大口黑鲈ghrelin基因SNPs的筛选及与生长性状关联性分析

ghrelin是脊椎动物的一种脑肠肽,有促进摄食的功能,并能促进生长激素(GH)释放,参与能量平衡调控和糖类代谢。为探索ghrelin基因多态性与大口黑鲈生长性状的相关性,针对大口黑鲈ghrelin基因的启动子序列,实验采用直接测序法获得了2个SNPs位点:S1(A-642C)和S2(A-639C)。随机选取同批繁殖、同塘养殖的大口黑鲈采用Sna Pshot方法进行SNPs位点检测和分型。结果显示,实验群体在ghrelin基因2个SNPs位点上基本处于哈温平衡,S1和S2共组成5种双倍型(D1、D2、D3、D4和D5)。基因型与生长性状的相关性分析结果表明,S1位点AC型个体在体高与全长上显著高于CC型个体,S2位点AA型个体在头长上显著高于AC型个体。双倍型D1在体质量、全长和体高方面显著高于双倍型D3,在体质量、体宽和体高方面显著高于双倍型D4。本研究在大口黑鲈ghrelin基因启动子区域获得的与生长性状相关的SNPs标记,可为大口黑鲈分子标记辅助育种提供帮助。     

大口黑鲈形态性状对体重的影响效果分析

对大口黑鲈全长、体长、体高、体宽、眼间距、头长、吻长、尾柄长、尾柄高和体重共 10个性状进行测定，运用相关分析、通径分析和多元回归分析，剔除与体长存在显著共线性的全长、体高、头长，尾柄高及回归方程中不显著的吻长和尾柄长，计算以体宽、体长、眼间距 3个形态性状为自变量，体重为依变量的相关系数、通径系数、决定系数及相关指数，定量分析大口黑鲈形态性状对体重的影响效果。结果显示，3个形态性状与体重的相关系数（0.942，0.979，0.928）均达到极显著水平（P＜0.01）；通径分析中，3个形态性状对体重的通径系数亦达到极显著水平（P＜0.01）,它们是直接影响体重的重要指标,其中体宽（P⁴=0.599）对体重的直接影响最大。所选形态性状与体重的相关指数R²＝0.980 ,说明所选性状是影响体重的主要形态性状。应用逐步多元回归分析建立了以体重为依变量(Y),体宽(X⁴)、体长(X²)和眼间距(X⁵)为自变量的回归方程：LgY=1.065 + 0.765 LgX²＋1.441 LgX⁴+0.543 LgX⁵。以上形态性状对体重影响效果相关数据的获得为大口黑鲈选育测量指标的确定提供了理论依据。     

大口黑鲈POU1F1启动子区域SNPs对生长的影响

采用基因组步移技术克隆了大口黑鲈垂体特异性转录因子POU1F1启动子序列,该序列全长1 629 bp,预测存在4个八聚体转录因子1(Oct-1)结合位点和TATA框等基本转录元件,1个Homeobox转录因子结合位点和2个cAMP反应元件结合蛋白(CREB)结合位点。采用直接测序法研究发现,大口黑鲈POU1F1启动子序列的-18和-183两个位点存在SNPs,该两个突变位点只检测到A、B两种单倍型。对养殖群体进行不同单倍型的基因频率和生长关联分析,结果表明,单倍型A的等位基因频率为0.246,单倍型B的等位基因频率为0.754;群体关联分析表明,基因型为AA型和AB型的个体在体质量、体长、全长、体高和体宽等方面明显高于BB型个体,但基因型为AB型和AA型的个体之间在体质量方面没有明显差异。该研究结果为下一步进行分子标记辅助选择提供了基础。     

