本文選題：波氏桿菌 + 分離鑒定　； 參考：《山東農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：兔波氏桿菌病是由支氣管敗血波氏桿菌引發(fā)的一種常見兔呼吸道疾病,主要引起成年兔的支氣管炎、鼻炎和膿皰性肺炎,斷乳仔兔和哺乳仔兔的急性死亡等。近年來,伴隨著我國規(guī)�；脴I(yè)養(yǎng)殖的迅速發(fā)展,支氣管敗血波氏桿菌的感染率明顯升高,并常與葡萄球菌、多殺性巴氏桿菌等混合感染,已成為導(dǎo)致家兔發(fā)病率和死亡率增加的重要原因之一,給養(yǎng)兔業(yè)造成重大經(jīng)濟損失�？股刂委熢谛笄蒺B(yǎng)殖中是一種重要的防制手段,但由于藥物濫用,細菌耐藥性日趨嚴重,常造成藥物治療失敗。所以增加對當(dāng)?shù)丶毦退幮约八幬锩舾行誀顩r的認知,對于正確分析耐藥性的發(fā)展趨勢及對臨床上指導(dǎo)合理用藥意義重大。針對當(dāng)前嚴重的多重耐藥現(xiàn)象,探究耐藥機制,針對耐藥原因找出對應(yīng)方法來延長已有抗生素的敏感期和研究新藥同為重要。本實驗在分離鑒定細菌基礎(chǔ)上,對藥物敏感性進行了檢測,并對耐藥基因進行了鑒定,對耐藥傳播機制進行了初步研究。研究主要包括以下內(nèi)容:1.本實驗2012~2014年自浙江省部分兔場采集病兔的鼻腔分泌物,根據(jù)細菌形態(tài)、培養(yǎng)特性、生化試驗,并結(jié)合PCR方法對分離菌株進行鑒定,分離鑒定出22株兔波氏桿菌;K-B氏紙片法檢測細菌對18種常用抗生素的藥物敏感性,藥敏結(jié)果顯示,分離菌多重耐藥嚴重,對青霉素G、頭孢拉定、鏈霉素、林可霉素、克林霉素、甲氧嘧啶耐藥嚴重,而對多黏菌素B、左氟沙星、強力霉素、四環(huán)素等藥物敏感。2.針對耐藥表型對應(yīng)的耐藥基因,設(shè)計特異性引物,使用PCR法對22株兔波氏桿菌分離株進行耐藥基因檢測,并進行測序分析。其中所有分離株均檢測到β-內(nèi)酰胺類耐藥基因blaTEM,檢測率為100%;17株檢測到磺胺類藥物耐藥基因sul1,檢測率為77%;分離菌株檢測到的這兩種耐藥基因與耐藥表型相符。17株檢測到Ⅰ類整合子的整合酶基因int1,檢測率為77%,說明整合子在兔波氏桿菌耐藥傳播機制中發(fā)揮作用。20株檢測到多重耐藥基因marC,檢測率為91%,因marC基因檢出率較高,推測波氏桿菌耐藥性與染色體基因突變介導(dǎo)的主動外排系統(tǒng)有關(guān)。9株檢測到轉(zhuǎn)座子遺傳標志基因merA,檢測率為41%,merA基因的存在及反向PCR結(jié)果顯示TEM耐藥基因的傳播轉(zhuǎn)座子存在相關(guān)聯(lián)系。3.對本研究所分離的22株兔波氏桿菌進行了質(zhì)粒提取及消除實驗,結(jié)果表明,所有22株分離菌均攜帶質(zhì)粒,質(zhì)粒消除后細菌部分耐藥性狀發(fā)生變化,對氨芐青霉素、頭孢唑林、四環(huán)素、多粘菌素B等藥物的敏感性增強。證明了質(zhì)粒介導(dǎo)了兔波氏桿菌分離菌株的部分耐藥性。本研究結(jié)果表明,兔波氏桿菌是引起兔呼吸道疾病的主要病原菌;分離菌耐藥率較高,并呈現(xiàn)多重耐藥;檢測到的耐藥基因與耐藥表型相符;質(zhì)粒、整合子、轉(zhuǎn)座子這三種轉(zhuǎn)移因子的檢出說明兔波氏桿菌存在多種耐藥傳播機制。
[Abstract]:Bordetella rabbit disease is a common respiratory tract disease caused by Bordetella bronchus. It mainly causes bronchitis, rhinitis and pustular pneumonia in adult rabbits, acute death of weaning rabbits and lactation rabbits, and so on.In recent years, with the rapid development of large-scale rabbit farming in China, the infection rate of Bordetella bronchus has increased significantly, and it is often mixed with Staphylococcus, Pasteurella multocida and so on.It has become one of the important reasons for the increase of morbidity and mortality of rabbits, and has caused great economic losses to rabbit farming.Antibiotic therapy is an important means of prevention and control in livestock and poultry breeding. However, drug abuse and drug resistance of bacteria are becoming more and more serious, which often lead to drug treatment failure.Therefore, it is of great significance for correctly analyzing the development trend of drug resistance and guiding rational drug use in clinic to increase the understanding of drug resistance and drug sensitivity of local bacteria.In view of the serious phenomenon of multidrug resistance at present, it is important to explore the mechanism of drug resistance, to find corresponding methods to prolong the sensitive period of existing antibiotics and to study new drugs.On the basis of isolation and identification of bacteria, the drug sensitivity was detected, the resistance gene was identified, and the mechanism of drug resistance transmission was studied.The research mainly includes the following contents: 1: 1.The nasal secretions of infected rabbits were collected from some rabbit farms in Zhejiang Province from 2012 to 2014. The isolated strains were identified by bacterial morphology, culture characteristics, biochemical tests and PCR method.A total of 22 strains of Bacillus vulgaris were isolated and identified by K-B disk method to detect the susceptibility of bacteria to 18 common antibiotics. The results of drug sensitivity showed that the isolates were multidrug resistant to penicillin G, cefradine, streptomycin, lincomycin, clindamycin, lincomycin, clindamycin, penicillin, cefradine, streptomycin and clindamycin.Methoxypyrimidine is highly resistant to polymyxin B, levofloxacin, doxycycline, tetracycline and so on.A specific primer was designed to detect the drug resistance genes of 22 isolates of Boxerobacterium rabbit by PCR method, and the results were analyzed by sequencing.Among them, 尾 -lactam resistance gene blaTEM was detected in all isolates, the detection rate was 100% and 17 strains detected sulfan resistance gene sul1, the detection rate was 770.The two resistant genes detected by the isolated strains were consistent with the phenotype of drug resistance.The integrase gene int1 of class I integron was detected, and the detection rate was 77. It indicated that integron played an important role in the transmission mechanism of drug resistance of Bacillus pipii. 20 strains of integron detected multidrug resistance gene marC.The detection rate was 91%, because the detection rate of marC gene was high.It is speculated that resistance of Bacillus bovis is related to the active efflux system mediated by chromosome mutation. A transposon genetic marker gene merA has been detected in 9 strains. The detection rate is 41% and the result of reverse PCR shows that the transmission of TEM resistance gene is transmissible.There is a correlation between the transposons.The plasmid extraction and elimination experiments were carried out on 22 strains of Bordetella rabbit isolated in this study. The results showed that all the 22 isolates carried plasmids. After the elimination of plasmids, some drug resistance characters of bacteria were changed, and ampicillin and cefazolin were detected.The sensitivity of tetracycline and polymyxin B was enhanced.It was proved that plasmid mediated partial drug resistance of the isolated strains.The results showed that Bacillus vulgaris was the main pathogen causing respiratory tract diseases in rabbits. The drug resistance rate of isolates was high and showed multidrug resistance. The detected resistant genes were consistent with the phenotype of drug resistance; plasmids, integrons, plasmids, integrons, plasmids, integrons, plasmids, integrons, plasmids, integrons,The detection of transposon these three transfer factors showed that there were many mechanisms of drug resistance transmission in B. rabbit.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S852.61

