云纹石斑鱼精子冷冻保存

以云纹石斑鱼精液为实验材料,对精子稀释液、抗冻剂种类和适宜浓度、冷冻保存液进行了筛选。结果表明,利用9g/L NaCl、10g/L KHCO3和10%小牛血清配制而成的稀释液EM1-2适宜于云纹石斑鱼精子冷冻保存,以2ml冷冻管为精子容器,在60L液氮生物保存罐中冷冻保存精子,冷冻解冻精子活力可达56.67%±5.77%,要优于TS-2、ES1-3和其他EM系列稀释液冷冻保存精子活力。利用EM1-2为基础液对抗冻保护剂进行筛选,结果显示,10%~20%的二甲基亚砜(DMSO)和1-2-丙二醇(PG)冷冻保存后精子活力无显著差异(P0.05),其中15%的DMSO和10%PG冷冻保存精子效果最优,解冻后精子活力分别可达54.52%±7.81%和57.24%±3.69%。利用冷冻保存1年的精液与云纹石斑鱼卵进行受精,受精率和孵化率均达到80%以上,与新鲜精子无显著性差异(P0.05)。本研究表明,利用EM1-2配制15%的DMSO或10%的PG可用于冷冻保存云纹石斑鱼精液。在此基础上,建立了精子冷冻库,保存精子130ml,为人工繁育和杂交育种提供了丰富的精子源。     

云纹石斑鱼精子冷冻保存

以云纹石斑鱼精液为实验材料,对精子稀释液、抗冻剂种类和适宜浓度、冷冻保存液进行了筛选。结果表明,利用9g/L NaCl、10g/L KHCO3和10%小牛血清配制而成的稀释液EM1-2适宜于云纹石斑鱼精子冷冻保存,以2ml冷冻管为精子容器,在60L液氮生物保存罐中冷冻保存精子,冷冻解冻精子活力可达56.67%±5.77%,要优于TS-2、ES1-3和其他EM系列稀释液冷冻保存精子活力。利用EM1-2为基础液对抗冻保护剂进行筛选,结果显示,10%~20%的二甲基亚砜(DMSO)和1-2-丙二醇(PG)冷冻保存后精子活力无显著差异(P0.05),其中15%的DMSO和10%PG冷冻保存精子效果最优,解冻后精子活力分别可达54.52%±7.81%和57.24%±3.69%。利用冷冻保存1年的精液与云纹石斑鱼卵进行受精,受精率和孵化率均达到80%以上,与新鲜精子无显著性差异(P0.05)。本研究表明,利用EM1-2配制15%的DMSO或10%的PG可用于冷冻保存云纹石斑鱼精液。在此基础上,建立了精子冷冻库,保存精子130ml,为人工繁育和杂交育种提供了丰富的精子源。     

鱼类精液超低温冷冻保存研究进展

鱼类精液超低温冷冻保存研究是从五十年代开始的,经过三十年的努力已取得很大成就。国外的工作主要是在海水鱼类和鲑科鱼类中开展的,并在某些鱼类,如鳕鱼、虹鳟、鲑鱼,大马哈鱼等的精液冷冻保存中获得成功。不少学者还对精液冷冻的原理,冷冻保存的技术环节如稀释液的配制、抗冻剂的种类及浓度、降温平衡、冷冻速度及保存方法等进行了较为详细的研究。我国在鱼类精液超低温冷冻保存方面的工作开展得比较晚。近年来,广东、广西和新疆等地对草、鲢、鳙、鲮等鱼的精液冷冻保存作了一些     

