Cryopreservation of carp sperm

About 70% of eggs of Cyprinus carpio reached the eyed stage in fertility tests with 1 ml of cryopreserved semen added to 1 g of eggs. When 1 ml of semen was added to 100 g of eggs, few eggs reached the eyed stage. In two fertility tests with sperm preserved for 342 days, using 5 g of eggs inseminated with 1 ml of semen, percentages of eyed eggs were 31.5 and 25.5, respectively. In a series of fertility tests, the chemical constituency of the medium in which fertilization was attempted influenced the fertility of cryopreserved sperm.     

Cryopreservation of sperm of common carp, Cyprinus carpio L.

Milt of common carp, Cyprinus carpio L. was cryopreserved in pellet form with the use of 12 extenders. Most efficient were: BE2 original extender (containing 85 mM NaCl, 50 mM KCl, 3 mm CaCl₂, 1 mm MgCl₂ with 10% dimethyl-acetamide (DMA) and 10% addition of hen's egg yolk) and Kurokura et al.'s extender with 15% DMA and 10% yolk (about 73% and 69% of eyed eggs, about 61% and 52% of swim-up larvae, respectively). Within the most effective treatments, survival from the eyed-egg stage to the swim-up stage was similar to that observed in the control group. Survival from the eyed-egg stage to the swim-up stage (percentage of eyed eggs was considered as 100%) was highly significantly and positively correlated with the actual rate of swim-up larvae.     

Cryopreservation of sperm in marine fish

Since the first work of Blaxter in 1953, fish sperm cryopreservation has been attempted on about 30 marine species. The present paper reviews the techniques used and the results published in these species. Particular attention is paid to the handling procedure of sperm before freezing, the problems of semen ageing and semen contamination with urine. The quality of frozen–thawed semen was evaluated using previously standardized biotests, such as a two‐step motility activation technique adapted for the different species and fertilization assays using a discriminating insemination technique. Most extenders used in marine fish are saline or sugar solutions. From the investigated cryoprotectants, dimethyl sulphoxide (DMSO) generally leads to the best results. Cooling rates range from 8 °C to 99 °C min^?1; the thawing rate is generally high. Compared with freshwater species, a high percentage of spermatozoa survives cryopreservation. Therefore, and because of the simplicity of the techniques, the cryopreservation of marine fish sperm is suited for application in aquaculture.     

Cryopreservation of sperm from spotted wolffish

The effects of different concentrations of cryoprotectant (dimethyl sulfoxide; DMSO), cooling rate and straw size on the post-thaw motility of frozen sperm from spotted wolffish, Anarhichas minor, were studied. There was no significant difference in the post-thaw motility of sperm treated with three different concentrations of DMSO (10, 20 and 30%). Similarly, there was no significant difference in the post-thaw motility of spermatozoa when using different freezing rates (i.e. distance of straws from the surface of liquid N₂, 4.7, 5.5 and 7.1°C min⁻¹) and the straw size (0.5 and 1.0 ml) did not affect survival. The cryopreservation of sperm can be used to make up for the frequent lack of sperm and/or the unsynchronised timing of sperm production in spotted wolffish males and the ovulation time in females. The results show that sperm from spotted wolffish can be frozen to secure access to viable sperm, but further experiments are needed in order to reveal the effect of different parameters on the post-thawing mortality and define the optimum conditions for cryopreservation.     

中华人民共和国水产行业标准黄河鲤鱼

本标准规定了黄河鲤[Cyprinus （Cyprinus)carpio haematopterus Temminck et Schlegel]的主要生物学性状、生长与繁殖、生化指标、遗传学特征及其检测方法。     

半滑舌鳎精子冷冻保存

半滑舌鳎精子冷冻保存对于人工繁殖育苗、杂交育种、雌核发育及其性别控制研究具有重要的意义,为此,本文对半滑舌鳎精子冷冻保存方法进行了研究。分别利用2．8mol／L的二甲基亚砜（DMSO）、甘油（Gly）和1,2-丙二醇（PG）冷冻保存该鱼精子。结果显示,DMSO冷冻保存精子的活力较高。利用MPRS＋2．8mol／L DMSO以1：0．5、1：1、1：1．5和1：2的比例稀释并冷冻精子,1：1比例在冻前能够抑制精子的运动,冻后活力可达82．50±3．54％,显著高于其他稀释比例（P〈0．05）。分别利用冷冻保存液A（MPRS＋2．8mol／L DMSO）和B（TS-2＋2．8mol／LDMSO）稀释平衡精子,精子在A中的冻前快速运动时间、寿命分别为37．75±6．45S和145．00±78．98S,与鲜精无显著差异（P〉0．05）。利用以上两种冷冻稀释液冷冻保存精子,精子在A液中的冻后活力和寿命分别可达53．50±6．69％和98．00±13．51s,冷冻效果优于B液（P〈0．05）。冷冻后精子的受精率和孵化率分别为55．00±5．00％和35．00±13．23％,受精率与鲜精无显著性差异（P〉0．05）,因此认为MPRS＋2．8mol／L DMSO可用于半滑舌鳎精子的冷冻保存。     

