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题名	大黄鱼刺激隐核虫虫株鉴定及“白点病”的防治探讨; Indentify a strain of Cryptocaryon irritans in Pseudosciaena crocea and the research on prevention and cure the “White-spot Disease”
作者	邬阳
答辩日期	2013 ; 2012
导师	苏永全
关键词	大黄鱼 Pseudosciaena crocea 刺激隐核虫 Cryptocaryon irritans 鉴定 Identify 生活史 Life history 白点病防治 Prophylaxis and treatment of White-spot disease.
英文摘要	大黄鱼（Pseudosciaena crocea，Richardson）是我国东海重要的养殖经济鱼种，2010年的年产量在海水养殖鱼类中排名第二，是我国六大优势出口水产品之一。 本文对目前大黄鱼养殖产业中存在的种质、养殖、饲料、水质及病害方面的问题进行了简要分析，重点研究了在大黄鱼养殖过程中危害最大的“刺激隐核虫白点病”，探讨了其病原、流行特性及控制措施等。 主要成果如下： 1、刺激隐核虫的鉴定 于2011年6月初从福鼎市八尺门大黄鱼养殖网箱采集寄生虫株，通过形态学初步鉴定为刺激隐核虫（Cryptocaryon irritans）。提取寄生虫DNA，扩增ITS序列并测序，用CLUSTAL-X软件编辑、校对和排序，以淡水小瓜虫（Ichthyophthirius multifiliis）的同源序列（GenBank登录号：DQ270015）作为外类群，利用MEGA软件，基于Kimura双参数模型构建系统发育树，重复抽样（Bootstrap）1000次评估系统树的可靠性。结果显示，分离虫株先与中国南海刺激隐核虫虫株聚合在一起，后与台湾、澳大利亚南部、以色列和美国刺激隐核虫虫株聚合在一起，并相对于淡水小瓜虫独立分支。结果表明所取样品为刺激隐核虫，该虫株表现出具适应高温的特性。 2、刺激隐核虫生活史 成功进行了刺激隐核虫传代培养，对其生活史进行了详细的观察和描述，阐明了该虫滋养体从宿主上脱落、包囊前体外膜逐渐变化，形成包囊壁及包囊发育阶段内部变化等过程。 3、刺激隐核虫的药物防治实验 分别使用急性毒性试验（acute toxicity assay）、阻动试验（immobilization assay），探讨了冰醋酸、福尔马林、醋酸铜对大黄鱼幼鱼的安全性及对刺激隐核虫幼虫、包囊的杀灭效果。结果显示：以100 mg/L冰醋酸处理幼鱼8 h或150 mg/L处理2 h，均能有效杀灭刺激隐核虫幼虫而不对大黄鱼幼鱼造成伤害；使用福尔马林以30、50、100 mg/L的量分别处理4、1、0.5 h也能起到同样的杀灭效果。值得指出的是，醋酸铜对大黄鱼的刺激作用高于刺激隐核虫，故不适合治疗大黄鱼幼鱼感染的“白点病”。 本文为刺激隐核虫鉴定及生活史研究提供新的参考依据，探讨了冰醋酸、福尔马林、醋酸铜三种药物防治白点病的有效性和安全性，为刺激隐核虫病的预防与控制提供了新的渔药。; Large yellow croaker （Pseudosciaena crocea, Richardson）, is an economically important fish species in eastern and southern part chinese coasts, ranking in the second place of mariculture fishery annual yield. It is one of the top 6 dominance export aquatic products. In present study, we made a brief analysis on the main problems in large yellow croaker mariculture industry, such as:germplasm resource, stocking density, feedstuff administration, water environment, diseases etc. We focused on the most destructive parasitical disease in the process of large yellow croaker mariculture, that is:“white-spot disease” caused by Cryptocaryon irritans. Investigated it`s pathogen, epidemical characteristic and methods to control. The main results as follow: 1. Morphology of C.irritans and DNA sequence analysis Sampled one isolate of Cryptocaryon irritansin Bachimen large yellow croaker mariculture area in Fuding of Fujian province. As this isolate performanced higher water temperature adaptability, we want to know whether the sampled pathogen is Cryptocaryon irritans. We primary observed the morphological features of this parasite, later extracted DNA from pooled protozoan, amplified the ITS （internal transcribed spacers）sequences by PCR（polymerase chain reaction） and sequencing, edited, checked, aligned the sequence using the program CLUSTAL-X. a Ichthyophthirius multifiliis isolate was included for comparision. The phylogenetic trees were reconstructed using MEGA version 3.0. Bootstrap values are based on 1000 replicates for the NJ. The result show that sampled isolate were clustered with South Chinese sea C.irritans strain, then clustered with Taiwan, South Australia, Israel and USA strains, and distinct from Ichthyophthirius multifiliis isolate. Suggest that it is a new strain of C.irritans, adapted to higher water temperature. 2. The life history of C.irritans Successfully propagated C.irritans in vivo. Observed and described the life history of C.irritans, clarifed the process of trophonts falling-off from the hosts, the process of cyst wall formation, and the internal change of cysts in developmental stage. Using acute toxicity assay on juvenile fish of P.crocea, immobilization assay to C.irritans theronts, and killing experiment to C.irritans tomonts, investigated security on juvenile fish of P.crocea, killing effect on C.irritans theronts, tomonts of three chemical drugs:Acetic acid, Formaldehyde solution, Coper（Ⅱ）acetate monohydrate. 3. Prophylaxis and treatment against C.irritans The result show that:exposing juvenile fish of P.crocea to 100 mg/L, 150 mg/L acetic acid for 8, 2 h, respectively, can kill C.irritans effectly and made no damage to P.crocea, the same effect can also achieved by exposing juvenile fish of P.crocea to 30 ,50, 100 mg/L Formaldehyde solution for 4、1 、0.5 h. Moreover, since Coper（Ⅱ）acetate monohydrate are of moretoxicity to juvenile fish of P.crocea than to C.irritans, so we don’t recommend using this drug as a method to control“white-spot disease”. This study provided a reference for the identification and life-history research to Cryptocaryon irritians. Investigated the effectiveness and safety of three chemical drugs against “white-spot disease”, provided new methods to prevention and control the disease.; 学位：理学硕士; 院系专业：海洋与环境学院海洋学系_海洋生物学; 学号：22420091151161
语种	zh_CN
出处	http://210.34.4.13:8080/lunwen/detail.asp?serial=37975
内容类型	学位论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/53242]
专题	海洋地球－学位论文
推荐引用方式 GB/T 7714	邬阳. 大黄鱼刺激隐核虫虫株鉴定及“白点病”的防治探讨, Indentify a strain of Cryptocaryon irritans in Pseudosciaena crocea and the research on prevention and cure the “White-spot Disease”[D]. 2013, 2012.

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