本文選題：解淀粉芽胞桿菌 + 分離鑒定��； 參考：《吉林農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：輪狀病毒病是由輪狀病毒感染引起的初生動物消化機(jī)能紊亂的一種腸道傳染病,臨床上以腹瀉、脫水為主要特征,給我國的畜牧業(yè)生產(chǎn)造成重大損失。目前對于輪狀病毒病的治療沒有特效藥物,一般以預(yù)防為主。因此,尋找安全、有效的預(yù)防和治療輪狀病毒病的制劑顯得尤為重要。研究表明,芽胞桿菌中的個別種類可以產(chǎn)生抗病毒物質(zhì)。所以本試驗旨在獲得既具有抗菌又具有抗病毒活性的益生芽胞桿菌,為拓展微生態(tài)制劑的功能奠定一定基礎(chǔ)。試驗采集20份不同地點的土壤樣品,分別以金黃色葡萄球菌和大腸桿菌為指示菌,采用平板涂布法,篩選出具有抗菌作用的芽胞桿菌4株,分別命名為YI-1、YI-3、YI-7、YI-8。經(jīng)過小鼠毒性試驗及耐人工胃酸、人工腸液試驗對這4株菌進(jìn)行復(fù)篩,結(jié)果發(fā)現(xiàn)菌株YI-1具有安全、可抗人工胃酸、人工腸液的特點。經(jīng)分子生物學(xué)和檸檬酸鹽試驗鑒定該菌為解淀粉芽胞桿菌。用MA-104細(xì)胞培養(yǎng)輪狀病毒,測定病毒滴度。試驗乳鼠隨機(jī)分為5組,分別為陰性對照組、陽性對照組、治療組、預(yù)防組、細(xì)菌與病毒體外作用組,6日齡灌服病毒,每日觀察乳鼠精神狀態(tài)、排便情況,并在接種病毒后的48h、96h、168h,每組隨機(jī)取2只脫頸處死后,剖檢觀察消化道病變。同時分別取空腸、回腸組織,用甲醛固定,做石蠟切片,用HE染色后測定各組的空腸、回腸絨毛高度。結(jié)果顯示在接種病毒后的96h,5組乳鼠的腹瀉率分別為0%、100%、66.7%、28.5%和0%;剖檢可見發(fā)病鼠空腸、回腸內(nèi)充滿氣體,腸壁變薄,腸黏膜脫落;制備的病理切片經(jīng)HE染色可見發(fā)病乳鼠空腸、回腸部分腸壁水腫,腸上皮細(xì)胞間連接被破壞;腸絨毛腫脹、變短;微絨毛排列紊亂。用SPSS 17.0統(tǒng)計軟件對測得的空腸、回腸絨毛高度進(jìn)行分析,結(jié)果顯示各組間腸絨毛高度差異顯著,具有統(tǒng)計學(xué)意義。
[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.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S855.3

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