鱼类精子超低温冷冻保存技术及其应用

鱼类精子的超低温冷冻保存对鱼类种质资源保护、低温生物学、遗传育种和水产养殖业都具有重要的意义。鱼类精子超低温冷冻保存技术主要包括冷冻保存方法、抗冻保护液、降温、解冻速率、冷冻保存温度、抗冻剂去除等过程。本文就鱼类精子超低温冷冻保存技术及其应用进行了综述。     

pH值、温度、渗透压对鲤鱼精子低温保存效果的影响

就pH值、温度、渗透压等理化因子对鲤鱼精子液态低温保存效果的影响进行了初步研究。结果表明,保存液的pH值为弱酸性(pH6.5)时保存效果最好;低温(0~4℃)条件下,鲤鱼精子的保存效果明显比室温(18~25℃)条件下的保存效果好;在1.2%的NaCl溶液中,鲤鱼精子保存的时间较长。     

移动声源对鲤、草鱼的诱引效果

目前我国淡水水域的养殖对象大多为中、底层鱼类 ,仍主要采用传统渔具捕捞。其缺点是受底质、水深、环境以及水流等因素限制 ,要求鱼群较集中 ,劳动作业强度大。为了改进传统的捕捞作业方式 ,本研究利用鱼类的生物学特性 ,参考音响驯化技术的有关参数 ,结合投饵用 4 0 0Hz正弦波连续音移动声源对鲤鱼 (Cyprinuscarpio)、草鱼 (Ctenopharyngodonidellus)进行诱引驯化试验。结果表明 ,最初放声时 2种鱼均产生惊愕反应 ,迅速离声源而去 ;但驯化 6d后 ,鲤、草鱼在放声后迅速游向声源 ,聚集率均为 10 0 %。经移动声源驯化 6d后 ,鲤、草鱼能够跟随声源迅速移动 ,跟随率均为 10 0 % ,平均跟随时间分别为 19.8s和 2 0 .2s。t检验证实 ,2种试验鱼跟随时间无显著差异 (a =0 .0 5 ) ,说明鲤、草鱼对移动声源的诱引反应敏感。本研究成果旨为音响驯化技术在淡水渔业中的应用及新渔具渔法的研制开发提供更进一步的科学依据     

Cryopreservation of summer flounder,Paralichthys dentatus L., sperm

The summer flounder, Paralichthys dentatus L., is a high‐value species and considerable research has been conducted to determine practices conducive for its culture. As milt can be limited in this species, experiments were conducted to develop a practical sperm cryopreservation protocol for hatchery use. Two dilution ratios (1:2 and 1:4; sperm:extender), 2 diluents (saline and sucrose‐based), 2 cryoprotectants (10% DMSO and 12% glycerol) and 3 freezing rates (?5, ?10 and ?15°C min^?1) were evaluated using differential staining to assess post‐thaw sperm survival. Seven combinations of the factors examined reduced post‐thaw viability by less than 30%. The average viability of sperm from fresh, pooled flounder milt (67.2 ± 2.9%) was not different from that of thawed milt diluted 1:4 with sucrose diluent (10% DMSO) frozen at ?5°C min^?1 (38.4 ± 7.7%) and fertilization and hatch success were not different in trials using fresh or thawed, cryopreserved sperm. From these experiments a practical sperm cryopreservation method was developed, but further refinement of the freezing protocol is necessary to optimize results.     

Cryopreservation of sea cucumber Apostichopus japonicus (Selenka) sperm

A simple and convenient method for the cryopreservation of sea cucumber Apostichopus japonicus (Selenka) sperm was tested in the present study. The highest motility (76.7±2.9%) of post‐thawing sperm was obtained in 15% dimethyl sulphoxide (DMSO) with a 1:9 dilution (semen volume to DMSO volume) when 0.5 mL semen–DMSO mixture was frozen at 6 cm above liquid N₂ in a closed styrofoam box. After thawing, sperm cryopreserved in glycerol almost lost motility entirely. Although there was no significant difference in percentage of motile sperm between 15% and 20% DMSO, the duration of sperm motility of 15% DMSO group was longer than that of 20% DMSO group. The motility of post‐thawing sperm enhanced when the dilution ratio of semen increased from 1:1 to 1:9. Morphological changes such as the loss of mitochondria, swollen plasma membrane and broken or rolled‐up tails were observed in post‐thawing sperm using an eosin–nigrosin staining. The fertility of cryopreserved sperm was significantly lower than that of unfrozen sperm. The 10‐fold increase in sperm to egg ratio resulted in double fertility for cryopreserved sperm, and about 70% fertility relative to the control.     

