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     

鱼类精液超低温冷冻保存的基本原理、研究现状及应用前景

<正> 利用低温保存生命是一项新兴的科学手段。自1949年英国Polge用甘油冷冻保存动物精液成功以来,这项技术突飞猛进。目前冷冻配子及胚胎已经应用在医学研究和畜牧业生产上。而水产生物配子(主要是精子)的冷冻保存,经过三十年的努力也取得很大成就。从现有资料看,鱼类精液冷冻保存的研究大多集中在冷水性的鲑鳟鱼类以及某些海水鱼类上,结果较为理想。而有关鲤科鱼类精液冷冻保存的研究报道尚不多。冷冻配子具有超越时空的功效,可以使不同生殖期或地理间隔的品系得以交配,性转换的个体得以自交;可以不间断地为水产遗传学及生物技术研究提供配子材料;还可     

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。     

白鲢冻精后代的生长观察

鱼类精液是鱼类自然繁殖及人工授精的基础，其品质的优劣直接关系到后代的性状。近年来，鱼精冷冻保存作为鱼类选种育种的一项新手段为水产科技界所重视，并取得可喜的进展。实验证明，鱼类精液经超低温(-196℃)冷冻保存之后，解冻复苏的精子具有正常的授精能力且胚胎发育正常，     

白鲢冻精后代的生长观察

鱼精冷冻保存作为鱼类选种育种的一项新手段，已为人们所重视，并取得了可喜的进展。实验证明，鱼类精液经超低温(-196℃)冷冻保存之后，解冻复苏的精子具有正常的受精能力，且胚胎发育正常，孵出正常鱼苗。但随着研究工作的深入，人们十分自然地关注到这样一个问题，     

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…     

鱼精低温冷冻贮存、受精与孵化的实验研究

<正> 引言家畜低温冷冻精液的技术与理论已达高度水平,并且走向实用化,参考的资料也比较丰富,而有关鱼类的精液、卵子的低温冷冻,在国内外正处于研究发展中,其目的都是为育种工作提供保存优良品种鱼精。有关精子低温冷冻保存,1954年以鳟鱼、鲤鱼等为材料进行了研究,至今在冷冻方法、稀释保护液、温热解冻以及复苏率,受精孵化等仍存在着许多问题。最近有些日本淡水鱼类养     

鱼类精子冷冻保存研究进展

鱼类精子的长期保存对于遗传育种、生物多样性保护、濒危物种保护、渔业可持续发展具有重要意义,引起了人们越来越广泛的关注和重视.低温生物学的不断研究和发展为精子的长期稳定保存提供了理论和技术方面的支持.近年来鱼类精子的冷冻保存在保存原理和技术方面都取得了极大的成就,实现了多种鱼类精子冷冻保存的技术突破,并对冷冻保存的原理、冷冻损伤机制等方面有了更深刻的认识,在冷冻精子的生产应用方面也越来越趋于成熟稳定.本文仅针对鱼类精子冷冻保存的研究现状及发展前景作以阐述,以期对更加深入广泛的研究工作有所帮助.     

养殖鲑鳟鱼类鱼肉着色技术

1　前言作为鲑鳟鱼类海水网箱养殖实用技术之一 ,我们所实施的海水养殖鲑鳟鱼类鱼肉着色因适应国际市场需求 ,提高养殖生产效益 ,引起企业广泛关注。天然洄游鲑鳟鱼类依赖于来自饵料生物的类胡萝卜素色素虾青素 ,呈现深受消费者喜爱的橙红肉色。要想满足消费者需求 ,养殖洄游鲑鳟鱼类鱼肉必须着色成天色鲑鳟鱼类肉色。因而 ,在洄游鲑鳟鱼类养殖生产中 ,鱼肉着色至关重要。世界主要鲑鳟鱼类养殖国均对这一问题予以高度重视 ,并进行大量研究。我们所做尝试也主要依赖于借鉴世界各国 ,尤其是挪威、美国、日本、智利先进经验。由于鱼类不能合成…     

鲑鳟鱼类低温麻醉运输技术

人们通常所说的鲑鳟鱼类,是一个笼统的概念,它不是特指某一种鱼,而是鲑科鱼类的统称,在我国主要包括虹鳟、大西洋鲑、银鲑等。鲑鳟鱼类只能在低温水域中繁衍、生长,正常生长的上限水温很低,没有明显的下限温度,因此被称为冷水鱼。其中降海型鲑鳟鱼类是指能适应各种盐度,在淡水和海水中均能成活、生长的鲑鳟鱼类。     

气候变化对鲑鳟鱼类栖息环境及相关生物学的影响

鲑鳟鱼类作为对温度和周围环境比较敏感的冷水性鱼类,气候变化尤其是全球温度升高引起环境的相关变化对其影响明显。本文针对国内外有关气候变化对鲑鳟鱼类栖息地及相关生物学影响的研究情况,先从从栖息地环境方面阐述了气候变化对水生生物的影响,又从温度、CO2浓度、降雨量及酸雨的增加、紫外线辐射增强和鱼病的传播等方面探讨气候变化对鲑...     

