【摘要】：子宮微生物菌群在防治病原菌感染生殖道中起著重要作用,目前對于子宮內膜炎的治療普遍使用抗生素療法,但抗生素會在牛奶中殘留以及長期使用抗生素會造成耐藥菌株的產(chǎn)生,因此生產(chǎn)中急需抗生素替代品治療奶牛子宮內膜炎。本研究采用傳統(tǒng)培養(yǎng)方法分離產(chǎn)后健康奶牛陰道乳酸桿菌,為防治奶牛子宮內膜炎的微生態(tài)制劑開發(fā)提供菌株。采用乳酸桿菌培養(yǎng)基分離陰道乳酸桿菌,通過抑菌實驗篩選乳酸桿菌,通過16S rDNA序列分析方法對乳桿菌分離株進行鑒定,并在體外測試乳酸桿菌的產(chǎn)酸、產(chǎn)H2O2能力,采用藥敏紙片法評估乳酸桿菌分離株的安全性,分析乳酸桿菌抗菌物質。通過以上益生功能篩選出最佳益生菌候選,并通過建立家兔病理模型研究乳酸桿菌治療子宮內膜炎臨床效果。本研究共分離獲得30株乳酸桿菌分離株,通過抑菌能力測試共篩選出7株對病原菌具有抑菌能力的乳酸桿菌,經(jīng)鑒定分別為2株為唾液乳桿菌(Lactobacillus salivarius)、2株黏膜乳桿菌(Lactobacillus mucosae)、1株羅伊氏乳桿菌(Lactobacillus reuteri)、1株格氏乳桿菌(Lactobacillus gasseri)和1株約氏乳桿菌(Lactobacillus johnsonii)。7株乳酸桿菌具有良好的產(chǎn)酸性能,并且至少對7種抗生素中的3種以上具有敏感性,大部分乳酸桿菌在一定程度上能夠產(chǎn)生H2O2。有機酸是乳酸桿菌抑制金黃色葡萄球菌、沙門氏菌和大腸桿菌的主要抑菌物質。唾液乳桿菌LWT1和魏斯氏菌組合治療家兔病理模型的有效率為80%,治愈率為60%。綜上所述,唾液乳桿菌LWT1和魏斯氏菌組合制備微生態(tài)制劑能夠用于治療子宮灌注大腸桿菌引起的家兔子宮內膜炎,可用于進一步臨床防治奶牛產(chǎn)后子宮內膜炎。
[Abstract]:The microflora of uterus plays an important role in the prevention and treatment of genital tract infection. At present, antibiotic therapy is widely used in the treatment of endometritis. However, antibiotic residues in milk and long-term use of antibiotics will lead to the production of drug-resistant strains. Therefore, antibiotic substitutes are urgently needed to treat endometritis in dairy cows. In this study, lactobacillus vaginalis was isolated from postpartum healthy dairy cows by traditional culture method, which provided a strain for the development of microecological preparation for the prevention and treatment of endometritis in dairy cows. Lactobacillus vaginalis was isolated on Lactobacillus culture medium, Lactobacillus was screened by bacteriostatic experiment, Lactobacillus was identified by 16s rDNA sequence analysis method, and the acid-producing and H2O2 production ability of Lactobacillus was tested in vitro. The safety of Lactobacillus isolates was evaluated by drug sensitive disk method, and the antimicrobial substances of Lactobacillus were analyzed. The best candidate for probiotics was screened by the probiotics above, and the clinical effect of Lactobacillus on endometritis was studied by establishing rabbit pathological model. In this study, 30 strains of Lactobacillus were isolated, and 7 strains of Lactobacillus with bacteriostasis ability were screened by the test of bacteriostatic ability. Two strains of Lactobacillus saliva, two strains of Lactobacillus (Lactobacillus salivarius), two strains of Lactobacillus mucosal, one strain of Lactobacillus Roy, one strain of (Lactobacillus reuteri), one strain of Lactobacillus gravis, one strain of Lactobacillus, (Lactobacillus johnsonii) .7, were identified as Lactobacillus spp. And at least 3 of the seven antibiotics are sensitive to them, and most Lactobacillus can produce H _ 2O _ 2 to some extent. Organic acids are the main bacteriostatic substances of Lactobacillus against Staphylococcus aureus, Salmonella and Escherichia coli. The effective rate and cure rate of lactobacillus saliva LWT1 combined with Weiss's bacillus in treating rabbit pathological model were 80 and 60 respectively. To sum up, the combination of Lactobacillus saliva LWT1 and Weiss's bacillus can be used to treat the rabbit endometritis caused by Escherichia coli, and can be used to prevent and treat postpartum endometritis in dairy cows.
【學位授予單位】：吉林農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S858.23

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