魁蚶稚贝的底质选择性及其潜沙能力评价

底质是魁蚶稚贝潜沙、水平移动、存活与生长的重要环境因素,选择底质适宜的海区开展底播增殖,可以有效提高魁蚶稚贝的增殖效果,但目前仍缺乏相关研究。因此,在实验室条件下,研究了10 mm<壳长<15 mm、15 mm<壳长<20 mm、20 mm<壳长<25 mm、25 mm<壳长<30 mm 4种规格魁蚶稚贝对4种底质粒径(细砾、粗砂、中砂、细砂)的选择模式,并量化分析了20 mm<壳长<30 mm稚贝在5种底质粒径(增设泥底质)条件下的潜沙能力。结果表明:(1)10 mm<壳长<15 mm稚贝的足丝较发达,以附着生活为主,明显偏好粗砂底质,其在两种底质交界位置的分布率、水槽的贴壁率均最高;15 mm<壳长<30 mm稚贝转入营埋栖生活,随着规格的增大,稚贝选择的底质粒径亦增大,而在2种底质交界位置的分布率、水槽贴壁率也随之减小。其中15 mm<壳长<20 mm稚贝偏好选择中砂,分布率为50.50%,其次为细砂;20 mm<壳长<25 mm稚贝对不同粒径底质的适应性均较强;25 mm<壳长<30 mm稚贝偏好选择粗砂底质,分布率达到65%,其次为中砂。(2)20 mm<壳长<30 mm稚贝在淤泥底质中的初潜时间最短,为2.74 min,其次为中沙、细沙;在细砂底质中的潜沙率最高,达到85%,其次为中沙。综合分析认为,20 mm<壳长<30 mm稚贝是较适宜的底播苗种规格,而以中砂(0.25 mm<粒径<0.5 mm)和细砂(粒径<0.25 mm)为主要粒径组成的海区宜进行魁蚶稚贝的底播增殖。     

魁蚶生殖生物学初步研究

通过组织学观察,结合人工育苗实践,魁蚶的生殖周期可分为5期,即形成期、生长期(小生长期和大生长期)、成熟与排放期、退化期和休止期。魁蚶性腺在春天水温回升至4℃以上时开时发育;性腺发育的最适温度为14～20℃;产卵季节是6月下旬～8月下旬,7月为产卵盛期;产卵适温20～25℃。     

魁蚶常温育苗技术

魁蚶（Scapharcabroughtonii），俗称赤贝，是一种大型、经济价值较高的贝类，其软体部的加工品“赤贝肉”，具有较高的出口创汇价值。近几年，我们利用夏秋季育苗场相对空闲时间进行魁蚶常温育苗，取得了较好效果。一、亲贝选择１．亲贝的采捕时间因不同海区水温和饵料条件的差异，魁蚶的性腺发育程度也不尽相同，一般在海区底层水温升至２０～２５℃时性腺逐渐进入成熟排放期。我国的黄海北部海区７～８月份为产卵盛期，故常温育苗多在７～８月份采捕亲贝。２．亲贝的选择方法选择壳长７～１０厘米、外形完整无损伤的魁蚶作亲贝。解剖检…     

盐度胁迫对魁蚶稚贝血淋巴渗透压及鳃Na~+/K~+-ATP酶活力的影响

文章研究了盐度胁迫下魁蚶(Anadara broughtonii)稚贝(壳长25~35 mm)血淋巴渗透压和鳃Na+/K+-ATP酶活力的变化。结果显示,突变胁迫下,盐度20、25、35、40组的稚贝血淋巴渗透压均在48 h内显著降低或波动升高后趋于稳定,盐度15组则需96 h完成波动调整。以日变幅5渐变至高盐40过程中,稚贝血淋巴渗透压在48 h内趋于稳定;低盐15组仅需72 h完成渗透压调节,调整时间显著小于突变组。盐度突降胁迫下,各处理组稚贝鳃Na+/K+-ATP酶活力12 h后显著升高,24 h分别达到最大值,随后快速回落并稳定在高于对照组水平;盐度骤升胁迫下,鳃ATP酶活力波动降低,在12~24 h达到最小值后稳定在低于对照组水平。低盐(15)和高盐(40)渐变组的鳃ATP酶活力变幅均小于突变组。盐度胁迫下鳃Na+/K+-ATP酶活力的响应方式不同,即低盐胁迫时活力增强,高盐胁迫时活力降低。采用盐度渐变的驯化处理更有利于魁蚶稚贝应对盐度胁迫。     