大口黑鲈HSC70-1基因多态性及其双倍型与生长性状的关联分析

为了探讨HSC70-1基因对大口黑鲈(Micropterus salmoides)生长性状的影响作用,开发与生长性状相关的分子标记,该研究采用PCR技术扩增得到HSC70-1基因的cDNA和DNA全序列,编码区cDNA序列长1 953 bp,编码650个氨基酸,其DNA全序列长3 390 bp,包含8个外显子和7个内含子。采用直接测序法在该基因上筛选到4个单核苷酸多态性(SNP)位点,分别位于核苷酸序列中的第821、第1 105、第1 200和第2 804碱基处,且处于内含子上。SNP标记与性状的关联分析结果表明,C-821G位点上CC基因型个体的体质量和全长均显著高于CG和GG基因型(P0.05)。A-2804T位点上TT基因型个体的体质量和全长均显著高于AT和AA基因型(P0.05)。其他位点不同基因型个体间的生长性状均不存在显著差异(P0.05)。通过合并基因型发现,双倍型D1 (CC--CCTT)的各个生长性状表现最优,在体高和全长上显著高于D5。C-821G、A-2804T和D1与生长性状显著相关,可作为大口黑鲈分子标记辅助育种的候选标记。     

大口黑鲈EST-SNP标记开发及其与生长性状的相关性分析

为探讨EST-SNP标记的有效性及其与大口黑鲈(Micropterus salmoides)生长性状间的相关性,本研究采用RNA-seq技术进行大口黑鲈生长快个体和生长慢个体肌肉组织的转录组分析,分别检测到15 273和17 376个EST-SNP标记。采用Sna Pshot技术对其中75个EST-SNP标记在大口黑鲈生长性状极端群体中的EST-SNP进行多态性检测,结果显示有37个EST-SNP标记具有多态性,准确性为49.3%。进一步采用一般线性模型分析37个EST-SNP标记与大口黑鲈生长性状的相关性,结果表明:CL1452.Contig9_All-847位点的CC基因型在体质量、全长和尾柄长3个性状的均值都显著高于TT基因型(P0.05),CC型在体高、头长中的均值都比CT和TT型的均值高,但差异未达到显著水平(P0.05),其它位点不同基因型个体间的生长性状均不存在显著差异(P0.05)。其中10个EST-SNP标记在大口黑鲈"优鲈1号"群体中的平均观测杂合度(Ho)和平均多态(PIC)信息含量分别为0.459和0.356,表明大口黑鲈"优鲈1号"群体具有较丰富的遗传多样性。CL1452.Contig9_All-847位点与生长性状显著相关(P0.05),可用于大口黑鲈分子标记辅助育种。     

大口黑鲈肌球蛋白重链基因SNPs的筛选及与生长性状的关联

肌球蛋白是肌肉细胞的重要组成成分,影响肌纤维的组成和肌肉生长。为了研究肌球蛋白重链(MYH)基因多态性与大口黑鲈生长性状的相关性,本研究采用PCR技术扩增得到编码区序列全长为9759 bp的大口黑鲈MYH基因,该基因包含37个外显子和36个内含子,编码1940个氨基酸。采用直接测序法在MYH基因上筛选到8个单核苷酸多态性标记(SNP)位点(A-305G、G-558C、A-2784C、A-2816G、T-4765A、C-6206T、C-6811T和G-6935T),有4个位于外显子上,其中2个属于同义突变。用SNa Pshot方法对从同批繁殖、同塘养殖的大口黑鲈"优鲈1号"群体中随机选取的430尾个体中各位点的基因型进行检测。结果显示,内含子上的A-2784C和A-2816G位点完全连锁,所有位点在大口黑鲈"优鲈1号"群体中的平均有效等位基因数、平均观测杂合度和平均期望杂合度分别为1.635、0.406和0.373,仅C-6206T、C-6811T和T-4765A位点的基因型频率分布符合Hardy-Weinberg定律。采用一般线性模型分析各位点与大口黑鲈生长性状之间的相关性,研究发现,C-6811T位点CC基因型个体的体质量和全长显著大于TT基因型,CC基因型个体的体高和尾柄长显著大于CT和TT基因型,其余位点不同基因型个体间的生长性状均不存在显著差异。C-6811T位点与生长性状显著相关,可作为大口黑鲈分子标记辅助育种的候选标记。     

