本文選題：奶牛乳房炎　切入點(diǎn)：調(diào)查　出處：《揚(yáng)州大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：奶牛乳房炎是影響奶牛養(yǎng)殖業(yè)最重要的疾病之一,同時(shí)也是奶牛常發(fā)病之一,給乳品生產(chǎn)和奶牛養(yǎng)殖造成巨大經(jīng)濟(jì)損失。不同種類的病原菌感染是引起奶牛乳房炎的最重要原因。為了解江蘇地區(qū)奶牛乳房炎流行病學(xué)以及病原微生物感染現(xiàn)狀,本研究通過對(duì)江蘇省4個(gè)地區(qū)的5個(gè)規(guī)�；膛ｐB(yǎng)殖場進(jìn)行采樣并進(jìn)行病原菌的分離,同時(shí)對(duì)分離的病原菌的藥物敏感性進(jìn)行檢測并針對(duì)四環(huán)素嚴(yán)重的耐藥現(xiàn)狀進(jìn)行四環(huán)素相關(guān)耐藥基因的檢測與分析。研究內(nèi)容一、江蘇地區(qū)奶牛乳房炎流行病學(xué)調(diào)查通過BMT檢測試劑對(duì)江蘇省4個(gè)地區(qū)的奶牛養(yǎng)殖場的奶牛進(jìn)行奶牛乳房炎檢測,根據(jù)檢測結(jié)果采集隱形及臨床奶牛乳房炎乳樣,并對(duì)這些乳樣病原菌的分離、純化并利用MALID-TOF-MS方法對(duì)江蘇地區(qū)乳房炎源鏈球菌進(jìn)行種屬鑒定。從2014年10月30日至2016年8月17日,共在江蘇地區(qū)四個(gè)城市的牛場(A、B、C、D)中采集臨床和隱形奶牛乳房炎奶樣,一共采集奶樣536份,共分離葡萄球菌382株(47.75%),鏈球菌207株(25.88%),桿菌211株(26.38%)。在受鑒定的207株鏈球菌中共鑒定奶牛乳房炎相關(guān)鏈球菌無乳鏈球菌165株(79.71%),停乳鏈球菌3株(1.45%),乳房鏈球菌3株(1.45%)。研究結(jié)果表明引起江蘇地區(qū)奶牛乳房炎的鏈球菌主要為無乳鏈球菌。研究內(nèi)容二、乳房炎源性鏈球菌耐藥性檢測根據(jù)CLSI規(guī)定的K-B紙片法對(duì)256株乳房炎源性鏈球菌進(jìn)行8類20種藥物進(jìn)行藥物敏感性檢測,結(jié)果顯示鏈球菌對(duì)四環(huán)素、苯唑西林、大觀霉素和多粘菌素B均表現(xiàn)出非常嚴(yán)重的耐藥現(xiàn)象,對(duì)青霉素、諾氟沙星、環(huán)丙沙星等藥物的中介率較高。多重耐藥方面主要的多重耐藥表現(xiàn)為三重、四重、五重和六重耐藥,其中四重耐藥菌株數(shù)量最高(19.92%),十重以上耐藥現(xiàn)象占總受檢數(shù)的10.33%。萬古霉素、頭孢菌素和紅霉素、克拉霉素和克林霉素以及呋喃妥因敏感性較高。研究結(jié)果表明江蘇地區(qū)奶牛乳房炎源鏈球菌耐藥現(xiàn)狀嚴(yán)重,大環(huán)內(nèi)酯類藥物和萬古霉素以及呋喃妥因仍可作為臨床用藥。研究內(nèi)容三、乳房炎源性鏈球菌四環(huán)素耐藥基因檢測本研究對(duì)四環(huán)素耐藥基因(tetM、tetO、tetKA、tetL)等基因進(jìn)行PCR擴(kuò)增。參考文獻(xiàn)設(shè)計(jì)相關(guān)引物,從鏈球菌基因組中擴(kuò)增出tetM、tetO、tetK tetL等耐藥基因,經(jīng)BLAST分析表明擴(kuò)增的片段與GenBank中參考序列同源性均在99%以上。對(duì)256株鏈球菌進(jìn)行四環(huán)素耐藥基因的分子流行病學(xué)檢測,根據(jù)檢測結(jié)果顯示tetM基因的檢出率最高,為39.06%,tet0的檢出率為15.63%,tetK的檢出率為0.78%,tetL未在本次檢測中發(fā)現(xiàn)。研究結(jié)果表明江蘇地區(qū)乳房炎源性鏈球菌對(duì)四環(huán)素的嚴(yán)重耐藥現(xiàn)狀主要由相關(guān)耐藥基因tetM和tetO引起。同時(shí)四環(huán)素耐藥率高于耐藥基因的檢出率,分析原因或與其他同機(jī)制耐藥基因編碼或其他耐藥機(jī)制導(dǎo)致。本研究表明四環(huán)素耐藥現(xiàn)狀嚴(yán)重、耐藥機(jī)制十分復(fù)雜,臨床管理和治療十分困難。
[Abstract]:Cow mastitis is one of the most important diseases affecting dairy cattle breeding, and it is also one of the common diseases in dairy cattle. In order to understand the epidemiology of dairy cow mastitis and the present situation of pathogenic microorganism infection in Jiangsu province, the most important cause of dairy cow mastitis was caused by different kinds of pathogenic bacteria. In this study, five large-scale dairy cattle farms in 4 regions of Jiangsu Province were sampled and the pathogens were isolated. At the same time, the drug sensitivity of isolated pathogenic bacteria was detected and the tetracycline related drug resistance gene was detected and analyzed according to the current situation of tetracycline serious drug