不同饵料对中国对虾幼虾生长及感染WSSV存活率的影响

采用4种饵料投喂中国对虾（Fenneropenaeus chinensis）幼虾，用投喂感染的方法人工感染WSSV。测定体长、体重以及各组的攻毒存活率，实验周期15d。ANOVA分析结果表明，投喂鲜活卤虫组体长、体重的增长明显优于其他各组，差异达极显著水平（P〈0．01）；投喂人工配合饵料实验组的体长、体重增长量最小；投喂鱼肉组体长增长慢于投喂蛤蜊肉组，而体重的增长快于投喂蛤蜊肉组，差异均不显著（P〉0．05）。投喂卤虫成体和投喂鱼肉两组的攻毒存活率最高，明显高于投喂配合饵料和蛤蜊肉两实验组，差异达极显著水平（P〈0．01）；投喂卤虫组和投喂鱼肉组之间存活率无显著差异（P〉0．05），投喂人工配合饵料组和蛤蜊肉组差异不显著（P〉0．05）。巢式PCR检测表明，人工感染前的中国对虾幼虾少数携带WSSV，人工感染后全部个体检测到病毒特征片段。     

卤虫在不同饵料培养介质中的生长规律

以啤酒酵母和单胞藻为投喂饵料，新疆艾比湖卤虫（Artemia parthenogenetica)为投喂对象，建立三个实验组分别进行研究，观察在不同饵料条件下艾比湖卤虫的生长，探讨不同饵料对卤虫的生长、成活的影响。通过对卤虫体长和存活率的测量，建立了卤虫在不同饵料情况下的生长方程、生长曲线和存活率曲线，并通过对方程和曲线的分析获得以下结论：投喂单胞藻组卤虫的生长最慢，存活率最低25表面以后全部死亡；投喂酵母组卤虫生长最快，在0≤t≤42.6时体长值最高，存活率较低，投喂单胞藻和酵母的混合饲料组卤虫的前期生长次于酵母组，后期生长超过之，存活率较高且成熟个体产虫量和产卵量也多于酵母组。     

鳗弧菌和溶藻弧菌二联疫苗对大菱鲆的免疫效果

将鳗弧菌（Vibrio anguillarum）和溶藻弧菌（V.alginolyticus）分别用0.3%福尔马林灭活,制备成鳗弧菌和溶藻弧菌2种单苗,并将2种菌等量配制成二联疫苗,单次腹腔注射免疫大菱鲆（Scophtalmus maximus）.通过检测免疫后替代途径补体活力、溶菌酶活力、抗体凝集效价的变化并分别进行攻毒实验比较疫苗的免疫效果.结果显示,3种疫苗对所测定的各免疫指标都有一定的促进作用,但二联疫苗后期效果低于2种单苗.二联疫苗对鳗弧菌和溶藻弧菌的免疫保护率分别为83.52%和83.24%,而鳗弧菌单苗对鳗弧菌攻毒、溶藻弧菌单苗对溶藻弧菌攻毒的免疫保护率分别为80.37%和74.89%.可以认为2株菌具有制备二联疫苗的可能性.[中国水产科学,2006,13（3）：397-402]     

云斑尖塘鳢转饵期仔鱼生长和存活率的研究

在水温(30±1)℃条件下对人工孵化的云斑尖塘鳢(Oxyeleotris marmorata)仔鱼进行转饵期不同投喂策略实验,对仔鱼生长和成活情况进行了研究,设计5个投喂组:轮虫投喂组(A组);70%轮虫+30%卤虫投喂组(B组);50%轮虫+50%卤虫投喂组(C组);30%轮虫+70%卤虫投喂组(D组);卤虫(幼虫)投喂组(E组)。结果显示:云斑尖塘鳢仔鱼对卤虫的摄食强度随卤虫在饵料中比例的升高而增大,其对混合饵料中轮虫和卤虫的摄食强度的比值(Ir/Ia)随食物组成的变化发生相应的变化。以轮虫-卤虫不同比例的混合饵料投喂时,D组仔鱼的增长率、增重率最高,可存活率不足50%,高于E组,显著低于其它各组;C组仔鱼生长指标略低于D组,但存活率为(74.00±2.29)%,综合指标显著好于其它各组。当混合饵料中轮虫和卤虫各占50%时,10日龄开始投喂组仔鱼的全长(13.07±0.13)mm,特定生长率(10.25±0.37)%/d,存活率(82.14±0.72)%,综合生长指标最好。结果表明,云斑尖塘鳢仔鱼转饵期进行联合投喂的最佳饵料比例为50%轮虫+50%卤虫,最佳转饵时间为10日龄。     

