牙鲆胚胎程序化冷冻保存研究

以牙鲆胚胎为材料,对胚胎程序化冷冻保存的主要环节进行了探讨,以优化牙鲆胚胎的程序化超低温冷冻保存法。单一抗冻剂的毒性试验表明甲醇(Methanol)、1,2-丙二醇(PG)的毒性低于其他抗冻剂的毒性。混合抗冻剂的毒性试验表明,20%~25%PM抗冻剂的毒性很低,经其平衡处理的胚胎,冷冻至-30℃,成活率可达80%以上。植冰前采用2℃/min的降温速率,从植冰后到入液氮前温度采用8℃/min的降温速率效果较好。入液氮前温度的筛选实验表明,入液氮前的温度以-45℃较好。利用20%PM和22%PMP,采取优化的降温程序冷冻保存尾芽期的牙鲆胚胎,分别获得5枚和7枚复活胚胎,并全部孵化出膜。     

鱼类胚胎低温冷冻保存降温速率研究

在低温下采用慢速降温速率，以０．２－０．５℃／分降至－４０℃以上温度，胚胎获得２０％以上成活率。在超低温下采用快速降温速率，以２℃／分降至－４０℃，再以１０℃／分降至－１９６℃，胚胎获得了９０％以上复活率。     

大菱鲆胚胎的玻璃化冷冻保存

Vitrification solutions were examined for their suitability of cryopreservation of turbot (Seophthulnms maximus) embryos. PMPI gave higher survival rate (75.68%) than other vitrification solutions and was considered to be suitable liar the cryopreservation of turbot embryos. The freezing point of the vitrification solutions consisted of PG and MeOH in the proportion of 3:2 was measured when the vitrification solutions were cryopreserved. It showed that the vitrification solutions with a cryoprotectant concentration of over 41% have no freezing point and the freezing points of vitrification solutions containing 33.33%-40% cryprotectants were between -32.4℃ and -65.4℃, The freezing point decreased with the increase of cryoprotectant concentration. The resistance of turbot embryos at different stages to the cryoprotectants was studied, It was found that the turbot embryos from 4-5 pairs somite to tail bud were more resistant to cryoprotectants and suitable to vitrify. A live embryo was obtained after cryopreservation in liquid nitrogen for 14h and was hatched out。     

泥鳅胚胎玻璃化液超低温冷冻保存研究

泥鳅胚胎（胚孔封闭期）用10＃玻璃化液作为低温保护剂，经低温（－196℃）冷冻保存17h后，38℃水浴解冻，解冻后的胚胎获得了成活。在稀释液B1、B2、B3中分别有26．7％，15．0％，3．0％成活的胚胎。解冻后的胚孔封闭期胚胎经过50h的培养，胚胎从胚孔封闭期发育至尾鳍出现期。     

超低温冷冻保存卤虫胚胎的损伤机理↑（*）

通过扫描电镜观察超低温冷冻保存的卤虫胚胎膜结构,查明造成卤虫胚胎损伤的主要机理是：（１）玻璃化液使胚胎膜受到损伤,改变了膜的通透性,玻璃化液进入胚胎内部造成胚胎死亡;（２）在超低温冷冻保存过程中,冷冻物理损伤造成胚胎膜的破裂,导致胚胎死亡。     

哺乳动物 胚胎冷冻研究进展

哺乳动物胚胎冷冻研究始于50年代。该方法的研究进展速度很快。部分家畜的冷冻胚胎已走向商品化阶段。本文着重阐述了胚胎冷冻的方法及其研究进展。     

牛胚胎分割技术研究进展

牛胚胎冷冻的成功,使胚胎移植技术不受时间、地域的限制,并且可以做到对受体进行选择,从而可提高移植妊娠率;牛胚胎分割的成功,是增加胚胎数目和生产同卵双胎或多胎,获得更多牛犊的一个有效方法。如果将胚胎冷冻和胚胎分割技术结合起来,那末就可以达到既增加胚胎数目,降低胚胎移植成本,又利于胚胎移植技术在生产中推广应用的双重目的。     

滩涂地区人工繁殖加州鲈鱼的试验研究

滩涂地区人工繁殖加州鲈鱼的试验研究为了充分发挥江苏省沿海滩涂的土地、水利、技术、人才等优势，大力发展鲈鱼养殖业，我们于１９９５年初开始进行加州鲈鱼在沿海滩涂地区的人工繁殖试验，寻求沿海滩涂地区发展特种水产品养殖的新路子。一、试验材料（一）试验地点及池...     

