發(fā)布時(shí)間：2018-03-23 00:27

  本文選題：禽沙門菌　切入點(diǎn)：抗體檢測(cè)　出處：《揚(yáng)州大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：沙門菌病(Salmonellosis)是不同血清型沙門菌感染人類和動(dòng)物所引起的疾病的總稱。由沙門菌引起的食物中毒在世界各國的細(xì)菌性食物中毒中常位居前列,其導(dǎo)致的畜禽生產(chǎn)能力的降低更對(duì)經(jīng)濟(jì)造成了嚴(yán)重影響。不僅嚴(yán)重阻礙了畜禽業(yè)的發(fā)展,而且威脅人類健康,己經(jīng)成為公共衛(wèi)生重點(diǎn)關(guān)注的問題。本研究主要調(diào)查了江蘇省海安縣部分雞場(chǎng)和揚(yáng)州大學(xué)禽病門診臨床沙門菌污染狀況,對(duì)血液樣品樣進(jìn)行抗體檢測(cè),分離臨床肝臟樣品,規(guī)模場(chǎng)活禽泄殖腔棉拭子、雞蛋和環(huán)境樣品中的沙門菌及血清型鑒定,并對(duì)分離到的沙門菌進(jìn)行了抗生素敏感性測(cè)定、分子分型和毒力基因檢測(cè),為制定防控措施提供了重要參考。2014-2015年間,對(duì)江蘇省海安縣4個(gè)不同飼養(yǎng)模式雞場(chǎng)分別進(jìn)行4次血樣采集,采集血樣共1600份,進(jìn)行血清玻板凝集試驗(yàn),檢測(cè)得到陽性血清共165份,陽性率為10.3%。其中,開放飼養(yǎng)模式雞場(chǎng)陽性率為22%,明顯高于其他飼養(yǎng)模式(半封閉飼養(yǎng)模式7.4%,全封閉飼養(yǎng)模式4.5%)；夏季血樣檢測(cè)到的陽性率為20.3%,明顯高于其他季度(冬季8.3%,春季7.5%,秋季5.3%)。共采集泄殖腔棉拭子1600份,分離出沙門菌12株,陽性率為0.75%。其中,開放飼養(yǎng)模式分離率為1.5%(6/400),高于半封閉飼養(yǎng)模式0.63%(5/800),高于全封閉飼養(yǎng)模式0.25%(1/400)。采集的雞蛋、飼料和環(huán)境樣品中未分離到沙門菌。從揚(yáng)州大學(xué)禽病門診共采集臨床樣品161份,分離獲得18株沙門菌,陽性率為11.2%。揚(yáng)州地區(qū)沙門菌分離率最高,為13.6%(11/81)；其次是鎮(zhèn)江和萊安,均為12.5%；淮安、南通、南京依次為8.7%、7.4%、7.1%。對(duì)分離到的30株沙門菌進(jìn)行血清學(xué)試驗(yàn)和藥物敏感性檢測(cè)。血清學(xué)試驗(yàn)結(jié)果顯示,其中20株為1,9,12：-:-(雞沙門菌或雛沙門菌)、5株為1,4,[5],12：i：1,2(鼠傷寒沙門菌)、3株為1,4,[5],12：b：1,2(乙型副傷寒沙門菌)、2株為1,9,12：g,m：[1,7](腸炎沙門菌)。12種抗菌藥物的敏感性試驗(yàn)結(jié)果顯示,所有分離株對(duì)丁胺卡那霉素、恩諾沙星、環(huán)丙沙星、氨曲南、卡那霉素均高度敏感(100%)：對(duì)四環(huán)素耐藥率最高,為40%；對(duì)其余抗生素均有不同程度的耐藥。有18株沙門菌至少對(duì)2種抗菌藥物耐藥。不同地區(qū)沙門菌耐藥率有所不同,南通地區(qū)沙門菌多重耐藥情況最為嚴(yán)重,耐4種及以上抗菌藥物的菌株比例達(dá)16.7%。不同血清型的沙門菌耐藥情況也有一定的差異,雞沙門菌或雛沙門菌耐藥率最高,鼠傷寒沙門菌次之,腸炎沙門菌耐藥率最低。為了進(jìn)一步了解分離株的遺傳特性,選取沙門菌7個(gè)管家基因aroC、dnaN、hemD、 hiaC、purE、sucA和thrA,對(duì)30株分離株和26株模板進(jìn)行MLST試驗(yàn),共獲得8個(gè)不同的ST型,分別為ST11、ST19、ST86、ST92、ST99、ST1690、ST2039和ST2151,其中ST2151為新發(fā)現(xiàn)的ST型。運(yùn)用eBURST和MEGA軟件進(jìn)行聚類分析,研究其進(jìn)化的相關(guān)性。選取沙門菌主要毒力島代表性基因,包括sitC (SPI-1)、invA (SPI-1)、spiC (SPI-2)、 sifA (SPI-2)、misL (SPI-3)、orfL (SPI-4)、pipD (SPI-5),及其他重要結(jié)構(gòu)性毒力基因(pefA、 msgA和lpfC)進(jìn)行檢測(cè)。其中,sitC、spiC、orfL、pipD陽性檢出率為100%,iroN、 msgA、lpfC以及sifA作為毒力島保守基因檢出率也很高,分別為92.5%、95%、97.5%、97.5%,檢出率最低的是misL基因,僅為45%。
[Abstract]:Salmonella disease (Salmonellosis) is a general term for different serotypes of Salmonella infection in human and animal disease caused by Salmonella. Food poisoning caused by many countries in the world of bacterial food poisoning often forefront, decreasing the number of livestock and poultry production capacity of more economic producedserious effect. Not only seriously hindered the development of animal husbandry moreover, a threat to human health, has become a public health concern. This study mainly investigates the part of Haian County of Jiangsu province and the Yangzhou University poultry farms outpatient clinical Salmonella contamination, antibody detection of blood sample and liver samples from clinical field, the scale of the live poultry cloacal swab, and Salmonella serotype identification egg and in environmental samples, and Salmonella were antibiotic sensitivity determination, molecular detection and virulence genes, set the prevention and control of system Provides an important reference for.2014-2015 years, in Haian County of Jiangsu Province, 4 farms of different feeding patterns were conducted 4 times blood collection, blood samples were collected 1600 samples, serum agglutination