卤虫在WSSV病毒病传播中的媒介作用

2003年8月对卤虫进行投喂(浸泡)感染，并进行中国明对虾投喂实验。各组对虾阳性感染率的单因素方差分析比较结果表明，卤虫可以携带有活性的WSSV病毒粒子，并可通过摄食导致对虾间接地携带病毒，但病毒感染能力有限。卤虫病理切片观察结果显示，攻毒后卤虫细胞核没有显著深染及核肿大现象，但无论是横切还是纵切均显示有上皮脱落、部分组织结构松散、细胞结构不完整现象。本试验确定卤虫成体可以携带WSSV并通过投喂感染对虾，使其潜在携带病毒，仔虾阶段投喂成体卤虫应经过严格检疫，制作含有经过检疫卤虫的微囊饵料作为卤虫及其他鲜活饵料的替代品或许是有效的防病措施。     

盐度因子对卤虫生长的影响

目前卤虫自然资源破坏严重、濒临枯竭，对卤虫高密度人工养殖的要求也就应运而生。而卤虫的生长和存活均受到盐度因子的影响，王睿等发现卤虫的生长和存活受到盐度的影响，证实卤虫的高耐盐性，Browne等人探讨了盐度和温度对卤虫生存和繁殖的联合作用，但他们均没有报道卤虫生长和孵化的最适宜盐度。本文应集约化培养技术研究之需，通过研究，报道盐度对卤虫生长发育及其存活率的影响，为卤虫集约化培养中盐度因子的控制提供参考。一、材料和方法１．材料大盐湖卤虫及休眠卵。２．饵料盐藻（DunaliellA．sp）２０％，小球藻（Chlorellasp…     

饵料和养殖密度对中国对虾幼虾生长及存活率的影响

在实验用虾遗传背景一致的情况下,分析了3种饵料(配合饲料、冰冻鲜鱼肉和活卤虫)和4个不同的养殖密度对小水体中国对虾幼虾生长和存活率的影响。结果表明,饵料和饲养密度对中国对虾幼虾生长及存活率有显著影响。在饵料、养殖密度单因子实验及饵料和养殖密度相结合的双因子实验中,幼虾的生长均表现出极其显著的差异(P<0.01),活卤虫对幼虾生长的效应尤为突出。而养殖密度对中国对虾的行为生物学、个体间体重增量均有影响。随着养殖密度的提高,中国对虾增重变慢;同时,个体间体重增量差异变大。随着养殖密度的增加,中国对虾幼虾的存活率呈下降趋势,但不同饵料对存活率影响变化幅度较大,波动在58.1%～85.2%之间,其中投喂活卤虫养殖密度为50尾/桶的存活率最高(85.2%);投喂配合饵料4个养殖密度梯度的存活率变化不明显;而投喂冰冻鱼肉4个养殖密度梯度的存活率变化较大。因此,采用合适的饵料和养殖密度,能够提高中国对虾的生长速度,提高对虾养成存活率,改善生长过程中对虾群体体长、体重的整齐度。     

