山东省不同动物源沙门菌生物学特性的比较与分析

发布时间：2018-04-04 19:38

  本文选题：多位点序列分型　切入点：沙门菌　出处：《山东农业大学》2016年硕士论文

【摘要】：沙门菌属是肠杆菌科中的一个大属,血清型繁多,是危害人类与畜禽的一类重要病原菌。近年来,沙门菌污染畜禽产品的问题已引起养殖人员及公共卫生界的极大重视。近年来随着抗生素的使用甚至滥用,使得沙门菌的耐药性越来越严重,对养殖业构成极大的威胁。本试验先后从山东省不同地市采集不同动物源的样品,通过选择培养基、革兰氏染色和29,.分离沙门菌,并做生化试验查看其生化特性。通过玻片凝集试验,确定分离株的血清型。通过微量肉汤稀释法对分离的沙门菌进行多种药物的药敏试验,来检验其耐药性,同时检测分离菌株的2:种耐药基因的存在情况。测定沙门菌常见的四种毒力基因,了解分离沙门菌毒力基因的变异特点。通过多位点序列分型技术对分离株进行分型,了解沙门菌的基因序列型。通过鉴定总共分离获得了:;株鸡源沙门菌、89株鸭源沙门菌和=株猪源的沙门菌。不同动物源沙门菌血清型实验结果为:鸡源检出6种血清型,其中血清型为肠炎沙门菌最多,占88.5%;鼠伤寒沙门菌占1.3%;山夫登堡沙门菌占1.3%;印第安纳沙门菌占5.1%;汤卜逊沙门菌占2.6%;阿格玛沙门菌占1.3%。鸭源检出2种血清型,其中血清型为肠炎沙门菌最多,占67.9%;鼠伤寒沙门菌占32.1%。猪源共检测出3种血清型,其中血清型为鼠伤寒沙门菌最多,占65%;肠炎沙门菌占15%;德贝儿沙门菌占20%。通过MLST技术,鸡源检出7种ST型:ST11、ST19、ST26、ST128、ST14、ST17和New1,鸭源检出3种ST型:ST11、ST19和New2,猪源检出3种:ST34、ST40和ST3007,整体来看,分离菌的血清型和ST型数量比较接近,二者分型能力相近。通过药敏试验,鸡源菌株中,氨苄西林和多粘菌素的耐药率最高,达到了;=以上,对环丙沙星和头孢噻呋的耐药率也达到了8"=和8:":。鸭源菌株中,对环丙沙星和多粘菌素耐药率为2==,恩诺沙星为;3",对多西环素、庆大霉素和新霉素耐药率高于9=。猪源菌株中,对多西环素的耐药率最高为8,其次为氨苄西林为;=和氟苯尼考为48,在统计学分析中发现,不同动物源之间的耐药率差异显著现象普遍存在。耐药基因中的*和*%在三种动物源中检测中均高于:=,其次是四环素类的耐药基因,在鸭源菌中为4",在其他两种动物源中均高于8=,β'内酰胺类的耐药基因!)的检出率在三种动物源中均高于3=,此外?、53@7'?、%、*和*基因在三种动物源中的检出率为零,但三种动物源耐药基因检出率差异显著的情况比较少。毒力基因检测发现四种毒力基因同源率很高,但部分存在碱基突变,猪源均未发生变异。/基因:鸡源和鸭源主要有两个位点发生变异,但在氨基酸水平上并没有发生变异。基因:仅鸡源在两个位点发生变异,氨基酸比对发现,在碱基2位的变异导致了氨基酸改变。!基因:仅鸡源发生了四处碱基位点变异,突变均在开放阅读框内,并没有发生氨基酸水平上的变异。/基因序列没有发现碱基变异。本实验从多个层面分析了不同动物源的沙门菌的特性,从而使得我们对山东省部分地区不同动物源沙门菌有了初步认识,有助于更好地防治该菌引起的感染,从而有针对性的做出预防对策。
[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，

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