中华鲟胚胎细胞的冷冻保存及其核移植

为了保存濒危鱼类中华鲟种质资源,对其囊胚细胞和原肠细胞进行了冷冻保存和核移植试验。用二甲亚砜(DMSO)、1,2-丙二醇(PG)、羟乙基淀粉(HES)3种抗冻剂,配制4种冷冻保护液:CP1(12%D)、CP2(10%P)、CP3(8%D 6%E)、CP4(7%P 6%E),冷冻细胞存活率分别为47.4%±4.7%、64.4%±3.6%、54.7%±4.7%、76.7%±5.7%,CP4保存效果最好,说明添加非渗透型抗冻剂羟乙基淀粉能提高冷冻保存细胞的存活率。比较了两种冷冻方法,发现细胞在-7℃平衡30min之后直接投入液氮的一步冷冻降温法是可行的。对不同发育时期的胚胎细胞冷冻存活率进行了比较,结果表明原肠细胞冷冻存活率(64.4%±11.8%)明显比囊胚细胞的(57.1%±11.2%)高(P<0.05)。用冷冻复苏后的囊胚细胞和原肠细胞作供体,以中华鲟未受精卵作受体进行核移植,各移植了469和392枚卵,分别获得了5尾和2尾克隆鱼,核移植成功率分别为1.1%和0.5%。表明可通过冷冻保存胚胎细胞结合核移植技术保存鱼类种质资源。     

应用间接荧光抗体技术快速检测花鲈病原菌——鳗弧菌

以花鲈（Lateolabrax japonicus)弧菌病的病原菌-鳗弧菌（Vibrio anguillarum)W-1为抗原，制备兔抗血清；利用异硫氰酸荧光素标记的羊抗兔免疫球蛋白（FITC-IgG）为荧光标记二抗，并以罗丹明标记的牛血清白蛋白为背景染色，建立检测鳗弧菌的间接荧光抗体快速检测技术。应用该技术对人工感染后的花鲈组织（肌肉、鳃、肠、肾）样品和养殖水体样品进行了鳗弧菌检测，结果显示间接荧光抗体技术不仅可以用于诊断发病的感染花鲈，也可用于检测带菌状态或未发病的感染花鲈。     

Evaluation of silver perch (Bidyanus bidyanus, Mitchell) nutritional requirements during grow-out with high and low protein diets at two feeding levels

Silver perch (Bidyanus bidyanus, Mitchell) of 65 g average weight were reared at high density under controlled conditions on diets containing 24.8% and 40.6% protein. Diets were fed at 2% or 4% of the fish biomass day^?1. Both protein concentrations and feeding level of the diet influenced growth and proximate composition of the fish at the end of the 50‐day growth trial. Energy requirement for maintenance for these fish was found to be 77 cal BW^?0.8 and for each unit of energy retained 3.78 cal BW^?0.8 needed to be supplied, and for each gram of protein retained per metabolic body weight 5.04 g were required. The results indicate that it is possible to obtain the same weight increment when feeding a 24.8% protein diet at a level of 4% body weight day^?1 as compared with only 2% body weight day^?1 of a 40.6% protein diet. This study indicates that the nutritional requirements for early grow‐out are similar to those found in a previous study with juvenile silver perch and adds information concerning the required relationships between dietary amino acids. The results provide information required for further refinement of nutrition for this species.     

