本文選題：牛支原體 + 氟喹諾酮類抗生素��； 參考：《吉林農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：牛支原體(Mycoplasma bovis,M.bovis)是引起牛呼吸系統(tǒng)疾病的(Bovine respiratory disease,BRD)主要病原之一,該病原引發(fā)的疾病主要采用大環(huán)內(nèi)酯類、氟喹諾酮類、氨基糖苷類抗生素進(jìn)行治療,但隨著抗生素在獸醫(yī)臨床上的廣泛使用,M.bovis對上述藥物逐漸產(chǎn)生了耐藥性。近年來,M.bovis對其主要治療藥物的耐藥性所引起的抗感染失敗給我國畜牧養(yǎng)殖業(yè)造成了較嚴(yán)重的經(jīng)濟(jì)損失。本研究采用改良微量稀釋法檢測臨床分離的32株M.bovis對氟喹諾酮及氨基糖苷類抗生素的最低抑菌濃度(Minimal inhibitory concentration,MIC),結(jié)果表明,部分M.bovis已對氟喹諾酮類抗生素產(chǎn)生了耐藥性(其MIC為4或8μg/mL),分離的耐藥菌株占此次臨床分離M.bovis的19%;臨床分離的32株M.bovis均對氨基糖苷類抗生素敏感或中度耐藥,可能存在耐藥風(fēng)險(xiǎn)。分別成功誘導(dǎo)了對氟喹諾酮類抗生素高度耐藥的M.bovis和對氨基糖苷類抗生素高度耐藥的M.bovis,其中,經(jīng)體外誘導(dǎo)獲得的對一種氟喹諾酮類抗生素高度耐藥的M.bovis對其它兩種受試氟喹諾酮類抗生素均存在交叉耐藥現(xiàn)象,經(jīng)體外誘導(dǎo)獲得的對一種氨基糖苷類抗生素高度耐藥的M.bovis對其它兩種受試氨基糖苷類抗生素存在部分交叉耐藥現(xiàn)象。在此基礎(chǔ)上,本研究對臨床分離的M.bovis氟喹諾酮類抗生素敏感株、耐藥株及體外誘導(dǎo)獲得的高度耐藥株進(jìn)行氟喹諾酮類抗生素靶位突變分析,結(jié)果表明,臨床分離的耐藥菌株其氟喹諾酮類抗生素耐藥決定區(qū)(Quinolone resistance determining region,QRDR)均存在gyrA(Ser83Phe)或parC(Ser80Ile)的氨基酸突變;體外誘導(dǎo)的高度耐藥株中QRDR突變類型則以gyrA(Ser83Phe/Tyr或Glu87Lys)和parC(Ser80Ile或Asp84Asn/Tyr)的氨基酸發(fā)生突變?yōu)橹?耐藥性逆轉(zhuǎn)試驗(yàn)結(jié)果表明,1/4 MIC外排泵抑制劑羰基氰氯苯腙(carbonyl cyanide m-chlorophenyl hydrazone,CCCP)和維拉帕米(verapamil,VP)作用前后受試菌對環(huán)丙沙星、恩諾沙星、諾氟沙星的MIC值未發(fā)生變化,推斷M.bovis中不存在過量表達(dá)的以氟喹諾酮類抗生素為底物的主動外排系統(tǒng)。本研究對臨床分離的M.bovis氨基糖苷類抗生素敏感株、中度耐藥株及體外誘導(dǎo)獲得的高度耐藥株進(jìn)行氨基糖苷類抗生素靶位突變分析,結(jié)果表明,臨床分離的氨基糖苷類抗生素敏感株及中度耐藥株在氨基糖苷類抗生素耐藥決定區(qū)(Aminoglycosides resistance determining region,ARDR)均不存在堿基突變;經(jīng)體外誘導(dǎo)獲得的對慶大霉素、卡那霉素、大觀霉素高度耐藥的M.bovis在其16S rRNA(rrs3和rrs4)的ARDR中分別存在A1409T、A1408G、C1192A的堿基突變,但上述經(jīng)體外誘導(dǎo)獲得的氨基糖苷類抗生素高度耐藥株在編碼核糖體蛋白S5的rpsE基因上未檢測到有意義的堿基突變;對臨床分離的32株M.bovis進(jìn)行16S rRNA甲基化酶基因及氨基糖苷類抗生素耐藥基因檢測,結(jié)果表明,臨床分離的M.bovis均攜帶aphA1、strB磷酸轉(zhuǎn)移酶基因及aacA4乙酰轉(zhuǎn)移酶基因。此外,耐藥性逆轉(zhuǎn)試驗(yàn)結(jié)果表明,1/4 MIC外排泵抑制劑CCCP和VP作用前后受試菌對慶大霉素、卡那霉素、大觀霉素的MIC值亦未發(fā)生變化,推斷M.bovis中不存在過量表達(dá)的以氨基糖苷類抗生素為底物的主動外排系統(tǒng)。
[Abstract]:Mycoplasma bovis (M.bovis) is one of the main pathogens causing Bovine respiratory disease (BRD). The disease caused by this pathogen is mainly treated with macrolide, fluoroquinolone, aminoglycoside antibiotics, but with the extensive use of antibiotics in veterinary clinic, M.bovis against the above drugs In recent years, the anti infection failure caused by M.bovis's resistance to the main therapeutic drugs has caused serious economic loss to the livestock breeding industry in China. In this study, the minimal inhibitory concentration of 32 clinical isolates of M.bovis against fluoroquinolone and aminoglycoside antibiotics was detected by modified microdilution method. Minimal inhibitory concentration, MIC), the results showed that partial M.bovis had produced resistance to fluoroquinolone antibiotics (its MIC was 4 or 8 mu g/mL), and the isolated drug resistant strains accounted for 19% of the clinically isolated M.bovis; 32 strains of clinical isolates were both sensitive to aminoglycoside antibiotics or moderately resistant, and may have the risk of resistance. M.bovis