山东省贝类弧菌流行病学调查、药敏试验及毒力基因检测

发布时间：2018-06-11 14:35

本文选题：贝类 + 弧菌　；参考：《山东农业大学》2017年硕士论文

【摘要】：近年来,我国贝类养殖种类和养殖模式越来越多样化,养殖技术逐渐成熟,养殖面积不断扩大,贝类产业已成为我国水产业的重要支柱。但是,以弧菌为主要致病菌的微生物对贝类养殖产业带来严重危害。夏季的7、8月份是贝类弧菌病的高发季节,由弧菌引起的水生动物疾病流行面积广,发病率高,对水产养殖业造成严重的经济损失,一直是水产领域研究的一大热点。本试验对山东省主要贝类养殖区的养殖贝类和野生贝类进行了弧菌的流行病学调查。通过细菌分离、生化鉴定、16s-rDNA的克隆以及序列分析比对等方法,对分离的弧菌的生物学特性进行了初步研究。掌握了贝类弧菌的流行特点及规律,为预防疾病的发生提供了依据。自潍坊寿光,威海乳山、文登,青岛胶南,日照,滨州无棣,东营河口等地区采集了不同的贝类,包括青蛤、文蛤、扇贝、椭螺、尖螺、竹蛏、织纹螺、四角蛤、牡蛎、沙蛤、蓝蛤、花蛤、毛蚶、魁蚶、扁玉螺、贻贝等。从样品中分离出27种弧菌,8种其它细菌,共222株。经测序分析比对,主要有哈氏弧菌(Vibrio harveyi),溶藻弧菌(Vibrio alginolyticus),副溶血弧菌(Vibrio parahaemolyticus)、Vibrio chagasii、Vibrio azureus、需钠弧菌(Vibrio natriegens)、创伤弧菌(Vibrio vulnificus)、杭州弧菌(Vibrio hangzhouensis)等。结果显示,哈氏弧菌、溶藻弧菌、副溶血弧菌的检出率较高,分别占20%、18%、21%;其他弧菌相对较少。胶南地区副溶血弧菌和溶藻弧菌检出率较高;无棣哈氏弧菌和溶藻弧菌检出率较高,而副溶血弧菌检测率很低;河口、日照、寿光三个地区的三种弧菌的检出率都比较高,均在15%-35%之间。结果表明,弧菌是导致贝类发病的主要病原菌之一,对青蛤、文蛤、毛蚶、竹蛏、沙蛤的危害相对严重。其中哈氏弧菌、副溶血弧菌、溶藻弧菌分离出的菌株较多;哈氏弧菌主要分离自青蛤、文蛤、竹蛏、沙蛤、毛蚶;副溶血弧菌主要分离自文蛤、竹蛏、沙蛤、花蛤、毛蚶,沙蛤最多。溶藻弧菌在青蛤、文蛤检出率较高,其次为毛蚶。不同弧菌在不同贝类中的检出率有很大差异,在不同地区中的检出率也存在很大差异。对分离的哈氏弧菌、副溶血弧菌及溶藻弧菌进行了药敏试验。抗菌药物包括:阿米卡星、庆大霉素、萘定酸、环丙沙星、恩诺沙星、诺氟沙星、红霉素、阿奇霉素、四环素、多西环素、头孢噻吩、头孢呋辛、氨苄西林、哌拉西林、阿莫西林、氯霉素、复方新诺明、氟苯尼考,共17种。结果表明,三种弧菌都对氨苄西林和阿莫西林的耐药率较高,对其他药物的敏感程度不同,也有耐药菌出现。本试验对哈氏弧菌和副溶血弧菌主要毒力基因进行检测,哈氏弧菌主要毒力基因为luxR、toxR、vhhA、vhhB、vhpA、vhpB、toxS、flaA、pap6;副溶血弧菌主要毒力基因为tdh、trh、ureC、vscC2、vcrD2。结果表明,80%的哈氏弧菌检测到luxR、toxR、vhhA、vhhB;副溶血弧菌毒力基因tdh、trh和ureC的检出率在20%左右。本研究查明了2016年度山东省贝类弧菌病的流行情况,为贝类弧菌病的预警提供了依据;通过对哈氏弧菌、副溶血弧菌、溶藻弧菌的药敏试验,探明了贝类弧菌的抗菌谱,为贝类弧菌病的防控提供了用药依据。
[Abstract]:In recent years, the species and pattern of shellfish culture in China are more and more diversified, the culture technology is gradually mature, the area of culture is expanding, and the shellfish industry has become an important pillar of China's aquaculture. However, the microorganisms with Vibrio as the main pathogenic bacteria have brought serious harm to the shellfish culture industry. In the summer of 7,8, the height of the shellfish Vibrio was high. The epidemic area of aquatic animal diseases caused by Vibrio is wide, high incidence and serious economic loss to aquaculture industry, which has always been a hot spot in the field of aquatic products. The epidemiological investigation of Vibrio in the culture shellfish and wild shellfish in the main shellfish culture areas of Shandong province. Identification, 16s-rDNA cloning and sequence analysis compared with the peer-to-peer method, the biological characteristics of the isolated Vibrio were preliminarily studied. The epidemic characteristics and laws of the Vibrio shellfish were mastered to provide a basis for the prevention of the disease. From Weifang Shouguang, Weihai Rushan, Wendeng, Jiaonan, Qingdao, sunshine, Binzhou Wudi, Dongying estuary and other areas. Different shellfish, including clam, clam, scallop, ellipsoid, cusp, clam clam, clam, clam, clam, blue clam, clam, clam, clam, clam, Vibrio, and mussels. 