鲤鱼的故事和传说

鲤鱼是淡水鱼中品种最多、分布最广、养殖历史最悠久、技术最成熟、产量最高的鱼类.我国是鲤鱼的发源地,也是最早开展养殖的国家.     

禾花鲤池塘养殖技术

禾花鲤属鲤科温水性鱼类,其体型粗壮,全身略带紫色(乌褐),背部黑色,色彩亮丽,因以稻田禾花为食而得名。为了探索禾花鲤高效养殖新模式,笔者于2001年在广西区桂林市水产研究所九美桥基地进行了池塘养殖试验,结果表明:禾花鲤池塘养殖周期短,池塘利用率高,鱼种投资少,养殖效益较好。现将禾花鲤池塘养殖技术总结如下,以飨读者。一、池塘条件池塘面积5亩~10亩为宜,水深1.5m~2.0m,水源充足,水质好,排灌方便,进水口设有拦野杂鱼设施,池底平坦,并备有增氧机。二、池塘准备用生石灰75kg/亩~100kg/亩,池塘水深10cm时干法清塘,以杀死池塘内的病原体、野…     

瓯江彩鲤网箱养殖技术

瓯江彩鲤原产于浙西南山区的稻田、坑塘中,俗称田鱼,体色丰富多彩,有红、花、白、粉、青等多种。瓯江彩鲤在当地有悠久的养殖历史,它食性广,生长速度快,同时,它又是一种广温性鱼类,最适生长温度在23℃~28℃之间,全国各地均能养殖;此外,该鱼具有抗病力强、耐低溶解氧、性情温顺、喜集群等特点,非常适合于小体积网箱高密度养殖。瓯江彩鲤与普通鲤鱼的网箱养殖投入相当,其市售价格却是普通鲤鱼的两倍,且市场畅销,经济效益十分可观,极具推广价值。现将有关瓯江彩鲤的网箱养殖技术介绍如下:一、网箱的制作及设置网箱用3×3聚乙烯网线编织的网片,…     

鲤鱼养殖技术之四 瓯江彩鲤池塘养殖技术

瓯江彩鲤俗称田鲤,具有较高的经济价值和观赏价值,仅浙江省丽水地区为其主要产区,且以稻田养殖或池塘混养为主要养殖模式.近年来,浙江省杭州市余杭区水产技术推广站在杭州市余杭区五常乡开展了瓯江彩鲤的池塘精养试验,通过低、中、高的三种放养密度模式对比试验,摸索出了一套较有实用性的瓯江彩鲤池塘养殖技术.     

鲤鱼的遗传连锁图谱（初报）

建立了鲤鱼的遗传连锁图谱。图谱有ＲＡＰＤ分子标记５６个,鲤鱼的ＳＳＬＰ标记２６个,鲤鱼ＳＳＬＰ标记１９个,斑马鱼的ＳＳＬＰ分子标记７０个,鲤鱼基因标记９１个,共有标记２６２个;图谱有５０个连锁组,连锁图给出鲤鱼的基因组大小在５７８９ＣＭ左右。     

Cryopreservation of scaly carp (Cyprinus carpio) sperm: effect of different cryoprotectant concentrations on post-thaw motility, fertilization and hatching success of embryos

The aim of the present study was to determine the effect of various cryoprotectants on post-thaw sperm quality and fertilizing capacity of cryopreserved scaly carp (Cyprinus carpio) semen. The present study focused on freezing of scaly carp sperm utilizing a practical and inexpensive protocol for aquaculture. Semen was diluted with Kurokura’s extender composing 3.6 g/l NaCl, 10 g/l KCl, 0.22 g/l CaCl₂, 0.08 g/l MgCl₂ and 0.2 g/l NaHCO₃. The extender contained three different cryoprotectants (DMSO, DMA and egg yolk) at ratios of 5, 10 and 15 %. Semen was placed into 0.25-ml straws and exposed to liquid nitrogen vapor (?120 °C) using an insulated box with an adjustable tray for 10 min and then plunged into liquid nitrogen (?196 °C) tank. The thawing process was performed in a water bath at 40 °C for 10 s. The results indicated that type of cryoprotectants and their concentrations are rather effective in scaly carp sperm cryopreservation on post-thaw sperm quality, while they are very important in order to obtain high fertilization rates. The highest fertilization rate was determined as 96.4 ± 0.15 % with 15 % egg yolk, while the highest hatching rate was determined as 99.3 ± 0.80 with 15 % DMA. In conclusion, the applied cryopreservation method for scaly carp sperm is suitable to fertilize high amounts of eggs.     