and M.bovis, highly resistant to fluoroquinolones, were not successfully induced, in which the highly resistant M.bovis of a fluoroquinolone antibiotic induced in vitro was cross resistant to two other fluoroquinolones, and was induced in vitro. M.bovis, which is highly resistant to an aminoglycoside antibiotic, is partially cross resistant to the other two kinds of aminoglycoside antibiotics. On the basis of this, the clinical isolates of M.bovis fluoroquinolone sensitive strains, drug-resistant strains and in vitro induced highly resistant strains were carried out with fluoroquinolone antibiotics. The results of target mutation analysis showed that the Quinolone resistance determining region (QRDR) in the drug-resistant isolates of the clinical isolates had the amino acid mutations of gyrA (Ser83Phe) or parC (Ser80Ile), and the QRDR mutation types of the highly resistant strains in vitro were gyrA. The amino acid mutation of er80Ile or Asp84Asn/Tyr was dominant, and the resistance reversal test showed that the MIC values of ciprofloxacin, enrofloxacin and norfloxacin were not changed before and after the effect of the carbonyl cyano-chlorobenzene hydrazone (carbonyl cyanide m-chlorophenyl hydrazone, CCCP) and Vera Pammy (verapamil, VP) on the pump inhibitor of 1/4 MIC. There was no excessive expression of the active efflux system with fluoroquinolone as the substrate. In this study, a clinically isolated M.bovis aminoglycoside antibiotic sensitive strain, moderately resistant strain and highly resistant strains in vitro were analyzed for the aminoglycoside antibiotic target mutation analysis. The results showed that the clinical isolates were separated from ammonia. There was no base mutation in the aminoglycoside antibiotic resistant determinant region (Aminoglycosides resistance determining region, ARDR) in the amino glycoside antibiotic sensitive and moderately resistant strains, and the highly resistant M.bovis of gentamicin, kanamycin and macromycin in the ARDR of 16S rRNA (rrs3 and rrs4) in vitro, respectively, respectively. There was a base mutation in A1409T, A1408G, and C1192A, but the above - mentioned aminoglycoside resistant highly resistant strains of the above - induced aminoglycoside resistant strains did not detect significant base mutations in the rpsE gene encoding ribosomal protein S5, and 32 strains of clinical isolates of M.bovis were detected by 16S rRNA methylase gene and aminoglycoside antibiotic resistance gene. The results showed that the clinical isolates of M.bovis all carried aphA1, strB phosphotransferase gene and aacA4 acetyltransferase gene. In addition, the drug resistance reversal test showed that the MIC values of the tested bacteria to gentamicin, kanamycin and mycophenycin before and after the action of CCCP and VP of the 1/4 MIC pump inhibitor were also not changed. It was inferred that there was no existence in M.bovis. Over expression of the active efflux system based on aminoglycoside antibiotics.

