本文選題：雞　切入點：致病性沙門氏菌　出處：《廣西大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：沙門氏菌具有廣泛的宿主,是一種重要的人畜共患病的病原菌。近年來,抗菌藥物在臨床上的普遍使用甚至濫用,導(dǎo)致沙門氏菌的耐藥問題越來越嚴重。沙門氏菌能夠入侵宿主體內(nèi)增殖和致病,與其毒力基因密切相關(guān)。本研究對雞源致病性沙門氏菌進行分離鑒定,對其耐藥性、致病性進行測定,對其耐藥基因、毒力基因進行檢測,分析耐藥基因與耐藥表型、毒力基因與致病性的關(guān)系,為沙門氏菌病的防治提供科學(xué)依據(jù)。采集疑似病雞的組織病料,通過SC增菌液對沙門氏菌進行增菌培養(yǎng)。用XLD瓊脂平板對沙門氏菌進行分離培養(yǎng)。選取純培養(yǎng)物的單個菌落,分別進行生化鑒定、血清學(xué)鑒定和PCR鑒定等,共獲得55株沙門氏菌分離株。這些菌株包含4個血清群,A群6株,B群23株,C群1株,D群25株。其中D群約占45%,為沙門氏菌優(yōu)勢血清群。采用CSLI推薦的K-B瓊脂法測定55株沙門氏菌分離株對18種抗菌藥物的敏感性。結(jié)果顯示,55株沙門氏菌分離株對利福平、阿莫西林、青霉素G、苯唑西林以及頭孢噻吩的耐藥率達到100%;對紅霉素、林可霉素、多粘菌素B、復(fù)方新諾明及呋喃妥因耐藥率達到90%以上;對強力霉素、鏈霉素及諾氟沙星的耐藥率在50%～90%之間:對慶大霉素及大觀霉素的耐藥率在10%以下,分別為7.27%、3.64%。利用PCR方法檢測55株沙門氏菌分離株的16種耐藥基因blaTEM、blaPSE、blacMY、aadA1、Aph(3)-IIa、aadA2、tetB、tetG、tetM、tetX、Sul1、Su13、qnrA、qnrB、qepA、qnrS。結(jié)果顯示,55株沙門氏菌分離株中,含有3種耐藥基因的菌株比例為25.45%,含有5種耐藥基因的菌株比例為21.82%,含有2種耐藥基因的菌株比例為21.82%,含有4種耐藥基因的菌株比例為20.00%,含有1種耐藥基因和7種耐藥基因的菌株比例分別為3.64%和1.82%。在所檢測耐藥基因中,tetX的檢出率最高,達到78.18%,而blaPSE、Aph(3)-Ⅱa、aadA2、qnrA、qnrB則未被檢出。利用PCR方法檢測55株沙門氏菌分離株的31種毒力基因(質(zhì)粒毒力基因spvR、spvA、spvB、spvC、spvD;毒力島基因invH、sipA、sipB、sopA、sopD、avrA、hilA、iacP、prgK、ssaB、ssaQ、sifA、sseL、ssrA、ttrB、mgtC、misL、rmbA、rhuM、sugR、orf319、siiD(spi4D)、siiE、spi4H、sopB、pipC)。結(jié)果顯示,質(zhì)粒毒力基因的檢出率均在60%以上;毒力島基因ttrB和orf319檢出率最高(92.73%),而ssaQ以及sifA檢出率均在10%以下。通過小鼠致病性試驗檢測55株沙門氏菌分離株的致病性。結(jié)果顯示,所有分離株對小鼠均具有致病性,但致死的時間和劑量有所差異。隨機選取其中10株進行測定,其半數(shù)致死量(LD50)介于3.18×108cfu/mL和4×107cfu/mL之間。以上研究結(jié)果表明,本研究分離鑒定獲得的55株雞源致病性沙門氏菌分離株中,D群血清型比例約為45%,為優(yōu)勢血清群;沙門氏菌普遍存在耐藥性,多重耐藥現(xiàn)象嚴重;分離菌株對小鼠具有致病性;其耐藥表型與耐藥基因、致病性與毒力基因之間均呈現(xiàn)正相關(guān)。
[Abstract]:Salmonella has a wide range of hosts and is an important pathogen of zoonosis. In recent years, antimicrobial agents have been widely used and even abused in clinic. The drug resistance of salmonella is becoming more and more serious. Salmonella can invade the host body to proliferate and cause disease, which is closely related to its virulence gene. The pathogenicity was determined, drug resistance genes and virulence genes were detected, and the relationship between drug resistance genes and phenotypes, virulence genes and pathogenicity were analyzed, which provided scientific basis for the prevention and treatment of salmonella disease. Salmonella was cultured in SC medium. Salmonella was isolated and cultured with XLD Agar plate. Single colony of pure culture was selected for biochemical identification, serological identification and PCR identification. A total of 55 Salmonella isolates were obtained, including 4 serogroup A, 6 strains, 23 strains of group B, 1 strain of group C, 25 strains of group D. among them, group D is about 45%, which is the predominant serotype of salmonella. K-B Agar method recommended by CSLI was used to determine the results. The sensitivity of 55 Salmonella isolates to 18 antimicrobial agents was determined. The results showed that 55 Salmonella isolates were resistant to rifampicin. The resistance rates of amoxicillin, penicillin G, oxacillin and cefthiophene to 100 kum; to erythromycin, lincomycin, polymyxin B, compound sulfamide and furantoin were over 90%; to doxycycline, to doxycycline, to doxycycline, to doxycycline, to doxycycline, to doxycycline, to doxycycline. The resistance rates of streptomycin and norfloxacin were between 50% and 90%. The resistance rates to gentamicin and spectinomycin were below 10%. 