罗非鱼 MC4R 基因克隆及与其生长相关的 SNPs 位点

黑素皮质素受体-4(Melanocortin receptor-4,MC4R)是5种已发现的黑素皮质素受体家族成员之一,属于跨膜G-蛋白耦联受体超级家族,对于调节动物的采食量、体质量及能最稳态有着重要的作用.本研究采用基因组步移技术获得了尼罗罗非鱼(Oreochromis niloticus)MC4R基因组DNA序列.该基因全长2547 hp,其中5'侧翼序列长为1481 bp,3'侧翼序列长为82 bp,编码区序列长为984 bp,只有1个外显子.采用PCR产物直接测序法、PCR-RFL和CRS-PCR技术对尼罗罗非鱼MC4R基因启动子和外显子区域进行了SNPs位点的检测和分犁,结果共检测到5个SNPs位点(-G1000C、-T974A、C276T、C561T和C708T),其中外显子上的3个SNPs位点均为同义突变.利用一般线性模型分析5个SNPs位点与尼罗罗非鱼生长性状的关系,结果表明,5个SNPs位点不同基因型之间体质量均值差异最大值分别为9.5%、23.2%、14.4%、18.6%和13.5%,没有达到显著水平(P＞0.05).将5个SNPs位点不同基因型组合成7种双倍型(去掉频率小于3%的组合),关联分析表明,在体质量和体长这2个主要生长性状方面,双倍型D4与D1、D3、D6存在显著性差异(P＜0.05),双倍型D2与D3、D6也存在显著性差异(P＜0.05).因而可以考虑将MC4R基因作为影响罗非鱼体质量等生长性状的候选基因,应用于罗非鱼分子标记辅助育种.     

不同大口黑鲈群体形态性状与体重和出肉率的相关性及通径分析

为探究大口黑鲈(Micropterussalmoides)形态性状与体重和出肉率的相关关系,便于通过形态性状分析筛选高体重和高出肉率的大口黑鲈群体,本研究选择国内广泛养殖或选育的4个大口黑鲈品种(美国原种、优鲈1号、优鲈3号、台湾群体)为实验对象,测定了体重(Y)、出肉率(YF)、全长(X1)、体长(X2)、体高(X3)、体厚(X4)、头长(X5)、眼间距(X6)、眼径(X7)、吻长(X8)、尾柄长(X9)、尾柄高(X10),使用相关分析、通径分析、决定系数分析和多元线性回归分析,筛选影响体重和出肉率的主要形态性状,并构建了多元回归方程。结果显示,对4个大口黑鲈群体体重影响最大的性状均为全长(X1)或体高(X3),而对4个大口黑鲈群体出肉率影响最大的性状分别为尾柄高(X10)、尾柄长(X9)、体长/尾柄长(X2/X9)、体重(Y)。美国原种形态性状与体重(Y)、出肉率(YF)的回归方程为:Y=–622.778+21.21X1+45.305X3+16.633X8+17.431X10,Y_F=12.394+3.683X10+24.152X1/X2;优鲈1号形态性状与体重...     