test, positive serum was detected by a total of 165, the positive rate was 10.3%. the open breeding mode of chicken was significantly higher than that of the positive rate was 22%. Other feeding mode (semi closed feeding mode 7.4%, closed feeding mode 4.5%); the positive rate of blood samples is detected by the summer of 20.3%, significantly higher than in other seasons (winter 8.3%, spring 7.5%, fall 5.3%) were collected. The cloacal swab cavity 1600, isolated 12 strains of Salmonella, the positive rate was 0.75%. the separation rate of feeding mode, open 1.5% (6/400), higher than the semi enclosed breeding mode 0.63% (5/800), higher than the full closed feeding mode 0.25% (1/400). The egg collection, feed and environmental samples were not isolated from avian salmonella. Yangzhou University The clinical disease clinic were collected 161 samples, isolated 18 strains of Salmonella, the positive rate of Salmonella 11.2%. isolated in Yangzhou was the highest, was 13.6% (11/81); followed by Zhenjiang and Ryan, were 12.5%; Huaian, Nantong and Nanjing were 8.7%, 7.4%, 7.1%. of 30 isolated strains of Salmonella by serological detection test and drug sensitivity. The results of serological tests showed that 20 of them were 1,9,12:-: - (Salmonella or Salmonella pullora), 5 strains were 1,4, [5], 12:i:1,2 (Salmonella typhimurium), 3 strains were 1,4, [5], 12:b:1,2 (paratyphoid salmonella), 2 strains were 1,9,12:g, m:[1,7] (enteritis Salmonella).12 antimicrobial susceptibility test showed that all isolates were susceptible to amikacin, enrofloxacin, ciprofloxacin, aztreonam, kanamycin was highly sensitive (100%): the tetracycline resistance rate was 40%; for the rest, antibiotics are not The same resistance degree. There are 18 strains of Salmonella at least 2 kinds of antimicrobial resistance. The resistance rates vary in different regions of Salmonella, Salmonella multi drug resistance in Nantong area is the most serious, resistant to 4 or more antimicrobials proportionreaches 16.7%. different serotypes of Salmonella resistant conditions also have some differences, or chicken Salmonella Salmonella pullora the highest resistant rate of Salmonella typhimurium and Salmonella enteritidis resistance rate is the lowest. In order to further understand the genetic characteristics of Salmonella isolates, selected 7 housekeeping genes aroC, dnaN, hemD, hiaC, purE, sucA and thrA, MLST test of 30 isolates and 26 strains of template, obtained a total of 8 different type ST, respectively ST11, ST19, ST86, ST92, ST99, ST1690, ST2039 and ST2151, wherein ST2151 is ST newly discovered. Cluster analysis was performed using eBURST and MEGA software, the correlation of evolution. The main selection of Salmonella Pathogenicity island representative genes, including sitC (SPI-1), invA (SPI-1), spiC (SPI-2), sifA (SPI-2), misL (SPI-3), orfL (SPI-4), pipD (SPI-5), and other important structural virulence genes (pefA, msgA and lpfC) were detected. Among them, sitC, spiC. OrfL, the positive rate of pipD was 100%, iroN, msgA, lpfC and sifA as pathogenicity island gene conservative detection rate is very high, respectively 92.5%, 95%, 97.5%, 97.5%, the lowest detection rate is misL gene, only 45%.

【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S852.61

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