魁蚶亲贝促熟与健康苗种培育技术

采用常温和升温促熟两种方式培育魁蚶亲贝,经产卵、孵化、选幼,筛选出健康苗种,投放附着基让其附着并进行中间培育.在550 m3水体中两个批次共孵出正常发育的D型幼虫42亿粒,附着变态后,获得健康稚贝约21亿粒.试验结果表明:魁蚶苗种培育过程中的D型幼虫孵化率、幼虫成活率及变态率与卵子的质量相关;亲贝在性成熟时间上存在一定程度的地理差异;魁蚶幼体的发育速度与温度成正比;魁蚶苗种挂到海区进行中间培育后,生长速度较快.     

施氏獭蛤稚贝对高温和干露的耐受性研究

针对南方沿海夏季高温对贝类种苗存活的影响,研究了北部湾海域施氏獭蛤自然群体人工繁育的稚贝对高温和干露的耐受性,以期为高温期稚贝培育和运输提供参考。试验结果表明:(1)施氏獭蛤稚贝的成活率在水温30~35℃时,存在显著的温度效应,随温度升高,稚贝存活率快速下降,组间差异显著(P0.05),高温敏感起始点为31℃,高起始致死温度为31.87℃,温度为33、34、35℃时,100%致死的时间分别为6d、4d和2d,7d后,稚贝不再死亡;(2)无沙干露组各温度维持100%存活的最长时间分别为20℃组18h,30℃组9h,10℃组6h,有沙干露组只有20℃组在15h内可维持100%存活,30℃组和10℃组在6h内已出现死亡。研究结果表明,无沙干露和20℃较适合稚贝的长途干露运输。研究结果为施氏獭蛤养殖海域的选取、稚贝的长途运输提供科学依据考。     

魁蚶人工苗种控温培育技术

详细介绍了魁蚶苗种的春季控温培育技术,其中包括亲蚶的选择、运输、促熟蓄养、采卵孵化、孵化期管理、选幼、幼体培育、水温和水质控制、投饵、日常管理、附着基投放和投放后的培育管理、稚贝出池.     

温度和盐度同缢蛏稚贝存活及生长的关系

本文阐述在室内控制的温度和盐度条件下，采用单因子和结合的方法，研究温度和盐度同缢蛏稚贝存活及生长的关系。缢蛏稚贝的适盐范围为4．50‰至28．30‰，最适盐度为12．40‰至16．30‰。在25℃时，它对低盐(1.80‰以下)有较强的忍耐性，对高盐度(33．50‰至40．00‰)也有一定的忍耐性。缢蛏稚贝的适温范围为10℃至35℃。但最适温度随盐度的不同而异，若在最适盐度的生境中，它的最适水温为27℃至30℃；如果稚贝在最适盐度以外的适盐范围内，经方差分析表明，它的最适温度是22℃。稚贝对低温(5℃至2℃)有很强的忍耐性，致死温度上限是40℃。缢蛏稚贝对盐度的敏感性，比它对温度的敏感性来得强。在22℃以下的温度组中和最适盐度范围内，盐度与温度的相关性不太明显；而在27℃以上的较高温度和适盐范围的上下极限时，其相关性显著，它们相应的适盐或适温范围明显地变窄。     