气候变化对鲑鳟鱼类栖息环境及相关生物学的影响

鲑鳟鱼类作为对温度和周围环境比较敏感的冷水性鱼类,气候变化尤其是全球温度升高引起环境的相关变化对其影响明显。本文针对国内外有关气候变化对鲑鳟鱼类栖息地及相关生物学影响的研究情况,先从从栖息地环境方面阐述了气候变化对水生生物的影响,又从温度、CO2浓度、降雨量及酸雨的增加、紫外线辐射增强和鱼病的传播等方面探讨气候变化对鲑鳟鱼类生物学方面的影响。同时指出目前研究中存在的地域局限性和缺乏评价标准的相关问题,并就存在的问题给出建议,提出了今后重点关注和尚待研究领域。     

乌克兰鳞鲤精子超低温冷冻保存方法研究

为研究适用于乌克兰鳞鲤精子的超低温冷冻保存方法,分析比较3种稀释液[Hank′s、Cortland、Freezefish冻精稀释液,精子与每种稀释液均设置3种比例(1∶1、1∶3、1∶5)]及3种体积分数为10%的抗冻剂(二甲基亚砜、1,2-丙二醇和丙三醇)对乌克兰鳞鲤精子低温(4 ℃)保存活力的影响;运用筛选出的冷冻保护液及稀释比例,分析比较3种“3步冷冻法”以及3种解冻温度(20、30、40 ℃)对乌克兰鳞鲤精子活力的影响。试验结果表明,采用Hank′s作为稀释液,10%二甲基亚砜为抗冻剂,精子与稀释液比例为1∶3,平均降温速率为12 ℃/min,解冻温度为30 ℃时,精子活力最高(>68%)。通过对稀释液、抗冻剂、稀释比例、降温速率和解冻温度的层层筛选,建立了适宜乌克兰鳞鲤精子超低温冷冻保存的方法,在其种质保护方面具有重要意义,为开展其他鱼类精子超低温冷冻保存提供参考。     

Tetraploid rainbow trout produced by cytochalasin B

At the Research Station for Salmonids, Sunndalsøra Unit, tetraploid rainbow trouts have been produced by keeping newly fertilized eggs for a short time in a solution containing cytochalasin B. The purpose of this work is to produce triploid sterile fish by fertilizing normal eggs with sperm from tetraploid males.     

Evaluation of the potential source of bacterial contamination during cryopreservation process of silver barb (Barbodes gonionotus) sperm

Application of good sanitation practice in cryopreservation process is the key issue to improve the quality of cryopreserved fish sperm. This study implemented standard sanitation protocol with minimal contamination and evaluated the source of bacterial contamination associated with laboratory equipment and related materials across a series of cryopreservation process of silver barb (Barbodes gonionotus) semen. The use of 16s rRNA sequencing and traditional biochemical methods were performed for bacterial identification. Animal origin (anal fin, culture water and semen contaminated with faeces and urine) and non‐animal origin (liquid nitrogen from liquid nitrogen dewar, outer surface of straw, air circulation in cryopreservation laboratory and latex gloves used during cryopreservation procedure) were determined for bacterial contamination. Aeromonas punctata subsp. caviae was the most abundant species in anal fin, latex groves and semen contaminated with faeces and urine. Bacillus safensis and Bacillus sp. were found as frequently recovered species from liquid nitrogen dewar. Aeromonas hydrophila subsp. hydrophila and Pseudomonas fluorescens, fish pathogenic bacteria, were isolated from all animal origin samples. This was the first report indicating that standard sanitation and hygiene methodologies are recommended for sperm cryopreservation of B. gonionotus to prevent bacterial contamination.     

Simulation modelling of high‐throughput cryopreservation of aquatic germplasm: a case study of blue catfish sperm processing

Emerging commercial‐level technology for aquatic sperm cryopreservation has not been modelled by computer simulation. Commercially available software (ARENA, Rockwell Automation, Inc. Milwaukee, WI) was applied to simulate high‐throughput sperm cryopreservation of blue catfish (Ictalurus furcatus) based on existing processing capabilities. The goal was to develop a simulation model suitable for production planning and decision making. The objectives were to: (1) predict the maximum output for 8‐h workday; (2) analyse the bottlenecks within the process, and (3) estimate operational costs when run for daily maximum output. High‐throughput cryopreservation was divided into six major steps modelled with time, resources, and logic structures. The modelled production line processed 18 fish and produced 1164 ± 33 (mean ± SD) 0.5‐mL straws containing one billion cryopreserved sperm. Two such production lines could support all hybrid catfish production in the United States and 15 such lines could support the entire channel catfish industry if it were to adopt artificial spawning techniques. Evaluations were made to improve efficiency, such as increasing scale, optimizing resources, and eliminating underutilized equipment. This model can serve as a template for other aquatic species and assist decision making in industrial application of aquatic germplasm in aquaculture, stock enhancement, conservation and biomedical model fish.     