轮虫、卤虫无节幼体及蝇蛆三种开口饵料对黄颡鱼仔鱼生长、存活及免疫酶活性的影响

在水温(25±0.5)℃下,将10 000尾4日龄的体长(0.25±0.004)cm的全雄黄颡鱼Pelteobagrus fulvidraco仔鱼放入长2m×宽1.4m×水深30cm的孵化槽中,投喂轮虫、卤虫无节幼体和蝇蛆三种开口饵料,研究不同开口饵料对黄颡鱼仔鱼生长、存活率以及免疫相关酶活性的影响。结果表明,饲养3d时,投喂卤虫无节幼体组仔鱼的体质量显著大于轮虫组(P0.05),而投喂蝇蛆组仔鱼的体质量介于二者之间,且与两组无显著差异(P0.05);7d后,投喂蝇蛆组仔鱼体质量最大,显著大于后两组(P0.05),卤虫无节幼体组次之,轮虫组最小;投喂蝇蛆组仔鱼的存活率最高,投喂蝇蛆组仔鱼酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和溶菌酶(LZM)活力显著高于其他两组(P0.05),而其他两组仔鱼这三种酶活力没有显著影响。认为与轮虫、卤虫无节幼体相比,蝇蛆是一种更适宜黄颡鱼仔鱼培育的开口饵料。     

扁吻鱼仔鱼集约化培育技术研究

正2014年～2016年,在新疆水生野生动物救护中心工厂化鱼类繁育车间,使用循环水矩形槽进行扁吻鱼子二代仔鱼的集约化培育,对仔鱼发育的主要形态特征及生活习性进行了观察,对仔鱼开口饵料的选择及培育水环境进行了分析。结果表明,水温11℃～24℃时,初孵仔鱼发育至第6天～7天开口摄食,以卤虫无节幼体作为开口饵料,第10天～11天开始投喂卤虫无节幼体+鳗仔鱼饲料,经过5天～10天的投喂培育,仔鱼全长长至10mm～15mm,存活率为67.2%～84.3%,静水环境下存活率明显高于循环水环境。     

中国盐湖卤虫的生物学特性及其在对虾育苗中的应用

本文通过对中国部分盐湖卤虫的自然种群组成、卤虫卵卵径、孵化特性、无节幼体体长以及投喂对虾幼体的饵料效果等的测定,对照卤虫卵和无节幼体生化成分的文献资料,评估了中国内陆盐湖卤虫的内在质量及其开发利用的前景。     

鳗弧菌O1/O2二价灭活疫苗免疫大菱鲆的抗体持续期和免疫保护期

本研究分析了鳗弧菌(Vibrio anguillarum)O1/O2血清型二价灭活疫苗免疫大菱鲆后的抗体持续期和免疫保护期。以鳗弧菌O1血清型VAM003株和O2血清型VAM007株为抗原制备了福尔马林灭活二价疫苗,将疫苗按照三种剂量(10~7 cells/尾、10~8 cells/尾、10~9 cells/尾)以腹腔注射途径免疫大菱鲆,在免疫后3 d、7 d、14 d、30 d、60 d、90 d、120 d、150 d,用血清凝集实验检测了免疫鱼血清的VAM003和VAM007抗体效价,用攻毒实验检测了疫苗的免疫保护率(RPS)。结果显示,在免疫后7 d三个剂量组的大菱鲆均产生了特异抗体,并获得27%～60%的RPS。三个剂量组大菱鲆的O1血清型抗体持续期分别90 d (10~7 cells/尾组)、150 d (10~8 cells/尾组)、150 d (10~9cells/尾组),而三个剂量组大菱鲆的O2血清型抗体持续期均150 d。三个剂量组的大菱鲆获得的免疫保护持续期均150 d;以RPS75%为有效免疫保护,各剂量组大菱鲆抵抗O1血清型病原感染的有效免疫保护期为:14～120d(10~7 cells组)、14～120 d (10~8 cells/尾)、14～150 d (10~9 cells/尾),抵抗O2血清型病原感染的有效免疫保护期为:14～60 d (10~7 cells组)、14～120 d (10~8 cells/尾)、14～120 d (10~9 cells/尾)。研究结果表明鳗弧菌二价灭活疫苗可为大菱鲆提供有效而稳定的免疫保护,获得的抗体持续期和免疫保护期为该疫苗的临床中试研究提供了基础。     