大口黑鲈组织蛋白酶B基因SNP筛选及其与生长性状关联分析

在动物体内组织蛋白酶B(cathepsin B,CTSB)是以酶原形式存在于溶酶体中,能水解多种蛋白,是调控动物生长发育的重要候选功能基因。为开发与大口黑鲈(Micropterus salmoides)生长性状相关的单核苷酸多态性(single nucleotide polymorphism,SNP)标记,本研究对大口黑鲈EST-SNP库中CTSB基因上的SNP位点进行筛选。经PCR扩增和测序分析,从该基因中筛选到1个SNP位点(命名为C+598G),该位点属于错义突变位点,编码的甘氨酸突变成为精氨酸。采用Sna Pshot方法对C+598G位点在165尾大口黑鲈养殖样本中的基因型进行检测,并分析其在群体中的遗传多样性。结果显示,C+598G位点在养殖群体中存在CC、GC和GG3种基因型,其频率分别是40.6%、47.3%和12.1%,C和G等位基因频率分别是64.2%和35.8%;该位点在群体中的观测杂合度(Ho)和多态信息含量(PIC)分别为0.473和0.354,符合哈代-温伯格平衡定律。进一步分析不同基因型与大口黑鲈生长性状相关性,结果表明,3种基因型个体的体质量、全长、体高、尾柄长和尾柄高性状平均值从小到大依次为GG型、GC型和CC型,其中GG型个体在体质量上显著小于CC型个体(P<0.05)。本研究获得的C+598G标记与大口黑鲈生长性状显著相关,可作为大口黑鲈分子标记辅助育种研究中的生长标记。     

刀鲚MSTN基因遗传多态性与生长性状的关联分析

为筛选刀鲚(Coilia ectenes)生长性状遗传改良的分子遗传标记,基于现有刀鲚转录组数据,利用基因克隆技术获得养殖刀鲚肌肉生长抑制素(myostatin,MSTN)基因的全长序列,采用PCR产物直接测序和SNaPshot技术分别进行基因多态性的筛选和分型,并将不同基因型与刀鲚的生长性状(体长、体高和体质量)进行关联分析。结果显示,养殖刀鲚MSTN基因的DNA序列主要由2个内含子和3个外显子组成,经SNP位点筛选,仅在第1内含子上筛选到2个颠换突变位点(g.564G>T和g.755G>T)和1个转换突变位点(g.786C>T);将3个SNP位点与生长性状进行关联分析发现,SNP位点g.786C>T与刀鲚的体长、体高和体质量呈显著性负相关,具有TT基因型个体的体长、体高和体质量显著低于CC型和CT型(P<0.05),而SNP位点g.564G>T和g.755G>T的不同基因型仅与刀鲚的体高具有一定程度的相关性;3个SNP位点对养殖刀鲚的群体遗传分析显示,刀鲚养殖群体的观测杂合度(H O)、期望杂合度(H E)和多态信息含量(PIC)分别为0.280~0.480、0.332~0.455和0.277~0.351,3个SNP位点均属于中度多态(0.25T有望作为剔除劣质刀鲚繁育亲本的候选分子标记。     

鲢、鳙对三角帆蚌池塘藻类影响的围隔实验

以浙江金华汤溪威旺养殖基地的三角帆蚌养殖水体为研究对象,通过围隔实验比较研究了单养鲢、鳙和三角帆蚌的池塘浮游植物密度、生物量和优势种(属)组成等的差异,以及养蚌池混养鲢鳙对水体浮游植物密度、生物量以及优势种变化的影响。结果表明,养蚌(10#)围隔的浮游植物平均密度和生物量均显著高于高密度鲢(12#)围隔(P<0.05),其蓝藻数量及生物量显著高于高密度鲢(12#)和低密度鳙(13#)围隔(P<0.05),绿藻数量则显著低于低密度鲢单养(11#)围隔(P<0.05)。在鱼蚌混养的情况下,单养蚌(10#)围隔浮游植物平均数量显著高于鲢-蚌混养(15#,16#)和鳙-蚌混养(17#,18#)围隔(P<0.05),其蓝藻数量及生物量极显著高于鲢-蚌混养(15#,16#)或鳙-蚌(17#,18#)围隔(P<0.01),其绿藻数量显著低于混养高密度鲢(16#)或低密度鳙(17#)的混养围隔(P<0.05)。研究结果充分说明,鲢、鳙和三角帆蚌三者对水体藻类组成的影响有别,三角帆蚌养殖池中适当混养鲢或鳙可以有效控制蓝藻(铜绿微囊藻)的生长,促进绿藻(四尾栅藻)的生长,并最终有利于三角帆蚌的养殖,混养鲢密度的增加有利于控制藻类生长,而鳙密度的增加促进了裸藻等中大型藻类的生长。     