16 resistant genes of 55 Salmonella isolates were detected by PCR method. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 strains of Salmonella spp. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 strains of Salmonella spp. The results showed that there were 55 strains of Salmonella spp. The results showed that there were 55 isolates of Salmonella spp. The results showed that there were 55 isolates of Salmonella. The proportion of strains containing three resistant genes was 25.45um, the proportion of strains containing five resistant genes was 21.822am, the proportion of strains containing two resistant genes was 21.822um, the proportion of strains containing four resistant genes was 20.00, the proportion of strains containing one drug resistance gene and seven drug resistance genes was 20.00. The proportion of resistant genes was 3.64% and 1.82, respectively. The detection rate of tetX was the highest among the tested resistant genes. The virulence genes of 55 Salmonella isolates were detected by PCR method. 31 virulence genes (plasmid spvRspvspvAspvBspvCspvDand virulence island gene invhophisiasopAsopAsopDavrAhila) were detected by PCR method, and the results showed that the virulence was higher than 60%. The results showed that the virulence was higher than that of the other three strains of Salmonella sphaesii. The results showed that the virulence of 31 virulence genes of 55 Salmonella isolates were detected by PCR method. The detection rate of the virulence genes of plasmid spvrrsspvspvspvCspvcspvDspvDspvDV DspvDvDvDvDvDspvDvDvDvDspvDvDv of the virulent island gene invHopsiaopsis Asopa Asopa DavrAhila was detected, and the results showed that the virulence gene was more than 60%. The results showed that the virulence was more than 60%. The results showed that the virulence was higher than that of the PSEP. The detection rate of the virulence gene was higher than that of the control group. The detection rate of 31 virulence genes of 55 Salmonella isolates was determined by PCR method. The detection rate of virulence island gene ttrB and orf319 was the highest, while the detection rate of ssaQ and sifA was below 10%. The pathogenicity of 55 Salmonella isolates was detected by mouse pathogenicity test. The results showed that all isolates were pathogenicity to mice. But the lethal time and dose were different. The LD50 of 10 strains was between 3.18 脳 10 8 cfur / mL and 4 脳 10 7 cfur / mL. The results showed that the LD50 was between 3.18 脳 10 8 cfur / mL and 4 脳 10 7 cfur / mL. In this study, 55 strains of chicken pathogenic Salmonella were isolated and identified, the proportion of serotypes of group D was about 455.It was the dominant serotype of Salmonella, the drug resistance of Salmonella was common, the phenomenon of multidrug resistance was serious, the isolated strains were pathogenicity to mice. There was a positive correlation between drug resistance phenotype and drug resistance gene, pathogenicity and virulence gene.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S852.61

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