissue were taken, fixed with formaldehyde, paraffin sections were made, jejunum and ileal villi were measured by HE staining. The results showed that the diarrhea rates of the 5 groups were 0 ~ 100% and 28.5%, respectively. The jejunum of the infected rats was found to be filled with gas, the intestinal wall became thinner, and the intestinal mucosa was shedding, and the pathological sections were stained with HE staining to show the jejunum of the infected rats, the ileum was filled with gas, the intestinal mucosa was shedding off, and the pathological sections were stained by HE. Part of ileum wall edema, intestinal epithelial intercellular junction was destroyed; intestinal villi were swollen and shortened; microvilli arranged in disorder. The height of jejunum and ileum villi was analyzed by SPSS 17.0 software. The results showed that there was significant difference in villi height among the groups.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S855.3

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