Cryopreservation of filefish (Thamnaconus septentrionalis Gunther, 1877) sperm

The present study examined the possibility of long‐term storage, by cryopreservation in liquid nitrogen, of the sperm of filefish (Thamnaconus septentrionalis). Changes in motility, survival rate, ultrastructure and fertilization rate of the sperm after freezing and thawing were tested. For selection of the immobilizing solution, artificial seawater (ASW) of 250, 350 and 450 mOsmol kg^?1 were tested. Sperm motility was significantly inhibited in 350 mOsmol kg^?1 ASW, and restored entirely after 100% ASW (1200 mOsmol kg^?1) was added. Two cryoprotectants, dimethyl sulphoxide and glycerol, were employed. The sperm was diluted at the ratio of 1:6 with the extenders, and frozen at a freezing rate of ?40°C min^?1 to ?100°C after equilibration for 10 min at room temperature, followed by plunging into liquid nitrogen. The highest post‐thawed sperm motility and survival rate were obtained with 5% glycerol. Afterwards, the effect of different freezing rates was examined using 5% glycerol as a cryoprotectant, and the rate of ?30°C min^?1 to ?100°C showed the best result.     

Effect of albumin and casein supplementation on the common carp Cyprinus carpio L. sperm motility parameters measured by CASA

Adhesion of sperm to a glass surface can be manifested by the low percentage of motile spermatozoa and lower speed. This crucial factor could disprove the sperm quality prediction from computer-assisted sperm analysis (CASA) results. In our study, this was particularly observed when solution L was used. However, protein supplementation was effective in the reduction in sperm adhesion. We found that albumin increased the sperm motility at concentrations of 0.25 % in solution L and 0.5 % in solution P. Casein was effective in both solutions at 0.25 % concentration. However, in solution L, the motility was lower than that measured in solutions P and L with the supplementation of BSA at 0.25 and 0.5 %. BSA supplementation was especially beneficial in terms of higher speed values regardless of an activation solution. Casein used with solution P allows us to obtain similar results as in the samples activated with BSA supplementation. Casein supplementation to solution L did not increase CASA results to the level observed in the samples activated with BSA added to the solution L. Our data suggest that the selection of an activation solution is one of the most crucial steps in the experiments requiring CASA data. Depending on the composition, different protein supplements should be used to reduce adhesion of sperm to a glass surface. We recommend the use of BSA at the 0.5–1 % concentration because at this level we obtained the highest CASA parameters in both activation media.     

草鱼与鲢、鲤不同混养模式系统的氮磷收支

采取陆基围隔实验法,选用草鱼(Ctenopharyngodon idellus)、鲢(Hypophythalmichthys molitrix)、鲤(Cyprinus carpio)研究不同养殖模式系统氮磷的收支。实验于2011年5月开始,10月结束,实验周期为6个月,每月定期采样,围隔面积为7 m×7 m。分别测定了草鱼单养(G组)、草鱼–鲢混养(GS组)、草鱼–鲤混养(GC组)、草鱼–鲢–鲤混养(GSC1,GSC2)系统氮磷的输入和输出,并分析了不同养殖模式水体和底泥氮磷的积累情况。主要实验结果为:(1)不同养殖模式下,饵料氮磷的输入是系统氮磷输入的主要途径,分别占总输入的85.54%~93.38%和82.60%~84.26%,其余为放养生物、降水和初始水层。(2)不同养殖模式下,系统氮的输出依次为养殖生物收获、底泥积累、水层积累、围隔布吸附和氨挥发,所占比例分别为62.80%~77.15%、15.19%~27.60%、5.04%~7.71%、1.54%~2.14%和0.22%~0.30%;系统磷的输出依次为底泥积累、水层积累、养殖生物收获、围隔布吸附,所占比例分别为76.46%~80.04%、13.04%~15.14%、4.09%~9.79%、0.71%~1.16%。(3)不同养殖模式下,底泥氮积累量GSC1组和GSC2组显著低于G组、GS组和GC组(P0.05),而磷积累量GSC1组和GSC2组显著低于G组和GS组(P0.05);水体氮磷积累量GSC2组显著低于G组(P0.05)。(4)系统氮磷利用率GSC2组显著高于G组、GC组和GSC1组(P0.05)。实验结果表明,GSC2组(草鱼0.38 ind/m2、鲢0.69 ind/m2和鲤0.55 ind/m2)能有效降低系统氮磷的积累,提高氮磷的利用率,是一种高效清洁的草鱼混养模式。