解淀粉芽孢杆菌和胶红酵母复合菌对虹鳟生长性能及胃黏膜、肠黏膜菌群结构的影响

为研究饲料中添加不同水平的复合菌剂(解淀粉芽孢杆菌,Bacillus amyloliquefaciens V4和胶红酵母,Rhodotorula mucilaginosa)对虹鳟(Oncorhynchus mykiss)幼鱼生长及消化道黏膜微生物菌群结构的影响,选用体重为(205.1±4.82)g的虹鳟幼鱼360尾,随机分为4组(每组3个重复,每个重复30尾),分别投喂基础饲料(C0)和3种添加水平为5×10~6/5×10~7 CFU/g(T1),1.5×10~7/1.5×10~8 CFU/g(T3),2.5×10~7/2.5×10~8 CFU/g(T5)的复合菌剂(B.amyloliquefaciens V4/R.mucilaginosa),实验周期42 d。研究结果发现饲料中添加复合益生菌对虹鳟的生长及存活有一定的促进和提高,T1比例的复合益生菌能够显著提高虹鳟的增重率和特定生长率、显著降低饲料系数(P0.05),同时T1和T3比例添加显著降低虹鳟的死亡率(P0.05);对其消化道黏膜细菌群落16S rDNA进行聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)指纹分析,结果表明:虹鳟胃黏膜和肠黏膜上微生物菌群种类存在差异;胃黏膜菌群DGGE图谱中分别检测到35.7±17.0(C0)、37.0±3.5(T1)、36.7±13.6(T3)、26.0±13.2(T5)条谱带,各处理组间谱带数目无显著差异(F=0.500,P=0.692),肠黏膜菌群DGGE图谱显示分别检测到23.3±5.8(C0)、22.3±3.2(T1)、16.7±8.0(T3)、24.7±7.4(T5)条谱带,各处理组间谱带数目也无显著差异(F=0.916,P=0.475);胃黏膜菌群多样性随着益生菌添加量增加,菌群多样性有升高趋势,但是在最高浓度组(T5)多样性降低,肠黏膜菌群多样性随着复合菌剂的添加,多样性指数持续降低,中浓度添加组(T3)多样性最低,但是随着添加浓度升高,呈现恢复和升高趋势(T5);基于所得PCR-DGGE指纹图谱中谱带丰度值数据的UPGMA聚类和PCA排序分析均显示胃黏膜微生物群落与肠黏膜微生物群落结构差异明显,大致分为两个不同的分支,胃黏膜和肠黏膜微生物菌群并没有按照不同处理组而有显著分化。以上结果表明解淀粉芽孢杆菌和胶红酵母复合添加能显著促进虹鳟的生长,提高存活率,外源益生菌添加对虹鳟消化道黏膜上优势菌群能产生一定影响,但并未对胃黏膜及肠黏膜菌群多样性产生显著影响,同时也未显著改变虹鳟肠黏膜微生物菌群结构,高比例添加降低消化道黏膜细菌数量及多样性风险。     

鱼类精液冷冻保存技术操作规程

经过十多年的努力，完成了我国主要淡水养殖鱼类，如青鱼、草鱼、鲢鱼、鳙鱼。兴国红鲤、镜鲤、团头动，以及大口鲶、长吻既和中华鳄等十余种鱼类基础生物学的多项研究，超低温冷冻保存技术和批量冷冻保存技术研究，达到生产上实际应用水平。在长江水产研究所建成我国第一座淡水渔类冷冻精液库，冷冻精液的解冻复活率、解冻后冻精受精率和孵化率分别达到（60－75％。80-95％和75－90％。为促进我国鱼类精液冷冻保存技术的进一步发展，使淡水养殖鱼类精液冷冻保存规范化、标准化，并逐步在渔业生产上推广应用，特制订本规程。l亲鱼的选择和精…     

三倍体虹鳟饲料虾青素添加策略探讨

选择17万余尾平均体重约1 kg的三倍体虹鳟(Oncorhynchus mykiss)进行饲料虾青素水平对三倍体虹鳟生长性能、出成率和肌肉着色影响的中试试验。试验设计3种饲料配方，分别在同一商业饲料配方基础上添加20、30 mg/L和40 mg/L的虾青素。在周长100 m的圆形商业网箱中进行为期10个月的养殖投喂，每种饲料投喂3个网箱。结果表明：添加40 mg/L虾青素饲料组三倍体虹鳟增重率和饲料系数方面均表现出优势；添加40 mg/L虾青素饲料组三倍体虹鳟肝脏小，肝色好，肥满度高；添加40 mg/L和30 mg/L虾青素饲料分别在商品鱼上市前七个月和九个月进行投喂，肉色即可达到标准(SalmoFanTM值>28)。     