不同饵料对美洲西鲱仔鱼生长、相关酶活力及体脂肪酸的影响

以美洲西鲱(Alosa sapidissima)仔鱼(孵化后10 d)为实验对象,在室内进行为期30 d的摄食生长试验,分别投喂不同的饵料[单独投喂卤虫(Artemia)、单独投喂微颗粒饲料、卤虫与微颗粒饲料混合投喂],分析不同饵料的投喂对存活、生长、消化酶活性、非特异性免疫相关酶活性以及体脂肪酸组成的影响。结果表明:(1)混合投喂组的存活与生长表现都要显著优于卤虫组和微颗粒饲料组(P0.05),微颗粒饲料组与卤虫组相比差异不大,而试验后期(20~30 d)微颗粒饲料组的存活与生长表现要优于卤虫组;(2)消化酶的结果显示,胃蛋白酶和淀粉酶活力在各组中无显著性差异(P0.05),胰蛋白酶、糜蛋白酶和脂肪酶在卤虫组中活力最低,显著低于其他两组(P0.05),其中糜蛋白酶和脂肪酶活性在微颗粒饲料组中要显著高于混合投喂组(P0.05);(3)碱性磷酸酶和超氧化物岐化酶活力在各组中无显著性差异(P0.05),过氧化氢酶和溶菌酶活力在微颗粒饲料组中最高,卤虫组中最低,两者之间差异显著(P0.05);(4)鱼体脂肪酸受饵料脂肪酸组成影响较大,微颗粒饲料中n-3HUFA和DHA含量较高,其投喂后在鱼体中的含量也高。综上所述,不同饵料投喂对美洲西鲱仔稚鱼存活、生长、消化酶活性、非特异性免疫相关酶活性以及体脂肪酸影响显著,在试验20 d之前使用卤虫与微颗粒饲料混合投喂,20 d后完全转食微颗粒饲料,可以降低成本,同时提高生产效率。     

Brachionus vs Artemia duel: Optimizing first feeding of Upogebia pusilla (Decapoda: Thalassinidea) larvae

Larval rearing of many marine organisms is dependent on the availability of live food. The aim of this study was to optimize larval first feeding for the mud shrimp Upogebia pusilla, by comparing the effectiveness of the two most commonly used live feeds: Brachionus plicatilis and Artemia sp. nauplii. Survival, larval duration, molt synchronism and megalop size were compared using five feeding treatments: Artemia from zoea I to IV (B0), Brachionus during zoea I and Artemia from zoea II to IV (B1), Brachionus during zoea I and II and Artemia during zoea III and IV (B2), Brachionus from zoea I to III and Artemia during zoea IV (B3) and Brachionus from zoea I to IV (B4). The proportion of larvae that reached the megalop stage was 0.00% in treatment B0, 3.33% in treatment B1, 33.33% in treatment B2, 66.67% in treatment B3 and 76.67% in treatment B4. Larvae fed on rotifers until zoea III or zoea IV stages had a higher survival but no differences were found either in time to reach megalop or in megalop size. This study demonstrates that rotifers are essential for the survival and development of U. pusilla early larval stages but that rotifers can be successfully replaced by Artemia nauplii in the zoea IV stage.     

Use of Microbacterium sp. and Exiguobacterium mexicanum to improve the survival and development of Artemia under xenic conditions

The effect of Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N was evaluated in the diet of Artemia under xenic conditions. Viable cultures of bacteria were provided to xenic cultures of Artemia in combination with Sacharomyces cerevisiae, cornflour or Spirulina, and the effect on the survival and growth was recorded. The use of these bacterial strains improves significantly the survival of Artemia independently of the used food (P < 0.05), and variable results were observed in the growth.     

