本文選題：多位點(diǎn)序列分型　切入點(diǎn)：沙門菌　出處：《山東農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：沙門菌屬是腸桿菌科中的一個(gè)大屬,血清型繁多,是危害人類與畜禽的一類重要病原菌。近年來,沙門菌污染畜禽產(chǎn)品的問題已引起養(yǎng)殖人員及公共衛(wèi)生界的極大重視。近年來隨著抗生素的使用甚至濫用,使得沙門菌的耐藥性越來越嚴(yán)重,對養(yǎng)殖業(yè)構(gòu)成極大的威脅。本試驗(yàn)先后從山東省不同地市采集不同動(dòng)物源的樣品,通過選擇培養(yǎng)基、革蘭氏染色和29,.分離沙門菌,并做生化試驗(yàn)查看其生化特性。通過玻片凝集試驗(yàn),確定分離株的血清型。通過微量肉湯稀釋法對分離的沙門菌進(jìn)行多種藥物的藥敏試驗(yàn),來檢驗(yàn)其耐藥性,同時(shí)檢測分離菌株的2:種耐藥基因的存在情況。測定沙門菌常見的四種毒力基因,了解分離沙門菌毒力基因的變異特點(diǎn)。通過多位點(diǎn)序列分型技術(shù)對分離株進(jìn)行分型,了解沙門菌的基因序列型。通過鑒定總共分離獲得了:;株雞源沙門菌、89株鴨源沙門菌和=株豬源的沙門菌。不同動(dòng)物源沙門菌血清型實(shí)驗(yàn)結(jié)果為:雞源檢出6種血清型,其中血清型為腸炎沙門菌最多,占88.5%;鼠傷寒沙門菌占1.3%;山夫登堡沙門菌占1.3%;印第安納沙門菌占5.1%;湯卜遜沙門菌占2.6%;阿格瑪沙門菌占1.3%。鴨源檢出2種血清型,其中血清型為腸炎沙門菌最多,占67.9%;鼠傷寒沙門菌占32.1%。豬源共檢測出3種血清型,其中血清型為鼠傷寒沙門菌最多,占65%;腸炎沙門菌占15%;德貝兒沙門菌占20%。通過MLST技術(shù),雞源檢出7種ST型:ST11、ST19、ST26、ST128、ST14、ST17和New1,鴨源檢出3種ST型:ST11、ST19和New2,豬源檢出3種:ST34、ST40和ST3007,整體來看,分離菌的血清型和ST型數(shù)量比較接近,二者分型能力相近。通過藥敏試驗(yàn),雞源菌株中,氨芐西林和多粘菌素的耐藥率最高,達(dá)到了;=以上,對環(huán)丙沙星和頭孢噻呋的耐藥率也達(dá)到了8"=和8:":。鴨源菌株中,對環(huán)丙沙星和多粘菌素耐藥率為2==,恩諾沙星為;3",對多西環(huán)素、慶大霉素和新霉素耐藥率高于9=。豬源菌株中,對多西環(huán)素的耐藥率最高為8,其次為氨芐西林為;=和氟苯尼考為48,在統(tǒng)計(jì)學(xué)分析中發(fā)現(xiàn),不同動(dòng)物源之間的耐藥率差異顯著現(xiàn)象普遍存在。耐藥基因中的*和*%在三種動(dòng)物源中檢測中均高于:=,其次是四環(huán)素類的耐藥基因,在鴨源菌中為4",在其他兩種動(dòng)物源中均高于8=,β'內(nèi)酰胺類的耐藥基因!)的檢出率在三種動(dòng)物源中均高于3=,此外?、53@7'?、%、*和*基因在三種動(dòng)物源中的檢出率為零,但三種動(dòng)物源耐藥基因檢出率差異顯著的情況比較少。毒力基因檢測發(fā)現(xiàn)四種毒力基因同源率很高,但部分存在堿基突變,豬源均未發(fā)生變異。/基因:雞源和鴨源主要有兩個(gè)位點(diǎn)發(fā)生變異,但在氨基酸水平上并沒有發(fā)生變異�；�:僅雞源在兩個(gè)位點(diǎn)發(fā)生變異,氨基酸比對發(fā)現(xiàn),在堿基2位的變異導(dǎo)致了氨基酸改變。!基因:僅雞源發(fā)生了四處堿基位點(diǎn)變異,突變均在開放閱讀框內(nèi),并沒有發(fā)生氨基酸水平上的變異。/基因序列沒有發(fā)現(xiàn)堿基變異。本實(shí)驗(yàn)從多個(gè)層面分析了不同動(dòng)物源的沙門菌的特性,從而使得我們對山東省部分地區(qū)不同動(dòng)物源沙門菌有了初步認(rèn)識,有助于更好地防治該菌引起的感染,從而有針對性的做出預(yù)防對策。
[Abstract]:Salmonella is a large genus of Enterobacteriaceae in a serum type, there is a kind of important pathogenic bacteria harmful to human and livestock. In recent years, the problem of Salmonella contamination of livestock and poultry products has attracted great attention of farmers and the public health community. In recent years, with the use of antibiotics and even abuse, the increasing resistance of Salmonella the more serious, a great threat to the aquaculture industry. This experiment has from different cities of Shandong province were collected in different animal samples, medium through selection, Gram staining and 29. Separation of Salmonella, and biochemical tests to view its biochemical characteristics. By slide agglutination test, determine the serotype isolates. The trace Salmonella broth dilution method on the separation of drug sensitive test of a variety of drugs, to test the drug resistance, simultaneous detection of presence of 2: drug resistance genes in isolates of Salmonella. The determination of the common four Virulence genes, understand the variation characteristics of isolated Salmonella virulence genes. The isolates were typed by multilocus sequence typing, gene sequence type understanding. Through the identification of a total of Salmonella isolated; chicken Salmonella isolates, 89 strains of Salmonella from ducks and Salmonella strains from swine = animal results. Experimental Salmonella serotype: chickens were detected in 6 serotypes, the serotype of Salmonella enteritidis, typhimurium accounted for 88.5%; 1.3%; Salmonella entericasubsp 1.3%; Salmonella Indiana Salmonella accounted for 5.1%; 2.6%; 1.3%. for detection of duck origin Salmonella AGEMA 2 serotypes, including serum for most type of Salmonella enteritidis, typhimurium accounted for 67.9%; porcine 32.1%. were detected in 3 serotypes, the serotype of Salmonella typhimurium, enteritidis accounted for 65%; 15%; de Belle Salmonella accounted for 20%. 閫氳繃MLST鎶，

本文編號：1711381