本文選題：奶牛 + 乳房炎��； 參考：《河北農(nóng)業(yè)大學》2015年碩士論文

【摘要】：奶牛乳房炎是奶牛養(yǎng)殖業(yè)中最常見的一種疾病,同時也是制約乳業(yè)發(fā)展的重要因素。本研究利用DHI數(shù)據(jù)監(jiān)測和LMT法相結合方法對河北省7個地區(qū)的420份乳樣診斷奶牛乳房炎的發(fā)病情況。結果,DHI數(shù)據(jù)監(jiān)測顯示從420份樣品中檢測出126份陽性乳樣,陽性率為30%,同時采用LMT診斷試劑進行檢測,有128份陽性乳樣,檢出率30.48%,即本研究中DHI數(shù)據(jù)監(jiān)測與LMT試劑診斷的相符率為98.42%。其中邢臺、滄州、廊坊、唐山、保定、石家莊、張家口地區(qū)的乳樣陽性檢出率分別為35%、28%、24.44%、28.89%、30%、27.14%、33.33%。本研究利用微生物學和分子生物學相結合的方法,對河北省7個地區(qū)乳樣進行乳房炎致病菌的分離培養(yǎng)鑒定�？偣卜蛛x出4種主要致病菌,362株。引起奶牛乳房炎的主要致病菌分別為金黃色葡萄球菌、無乳鏈球菌、大腸桿菌、綠膿桿菌,其中金黃色葡萄球菌167株,占總菌株數(shù)的46%,無乳鏈球菌94株,占總菌株數(shù)的25.97%,大腸桿菌84株,占總菌株數(shù)的23%,綠膿桿菌17株,占總菌株數(shù)的5%。在檢出的乳樣中,幾乎全部為金黃色葡萄球菌、無乳鏈球菌、大腸桿菌的混合感染,但其中檢出的17株綠膿桿菌全部由張家口地區(qū)乳樣中分離所得,為4種分離菌的混合感染。參照NCCLS推薦的K-B藥敏紙片法對奶牛乳房炎的四種分離菌進行15種藥物的敏感性試驗,試驗測定其抑菌圈大小。結果表明,七個地區(qū)致病菌對頭孢噻肟、呋喃唑酮、恩諾沙星、氟苯尼考及磷霉素的敏感性較高;對卡那霉素、阿奇霉素、四環(huán)素和環(huán)丙沙星耐藥嚴重。所有菌株對青霉素、鏈霉素幾乎完全耐藥。根據(jù)藥敏結果,河北省7個地區(qū)的362株菌中存在多重耐藥現(xiàn)象,約占全菌株數(shù)的86.46%。采用PCR方法,研究河北省奶牛乳房炎病原菌的耐藥基因的攜帶情況,評估耐藥表型與基因型的相關性。結果顯示:金黃色葡萄球菌分離株、無乳鏈球菌、大腸桿菌和綠膿桿菌的分離株檢測出的優(yōu)勢耐藥基因分別為gyr A、erm B、Qnr B、Qnr B。前3種細菌的耐藥表型與基因型的相符率高達85%以上。表明藥敏試驗結果準確,可以用于臨床指導用藥。用藥后各地區(qū)的DHI數(shù)據(jù)監(jiān)測9、10、11、12月份SCC水平均有顯著差異(P0.01),其下降幅度在14.01%-45%之間。選取部分致病菌的PCR擴增產(chǎn)物進行膠回收、測序,并與Gene Bank中已發(fā)表的相關序列進行同源性比對,從而確定病原菌所屬種類。綜上所述,本研究通過對分離鑒定的致病菌進行藥敏試驗和耐藥基因檢測,將試驗結果反饋回各奶牛場,指導其臨床用藥和飼養(yǎng)管理,同時為河北省不同地區(qū)奶牛乳房炎主要致病菌的防治提供了理論依據(jù)。
[Abstract]:Dairy cow mastitis is the most common disease in dairy farming, and it is also an important factor restricting the development of dairy industry. In this study, DHI data monitoring and LMT method were used to detect the incidence of Dairy Mastitis in 420 milk samples in 7 regions of Hebei province. Results, DHI data monitoring showed that 126 samples were detected from 420 samples. The positive rate of sex milk was 30%, and LMT diagnostic reagent was used to detect 128 positive milk samples. The detection rate was 30.48%. That is, the coincidence rate of DHI data monitoring and LMT reagent diagnosis in this study was 98.42%. in Xingtai, Cangzhou, Langfang, Tangshan, Baoding, Shijiazhuang and Zhangjiakou. The positive rates of milk samples were 35%, 28%, 24.44%, 28.89%, 30%, respectively. 27.14%, 33.33%. in this study, a combination of Microbiology and molecular biology was used to identify the isolation and culture of mammary mastitis pathogenic bacteria in 7 regions of Hebei province. A total of 4 main pathogens were isolated and 362. The main pathogenic bacteria causing cow mastitis were Staphylococcus aureus, Lactococcus lactis, Escherichia coli, and purulent purulent. Bacilli, including 167 strains of Staphylococcus aureus, accounted for 46% of the total number of bacteria, 94 strains of Streptococcus lactis, 25.97% of the total number of bacteria, 84 strains of Escherichia coli, 23% of the total number of bacteria, 17 strains of Pseudomonas aeruginosa, which accounted for 5%. in the total number of bacteria, almost all were Staphylococcus aureus, Streptococcus free Streptococcus and Escherichia coli. 17 strains of Pseudomonas aeruginosa isolated from the Zhangjiakou region were isolated from the milk samples of the Zhangjiakou region and were mixed infection of 4 isolated bacteria. The sensitivity tests of 15 kinds of drugs for four isolates of Dairy Mastitis in dairy cows were tested with reference to the K-B drug sensitive paper recommended by us, and the size of the bacteriostasis was tested. The results showed that seven regional pathogenic bacteria were to cefotaxime. Furazolidone, enrofloxacin, florfenicol and fosfomycin were more sensitive to kanamycin, azithromycin, tetracycline and ciprofloxacin. All strains were almost completely resistant to penicillin and streptomycin. According to the results of drug sensitivity, there were multiple resistance in 362 strains of 7 regions in Hebei Province, which accounted for the 86.46%. mining of the total number of bacteria. The PCR method was used to study the carrying status of the resistant genes of the pathogenic bacteria of dairy cow mastitis in Hebei Province, and to evaluate the correlation between the resistance phenotype and genotype. The results showed that the dominant resistance genes of Staphylococcus aureus isolates, Streptococcus lactis, Escherichia coli and Pseudomonas aeruginosa were Gyr A, ERM B, Qnr B, and Qnr B., respectively. The resistant phenotype of the bacteria was more than 85% of the genotype, indicating that the drug sensitivity test results were accurate and could be used for clinical guidance. The DHI data of 9,10,11,12 months after the drug use were significantly different (P0.01), and the decrease was between 14.01%-45%. The PCR amplification products of some pathogenic bacteria were selected for rubber recovery. Sequencing, and comparing the homologous sequences with the related sequences published in Gene Bank to determine the species of pathogenic bacteria. In summary, this study returns the results of the test to the dairy farms, directing the clinical medication and feeding management, and in Hebei province. It provides a theoretical basis for the prevention and treatment of the main pathogenic bacteria of dairy cow mastitis.
【學位授予單位】：河北農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.23

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本文編號：2114786