二倍体和三倍体虹鳟精液低温保存寿命的研究

与二倍体(2n组)精原液做比较,以不同密度为组别,研究了4℃下两组(3n-HD和3n-LD)三倍体虹鳟(Oncorhynchus mykiss)精原液的保存时限。结果发现,随着保存时间的延长,精液中精子的活力和寿命逐渐降低。2n组、3n-HD组和3n-LD组虹鳟精原液中精子分别在保存5d、4.5d和12.5d后失去活力;在7d、15d和14.5 d后死亡,即三倍体虹鳟精原液4℃保存时间约为二倍体的2倍,说明低温短期保存三倍体虹鳟精原液可以取得较好效果。     

条纹锯精液超低温冷冻保存研究

为建立条纹锯精液超低温冷冻保存方法,实验采用计算机辅助精子分析系统（CASA）分析了采用6种抗冻保护剂（GLY[甘油]、DMSO[二甲基亚砜]、PG[丙二醇]、EG[乙二醇]、METH[甲醇]、DMA[二甲基乙酰胺]）在4种浓度（5%、10%、15%、20%,v/v）下对条纹锯精液的冷冻保存效果。结果发现,以HBSS为稀释液,采用程序降温仪分步降温冷冻保存条纹锯精液,37℃水浴解冻后的精子中,15% PG 作为抗冻保护剂的精子运动率最高,达到（93.1±0.9）%,与鲜精差异不显著（P>0.05）,15% PG 作为抗冻保护剂的精子水浴解冻后精子的运动速度最高,平均直线速度、平均曲线速度、平均路径速度分别达到了（88.3±0.3）μm/s、（76.2±0.5） μm/s、（86.7±0.7） μm/s,与鲜精差异不显著（P>0.05）。在不同种类及不同浓度抗冻保护剂保护下,15% PG 作为抗冻保护剂的精子解冻后 1 min内运动率变化与鲜精差异不显著（P>0.05）。研究表明,15% PG为条纹锯最佳抗冻保护剂,可用于条纹锯精液的超低温冷冻保存。     

条纹锯鮨精液超低温冷冻保存研究

为建立条纹锯鮨精液超低温冷冻保存方法,实验采用计算机辅助精子分析系统(CASA)分析了采用6种抗冻保护剂(GLY[甘油]、DMSO[二甲基亚砜]、PG[丙二醇]、EG[乙二醇]、METH[甲醇]、DMA[二甲基乙酰胺])在4种浓度(5%、10%、15%、20%,v/v)下对条纹锯鮨精液的冷冻保存效果。结果发现,以HBSS为稀释液,采用程序降温仪分步降温冷冻保存条纹锯鮨精液,37℃水浴解冻后的精子中,15%PG作为抗冻保护剂的精子运动率最高,达到(93.1±0.9)%,与鲜精差异不显著(P0.05),15%PG作为抗冻保护剂的精子水浴解冻后精子的运动速度最高,平均直线速度、平均曲线速度、平均路径速度分别达到了(88.3±0.3)μm/s、(76.2±0.5)μm/s、(86.7±0.7)μm/s,与鲜精差异不显著(P0.05)。在不同种类及不同浓度抗冻保护剂保护下,15%PG作为抗冻保护剂的精子解冻后1 min内运动率变化与鲜精差异不显著(P0.05)。研究表明,15%PG为条纹锯鮨最佳抗冻保护剂,可用于条纹锯鮨精液的超低温冷冻保存。     