魁蚶育苗技术

魁蚶(Scapharca broughtonii)是一种大型、经济贝类,在我国主要分布于渤海及黄海北部。近几年由于过度捕捞,野生资源遭到破坏,自1989年以来,山东、辽宁沿海有关科研和生产单位开始了魁蚶室内人工育苗。笔者自1993年开始从事魁蚶苗种生产,积累了一些经验,现将采取的主要技术措施总结如下:1亲贝选择及采捕挑选壳长7￣12cm的魁蚶做为亲贝,将壳面有蚀孔和受伤的魁蚶剔除,视个体大小,一般每立方水体准备亲贝2￣6个。升温培育在3月中、下旬亲贝入池,室内强化促熟40￣50d,待海湾扇贝出池后产卵。常温培育在5月中、下旬以后进亲贝,亲贝室内促熟20￣…     

不同温度下多棘海盘车对菲律宾蛤仔的摄食选择性研究

为了解不同水温环境中(5~25℃)多棘海盘车(4~8 cm)对菲律宾蛤仔(20~40 mm)的摄食选择性,本实验利用饵料收益率、摄食规格选择性、摄食量与摄食率等指数,分析了温度、多棘海盘车规格及饵料规格对多棘海盘车摄食选择性的影响。结果显示,实验条件下,捕食者及被捕食者规格对饵料收益率影响显著,温度对饵料收益率影响不显著;根据饵料收益模型,4种规格多棘海盘车依次在摄食20、23、30和35 mm蛤仔时的饵料收益率最大,收益率分别为0.62、0.70、0.83、0.94 mg/min;5~15℃条件下,摄食量随水温升高而增大,15℃为多棘海盘车摄食蛤仔的最适水温,此时5~8 cm多棘海盘车摄食量均达最大值,分别为0.37、0.45和0.54 g/d,之后随水温升高摄食量减小;摄食率昼夜差异显著,夜间显著高于白天;青岛近海5—6月的水温(13~18℃)为蛤仔春季繁殖的适宜水温,也是多棘海盘车最适生长和摄食温度,此时应加强对多棘海盘车的防治和清除。     

Susceptibility by amberjack (Seriola dumerili), yellowtail (S. quinqueradiata) and Japanese flounder (Paralichthys olivaceus) to Neobenedenia girellae (Monogenea) infection and their acquired protection

This study investigated: 1) susceptibility differences to infection by Neobenedenia girellae (Capsalidae) between amberjack Seriola dumerili (Carangidae), yellowtail S. quinqueradiata and Japanese flounder Paralichthys olivaceus (Paralichthyidae); 2) growth and egg production of N. girellae on each fish species; 3) acquired protection of each fish species against this parasite. The number of N. girellae on S. dumerili was significantly higher than on S. quinqueradiata and P. olivaceus when these fishes were exposed to oncomiracidia in the same aquarium. Neobenedenia girellae growth on S. dumerili was fastest and, thus the number of eggs laid by parasites on S. dumerili was greater than on the other two species. Seriola dumerili and P. olivaceus, which were previously infected with N. girellae and treated by freshwater bath, acquired partial protection against re-infection by N. girellae. The relative re-infection of three S. dumerili individuals out of eleven individuals was markedly low compared with the initial infection, and the relative initial infection and re-infection on two P. olivaceus out of eleven individuals was markedly low. The results of this study could be useful to control N. girellae infections when cultivating S. dumerili, S. quinqueradiata and P. olivaceus.     

AFLP-based genetic linkage map of marine shrimp Penaeus (Fenneropenaeus) chinensis

This paper presents the genetic linkage map of the Chinese shrimp Penaeus (Fenneropenaeus) chinensis constructed with 472 AFLP markers. A hundred F₁ progeny from an intercross between a female from the new variety “Yellow Sea No. 1” and wild caught male used for the mapping study. Two separate maps were constructed for each parent. The female linkage map consisted of 197 marker loci forming 35 linkage groups and spanned a total length of 2191.1 cM, with an average marker space of 13.5 cM. The male map consisted of 194 marker loci mapped to 36 linkage groups and spanned a total length of 1737.3 cM, with an average marker spacing of 11.0 cM. The level of segregation distortion observed in this study was 12.2%. The estimated genome length of P. chinensis was 3150.3 cM for the female and 2549.3 cM for the male, respectively. The observed genome coverage was 69.6% for the female and 68.1% for the male map. The linkage maps constructed in this study provide basic information for further linkage studies on Chinese shrimp, and more importantly, the construction of the maps are part of the work of the genetic breeding programs which will be used for growth discovered in the QTL analysis of P. chinensis.     

