保存介质和温度对西伯利亚鲟卵子短期保存的影响

研究了不同保存介质(体腔液CF、Hepes液、RMS液)、温度(4℃、16℃)和保存时间(4 h、8 h、16 h、24 h)对西伯利亚鲟卵子短期保存的影响.结果显示:保存介质、温度和时间对卵子受精率、孵化率和初孵仔鱼畸形率有显著性影响(P<0.05),受精率、孵化率均随保存时间的延长而下降,畸形率上升.16℃条件下保存卵子受精率、孵化率和畸形率均高于4℃,但4℃下卵子的保存时间较16℃下长.研究表明,采用根据西伯利亚鲟体腔液生化成分配制的Hepes液作为保存介质,于16℃下保存4 h为西伯利亚鲟卵子的最佳保存条件,此时受精率、孵化率和畸形率分别为86.36%、94.74%和0.     

中华鲟胚胎的耗氧率

应用密闭定容装置 ,对中华鲟胚胎耗氧率进行了研究。结果对尾部接近心脏时期胚胎 ,在不同水温条件下耗氧率测定表明 ,随水温的升高胚胎耗氧率有上升的趋势。在 1 4℃时 ,耗氧率为 0 69mg 1 0 0eggs·min ;而 2 6℃时 ,耗氧率为 1 63mg 1 0 0eggs·min。中华鲟胚胎耗氧率随发育的进程而增大 ,在水源pH为 6 4、水温为 1 9℃条件下 ,受精卵的耗氧率为 0 2 8mg 1 0 0eggs·min ,而到胚胎心脏形成、搏动期开始耗氧率显著升高 ,为 0 87mg 1 0 0eggs·min,到胚体滚动、出膜期时又是一个高峰期 ,为 1 5 4mg 1 0 0eggs·min。     

中华鲟半胱氨酸蛋白酶抑制剂在毕赤酵母中的表达和活性分析

为研究鱼类半胱氨酸蛋白酶抑制剂（Cystatin）的功能并探索其在水产加工和病害防治中的应用潜力,将PCR改造后的编码成熟肽中华鲟（Acipenser sinensis）cystatin 基因亚克隆到毕赤酵母整合型表达载体pPICZαA,氯化锂法转化毕赤酵母菌株GS115,构建表达cystatin的酵母基因工程菌。经甲醇诱导、SDSPAGE检测培养基上清液,表明中华鲟cystatin在毕赤酵母中实现了高效表达,重组cystatin表达量约为215mg·L^-1。纯化后重组蛋白纯度达94.2%。生物活性检测结果表明,1μg重组中华鲟cystatin约能抑制15μg木瓜蛋白酶的水解活性。     

亚麻油替代鱼油对杂交鲟生长、脂肪酸组成及脂肪代谢的影响

为研究亚麻油替代不同水平的鱼油后对杂交鲟(Acipenser baeri Brandt♀×A. schrenckii Brandt♂)幼鱼[初均重(70.8±0.5) g]生长、脂肪酸组成、肝脏及肌肉脂肪沉积以及脂肪代谢的影响, 在油脂添加量为8%的饲料中用亚麻油分别替代0(LO0)、25%(LO25)、50%(LO50)、75%(LO75)和100%(LO100)的鱼油, 配制5种等氮(38.7%CP)等脂(10%CF)饲料。每组饲料随机设3个重复, 养殖周期为12周。结果表明,亚麻油替代100%的鱼油对杂交鲟幼鱼的生长没有显著影响, 而且随着饲料中亚麻油含量的上升, 饲料效率有所提高, 100%鱼油替代组的饲料效率明显高于100%鱼油组的(P<0.05); 但用亚麻油替代鱼油后, 肌肉和肝脏的粗脂肪含量以及血清中谷草转氨酶、谷丙转氨酶和乳酸脱氢酶活性明显升高(P<0.05); 肌肉亚麻酸和n-3多不饱和脂肪酸的含量与饲料中相应脂肪酸组成呈明显的线性相关关系(R2>0.69; P<0.05)。对于杂交鲟的脂肪代谢而言, 亚麻油的添加对血清中的游离脂肪酸、甘油三酯、高、低密度脂蛋白胆固醇的变化产生明显影响, 但亚麻油对血清总胆固醇和酮体影响不显著。考虑到亚麻油完全替代鱼油后, 肌肉中的EPA和DHA这两种长链高不饱和脂肪酸的含量仅下降了不到30%, 因此亚麻油应该是一种比较优质的鱼油替代品。     