Ascorbate dynamics in embryos and larvae of sea bass and sea bream, originating from broodstocks fed supplements of ascorbic acid

The transfer of L-ascorbic acid (AA) from broodstock to the fertilised eggs and it's dynamics in embryos and fasting larvae, has been studied in sea bass (Dicentrarchus labrax L.) and sea bream (Sparus aurata L.). Two types of diets were fed to the broodstocks: one containing sufficient ascorbate for normal growth and the other with an extra addition of a very high dose (2,000 mg/kg feed) of ethyl-cellulose-coated L-ascorbic acid.The concentration of AA and total ascorbate (L-ascorbic acid plus dehydroascorbic acid) was detected through several embryo and larval development stages, including larvae just before feeding. In sea bass and sea bream, the mean (SD) concentration of total ascorbate in fertilised eggs originating from broodstock fed AA supplemented diet, was 218.5 (17.7) and 122.4 (5.1) g/g wet weight respectively. This was significantly higher (P < 0.01) than the unsupplemented groups, which contained only 155.9 (6.9) and 103.9 (3.5) g/g wet weight respectively. A diet with a vitamin C content adequate for normal growth, may not be sufficient for broodstock when the goal is transfer of TAA to embryos. © Rapid Science Ltd. 1998     

The effect of predation on stunted and nonstunted white perch

Gosch NJC, Pierce LL, Pope KL. The effect of predation on stunted and nonstunted white perch.
Ecology of Freshwater Fish 2010: 19: 401–407. © 2010 John Wiley & Sons A/S Abstract – Predation is widely regarded as a means to prevent or minimise the establishment of a stunted (high density of slow growing individuals) population. We investigated the effect of predation on two different white perch Morone americana populations (stunted and nonstunted) by examining the stomach contents of piscivorous fishes. White perch and gizzard shad dominated piscivore diets in Branched Oak Lake, whereas white perch dominated piscivore diets in Pawnee Lake. White perch consumed in the stunted population (Branched Oak Lake) were larger and older than white perch consumed in the nonstunted population (Pawnee Lake). Many of the consumed white perch in the stunted population were sexually mature and had the opportunity to spawn at least once. In contrast, all of the consumed white perch in the nonstunted population were sexually immature. Predation may have reinforced the stunting of white perch in Branched Oak Lake through removal of the largest, oldest individuals.     

Oceanographic features that define the habitat of Pacific ocean perch, Sebastes alutus

This study investigates the possibility that oceanographic features could have effects on the variations observed in the apparent abundance of a demersal rockfish, Pacific ocean perch, Sebastes alutus. Simultaneous monitoring of physical variables and indices of fish abundance was achieved by attaching oceanographic equipment to the fishing gear of commercial vessels and sampling the fish catch. It was found that perch prefer a temperature range from at least 4.8C to 6.7C and that their movement patterns are linked to the movement of these temperatures by coastal wind patterns. Perch prefer areas with steep bathymetry, characterized by turbulent activity due to interactions between the local bathymetry and tidal currents.     

饥饿及恢复投饵过程中花鲈肌肉组成及非特异免疫水平的变化

探讨了饥饿及恢复投饵过程中花鲈（Lateolabrax japonicus）（196±38g）肌肉组成分及鱼体非特异免疫水平的变化。实验设S0组（对照组,持续正常投喂）,S5组（饥饿5d后恢复正常投喂）,S10组（饥饿10d后恢复正常投喂）,S15组（饥饿15d后恢复正常投喂）。S0组每隔5d取样,S5,S10和S15组分别在结束饥饿及恢复投饵后每隔5d取样测定。结果显示：饥饿程度对花鲈肌肉组成有显著影响。与对照组相比,S10组饥饿结束时肌肉的总脂含量显著降低;S15组饥饿结束时肌肉的总脂含量和粗蛋白含量都显著降,同时显著增加了肌肉的水分含量。表明饥饿过程中花鲈先动用肌肉脂肪,后动用肌肉蛋白。在恢复投饵过程中,S10和S15组总脂含量表现出先降后升的规律,而S15组蛋白表现为逐步回升的趋势。表明花鲈在饥饿后恢复投饵的过程中,先恢复肌肉蛋白含量,后恢复肌肉脂肪含量。饥饿15d时花鲈血清蛋白浓度显著降低。与对照组相比,S5组血清、脾脏和头肾溶菌酶活性均无显著变化;S10组头肾溶菌酶活性在恢复投饵10d时补偿性升高。S15组在饥饿结束时显著降低了花鲈头肾溶菌酶活性,但在恢复投饵5d时恢复到对照组水平。花鲈白细胞的吞噬活性也受饥饿程度及恢复投饵的影响。饥饿会降低花鲈血液白细胞的吞噬活性。在恢复投饵过程中,饥饿（5d）后恢复投饵会引起白细胞吞噬百分率和吞噬指数的补偿性升高。表明饥饿及恢复投饵也影响花鲈的非特异免疫水平。     