三种水生动物细胞系对两株蛙病毒敏感性的比较

利用3个不同物种的水生动物细胞系,包括爪蟾肾细胞系(A6)、大鲵胸腺细胞系(GSTC)和鲤上皮瘤细胞系(EPC),分别用沼泽绿牛蛙蛙病毒(RGV)和大鲵蛙病毒(ADRV)感染,进一步研究细胞病变显微形态、病毒滴度、细胞病变与不同感染时间的相关性等。结果显示,在光镜下可见感染病毒的细胞发生病变,A6和EPC细胞肿胀或破裂;GSTC细胞收缩或聚在一起形成多层。同种水产动物细胞系对不同蛙病毒的敏感性不同,在A6、EPC和GSTC细胞中,RGV的滴度分别为10~(3.6)、10~(5.9)和10~(6.6) TCID_(50)/m L;ADRV的滴度分别为10~(4.3)、10~(5.4)和10~(6.1) TCID_(50)/m L,表明GSTC细胞系对两种蛙病毒都更敏感。研究为后续蛙病毒致病机理提供了有用的信息和重要实验材料。     

Growth of rainbow trout fed a diet supplemented with earthworms,after chemical treatment

Whole frozen earthworms (Eisenia foetida) were evaluated as a partial replacement for commercial pellets for rainbow trout, Oncorhynchus mykiss. Earthworms were blanched and treated with concentrated sodium chloride (10%). Fish were submitted to four treatments: control fish were fed with a commercial diet and three groups of fish were fed with diets partially supplemented with earthworms. All fish showed the same appetite for the earthworm-supplemented diets as for the control diet during the course of the experiment (8 weeks). No significant differences (p > 0.05) were detected in the mean final body weights of all groups of fish. A significant decrease was found in the whole-carcass lipid content of fish fed diets containing 25%, 50% and 75% frozen earthworms. The results on growth rate and feed utilization efficiency of fish fed diets containing high levels of whole frozen worms suggested an adverse effect of worm incorporation, probably due to dietary energy/protein imbalance.     

南海大眼金枪鱼和黄鳍金枪鱼的群体遗传结构

测定了南海西沙和南沙群岛附近海区(11～12°N,15°N;110～112°E)黄鳍金枪鱼61尾(17尾成鱼、44尾幼鱼)和大眼金枪鱼26尾(22尾成鱼、4尾幼鱼)的线粒体基因组控制区部分序列(D-loop),结合GenBank数据库中印度洋、太平洋和大西洋群体的同源数据,分析结果:(1)黄鳍金枪鱼与大眼金枪鱼均具极高的单倍型多样性(Hd>99%),聚类树及群体间分化指数(FST和Snn)表明大眼金枪鱼群体分化程度明显高于黄鳍金枪鱼群体;(2)大眼金枪鱼和黄鳍金枪鱼的南海群体与印度洋群体之间基因流最强(Nm=51.638和261.280 10),其次为太平洋群体(Nm=10.868 8和-50.801 81);(3)黄鳍金枪鱼和大眼金枪鱼都基本服从群口扩张模型,而mismatch分布分别呈单、双峰,其中大眼金枪鱼的南海群体扩张较晚(Tau=7.902)且最为明显(θ1/θ0=99 999/14.752)。     

Growth and age at first maturity in turbot and halibut reared under different photoperiods