饲料中添加甘薯(Ipomoea batatas)块根与茎蔓对刺参(Apostichopus japonicus)生长和非特异性免疫力的影响

在刺参(Apostichopus japonicus)配合饲料中添加不同比例(10％、20％、30％、40％、50％、60％)的甘薯块根粉与甘薯茎蔓粉,测定了添加2种甘薯饲料成分对刺参生长和非特异性免疫力的影响.结果显示,在50 d实验期间,投喂添加甘薯块根粉饲料的实验组刺参平均体重随实验时间呈上升趋势,实验结束时,添加20％甘薯块根粉组终末体重显著高于对照组(P＜0.05),10％、30％组特定生长率(SGR)与对照组差异不显著(P＞0.05);投喂添加10％甘薯茎蔓粉的实验组刺参SGR与对照组差异不显著(P＞0.05),其余各组SGR均低于对照组(P＜0.05).投喂添加10％、20％甘薯块根粉饲料的实验组刺参体腔液中过氧化氢酶(CAT)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)均显著高于对照组(P＜0.05);投喂添加10％甘薯茎蔓粉饲料的实验组ACP显著高于对照组(P＜0.05),刺参体腔液中的溶菌酶(LZM)、总超氧化物歧化酶(T-SOD)、CAT和AKP与对照组差异不显著(P＞0.05).研究表明,当甘薯块根与茎蔓的添加比例分别在30％和20％以下时,可满足刺参的营养需求,促进或保证刺参的生长,提高刺参免疫能力.     

二倍体和三倍体虹鳟精液低温保存寿命的研究

与二倍体（2n组）精原液做比较,以不同密度为组别,研究了4℃下两组（3n-HD和3n-LD）三倍体虹鳟（Oncorhynchus mykiss）精原液的保存时限。结果发现,随着保存时间的延长,精液中精子的活力和寿命逐渐降低。2n组、3n-HD组和3n-LD组虹鳟精原液中精子分别在保存5d、4.5d和12.5d后失去活力;在7d、15d和14.5 d后死亡,即三倍体虹鳟精原液4℃保存时间约为二倍体的2倍,说明低温短期保存三倍体虹鳟精原液可以取得较好效果。     

Fertility of teleost semen as affected by dilution and storage in a seminal plasma-mimicking medium

Fertility trials were run with fish semen diluted in Fish Extender #6 and stored for up to 24 h at 10°C. After 30 min of storage, rainbow trout (Salmo gairdneri) and northern pike (Esox lucius) semen diluted 1:255 maintained control values of fertility. Brown trout (Salmo trutta) and pink salmon (Oncorhynchus gorbuscha) semen diluted 1:511 also maintained control values of fertility after 30 min of storage. As storage times increased, the dilution ratios that maintained control values of fertility decreased. After 24 h of storage, rainbow trout semen diluted 1:31 and brown trout semen diluted 1:63 maintained control values of fertility. A field trial with rainbow trout semen showed that semen diluted 1:8 maintained control values of fertility after 30 min of storage. Non-motile sperm cells did not significantly (P>0.05) interfere with the fertilizing ability of motile sperm cells. Calculations from the dilution trials on the number of inseminated sperm cells showed that approximately 4000–169 000 motile sperm cells/egg were required to maintain control values of fertility.     

Characterization of rainbow trout milt collected with a catheter: semen parameters and cryopreservation success