27 Vibrio, 8 other bacteria, 222 strains were isolated from the samples and were analyzed by sequencing and comparison, mainly Vibrio harveyi, Vibrio alginolyticus (Vibrio alginolyti) CUS), Vibrio parahaemolyticus (Vibrio parahaemolyticus), Vibrio chagasii, Vibrio Azureus, Vibrio natrium (Vibrio natriegens), Vibrio vulnificus (Vibrio vulnificus), Vibrio Hangzhou (Vibrio), etc. results showed that the detection rate of Vibrio Harris, Vibrio alginolyticus and Vibrio parahaemolyticus was 20%, 18%, 21% respectively, and the other Vibrio were relatively less. The detection rate of Vibrio parahaemolyticus and Vibrio alginolyticus in Jiaonan is high, and the detection rate of Vibrio Harris and Vibrio alginolyticus in Wudi is high, while the detection rate of Vibrio parahaemolyticus is very low; the detection rates of three Vibrio in three regions of the estuarine, sunshine and Shouguang are both high, both in 15%-35%. The result shows that Vibrio is one of the main pathogens causing shellfish disease. The harm of clam, clam, clam, clam, clam clam, clam clam is relatively serious. Among them, Vibrio Harris, Vibrio parahaemolyticus and Vibrio alginolyticus are more isolated. Vibrio Harris is mainly isolated from clam, clam clam, clam clam, clam, clam, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams, clams and clams. The detection rate is high, followed by clam. The detection rate of different Vibrio in different shellfish is very different, and the detection rate in different areas is also very different. The antimicrobial susceptibility tests of Vibrio Harris, Vibrio parahaemolyticus and Vibrio alginolyticus are carried out. The antimicrobial agents include: Amikacin, gentamicin, naphthyl acid, ciprofloxacin, enrofloxacin Norfloxacin, erythromycin, azithromycin, tetracycline, doxycycline, cefuroxime, cefuroxime, ampicillin, piperacillin, amoxicillin, chloramphenicol, compound neolin, and florfenicol were 17. The results showed that the resistance rate of three Vibrio to ampicillin and A Mo was higher, sensitive to other drugs and resistant to other drugs. The main virulence genes of Vibrio Harris and Vibrio parahaemolyticus were detected in this experiment. The main virulence genes of Vibrio Harris were luxR, toxR, vhhA, vhhB, vhpA, vhpB, toxS, flaA, pap6. The main virulence genes of Vibrio parahaemolyticus were TDH, TRH, ureC, and Vibrio parahaemolyticus. The detection rate of virulence genes TDH, TRH and ureC was about 20%. This study identified the epidemic situation of shellfish disease in Shandong province in 2016, provided the basis for the early warning of the shellfish Vibrio disease, and through the drug sensitivity test of Vibrio Harris, Vibrio parahaemolyticus and Vibrio alginolyticus, the antimicrobial spectrum of the Vibrio shellfish was explored and provided for the prevention and control of the shellfish Vibrio disease. The basis of drug use.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S944

【参考文献】