奶山羊乳房炎病原菌分离鉴定及多重PCR检测方法建立

发布时间：2018-11-16 12:59

【摘要】：奶山羊乳房炎是奶山羊养殖业中较为常见且危害巨大的疾病之一,它不仅给养殖者造成严重的经济损失,同时也威胁着羊奶及其相关奶产品消费者的健康,造成食品安全隐患。奶山羊乳房炎病因复杂,与病原微生物、动物自身、饲养管理等都有很大的关系。病原微生物种类繁多且存在较大的地域差异,给该病的诊治带来了极大的困难。目前,我国尚没有可以快速、准确诊断奶山羊乳房炎并且对病原微生物做出精确定位的诊断方法,本次研究主要针对陕西部分地区奶山羊乳房炎进行研究,以期能够为奶山羊乳房炎的诊断、治疗提供一定的理论依据。本次试验从陕西省富平县、陇县两个地区随机选取的11个养殖场采取的55份临床型奶山羊乳房炎乳样进行病原菌分离鉴定,针对主要病原菌进行药敏试验、建立多重PCR检测方法。研究结果如下:1.分离得到形态各异的奶山羊乳房炎病原菌8种54株,其中大肠杆菌18株、金黄色葡萄球菌14株、蜡状芽孢杆菌7株、绿脓杆菌6株、无乳链球菌4株、凝固酶阴性葡萄球菌3株、真菌2株。大肠杆菌分离率最高,达33.3%,其次为金黄色葡萄球菌,分离率为25.9%。2.分离得到的主要病原菌大肠杆菌、金黄色葡萄球菌、蜡状芽孢杆菌、无乳链球菌,均对氯霉素、四环素、左氟沙星和阿米卡星等药物表现出高度敏感,而对青霉素、万古霉素、环丙沙星、氨苄西林、头孢唑啉和庆大霉素等常用药表现中度或重度耐药。3.建立了一种直接以菌液作为模板,能够一次性检测蜡状芽孢杆菌、绿脓杆菌、金黄色葡萄球菌和大肠杆菌4种奶山羊乳房炎主要致病菌的多重PCR检测方法。
[Abstract]:Dairy goat mastitis is one of the most common and harmful diseases in the dairy goat breeding industry. It not only causes serious economic losses to the breeders, but also threatens the health of the consumers of goat milk and its related milk products, resulting in hidden dangers of food safety. The etiology of dairy goat mastitis is complicated, which is related to pathogenic microorganism, animal itself, feeding management and so on. There are many kinds of pathogenic microorganisms and there are great regional differences, which brings great difficulties to the diagnosis and treatment of the disease. At present, there is no diagnostic method for rapid and accurate diagnosis of dairy goat mastitis and accurate location of pathogenic microorganisms in China. This study mainly focuses on dairy goat mastitis in some parts of Shaanxi Province. In order to provide a theoretical basis for the diagnosis and treatment of mastitis in dairy goats. In this experiment, 55 milk samples of clinical dairy goat mastitis were isolated and identified from 11 farms randomly selected from Fuping County and Longxian County, Shaanxi Province, and drug sensitivity tests were carried out for the main pathogens. A multiplex PCR detection method was established. The results are as follows: 1. Among them, 18 strains of Escherichia coli, 14 strains of Staphylococcus aureus, 7 strains of Bacillus cereus, 6 strains of Pseudomonas aeruginosa and 4 strains of Streptococcus lactococcus were isolated. 3 strains of coagulase negative staphylococcus and 2 strains of fungi. The isolation rate of Escherichia coli was the highest, up to 33. 3%, followed by Staphylococcus aureus, and the isolation rate was 25. 9. 2. Escherichia coli, Staphylococcus aureus, Bacillus cereus, Streptococcus lactococcus were all highly sensitive to chloramphenicol, tetracycline, levofloxacin and amikacin, but highly sensitive to penicillin and vancomycin. Ciprofloxacin, ampicillin, cefazolin and gentamycin, and other commonly used drugs showed moderate or severe drug resistance. 3. A multiplex PCR method was developed for the detection of four major pathogens of mastitis in dairy goats, I. e., Bacillus cereus, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.27

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