Collection of fish milt by stripping risks the danger of milt contamination by urine. This may seriously influence milt characteristics and quality, including usefulness for cryopreservation. Urine contamination of milt may be avoided by using a catheter for sperm collection. The objectives of this study were to provide basic characteristics of milt collected with a catheter, to test the usefulness of this milt for cryopreservation, and to correlate characteristics of fresh and cryopreserved semen with sperm fertility rates. Milt from 25 rainbow trout Oncorhynchus mykiss (Walbaum) males were used. All samples were cryopreserved using the pellet method within 1 h of collection, using 0.6 m sucrose and 10% dimethyl sulphoxide (DMSO) as an extender. Catheterization resulted in semen of very good motility (> 90% motile spermatozoa) and high fertilization rates after cryopreservation (mean fertilization rate 81.8 ± 13.3% of control, at a sperm/egg ratio of 2.4 ± 0.3 × 10⁶). Osmolality of seminal plasma and concentrations of sodium, potassium and magnesium ions had low variability, which suggests that they are important for creating a stable environment for sperm storage in the sperm duct. Higher variability of certain seminal plasma characteristics, such as protein concentration and antiproteinase activity, suggests that these characteristics are related to individual semen features of particular males. A strong correlation of seminal plasma zinc concentration with protein concentration may reflect an importance of zinc in semen biology. Cryopreservation caused a significant release of protein and acid phosphatase from spermatozoa. Our results did not reveal any single characteristic of semen collected by catheter that could be used as a powerful predictor of cryopreservation success, presumably because all samples were of high quality.     

Effect of extender composition and freezing rate on post-thaw motility and fertility of Arctic char, Salvelinus alpinus (L.), spermatozoa

The effects of extender composition and freezing rate on motility and fertility of frozen‐thawed Arctic char, Salvelinus alpinus, spermatozoa were investigated. Three freezing rates, two semen diluents and three cryoprotectants were tested. Semen frozen in 0.3 mol L^?1 glucose diluent with 10% methanol as a cryoprotectant or in a diluent described by Lahnsteiner with 10%N,N‐dimethylacetamide (DMA) resulted in the highest sperm motility. Fertility was the highest for semen frozen in a glucose–methanol extender but was not significantly different than that for semen frozen in Lahnsteiner's diluent with 10% DMA. Dimethyl sulphoxide (DMSO) at 10% was a relatively ineffective cryoprotectant with either semen diluent. Semen frozen at 6 cm above the surface of liquid nitrogen resulted in a higher post‐thaw sperm motility and fertility than semen frozen at 5 cm. The addition of 7% fresh egg yolk to glucose diluent containing methanol or DMSO did not improve the fertility of frozen‐thawed spermatozoa. However, the addition of 7% fresh egg yolk to glucose–DMA extender significantly improved the fertilization percentages of frozen‐thawed spermatozoa. In conclusion, dilution of semen 1:3 in 0.3 mol L^?1 glucose with 10% methanol and freezing 6 cm above the surface of liquid nitrogen (freezing rate of 40±8°C min^?1, mean±SD from ?5 to ?55°C) is a promising protocol for cryopreservation of Arctic char semen.     

Fertility of rainbow trout (Salmo gairdneri) gametes: Gamete viability in artificial media

A series of experiments was performed to test the effect of potassium ions (K⁺) on the storage of rainbow trout (Salmo gairdneri) sperm cells and eggs. Storage of eggs for up to 10 min in saline solutions, extender with K⁺ concentrations of ≤3.4 mM and coelomic fluid did not affect fertility. Fish Extender #6 had a detrimental effect on eggs within 15 s of application and reduced fertility to near zero within 10 min. Variation in the K⁺ concentration indicated that high (≥6.8 mM) concentrations decreased egg fertility while low (≤3.4 mM) concentrations had no effect on fertility. Insemination of eggs before the addition of Fish Extender #6 allowed storage for 14 h. Semen diluted (1:10) in coelomic fluid maintained fertility for 45 s, whereas semen diluted (1:1) in coelomic fluid maintained fertility for 60 min. Fish Extender #6 at K⁺ concentrations between 0 mM and 68 mM had no effect on fertility of semen diluted 1:2.5 after 1 h of storage.     