太平洋鳕RAG1和IgM基因荧光定量PCR方法的建立及初步应用

为研究太平洋鳕发育早期特异免疫系统形成的机制,通过RAG1和IgM基因的转录水平衡量特异免疫系统的发育特点.根据GenBank中RAG1和IgM的序列信息,分别设计1对特异引物,从太平洋鳕头肾中扩增得到RAG1和IgM的基因片段.将所获基因片段分别插入到克隆载体pMD18-T中,从而构建太平洋鳕RAG1和IgM基因的质粒标准品.建立并优化太平洋鳕RAG1和IgM基因绝对荧光定量PCR方法.为进一步验证该方法的可靠性,分别利用绝对定量和相对定量检验目的基因在太平洋鳕早期发育过程不同组织内的表达差异.以优化后的绝对荧光定量PCR方法检测不同发育时期太平洋鳕RAG1和IgM的表达情况.结果显示,RAG1的回归方程为y=-3.266x+33.77,回归系数R2=0.996;IgM的回归方程为y=-3.119x +27.61,回归系数R2 =0.998.绝对定量和相对定量结果在基因转录趋势上显现出一致性,即RAG1基因在胸腺和头肾中表达,且在胸腺中的表达量显著高于头肾中的表达量,在肝脏和脾脏中无表达;IgM基因在胸腺、头肾、肝脏和脾脏中均有表达,其中脾脏中表达量最高,其次是头肾.RAG1基因在太平洋鳕发育早期的表达水平很低,到61日龄(days posthatching,dph)至95 dph表达量显著提高;IgM基因在早期表达水平同样很低,到33 dph至61 dph才有明显表达,在95 dph时表达量显著提高.研究表明,本实验方法可靠,特异性较强,可成功对目标基因转录水平进行检测.     

罗非鱼海豚链球菌病的病理学观察

采用光学显微镜和透射电子显微镜技术分别对患海豚链球菌病的罗非鱼的病理学变化进行了研究。分离的病原菌革兰氏染色呈阳性,透射电镜负染观察菌体球形或卵圆形,直径0.6~1.0μm,多数呈链状排列。组织学病变主要表现为全身多组织、器官水肿,出血、变性、坏死以及炎症反应,特别是肝、脾、肾和脑分别表现为肝炎,脾炎,间质性肾炎和脑膜炎。超微结构观察发现病鱼肝、脾、肾、脑、心肌和骨骼肌等器官的细胞超微结构都有较为严重的破坏,细胞核畸形,染色质浓缩或边集,粗面内质网囊泡化及脱粒,线粒体肿胀,嵴断裂或溶解消失。研究表明,海豚链球菌能造成罗非鱼全身性组织器官病变,致使器官功能障碍,正常生理代谢调节紊乱,最后导致死亡。     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

拟穴青蟹丝裂原活化蛋白激酶激酶（MAPKK）基因的克隆与表达分析

采用qRT-PCR、RACE等方法,获得了拟穴青蟹丝裂原活化蛋白激酶激酶(MAPKK)基因cDNA全长序列。该基因全长1 558 bp,开放阅读框长度为1 224 bp,编码407个氨基酸残基。同源分析显示,该基因编码的蛋白与昆虫的相似性高达70%,推测MAPKK基因在节肢动物具有较高的保守性。经荧光定量PCR检测,MAPKK基因在拟穴青蟹多个组织中有表达,且在脑神经节和卵巢中表达量较高。在拟穴青蟹卵巢发育过程中,MAPKK基因在卵巢发育期(Ⅲ期)表达量最高,发育期为卵母细胞快速生长期,推测MAPKK具有促进卵母细胞快速生长的作用。     