Carbon supplementation and microbial management to stimulate artemia biomass production in hypersaline culture conditions

Two Artemia culture experiments were conducted in a zero water exchange system at salinity 80 g L^?1. In Experiment I, different carbon sources including molasses, glucose, sucrose and corn flour were given to supplement suboptimal feeding with the microalgae Dunaliella viridis, whereas the control was fed only D. viridis. In Experiment II, molasses was supplemented to the culture medium, as well as the bacteria Alkalibacterium sp. or the archaea Halobacterium sp., isolated from the brine in the local saltponds. The control groups were fed only with D. viridis or with D. viridis and molasses. Our study shows that in laboratory culture biofloc development can be stimulated under hypersaline conditions. Supplementation of carbon sources, bacteria and archaea benefited biofloc development, water quality in terms of nitrogen load and enhanced Artemia production. Denaturing gradient gel electrophoresis analysis on the bioflocs also indicates an increased biodiversity and richness of the microbial community in the culture medium. These results need to be confirmed for large scale Artemia pond production, in which case, we recommend molasses as carbon source.     

牙鲆趋化因子基因CXCL9的克隆、鉴定与表达分析

趋化因子是一类具有化学趋化功能的小分子分泌性蛋白,能够引导白细胞到损伤或者感染的部位,参与免疫调节和病理反应。为丰富鲆鲽鱼类趋化因子的相关基础研究,本研究克隆了牙鲆(Paralichthys olivaceus)趋化因子基因CXCL9,其全长为910 bp,开放阅读框为408 bp,编码135个氨基酸。该基因具有趋化因子超家族的典型特征,在35、37、60和77个氨基酸位置具有4个保守的半胱氨酸残基,形成两个二硫键。氨基酸序列分析表明,该基因与大菱鲆(Scophthalmus maximus)、半滑舌鳎(Cynoglossus semilaevis)的CXCL9基因具有较高的同源性,分别为78.5%和71.0%。实时荧光定量PCR结果显示, CXCL9在正常牙鲆肝脏和脾脏中的表达量非常高,在皮肤和鳃等组织中表达量次之,说明CXCL9特异性地在免疫相关组织中表达。经迟缓爱德华氏菌(Edwardsiella tarda)感染后,牙鲆肝脏中CXCL9 mRNA表达量在感染后6 h即出现高峰值,而头肾和脾脏中的高峰值出现较晚。经鳗弧菌(Vibrio anguillarum)感染后6 h,牙鲆肝脏中CXCL9 mRNA的表达量升高近8倍,头肾中的表达高峰值出现在感染后24 h,而脾脏中CXCL9 mRNA的表达量在感染后6 h迅速升高10倍,逐渐降低后48 h又出现高峰,为正常水平的20倍。上述研究结果说明CXCL9基因在病原菌感染过程中有快速响应,可能参与了免疫防御过程,将有可能发展为一种牙鲆细菌性疾病的生物标记。本研究为牙鲆趋化因子超家族免疫机制的深入研究奠定了基础。     

Use of ongrownArtemia in nursery culturing of the tiger shrimp

Juvenile and adultArtemia produced in a semi flow-through culture system were used as food for postlarval shrimp. The growth performance of shrimp reared on such ongrownArtemia live prey is identical to the growth obtained when feeding newly hatchedArtemia. However, a significantly better stress resistance is obtained when the postlarvae are exposed to a low salinity in a stress test. Besides nutritional and energetic advantages, the use ofArtemia biomass for feeding postlarval shrimp also results in improved economics as expenses for cysts and weaning diets can be reduced.     

The use of Artemia sp. or mysids as food source for hatchlings of the cuttlefish (Sepia officinalis L.); effects on growth and survival throughout the life cycle