Extenders and cryoprotectants for cooling and freezing of piracanjuba (Brycon orbignyanus) semen, an endangered Brazilian teleost fish

The aim of this study was to develop a protocol for semen storage of piracanjuba (Brycon orbignyanus) by both cool storage at 4 °C and cryopreservation at − 196 °C. Semen was diluted in some fish semen extenders (Exp. 1) or in extenders combined with the antibiotic gentamycin sulfate (Exp. 2) and stored at 4 °C. Sperm motility was estimated every 24 h. Then, the effects of egg yolk (0 and 5%), cryoprotectants (dimethyl sulphoxide — DMSO, methanol, and methylglycol) and extenders (NaCl 154 mM, BTS™ Minitub and M III™ Minitub) on semen cryopreservation were evaluated (Exp. 3). Semen was added to each of eighteen cryosolutions (2 yolk concentrations × 3 cryoprotectants × 3 extenders), aspirated into 0.5-mL straws, frozen in nitrogen vapor (Taylor-Wharton, CP 300, “dry shipper”) and stored at − 196 °C. Sperm motility was evaluated after thawing at 60 °C-water bath for 8 s. The three cryosolutions that produced the highest post-thaw sperm motility were used again to freeze semen. Post-thaw semen quality was then evaluated under three tests: sperm motility, the percentage of live spermatozoa and hatching rate (Exp. 4). Piracanjuba semen diluted (1:10 total volume) in NaCl 200 mM or in Saad solution (NaCl 200 mM, Tris 30 mM) maintained motility above 35% for as long as 7 days, at 4 °C. Motility of only 7% was observed on undiluted semen after 3 days at 4 °C. There was neither beneficial nor detrimental effect of gentamycin on sperm motility at 250 μg/mL. Egg yolk addition to the cryosolution was beneficial in samples cryopreserved in NaCl 154 mM and in M III™, but detrimental for samples cryopreserved in BTS™. Methylglycol was the most effective cryoprotector compared to DMSO and methanol. Motility and percentage of live spermatozoa were similar among semen cryopreserved in NaCl–yolk, M III™–yolk and BTS™, all containing 10% methylglycol, but lower than fresh control. Hatching rates of eggs fertilized with sperm cryopreserved in NaCl–yolk or BTS™ were higher than for eggs fertilized with sperm cryopreserved in M III™–yolk, but lower than control fertilizations. The semen cryopreservation protocols developed here will be used to set up a gene bank for endangered piracanjuba populations.     

Physiological and Biochemical Determination of Rainbow Trout,Oncorhynchus mykiss,Semen Quality for Cryopreservation

In rainbow trout, Oncorhynchus mykiss, parameters to determine semen fitness for cryopreservation and quality control of cryopreserved semen were investigated. The following parameters can be used to evaluate semen fitness for cryopreservation as they are statistically significant (P < 0.01) correlated to the post-thaw fertilization rate: motility rate of fresh semen (y = 4.996x - 0.0958x² + 0.0006x³ - 5 1.7363); sperm velocity of fresh semen (y = 6.741x - 0.036x² - 268.37); seminal plasma osmolality (y = 0.539x - 125.59); seminal plasma pH (y = -82.768x + 728.133); seminal plasma triglyceride levels (y = 0.069x + 29.863); seminal plasma ß-D-glucuronidase activity (y = -1.112x + 0.0058x² + 82.229); seminal [lasma lactate dehydrogenase activity (y = -0.096x + 0.00006x² + 583.80); spermatozoan acid phosphatase activity (y = -132.51x + 126.38x² + 66.48); spermatozoan adenylate kinase activity (y = 3.474x + 4.925). Quality of deep-frozen semen can be evaluated by motility parameters (P < 0.01): frozen/thawed semen motility rate and post-thaw fertilization rate: y = 1.943x + 28.002; sperm velocity and post-thaw fertilization rate: y = 0.8812x - 0.0059x² + 24.9686.     

