饲料中胆汁酸对红鳍东方鲀脂肪酸组成及抗氧化能力的影响

为研究饲料中添加胆汁酸对红鳍东方鲀脂肪酸组成及抗氧化能力的影响，以鱼油为主要脂肪源，配制五种等氮饲料，以初始体重14g的红鳍东方鲀进行为期56天的投喂养殖实验。设置适宜脂肪含量(8%)的对照组，并在对照组添加0.02%及0.10%的胆汁酸，分别获得低胆汁酸添加组(BA)和高胆汁酸组(HBA)。另设一高脂组(脂肪含量12.5%，HL组)及高脂+0.02%胆汁酸组(HLBA组)。结果表明，添加胆汁酸的处理组肌肉和肝脏MUFA均低于对照组，其中HBA组显著低于对照组(P<0.05)。肌肉中n-6PUFA含量在HLBA和HL组显著低于其他组(P<0.05)。在适宜脂肪含量的三个处理组间，肝脏中的C20:0、C16:1n-7、C18:1n-9、C18:2n-6、C20:4n-6、C18:3n-3、C18:4n-3、C20:5n-3和C22:6n-3都呈现随着胆汁酸添加量增加而降低的趋势。血清超氧化物歧化酶活性和谷胱甘肽过氧化物酶活性都有随着胆汁酸添加而升高的趋势，且在高脂组显著高于对照组(P<0.05)；血清谷胱甘肽还原酶则呈现相反的趋势。肝脏中谷胱甘肽过氧化物酶1的mRNA相对表达量在对照组显著高于其他组，而BA组氧化还原酶1的基因表达量显著高于对照组；在高脂组添加胆汁酸显著升高了过氧化氢酶的基因表达量。综上所述，饲料中胆汁酸的添加降低了饲料及鱼体组织中一系列长链脂肪酸的含量；胆汁酸对血清抗氧化蛋白活性的影响与其对肝脏抗氧化相关基因表达的影响结果不一致。     

Dietary protein requirement of juvenile obscure puffer,Takifugu obscurus

A 10‐week feeding experiment was conducted to determine the optimum dietary protein requirement of juvenile obscure puffer (Takifugu obscurus). Six isoenergetic (20 MJ kg^?1 gross energy) diets were formulated to contain graded levels of 34%, 38%, 42%, 46%, 50% or 54% crude protein (as dry matter basis). The results showed final body weight, weight gain and specific growth rate (SGR) increased significantly with increasing protein levels up to 42% and then decreased thereafer. Second‐order polynomial regression analysis (y = ?0.0024x² + 0.1788x ? 1.3196, R² = 0.9032) indicated a maximum SGR at protein level of 37%. Feed conversion ratio (FCR) decreased with increasing levels of dietary protein up to 42% and increased thereafter. Second‐order polynomial regression analysis (y = 0.0054x² ? 0.4351x + 10.391, R² = 0.753) indicated a minimum FCR at protein level of 40%. Protein efficiency ratio (PER) of fish fed the 34%, 38% and 42% diets was significantly higher than that of fish fed the 46%, 50% and 54% diets, and broken‐line analysis indicated PER tended to decrease when dietary protein level was higher than 40%. Generally, whole body lipid content, total cholesterol, low‐density lipoprotein cholesterol and triacylglycerol decreased with increasing levels of dietary protein. Fish fed the 42% protein diet showed the highest essential amino acids (histidine, isoleucine, leucine, lysine and threonine) and non‐essential amino acids (aspartic acid and glutamic acid) in muscle. Based on the second‐degree polynomial regression analysis of SGR and FCR and broken‐line analysis of PER, the optimal dietary protein level of obscure puffer is estimated to be between 37% and 40% (% as dry matter basis).     

Molecular Characterization of the Von Willebrand Factor Type D Domain of Vitellogenin from Takifugu flavidus

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val¹¹⁵, ASP¹¹⁶, Val¹¹⁷, and Lys¹²²) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein–toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.     

池养无毒暗纹东方鲀的人工繁殖

研究了温室(18～25℃)池养暗纹东方鲀性腺发育、人工繁殖和河鲀毒素.试验结果表明池养暗纹东方鲀雄鱼成熟系数二、三月份达到最大值分别为0.1480±0.0100和0.1317±0.0l40;三、四月份雌鱼成熟系数达到最大值分别为0.2440±0.0400和0.3058±0.0460.与野生的暗纹东方鲀相比较,经统计检验,P＞0.05,成熟系数无显著性差异.池养暗纹东方鲀不经降海洄游,性腺完全能发育成熟,能进行人工催产并孵化得苗,并且证明活体各部分均无毒或弱毒,这对于大规模开展暗纹东方鲀的人工养殖具有重要意义.     