山东沿海魁蚶繁殖周期与生化成分的周年变化

自2010年10月至2011年9月,对山东省鳌山卫海区魁蚶的繁殖周期、生化成分的季节变化及其与环境因子的关系进行了研究。每月采集样品1次,测定采样点水温和叶绿素a含量,采用组织学方法分析性腺的季节变化,并分别测定外套膜、闭壳肌、性腺—内脏团和足组织的生化成分(糖原、脂肪和蛋白质)含量。结果表明,该海区魁蚶雌雄比例为1∶1,雌雄性腺发育同步,全年只有1个繁殖期;配子发生始于2月,分别有25.0%和53.9%的雌雄个体性腺进入形成期,随水温和叶绿素a含量的升高而发育,6月大部分个体成熟并有部分进入排放期,至8月配子集中排放(雌:58.3%;雄:69.2%)。生化分析显示,脂肪含量在性腺—内脏团中随性腺发育积累储存,产卵后显著降低;所有组织中的糖原含量在3—7月显著高于其它月份,并且性腺中的含量高于其他组织,于5月达最大值64.2%,表明糖原在魁蚶繁殖活动中具有重要作用;蛋白质含量在除闭壳肌外的其他3种组织中出现冬季和产卵盛期两个低谷,暗示蛋白质能够弥补糖原的供能不足,与繁殖活动存在密切联系。     

Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.     

氟苯尼考在鲫和草鱼体内的药代/药效动力学联合参数及其临床给药方案的研究

为研究氟苯尼考在鲫和草鱼体内的药代学、药效动力学联合参数,并制定氟苯尼考对鲫、草鱼的精确用药方案,本实验结合氟苯尼考对致病性嗜水气单胞菌CAAh01的体外药效学研究和口灌不同剂量的氟苯尼考在鲫、草鱼体内药代动力学研究,确定了氟苯尼考防治该致病菌引起的鲫和草鱼细菌性败血症的给药方案。研究结果显示,氟苯尼考对CAAh01菌株的最小抑菌浓度(MIC)为0.5μg/mL,最小杀菌浓度(MBC)为1.0μg/mL,防细菌耐药突变浓度(MPC)为6.0μg/mL,防耐药突变选择窗(MSW)为0.5~6.0μg/mL。按10、20、30 mg/kg体质量剂量对鲫、草鱼口灌氟苯尼考后,在鲫体内,血药浓度大于MPC的维持时间分别为5、8、24 h;AUC24/MIC分别为177.06、265.90、426.50;Cmax/MIC分别为15.59、21.32、31.24。在草鱼体内,血药浓度大于MPC的维持时间分别为0、0、3 h;AUC24/MIC分别为38.60、75.08、121.94;Cmax/MIC分别为4.75、10.08、19.99。研究表明,综合血药浓度维持MPC以上的时间、AUC24/MIC或Cmax/MIC指标,氟苯尼考适用于鲫细菌性疾病的防治,其防突变用药方案为剂量30 mg/kg,每日1次给药,休药期不低于20 d。对于草鱼细菌性疾病的防治,氟苯尼考不宜连续使用。     

溶藻弧菌诱导马氏珠母贝血淋巴cDNA差减文库的构建及分析

为了探讨马氏珠母贝免疫防御机制,筛选免疫防御相关功能基因,以致病性溶藻弧菌人工感染的马氏珠母贝血淋巴为材料,采用抑制性差减杂交(SSH)技术,构建了溶藻弧菌诱导的马氏珠母贝血淋巴cDNA差减文库;以马氏珠母贝管家基因β-actin作为差减指标检测该文库的差减效率;对选取的600个阳性克隆进行测序及生物信息学分析。结果显示,该文库的差减效率可达210倍;PCR阳性检测显示差减片段为100~750 bp,对随机挑取600个克隆的测序结果显示,共获得414个有效EST序列;通过BLAST同源性比对,有167个EST序列(占40.34%)与NCBI数据库中已知功能蛋白质具较高同源性,其中7个EST序列(占1.69%)与免疫防御相关基因同源;此外,204个EST(占49.28%)在数据库中未发现同源序列。研究表明,SSH技术能有效富集马氏珠母贝血淋巴差异表达基因,研究结果为探索马氏珠母贝免疫基因作用机理和调控机制奠定了基础。