Composition of the microbial communities in the gastrointestinal tract of perch (Perca fluviatilis L. 1758) and cestodes parasitizing the perch digestive tract

Using the approach of sequencing the V3-V4 region of the 16S rRNA gene, we have analysed the bacterial diversity associated with the distinct compartments of the gastrointestinal tract of perch (Perca fluviatilis) and cestodes (Proteocephalus sp.) parasitizing their digestive tract. The dominant microbiota associated with cestodes (Proteocephalus sp.) was represented by bacteria from the genera Serratia, Pseudomonas and Mycoplasma. By comparing the associated microbiota of perch and cestodes, a clear difference in bacterial composition and diversity was revealed between the community from the stomach content and other parts of the gastrointestinal tract of fish. Microbiota associated with cestodes was not significantly different in comparison with microbiota of different subcompartments of perch (mucosa and content of intestine and pyloric caeca) (ADONIS, p > .05) excluding microbiota of stomach content (ADONIS, p ≤ .05). PICRUSt-based functional assessments of the microbial communities of perch and cestodes indicated that they mainly linked in terms of metabolism and environmental information processing and could play an important role in the nutrition and health of host.     

Effects of larger fish and size grading on growth and size variation in fingerling silver perch

This study presents two experiments addressing growth and size variation in fingerling silver perch (Bidyanus bidyanus. In the first experiment, fish close to mean population size were raised either in the presence or absence of five larger fish for 60 days. Mean specific growth rate (SGR) and increases in the coefficient of variation and skewness were lower in the presence of larger fish, indicating a negative effect of large fish on the growth of smaller ones. In the second experiment, fingerlings were graded into groups smaller and larger than the median size of the population and raised in size-sorted groups of 60 large or small fish and mixed groups of 30 fish of each size category, for 60 days. There was no difference in mean SGR among groups, nor between the mixed group ad the weighted mean of the small and large groups. Biomass gain was higher in the mixed groups than in the weighted small-and-large groups, probably due to a slightly lower survival in the groups comprised of large fish. The fact that the effect of large fingerlings on the growth of smaller ones was evident in the first, but not the second, experiment may be attributed to higher size disparity between large and small fingerlings in the first experiment.     

Induced out-of-season and seasonal tank spawning and stripping of pike perch (Sander lucioperca L.)

Nine trials were carried out on the induced propagation of pike perch in out‐of‐season as well as in the natural reproduction period. From January till May, periodically, 16–20 pairs were transported to the hatchery, and acclimated to 15–16°C. The following hormonal treatments were used to induce reproduction: carp pituitary homogenate (CP) and human chorionic gonadotropin (hCG) alone or in combination; gonadotrop‐releasing hormone ‘Ovurelin’ (D‐Phe⁶‐LH‐RH) alone or with metaclopramid (MTC); and ‘Ovopel’ [containing des‐Gly¹⁰‐(D‐Ala⁶)‐LH‐RH‐ethylamide and MTC]. The effects of daylight regime, water temperature and the length of the pre‐spawning conditioning period on the reproduction performance were determined. From the 130 hormonally treated pairs, 122 either spawned in tanks or were stripped. There were no differences in the ovulation rate between out‐of‐season (93%) and seasonal (91%) propagation. The developmental stage of the eggs correlated with the latency period, which significantly declined from January till April. The latency period was the shortest in fish treated with hCG and, secondly, with CP. The germinal vesicle migration did not proceed without hormonal treatment, although in some trials breeders were kept at the spawning temperature for 1–2 weeks. The results demonstrate that the pike perch can be successfully induced to spawn about 3 months earlier than its natural spawning season, which allows a significantly prolonged yearly supply of fry.