四川部分地区山羊皮下脓肿的细菌学分析

发布时间：2018-11-12 15:36

【摘要】：山羊饲养历史悠久,具有很高的经济价值。我国改革开放30多年以来,山羊畜牧产业发展迅速,占全国畜牧产业总产值的6%左右,四川作为山羊养殖大省,山羊存栏总量占据全国前五,显得尤为重要。在畜牧生产中,山羊皮下脓肿是一种常见多发病,对养羊产业造成巨大的经济损失。近来对其的报道比较多,却没有很好的治疗办法,关键在于其病原菌来源的多样性。针对此,本实验展开对四川主要养羊地区的山羊皮下脓肿病原学调查,为判明四川省主要养羊区的山羊皮下或淋巴结脓肿的致病病原并为该病的预防治疗提供依据。本研究于2015年4月至9月期间在四川宣汉、广安、攀枝花、巴中、资阳几个重要养羊地区共采集47份山羊脓肿病料,分别在需氧和厌氧条件下培养分离纯化出山羊皮下脓肿的致病菌。对分离纯化的致病菌进行菌落形态观察和细菌生化特性分析,并结合16S rRNA基因序列测序分析进行细菌鉴定。选择致病菌测定其半数致死量(LDso)以判定其毒力强弱,并对鉴定的致病菌株进行药物敏感性试验,筛选出致病菌株的敏感性药物。1、对六个规模化养羊场共2440只存栏山羊进行调查,有230只发病,该病在各羊场的发病率从8.36%-11.27%不等,平均发病率为9.43%。采集的47份病料中共分离得到118株细菌,其中革兰氏阴性菌42株,占35.6%:包括大肠杆菌15株,占12.7%,弯曲假单胞菌27株,占22.9%：革兰氏阳性菌76株,占64.4%:包括伪结核棒状杆菌22株,占18.6%,金黄色葡萄球菌13株,占11.0%,芽孢杆菌41株,占34.7%。2、对分离得到的致病菌伪结核棒状杆菌和金黄色葡萄球菌进行毒力检测,其中伪结核棒状杆菌菌株对小鼠的致病力较强,LD50为107.5cfu/ml,而金黄色葡萄球菌的LD50为109.7cfu/ml。3、对试验分离得到的22株伪结核棒状杆菌和13株金黄色葡萄球菌采用美国临床和实验室标准协会(CLSI)推荐的Kirby-Bauer纸片法分别进行9种临床常用抗菌药物的耐药性试验,并选取标准菌株伪结核棒状杆菌ATCC19410和金黄色葡萄球菌ATCC25923作为质控菌株进行试验结果可靠性验证。药敏结果表明：试验菌株均表现出严重的耐药性,但对药物的敏感性不同。伪结核棒状杆菌对利福平、复方新诺明、环丙沙星和链霉素具有一定的敏感性,对青霉素和庆大霉素耐药。金黄色葡萄球菌对左氧氟沙星和利福平均具有较高的敏感性,对青霉素、氨苄西林、克林霉素和头孢他啶耐药。该研究发现伪结核棒状杆菌和金黄色葡萄球菌为发病羊场的共同致病菌,并进行了药物筛选,为该地区山羊皮下脓肿病的防治提供了参考。
[Abstract]:Goat breeding has a long history and has high economic value. The goat animal husbandry industry has developed rapidly in China since the reform and opening up for more than 30 years, accounting for about 6% of the total output value of the national livestock industry. Sichuan, as a large province of goat farming, occupies the top five in the total amount of goat stock in the country, which is particularly important. In livestock production, goat subcutaneous abscess is a common disease, which causes huge economic loss to sheep industry. There are many reports about it recently, but there is no good treatment method. The key is the diversity of pathogen source. In order to identify the pathogeny of goat subcutaneous or lymph node abscess in the main goat raising area of Sichuan province and provide the basis for the prevention and treatment of the disease, this study investigated the etiology of goat subcutaneous abscess in the main goat raising area of Sichuan province. From April to September 2015, 47 goat abscesses were collected in Xuanhan, Guangan, Panzhihua, Bazhong and Ziyang, Sichuan Province. The pathogenic bacteria of goat subcutaneous abscess were isolated and purified under aerobic and anaerobic conditions. The colony morphology and biochemical characteristics of the isolated and purified pathogenic bacteria were observed and identified by 16s rRNA gene sequencing. In order to determine the virulence of pathogenic bacteria, (LDso) was selected to determine its virulence, and the susceptibility test of pathogenic strains was carried out to screen the sensitive drugs of pathogenic strains. A total of 2440 sheep were collected from six large-scale sheep farms. The incidence of the disease ranged from 8.36% to 11.27%, and the average incidence rate was 9.43%. A total of 118 strains of bacteria were isolated from 47 samples, 42 of which were Gram-negative bacteria, accounting for 35.6k.There were 15 strains of Escherichia coli, 12.7m, 27 strains of Pseudomonas flexor, 22.910 strains of Gram-positive bacteria, 76 strains of Gram-positive bacteria, 15 strains of Escherichia coli, 27 strains of Pseudomonas bending and 76 strains of Gram-positive bacteria. Accounting for 64.4%: including 22 strains of pseudotuberculosis Corynebacterium tuberculosis (18.6B), 13 strains of Staphylococcus aureus (11.0%), 41 strains of Bacillus, 34.70.2. The virulence of the isolated pathogenic bacteria, Corynebacterium tuberculosis and Staphylococcus aureus, was tested. The virulence of the strain was stronger in mice, LD50 was 107.5 cfurml. The LD50 of Staphylococcus aureus was 109.7 cfuml.3. A total of 22 strains of pseudotuberculosis and 13 strains of Staphylococcus aureus were tested for drug resistance of 9 common clinical antimicrobial agents by Kirby-Bauer disk method recommended by (CLSI) of American Association of Clinical and Laboratory Standards. The standard strains ATCC19410 and ATCC25923 were selected as the quality control strains to verify the reliability of the test results. The results of drug sensitivity showed that all of the tested strains showed severe drug resistance, but the sensitivity to drugs was different. Corynebacterium pseudotuberculosis is sensitive to rifampicin, compound sulfamethoxine, ciprofloxacin and streptomycin, and resistant to penicillin and gentamicin. Staphylococcus aureus showed high average sensitivity to levofloxacin and rifampicin, and was resistant to penicillin, ampicillin, clindamycin and ceftazidime. This study found that pseudotuberculosis corynebacterium and Staphylococcus aureus were the common pathogenic bacteria in sheep farm, and the drug screening was carried out, which provided a reference for the prevention and treatment of goat subcutaneous abscess disease in this area.
【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S858.27

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