治疗奶牛子宫内膜炎益生乳酸杆菌筛选及临床效果研究

发布时间：2018-08-26 17:03

【摘要】：子宫微生物菌群在防治病原菌感染生殖道中起着重要作用,目前对于子宫内膜炎的治疗普遍使用抗生素疗法,但抗生素会在牛奶中残留以及长期使用抗生素会造成耐药菌株的产生,因此生产中急需抗生素替代品治疗奶牛子宫内膜炎。本研究采用传统培养方法分离产后健康奶牛阴道乳酸杆菌,为防治奶牛子宫内膜炎的微生态制剂开发提供菌株。采用乳酸杆菌培养基分离阴道乳酸杆菌,通过抑菌实验筛选乳酸杆菌,通过16S rDNA序列分析方法对乳杆菌分离株进行鉴定,并在体外测试乳酸杆菌的产酸、产H2O2能力,采用药敏纸片法评估乳酸杆菌分离株的安全性,分析乳酸杆菌抗菌物质。通过以上益生功能筛选出最佳益生菌候选,并通过建立家兔病理模型研究乳酸杆菌治疗子宫内膜炎临床效果。本研究共分离获得30株乳酸杆菌分离株,通过抑菌能力测试共筛选出7株对病原菌具有抑菌能力的乳酸杆菌,经鉴定分别为2株为唾液乳杆菌(Lactobacillus salivarius)、2株黏膜乳杆菌(Lactobacillus mucosae)、1株罗伊氏乳杆菌(Lactobacillus reuteri)、1株格氏乳杆菌(Lactobacillus gasseri)和1株约氏乳杆菌(Lactobacillus johnsonii)。7株乳酸杆菌具有良好的产酸性能,并且至少对7种抗生素中的3种以上具有敏感性,大部分乳酸杆菌在一定程度上能够产生H2O2。有机酸是乳酸杆菌抑制金黄色葡萄球菌、沙门氏菌和大肠杆菌的主要抑菌物质。唾液乳杆菌LWT1和魏斯氏菌组合治疗家兔病理模型的有效率为80%,治愈率为60%。综上所述,唾液乳杆菌LWT1和魏斯氏菌组合制备微生态制剂能够用于治疗子宫灌注大肠杆菌引起的家兔子宫内膜炎,可用于进一步临床防治奶牛产后子宫内膜炎。
[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.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23

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