Twoexperiments were conducted to determine the effects ofArtemia sp. or mysids on growth and survival ofS. officinalis hatchlings, and their effect throughout thelife cycle. For experiment I, for the first 20 days, one group was fed adultArtemia sp. and the other was fed mysid shrimp(Paramysis nouvelli). Eggs laid by females in both groupswere counted and weighed, and hatchlings were weighed, to determine differencesin both groups. For experiment II, during the first 10 days, one group was fedArtemia sp. and the other was fed mysids (P.nouveli). After the period of differentiated feeding, the 2 groupsinexperiment I were fed grass shrimp (Paleomonetes varians)to 70 days old, and dead crabs (Carcinus maenas)afterwards. Cuttlefish in experiment II were fed grass shrimp from day 10 untilthe end of the experiment. For both experiments, hatchlings fed mysids grewsignificantly bigger (p < 0.01) and survival was higher. For experiment I,eggs laid by females fed mysids and the hatchlings born from these eggs werebigger (p < 0.001) compared to the group fed Artemiasp.initially. Individual fecundity was slightly higher for females in the groupfedArtemia sp. (163 eggs female^–1) than forthe group fed mysids (144 eggs female^–1). Egg laying startedatthe age of 125 days and lasted 45 days in both groups. Time between first egglaying day and first hatchlings to be born was 21 days. The last female to die(after spawning) in both groups was 167 days (less than 6 months old).     

大弹涂鱼TLR基因分子进化研究及其在鳗弧菌胁迫后的表达分析

大弹涂鱼(Boleophthalmuspectinirostris)是一种生活在潮间带的淤泥滩和红树林等两栖环境中的鱼类,其免疫系统面临比水生生活更大的选择压力。Toll样受体基因(简称TLR)是重要的先天免疫成员,一直是鱼类分子免疫学的研究热点之一。为了探究大弹涂鱼TLR基因是否因为其独特的生活环境而产生适应性进化以及其TLR基因在受到细菌攻击后的免疫应答模式,本研究从大弹涂鱼皮肤转录组中获得了TLR5, TLR8和TLR9完整序列以及TLR3和TLR7部分序列,采用分子生物信息学对大弹涂鱼TLR5, TLR8和TLR9基因序列以及氨基酸序列进行了分析,并根据所构建的系统发育树对5个TLR基因进行了分子进化分析,采用荧光定量PCR方法对大弹涂鱼5个TLR基因的组织表达分布和鳗弧菌攻击后5个TLR基因的免疫应答模式开展了研究。结果显示, TLR5基因全长3071 bp,包括长度为2646 bp的编码区,共编码882个氨基酸; TLR8基因全长3175 bp,包括长度为3033 bp的编码区,共编码1011个氨基酸; TLR9基因全长3398 bp,编码区长度为3093 bp,共编码1031个氨基酸。大弹涂鱼3个TLR基因与其他物种的TLR基因结构相似,具有高度保守性。位点模型结果表明,鱼类TLR3, TLR5和TLR8是高度保守的,而TLR7和TLR9在长期进化过程中产生了适应性进化;而进化枝–位点模型结果表明,为了适应更加复杂多变的两栖环境,大弹涂鱼TLR9基因可能产生了适应性进化。大弹涂鱼5个TLR基因在8个健康组织(肠,眼,肾,肝,脑,肌肉,脾和皮肤)中均有表达,在肝脏和脾脏中的表达量较高。在受到鳗弧菌(Vibrio anguillarum)攻击后的免疫表达模式表明了大弹涂鱼5个TLR基因在应对细菌入侵时起到了重要作用。     

Evaluation of vitamin C-enriched Artemia nauplii for larvae of the giant freshwater prawn

The effect of high levels of ascorbic acid (AA) delivered through enriched live food has been verified through the successful culture of larval giant freshwater prawn, Macrobrachium rosenbergii. Two successive feeding trials were set up using a control (550 g AA g^–1 DW) and two different AA-enrichment levels in Artemia (1300 and 2750 g AA g^–1 DW). Under standard culture conditions, no differences in growth nor survival could be observed demonstrating that the nutritional requirements are below 550 g AA g^–1 DW, which is the normal level occurring in freshly-hatched Artemia. However, a significantly positive effect could be demonstrated on the physiological condition of the postlarvae, measured by means of a salinity stress test, when vitamin C-boosted live food was administered. Since the AA levels in the predator larvae are linked with the enrichment levels in the live prey, it may be assumed that a positive influence on stress resistance was caused by feeding vitamin C-enriched Artemia. It is expected that under suboptimal conditions, supplementation of high vitamin C levels might also enhance production characteristics.