微卫星标记对红鳍东方鲀繁殖的指导应用及遗传分析

利用已有设计的20个具有多态性的微卫星标记对16尾红鳍东方鲀亲鱼(10尾♂,6尾♀)的有效等位基因数、观望杂合度、期望杂合度、多态信息含量、遗传相似性系数、遗传距离等进行了检测.据此设计了10对亲鱼配组,获得10个子代家系.其中母本中16和8号的平均有效等位基因数和平均期望杂合度较其他亲本高,将其分别与3个父本配组,其他则参照遗传距离进行配组.于次年,对F110个家系的体质量、体长均值进行了比对,发现16♀与11♂配组得到的子代体质量、体长均值为最大.8♀和10♂配组得到的子代体质量、体长均值最小.结果表明,红鳍东方鲀子代杂种优势与亲本个体差异有关.     

红鳍东方纯越冬试验

１９９６年１１月２１日至１９９７年５月２日和１９９８年１１月至１９９９年４月１２日的两次试验结果表明，红鳍东方Ｔｕｎ越冬水温较低或日水温差变化较大会影响成活率，应控制在１６￣１７℃较好，日水温差变化小于４℃，并在自然水温不低于１１℃时就开始升温。体长１００￣１５０ｍｍ，体重６０￣１４０ｇ的个体越冬期放养密度为１８￣３０尾／ｍ＾３。饵料中添加维生素Ｃ和Ｅ、“多维”及抗生素等，成活率为９６．５％，日增     

暗纹东方鲀微卫星筛选和群体遗传多样性分析

为了解长江野生暗纹东方鲀群体遗传多样性现状,采用已发表的16对红鳍东方鲀微卫星引物对暗纹东方鲀基因组DNA进行PCR扩增,结果有10对微卫星引物(占总数62.5%)能扩增出特异性条带。利用这10对微卫星引物对采自长江常熟江段30尾暗纹东方鲀群体进行扩增分析,共获得51个等位基因,每个位点等位基因数在2到10之间,平均为5.1。10个位点平均观察杂合度(Ho)为0.820 0,平均期望杂合度(He)为0.692 8,平均多态信息含量(PIC)为0.630 0。8个位点表现为高度多态(PIC>0.5),2个位点表现为中度多态(0.25相似文献   

苯并芘和V_E对红鳍东方鲀抗氧化酶影响研究

试验结果表明,当水中苯并芘质量浓度为50μg/L时,红鳍东方鲀的超氧化物歧化酶和过氧化氢酶酶活性显著低于空白对照组,最多分别低93.1%和35.4%。说明其体内抗氧化体系受到明显的抑制,且两种酶活性均表现出先抑制后升高的趋势,证明红鳍东方鲀对于苯并芘的胁迫具有一定的自我调节机能。给鱼体注射VE(3个缓解组剂量分别为10mg/尾、20mg/尾和30mg/尾)后,红鳍东方鲀的超氧化物歧化酶和过氧化氢酶酶活性都有一定的提高,表明VE对苯并芘的影响有缓解作用。     

不同投饵率对暗纹东方鲀幼鱼生长的影响

比较投饵率水平分别为3%、4%、5%、6%时,对暗纹东方鲀幼鱼生长、饲料效率的影响,以确定暗纹东方鲀幼鱼阶段的适宜投饵率。在23℃下,对规格为3.45～3.56g的暗纹东方鲀幼鱼进行30d的生长实验,结果表明:不同投饵率对暗纹东方鲀幼鱼的特定生长率、质量相对增加率、饲料效率、存活率均有显著影响(p<0.05);暗纹东方鲀幼鱼的适宜投饵率为4%,在投饵率为4℅时,质量相对增加率为116.04±0.04%,饲料效率达83.70±0.02%,特定生长率为111.49±0.03%/d,存活率为84.44±0.02%。     

暗纹东方鲀sox9基因的克隆和组织表达分析

为初步探究暗纹东方鲀sox9基因在性腺发育和性别分化过程中的作用,通过设计兼并引物、RACE及荧光定量PCR技术,成功克隆出暗纹东方鲀sox9基因的cDNA全长序列,并分析其相应的生物信息学特征及其在雌雄个体的组织表达水平。结果表明:sox9a基因cDNA全长为1 248 bp(NCBI登录号:MH218818),包括684 bp的ORF,编码227个氨基酸,297 bp的5′UTR和267 bp的3′UTR。sox9b基因全长为1 941 bp(NCBI登录号:MH218819),包括1 470 bp的ORF,编码489个氨基酸,306 bp的5′UTR和165 bp的3′UTR。同源性和系统发育分析结果表明暗纹东方鲀sox9基因与红鳍东方鲀同源性最高,亲缘关系最近。氨基酸多重序列比对结果显示两个Sox9氨基酸序列的HMG box结构域在哺乳动物和鱼类中均高度保守。荧光定量PCR分析显示两个sox9基因普遍存在于雌鱼和雄鱼的各个组织中,且均在雌鱼下丘脑中表达量最高,在精巢中有少量表达,卵巢中几乎不表达。总体来说,除少数组织外,两个sox9基因在雌鱼组织中的表达量普遍高于雄鱼。本研究为了解暗纹东方鲀sox9的遗传特性及相关的生理功能,探索性别分化与性腺发育的分子调控机制提供理论依据。