【參考文獻】

相關(guān)期刊論文 前10條

1 劉玉芹;楊彩然;賈青輝;董淑珍;李蓉;鄭新宇;;豬源耐藥大腸桿菌質(zhì)粒及酶切圖譜分析[J];黑龍江畜牧獸醫(yī);2012年23期

2 張國勝;張克文;鹿明廣;;兔波氏桿菌病[J];中國畜禽種業(yè);2012年05期

3 劉燕;韋強;肖琛聞;鮑國連;季權(quán)安;龐安娜;;兔巴氏桿菌和波氏桿菌多重PCR檢測方法的建立[J];中國獸醫(yī)學(xué)報;2012年02期

4 陳鵬舉;張紅霞;魏曉莉;向忠菊;張光輝;;豬源支氣管敗血波氏桿菌的分離鑒定及藥敏試驗[J];中國畜牧獸醫(yī);2011年11期

5 王春梅;何啟蓋;操繼躍;;細菌多重耐藥泵的研究進展[J];畜牧獸醫(yī)學(xué)報;2011年04期

6 羅致;;淺談抗菌藥物研究進展[J];求醫(yī)問藥(下半月);2011年01期

7 左艷君;魏軍;賈偉;李剛;趙志軍;;血液中大腸埃希菌超廣譜β-內(nèi)酰胺酶耐藥基因型研究[J];寧夏醫(yī)科大學(xué)學(xué)報;2009年06期

8 耿光瑞;趙月平;李寸欣;趙文雙;;抗菌藥物的合理應(yīng)用研究進展[J];動物醫(yī)學(xué)進展;2009年08期

9 虞春華;李瓊;黃瑞;;多重耐藥傷寒沙門菌耐藥基因的研究[J];微生物與感染;2008年02期

10 楊詠潔;魯承;趙達卓;金美伶;;豬支氣管敗血波氏桿菌PCR診斷方法的建立及應(yīng)用[J];中國獸醫(yī)科學(xué);2006年01期

相關(guān)碩士學(xué)位論文 前2條

1 李科;兔波氏桿菌和巴氏桿菌LAMP快速診斷技術(shù)研究[D];浙江師范大學(xué);2013年

2 章春桃;兔支氣管敗血波氏桿菌fimN基因的克隆表達及快速檢測方法的建立[D];浙江師范大學(xué);2011年

，

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