resistance. Epidemiological investigation of dairy cow mastitis in Jiangsu province dairy cow mastitis was detected in dairy cattle farms in 4 areas of Jiangsu Province by using BMT reagent. According to the test results, invisible and clinical milk samples of dairy cow mastitis were collected. The strains of Streptococcus mastitis were isolated, purified and identified by MALID-TOF-MS from October 30th 2014 to August 17th 2016. A total of 536 milk samples were collected from clinical and invisible dairy cow mastitis in four cities of Jiangsu province. A total of 382 strains of Staphylococcus, 207 strains of streptococcus and 211 strains of bacilli were isolated. 165 strains of streptococcus associated with mastitis were identified, 3 strains of Streptococcus brevis and 1.45% of Streptococcus mastococcus were identified. The results show that the main streptococcus causing mastitis of dairy cattle in Jiangsu area is Streptococcus acuminata. Resistance of Streptococcus mastitis to tetracycline and oxacillin were detected by K-B disk method according to CLSI. The drug resistance to penicillin, norfloxacin, ciprofloxacin and other drugs is high. The main multidrug resistance in multidrug resistance is triple and quadruple. Among the five and six resistant strains, the four resistant strains were the highest in number (19.922), and the ten or more resistant strains accounted for 10.33% of the total tested drug resistance. Vancomycin, cephalosporin and erythromycin, Clarithromycin, clindamycin and furantoin were highly sensitive. Macrolides, vancomycin and furantoin can still be used as clinical drugs. PCR amplification of tetracycline resistance gene of Streptococcus mastitis. TetMmitt OttetetK tetL was amplified from the genome of Streptococcus mastitis by PCR. The results of BLAST analysis showed that the homology between the amplified fragments and the reference sequence in GenBank was above 99%. The results of molecular epidemiological analysis of tetracycline resistance gene in Streptococcus spp showed that the detection rate of tetM gene was the highest. The detectable rate of tetet0 was 15.63 tetK and 0.78 tetL was not found in this test. The results showed that the severe resistance of Streptococcus mastitis to tetracycline was mainly caused by tetM and tetO. The rate of tetracycline resistance was higher than that of resistance gene. This study shows that tetracycline resistance is serious, drug resistance mechanism is very complex, clinical management and treatment is very difficult.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S858.23

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