Characterization of the testicular semen of the African catfish, Clarias gariepinus (Burchell, 1822), and its short-term storage

Semen of the African catfish, Clarias gariepinus (Burchell, 1822), was investigated with respect to its cellular composition, sperm cell density, maturation grade, motility and fertility. Storage conditions were tested, whereby sperm viability was assessed by measurement of the motility after activation and by fertility tests. Testicular semen differed in its composition, i.e. the sperm density and numbers of spermatids, according to the maturity grade of the testis. Two semen types could be distinguished: semen type I was characterized by high sperm densities and low numbers of spermatids and semen type II had lower sperm densities and higher numbers of spermatids. Two semen types did not differ in motility and fertility (when adjusted for differences in sperm density). During storage, the sperm viability was influenced by the sodium concentration of the storage medium, temperature, membrane stabilizers as bovine serum albumen (BSA) or hen egg yolk, antibiotics and oxygen. Semen viability was maintained best when it was diluted at a ratio of 1:5 in storage solution (150 mmol L^?1 NaCl, 2.5 mmol L^?1 KCl, 1 mmol L^?1 CaCl₂, 1 mmol L^?1 MgSO₄, 20 mmol L^?1 Tris (pH 8.5) and 0.5% BSA or 0.5% hen egg yolk) and stored at 4 °C. Oxygen gassing and addition of antibiotics (1 mg mL^?1 gentamycine sulphate) to the storage solution affected the two semen types in different ways. Antibiotics had no effect on type I semen, but had a positive effect on type II semen. Oxygen gassing had a positive effect on type I semen but a negative effect on type II semen.     

Quality of eggs and spermatozoa of rainbow trout fed an n-3 essential fatty acid-deficient diet and its effects on the lipid and fatty acid components of eggs, semen and livers

ABSTRACT: The effect of an n-3 essential fatty acid (EFA) -deficient diet on spawning and on the lipid and fatty acid contents of eggs, semen and livers of rainbow trout Oncorhynchus mykiss was investigated. Fish were split into two groups and fed either of two diets for a period of 4 months prior to the start of the spawning season. The control group was fed a commercial diet, containing n-3 EFA, whereas the experimental group was fed an n-3 EFA-deficient diet. Fish were 3 years old at the time of spawning. Eggs and semen were stripped off five females and five males from each diet group and cross-fertilized. Two of the five males fed the deficient diet showed a lower sperm motility, resulting in slightly lower mean hatching rates when crossed with eggs of either group. Higher lipid contents in the EFA-deficient diet were reflected in the egg and semen lipid contents, whereas the lipid contents of male livers of both diet groups were higher than those of female livers. In livers and eggs, the main polar lipid was phosphatidylcholine accompanied by phosphatidylethanolamine in sperm polar lipids. The non-polar lipids of eggs were mainly triacylglycerols but in livers and semen, free fatty acids and free sterols were more abundant. Essential fatty acids, namely n-3 highly unsaturated fatty acids and linolenic acid, were generally lower in eggs, semen and livers sampled from the EFA-deficient diet group. These results indicate that the fertilization, eyed and hatching rates obtained from crossings with males fed the EFA-deficient diet were slightly lower because of the dietary effect on sperm motility. Moreover, lipids of eggs, semen and livers of male and female rainbow trout were influenced greatly by their dietary availability.     

Cryopreservation of rainbow trout spermatozoa: The influence of sperm quality,egg quality and extender composition on post-thaw fertility

The presence of 5% to 20% hen's egg yolk in a sucrose-based extender significantly improved post-thaw fertility of cryopreserved rainbow trout (Salmo gairdneri) spermatozoa compared to when the extender without hen's egg yolk was used. However, the degree of increased cryoprotection associated with hen's egg yolk was affected by the quality of the milt. Considerable variation was detected in the performance of various batches of trout eggs used to test post-thaw fertility and the composition of the extender was shown to affect fertilizations differentially with some of the eggs. Despite this variation, the extender containing 10% hen's egg yolk consistently gave high post-thaw fertility in samples of cryopreserved milt (67.3±3.0% S.E.M.) in thirty replicated trials. As such, the method described is reliable for cryopreserving rainbow trout milt and fertilizing small quantities of eggs.