The effect of extended photoperiods on growth and age at first maturity was investigated in 166 (79 females and 87 males) individually tagged Atlantic halibut Hippoglossus hippoglossus and in 114 (50 females and 64 males) individually tagged turbot Scophthalmus maximus. The halibut were reared at 11 °C on four different light regimes from 10 February to 6 July 1996: simulated natural photoperiod, (LDN), continuous light (LD24:0), constant 8 h light and 16 h darkness (LD8:16) and LD8:16 switched to continuous light 4 May 1996 (LD8:16–24:0). From 6 July 1996 to 9 February 1998 the LD24:0 and LD8:16–24:0 were reared together under continuous light and the LDN and LD8:16 together under natural photoperiod. The turbot were reared at 16 °C on three different light regimes: constant light (LD24:0), 16 h light:8 h darkness (LD16:8), or simulated natural photoperiod (LDN). After 6 months on the different photoperiods, the turbot was reared together on LDN for approximately 12 months until first maturation. Juveniles subjected to continuous light (halibut) or extended photoperiods (halibut and turbot) exhibited faster growth than those experiencing a natural photoperiod or a constant short day. Moreover, when the photoperiod increased naturally with day-length or when fish were abruptly switched from being reared on short-day conditions to continuous light, a subsequent increase in growth rate was observed. This growth enhancing effect of extended photoperiods was more apparent on a short time scale in Atlantic halibut than in turbot, but both species show significant long-term effects of extended photoperiods in the form of enhanced growth. In both species lower maturation of males was seen in groups exposed to extended or continuous light compared to LDN and this could be used to reduce precocious maturation in males leading to overall increase in somatic growth. This revised version was published online in August 2006 with corrections to the Cover Date.     

Profitability evaluation of resource allocation for growing redtail and Chinese shrimp

A farming survey was conducted to evaluate the profitability of resource allocation for producing redtail shrimp, Penaeus penicillatus (Alock) and Chinese shrimp, P. chinensis (Osbeck). Data on production costs and total revenue according to different harvest sizes of shrimp species were collected based on 108 individual ponds operated by 25 surveyed shrimp farmers. The results indicated that growing Chinese shrimp to a marketable size of 12 g was the most profitable management strategy. The profits could be maximized by integrating small-scale farms to a 12.41 ha optimal farming area when the available resources remained the same as those for a 10 ha farm.     

Comparative digestive enzyme ontogeny in two marine larval fishes: Pacific threadfin (Polydactylus sexfilis) and bluefin trevally (Caranx melampygus)

The specific activity of digestive enzymes; aspartic and serine protease, collagenase, lipase, acid and alkaline amylase, acid and alkaline phosphatase, and chitinase was assayed throughout early development in two species of marine fishes: the Pacific threadfin (Polydactylus sexfilis) and bluefin trevally (Caranx melampygus). Specific enzyme activities were determined on whole larval extracts sampled at selected stages of development, from day 0 to day 30 post-hatching. Similar developmental patterns of enzyme specific activity were observed in the two species, although differences in timing, amplitude and effects of first feeding were noted. Amylase activity increased prior to first feeding, peaking at the middle of the larval period, and becoming nearly undetectable by the time of larval-to-juvenile metamorphosis. Serine protease, collagenase, lipase and alkaline (and acid for threadfin) phosphatase activities increased gradually, followed by sharp increases to a plateau during the second half of larval development. Aspartic protease and chitinase activities, in addition to acid phosphatase (for trevally), were low to undetectable in the first half of development, increasing through metamorphosis. In the trevally only, this group of enzymes exhibited high activity levels at the time of hatching, followed by a precipitous drop. Species-dependent differences in enzyme specific activity at first feeding may have been a result of differences in yolk utilization. These results, taken in the context of earlier reports, lead us to conclude that carbohydrate utilization may play a significant role in the earlier phases of development among some marine larvae, followed by a shift to protein and lipid utilization.     

Reproductive biology,early ontogeny,and effect of temperature on development in wolffish: comparison with salmon

Peculiarities of reproduction, early ontogeny, methods of egg incubation, and influence of temperature on development are compared in wolffish (Anarhichas lupus L.) and in salmonids, mainly Atlantic salmon (Salmo salar L.). Reproductive indices of females are similar in both species, but differ in males due to the small production of sperm in wolffish, a difference explained by internal fertilization in the latter species. The rate of embryonal development until the beginning of differentiation of fin fold at the same temperature is similar in both species, but the period from activation to hatching is more than twice as long in wolffish as in Atlantic salmon. Wolffish hatch at a more advanced stage with a small remnant of yolk sac and begin to feed shortly thereafter. As in Atlantic salmon, eggs and larvae of wolffish have a wide range of temperature resistance except before and after hatching. Based on the difference of life cycles in wolffish and salmon, technologies for their artificial breeding are compared.     