Cryopreservation of sperm from natural and sex-reversed orange-spotted grouper (Epinephelus coioides)

The shortage of males and/or sperm has been an impediment to the aquaculture of orange-spotted grouper (Epinephelus coioides). This study reversed orange-spotted grouper females into males using hormone implants. A cryopreservation protocol for sperm was developed using normal males, and then using similar procedures the cryopreservation of sperm from sex-reversed males was compared. Immature, young and mature female fish were injected with 4 mg kg⁻¹ BW 17α methyltestosterone as implants and the gonad development stage was monitored over a 120-day period. All treated females converted into functional males within 120 days of the experimental period. Younger females (2Y) were all males within 30 days, although not all were capable of fertilizing fresh ova until day 60. The time after injection to sex reversal in immature fish was 50% shorter than in older females. Postthaw fertilization (81%, 82%) and hatching (45%, 47%) of cryopreserved sperm from natural males were the highest in trehalose (15–20%) with 150 mmol NaCl treatment; however, it was less than the control (89% fertilization and 69% hatch). There was no difference in the postthaw fertilization and the hatch percentages between sex-reversed male sperm (64% and 46% respectively) compared with natural male sperm (59% and 49%). The findings of this study suggest the potential use of sex-reversed males and cryopreserved sperm for commercial production of orange-spotted grouper seed for aquaculture.     

A morphological classification proposal for curimba (Prochilodus lineatus) sperm damages after cryopreservation

The present study evaluated some cryoprotectants and concentrations for curimba (Prochilodus lineatus) semen preservation from a simple methodology of analysis of sperm morphology. Semen of nine males was diluted (1:4) into eight cryosolutions, all of them containing 5% of BTS^? Minitüb, still comprising methanol or dimethyl sulphoxide (DMSO) at four concentrations: 5%, 7.5%, 10% and 12.5%. Morphological sperm analysis was performed on an optical microscope at × 1000 magnification. Damages on the head, midpiece and tail rest (the main and terminal portions) were investigated and classified as primary or secondary, according to the assumed influence on fertilization. There was a linear reduction in the total damages with an increase in the cryoprotectant concentration. Samples with DMSO showed the lowest percentage of fractured tails and tail stumps. The cryopreservation protocol retained the fertilizing potential of the sperm after freezing and both methanol 5% and DMSO 7.5% conferred adequate protection to the curimba sperm cells. Sperm morphology, as pointed out here, must be incorporated into routine fish sperm (cryopreserved or not) analysis once sperm defects influence fertilization directly. To the best of our knowledge, this was the first report on curimba sperm damages after cryopreservation by optical microscopy. The classification model presented in this study can be adequate to tackle fish sperm damages.     

Sperm quality and cryopreservation of Brazilian freshwater fish species: a review

The Brazilian freshwater fish diversity is the richest in the world. Only 0.7% of all Brazilian species have had any aspect of their sperm biology addressed up to this date. The majority of the fish species described in this review migrate during the spawning season (a phenomenon known as piracema). Urbanization, pollution, hydroelectric dams and deforestation are some of the causes of stock depletion or even local extinction of some of these species. The knowledge concerning sperm quality and minimum sperm:egg ratio is important to maximize the use of males without reducing hatching rates. Furthermore, sperm cryopreservation and gene banking can guarantee the conservation of genetic diversity and development of adequate breeding programs of native fish species. In this review, we present and evaluate the existing information on Brazilian fish species that have been subject to sperm quality and cryopreservation studies. The following parameters were evaluated: volume of extractable sperm, sperm motility, sperm concentration, freezing media, freezing methods, and post-thaw sperm quality. Although the existing protocols yield relatively high post-thaw motility and fertilization rates, the use of cryopreserved sperm in routine hatchery production is still limited in Brazil.     

Evaluation of Commercial‐scale Approaches for Cryopreservation of White Crappie,Pomoxis annularis,Sperm

Crappie, Pomoxis spp., are popular game fish throughout North America and are produced by public and private hatcheries. However, production is limited by a lack of information on tank culture and induced spawning methods. Development of techniques for storage of sperm and in vitro fertilization would increase flexibility in spawning. Therefore, techniques for sperm cryopreservation were examined in white crappie, Pomoxis annularis. Sperm from adult wild white crappie were used to evaluate sperm extender, cryoprotectant agent and concentration, and cooling technique based on post‐thaw sperm motility. Percent egg fertilization was also compared between sperm stored in the two best cryopreservation protocols and two different osmotic activator solutions. Sperm were cryopreserved using treatment combinations of two extenders (350 mOsmol/kg Hanks' balanced salt solution [HBSS] and 350 mOsmol/kg Ca²⁺free HBSS) and two cryoprotectants (dimethyl sulfoxide [DMSO] and methanol) at concentrations of 5, 10, and 15% that were cooled at four different rates: 5, 10, 20, and 40 C/min. Post‐thaw sperm motility and fertilization rates indicated white crappie sperm can be cryopreserved using either extender, cryoprotectants of either 5% DMSO or 10% methanol, and cooling at 40 C/min. A follow‐up experiment demonstrated sperm in suspensions on ice retained viability after overnight transport.