【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S852.62

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 賈博巖;趙立峰;高鐸;孔令聰;劉樹明;王春鳳;馬紅霞;;牛支原體氟喹諾酮類藥物耐藥靶位突變分析[J];中國獸藥雜志;2015年12期

2 王梓;孔令聰;賈博巖;劉樹明;馬紅霞;;氨基糖苷類抗生素耐藥機(jī)制研究進(jìn)展[J];中國獸藥雜志;2015年03期

3 高鐸;孔令聰;王梓;馬紅霞;;牛支原體的分離鑒定及其體外藥物敏感性分析[J];中國獸藥雜志;2014年03期

4 孔令聰;張春艷;高云航;馬紅霞;;牛支原體耐藥性研究進(jìn)展[J];中國獸藥雜志;2013年09期

5 張利;李玉平;黎曉敏;尼瑪倉木拉;;牛支原體藥物敏感性試驗(yàn)[J];動物醫(yī)學(xué)進(jìn)展;2012年02期

6 劉軼秋;吳聰明;沈建忠;李蓓蓓;張瑞婷;趙暉;;3種禽源支原體替米考星耐藥株的體外誘導(dǎo)及23S rRNA基因V域堿基突變分析[J];畜牧獸醫(yī)學(xué)報(bào);2011年07期

7 胡長敏;石磊;龔瑞;白智迪;郭愛珍;;牛支原體病研究進(jìn)展[J];動物醫(yī)學(xué)進(jìn)展;2009年08期

8 葉萍;鄧超干;王輝;王曉川;;體外誘導(dǎo)人型支原體對氟喹諾酮類藥物耐藥的實(shí)驗(yàn)研究[J];重慶醫(yī)學(xué);2009年06期

9 辛九慶;李媛;郭丹;宋念華;胡守萍;陳超;裴潔;曹培麗;;國內(nèi)首次從患肺炎的犢牛肺臟中分離到牛支原體[J];中國預(yù)防獸醫(yī)學(xué)報(bào);2008年09期

10 石磊;龔瑞;尹爭艷;周勇;裴潔;胡智斌;王利霞;胡長敏;劉濤;陳穎鈺;廖娟紅;趙俊龍;陳煥春;郭愛珍;;肉牛傳染性牛支原體肺炎流行的初步診斷[J];華中農(nóng)業(yè)大學(xué)學(xué)報(bào);2008年04期

相關(guān)博士學(xué)位論文 前1條

1 姜曉冰;大腸桿菌質(zhì)粒介導(dǎo)喹諾酮耐藥機(jī)制研究[D];華南理工大學(xué);2013年

相關(guān)碩士學(xué)位論文 前3條

1 高鐸;牛支原體的分離鑒定及對大環(huán)內(nèi)酯類抗生素耐藥機(jī)制研究[D];吉林農(nóng)業(yè)大學(xué);2015年

2 孔令聰;牛源莢膜血清A型多殺性巴氏桿菌的分離鑒定及對喹諾酮類抗生素耐藥機(jī)制研究[D];吉林農(nóng)業(yè)大學(xué);2013年

3 李琳琳;銅綠假單胞菌氨基糖苷類抗生素耐藥機(jī)制的研究[D];吉林大學(xué);2011年

，

本文編號：1807830