达氏鲟精子的主要生物学特性

达氏鲟(Acipenser dabryanus)属淡水定居性鲟鱼类,为我国特有种,主要分布在长江上游干流及金沙江下游。长期人为过度捕捞及其生存环境的持续污染和水利工程的影响,使得达氏鲟自然种群资源遭到严重破坏,其配子质量的下降己经成为限制其规模化人工繁殖成功的关键因素之一,因此为解决达氏鲟规模化人工繁殖过程中存在的关键性技术点,作者从达氏鲟精液基本特征、精浆元素组成以及不同水体和Na+、K+对达氏鲟精子活力的影响、精子超微结构方面入手,对达氏鲟精子的生理生态特性进行了研究。结果显示,达氏鲟精子平均密度为1.52×109个/ml;精浆元素以Na+含量最高,其次是K+,之后为Ca2+、Mg2+、Cu2+、Zn2+,其中Na+、K+、Zn2+在达氏鲟精浆中的含量有极显著性差异(P0.01),Ca2+、Cu2+、Mg2+差异不明显;精子在江水中的活力最高;在Na+浓度为20 mmol/L时,精子活力最高,精子快速运动时间(FT)和寿命(LT)分别为(66.7±7.1)s和(177.0±14.9)s;达氏鲟精子对K+浓度变化较为敏感,在K+浓度为0.05 mmol/L时,精子FT和LT最长,分别为(109.0±16.1)s和(189.3±12.4)s,K+浓度超过0.05 mmol/L后精子FT和LT急速下降,当K+浓度达到0.5 mmol/L以上时,精子活力立即受到抑制;达氏鲟精子细胞核长(5.67±0.20)μm,鞭毛长(63.16±2.79)μm,全长为(70.35±2.92)μm。     

Rapid HPLC analysis of retinoids and dehydroretinoids stored in fish liver: comparison of two lake sturgeon populations

All major retinoids (vitamin A) stored in the liver of lake sturgeon from two Québec lakes were analysed by high-performance liquid chromatography (HPLC). Four HPLC peaks represented approximately 90% of all liver retinoids. Their relative proportion was remarkably consistent in the two sturgeon populations with dehydroretinyl palmitate/oleate accounting for the largest proportion (50–60%). Most retinoids demonstrated significant positive correlations with age, but only in the Lake Berthelot sturgeon. The concentrations of all major retinoids were significantly ( P <0·0001) lower, as much as 40 times lower, in the Lake Saint-Louis population. Possible explanations for the extremely low liver stores in the Lake Saint-Louis sturgeon are discussed.     

How different rearing temperatures affect growth and stress status of Siberian sturgeon Acipenser baerii larvae

