江苏地区禽源沙门菌的临床检测及相关特性的分析

发布时间：2018-03-23 00:27

本文选题：禽沙门菌　切入点：抗体检测　出处：《扬州大学》2016年硕士论文　论文类型：学位论文

【摘要】：沙门菌病(Salmonellosis)是不同血清型沙门菌感染人类和动物所引起的疾病的总称。由沙门菌引起的食物中毒在世界各国的细菌性食物中毒中常位居前列,其导致的畜禽生产能力的降低更对经济造成了严重影响。不仅严重阻碍了畜禽业的发展,而且威胁人类健康,己经成为公共卫生重点关注的问题。本研究主要调查了江苏省海安县部分鸡场和扬州大学禽病门诊临床沙门菌污染状况,对血液样品样进行抗体检测,分离临床肝脏样品,规模场活禽泄殖腔棉拭子、鸡蛋和环境样品中的沙门菌及血清型鉴定,并对分离到的沙门菌进行了抗生素敏感性测定、分子分型和毒力基因检测,为制定防控措施提供了重要参考。2014-2015年间,对江苏省海安县4个不同饲养模式鸡场分别进行4次血样采集,采集血样共1600份,进行血清玻板凝集试验,检测得到阳性血清共165份,阳性率为10.3%。其中,开放饲养模式鸡场阳性率为22%,明显高于其他饲养模式(半封闭饲养模式7.4%,全封闭饲养模式4.5%)；夏季血样检测到的阳性率为20.3%,明显高于其他季度(冬季8.3%,春季7.5%,秋季5.3%)。共采集泄殖腔棉拭子1600份,分离出沙门菌12株,阳性率为0.75%。其中,开放饲养模式分离率为1.5%(6/400),高于半封闭饲养模式0.63%(5/800),高于全封闭饲养模式0.25%(1/400)。采集的鸡蛋、饲料和环境样品中未分离到沙门菌。从扬州大学禽病门诊共采集临床样品161份,分离获得18株沙门菌,阳性率为11.2%。扬州地区沙门菌分离率最高,为13.6%(11/81)；其次是镇江和莱安,均为12.5%；淮安、南通、南京依次为8.7%、7.4%、7.1%。对分离到的30株沙门菌进行血清学试验和药物敏感性检测。血清学试验结果显示,其中20株为1,9,12：-:-(鸡沙门菌或雏沙门菌)、5株为1,4,[5],12：i：1,2(鼠伤寒沙门菌)、3株为1,4,[5],12：b：1,2(乙型副伤寒沙门菌)、2株为1,9,12：g,m：[1,7](肠炎沙门菌)。12种抗菌药物的敏感性试验结果显示,所有分离株对丁胺卡那霉素、恩诺沙星、环丙沙星、氨曲南、卡那霉素均高度敏感(100%)：对四环素耐药率最高,为40%；对其余抗生素均有不同程度的耐药。有18株沙门菌至少对2种抗菌药物耐药。不同地区沙门菌耐药率有所不同,南通地区沙门菌多重耐药情况最为严重,耐4种及以上抗菌药物的菌株比例达16.7%。不同血清型的沙门菌耐药情况也有一定的差异,鸡沙门菌或雏沙门菌耐药率最高,鼠伤寒沙门菌次之,肠炎沙门菌耐药率最低。为了进一步了解分离株的遗传特性,选取沙门菌7个管家基因aroC、dnaN、hemD、 hiaC、purE、sucA和thrA,对30株分离株和26株模板进行MLST试验,共获得8个不同的ST型,分别为ST11、ST19、ST86、ST92、ST99、ST1690、ST2039和ST2151,其中ST2151为新发现的ST型。运用eBURST和MEGA软件进行聚类分析,研究其进化的相关性。选取沙门菌主要毒力岛代表性基因,包括sitC (SPI-1)、invA (SPI-1)、spiC (SPI-2)、 sifA (SPI-2)、misL (SPI-3)、orfL (SPI-4)、pipD (SPI-5),及其他重要结构性毒力基因(pefA、 msgA和lpfC)进行检测。其中,sitC、spiC、orfL、pipD阳性检出率为100%,iroN、 msgA、lpfC以及sifA作为毒力岛保守基因检出率也很高,分别为92.5%、95%、97.5%、97.5%,检出率最低的是misL基因,仅为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%.

【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S852.61

【相似文献】