致急性肝胰腺坏死病副溶血弧菌(VpAHPND)自然感染三疣梭子蟹

近年来包括急性肝胰腺坏死病(AHPND)在内的多种新发疫病的流行,使我国甲壳类养殖业遭受了严重的经济损失。为了筛查导致山东潍坊某养殖场中一虾蟹混养池塘内患病三疣梭子蟹感染的可能病原,本研究采用分子生物学检测方法,对三疣梭子蟹样品进行了白斑综合征病毒(WSSV)、传染性皮下及造血组织坏死病毒(IHHNV)、虾血细胞虹彩病毒(SHIV)、致急性肝胰腺坏死病副溶血孤菌(Vp_(AHPND))、虾肝肠胞虫(EHP)、偷死野田村病毒(CMNV)、黄头病毒(YHV)和肝胰腺细小病毒(HPV)等8种病原的检测,并对样品进行了组织病理和原位杂交分析。分子生物学检测结果显示,患病三疣梭子蟹样品呈Vp_(AHPND)阳性,而呈现WSSV、IHHNV、SHIV、EHP、CMNV、YHV和HPV阴性。对样品进行Vp_(AHPND)套式PCR第二轮扩增产物的序列测定、比对和进化树分析,结果显示,扩增产物序列与致病副溶血弧菌质粒上pirA~(vp)毒力基因片段具有99%的同源性,该序列与已报道的多个致病副溶血弧菌PirA聚在进化树的同一主分支上。组织病理学分析显示,患病三疣梭子蟹的肝胰腺小管上皮细胞坏死,心肌纤维呈溶解样病变,鳃丝上皮柱突细胞明显坏死,胸神经节的神经细胞损伤严重,并且这些组织中还可见大量的细胞核固缩现象;原位杂交结果显示,肝胰腺、心肌、鳃组织及胸神经节中的病变部位均存在Vp_(AHPND)探针的蓝紫色杂交信号。以上表明,虾蟹混养池塘中三疣梭子蟹在自然状态下感染了Vp_(AHPND),并导致肝胰腺、心肌、鳃和胸神经节发生了严重病理损伤。本研究首次在养殖三疣梭子蟹中检测到Vp_(AHPND)感染并揭示了感染所致的病理变化,相关结果为揭示Vp_(AHPND)自然宿主种类和养殖三疣梭子蟹病害防控提供了基础信息。     

铜铸熟若蟹中麻痹性毒素成分的分离和鉴定

在分析了日本珊瑚扇蟹铜铸熟若蟹甲壳组织和内脏组织中毒性含量的基础上,对毒素提取液进行分离纯化,并通过液相色谱法对其毒素种类进行分析。毒性分析结果显示,毒蟹个体的含毒量在37 500~116 000 MU范围,远远超过成人中毒致死量(5 000 MU),但个体间的毒含量有较大差异;毒蟹个体的甲壳组织的比毒性(909~1 610 MU/g)总体高于内脏组织(111~576 MU/g)。Bio-Gel-P₂凝胶柱层析纯化结果显示,粗毒素提取液通过反复纯化可以达到纯化目的,毒素的比毒性从4.8 MU/mg上升到30 MU/mg左右;毒素进一步通过Bio-Rex 70离子交换柱层析的分离纯化结果显示,毒素中约有9%为膝沟藻毒素群(GTXs,麻痹性毒素中的一类)。高效液相色谱分析结果显示,膝沟藻毒素群的主要成分为GTX3和GTX2,并含有少量的dcGTX2和dcGTX3。