Environmental temperature is one of the critical factors affecting fish development. The aim of this study was to examine the impact of three different rearing temperatures (16, 19 and 22°C) throughout the endogenous feeding phase of the Siberian sturgeon Acipenser baerii. This was performed by assessing (a) larval survival and growth; (b) immunofluorescence localization and expression of genes involved in muscle development and growth – myog and Igf1; and (c) stress status through the expression of thermal stress genes – Hsp70, Hsp90α and Hsp90β – and whole body cortisol. Overall survival rate and larval weight did not differ significantly across temperatures. Larvae subjected to 22°C showed faster absorption of the yolk-sac than larvae subjected to 19 or 16°C. Both at schooling and at the end of the trial, larvae reared at 16°C showed significantly lower levels of cortisol than those reared at 19 or 22°C. IGF-1 immunopositivity was particularly evident in red muscle at schooling stage in all temperatures. The expression of all Hsps as well as the myog and Igf1 genes was statistically higher in larvae reared at 16°C but limited to the schooling stage. Cortisol levels were higher in larvae at 22°C, probably because of the higher metabolism demand rather than a stress response. The observed apparent incongruity between Hsps gene expression and cortisol levels could be due to the lack of a mature system. Further studies are necessary, especially regarding the exogenous feeding phase, in order to better understand if this species is actually sensitive to thermal stress.     

Cell culture and karyotype of Sakhalin sturgeon Acipenser mikadoi

A cell culture of rare and threatened species of Sakhalin sturgeon Acipenser mikadoi was established from a fragment of the pectoral fin and neighboring tissues. Initially, the culture consisted of different cell types including typical fibroblasts as well as cells of epithelial origin, myofibroblasts, etc. After approximately five passages, the culture largely consisted of cells with fibroblast morphology. Under normal culture conditions, these cells grew for more than one year at a constant rate and passed about 80 population doublings. In the absence of serum, cells entered the state of proliferative quiescence (G₀-state). When cultured without medium replacement for a long period of time, cells fused to form myofibers of about 1 cm in length. These myofibers could branch and acquired cross striation with time. About forty days after myofibers emerged, they degenerated, lost their shape, detached from the substrate, and finally died. The induction of adipogenic differentiation arrested cell proliferation and introduced lipophilic inclusions formed in a minor fraction of cells. The number of these inclusions was low, and cells with inclusions demonstrated various morphology distinct from typical adipocytes. The induction of osteogenic differentiation gave rise to cells that produce mineralized extracellular matrix and bone nodules. Chromosome analysis revealed a set of chromosomes typical for “high chromosome” sturgeon species. The variation in the chromosome number was very high (mean, 247 ± 33; modal value, 248). The analysis involving AT- and GC-specific fluorochromes has demonstrated that the telomeric and centromeric regions of all chromosomes are enriched in GC content. The distribution of AT- and GC-rich sequences along the chromosomes was heterogeneous. Long chromosomes were preferentially stained by the AT-specific dye, whereas small chromosomes demonstrated brighter fluorescence after 7-amino-actinomycin D staining; in particular, several small chromosomes fluoresced extremely brightly. This work is the first report of cell culture and karyotype analysis of Sakhalin sturgeon.     

人工标志放流中华鲟幼鱼的降河洄游

1998— 2 0 0 2年期间 ,向长江放流人工繁殖中华鲟 2月龄稚鲟 (全长 7 5— 17 0cm) 17 5 2万尾 ,其中 7795 7尾用CWT进行标记 ;14月龄幼鲟 (全长 5 5 0— 98 0cm) 4 0 0尾 ,全部用外挂银牌和CWT双重标记。放流后沿长江及沿海收集中华鲟稚鲟和幼鲟样本 ,4年共回收稚鲟样本 6 4 0 0尾 ,幼鲟样本 13尾 ,检测到携带标记的稚鲟和幼鲟各 13尾。人工放流的幼鲟降海洄游的速度平均达到 2 8 6km/ 2 4h(7 1— 10 0 2km/ 2 4h) ,回捕时离放流点的距离从 346—2 4 5 9km ,平均 16 0 0km ,回捕的标志幼鲟有 4 6 2 %的个体来自海区。初步估算出 1999年和 2 0 0 0年人工放流个体在长江口幼鲟种群中的贡献率分别为 2 2 81%和 0 997%。结果表明 ,人工放流中华鲟稚鲟和幼鲟的生长、洄游及分布与自然种群没有明显差异。放流较大规格的幼鲟有利于提高成活率 ,而目前长江中华鲟种群的补充仍以自然繁殖种群为主。