不同年龄和性腺发育阶段圆口铜鱼营养成分组成分析

由于水电梯级开发和过度捕捞等人类活动影响,长江上游珍稀特有鱼类圆口铜鱼（Coreius guichenoti）资源量显著下降。增殖放流是保护该物种的有效途径之一,而人工繁育过程中为圆口铜鱼提供营养全面的饲料对其性腺发育至关重要。通过分析比较不同年龄和性腺发育阶段的圆口铜鱼肌肉营养成分、肌肉和性腺氨基酸组成,可为其人工饲料的开发提供基础数据。结果显示：(1)随着年龄的增长,圆口铜鱼肌肉粗蛋白、粗脂肪和氨基酸含量均呈先降低、后增加的趋势;(2)性腺从II 期到IV 期的发育过程中,圆口铜鱼肌肉粗蛋白、粗脂肪和氨基酸含量均升高;(3)性腺中谷氨酸和亮氨酸在所有氨基酸中的占比较高,分别为9.82%和6.74%,其中10 种氨基酸的含量、必需氨基酸总量和氨基酸总量均显著低于肌肉组织中的含（P<0.05）,而每100 g 蛋白质中脯氨酸（3.19±0.22）g 和丝氨酸（3.56±0.27）g 含量显著高于肌肉组织中的含（P<0.05）。研究表明,圆口铜鱼肌肉营养成分、氨基酸组成与其年龄和发育期有着密切联系。随着性腺发育,其肌肉粗蛋白、粗脂肪和氨基酸含量呈增加趋势,谷氨酸、亮氨酸和脯氨酸在圆口铜鱼性腺发育过程中可能起关键作用。     

长江朱杨江段几种鱼类体内重金属铅、镉和铬含量的研究

2010年7-10月和2011年4月在长江干流朱杨江段采集8种鱼样本,其中瓦氏黄颡鱼(Pelteobagrus vach-elli)、鲤鱼(Cyprinus carpio)、鲇(Silus asotus)、圆筒吻鮈(Rhinogobio cylindricus)、大眼鳜(Siniperca kneri)和鲫鱼(Cyprinus auratus)等6种鱼的3龄样本4～8尾,铜鱼1～4龄样本32尾,圆口铜鱼1～3龄样本38尾。对各样本体内铅(Pb)、镉(Cd)、铬(Cr)的含量进行了检测。结果显示,所有鱼体样本体内均检测到存在这3种重金属离子。其中3龄组鱼体内Pb的平均含量为(1.885～2.731)μg/g,Cd的平均含量为(0.107～0.236)μg/g,Cr的平均含量为(0.514～0.918)μg/g。8种鱼类体内所含Pb和Cd均超过我国食品安全标准,而Cr含量均未超标。对铜鱼和圆口铜鱼不同年龄组间的Pb、Cd和Cr含量进行了比较。Pb在铜鱼3龄组体内的含量与2龄组差异不显著(P>0.05),但显著高于1龄组(P<0.05);2龄组和1龄组之间差异不显著(P>0.05)。Pb在圆口铜鱼3龄组的含量显著高于2龄和1龄组(P<0.05),但2龄和1龄组之间差异不显著(P>0.05)。Cd和Cr在两种鱼各自不同年龄组间差异均不显著(P>0.05)。回归分析显示,Pb在铜鱼和圆口铜鱼体内的含量与体长或体重均存在着极显著正相关关系(P<0.01),Cd的含量与体长或体重之间仅在圆口铜鱼中存在显著正相关关系(P<0.05);而在这两种鱼中Cr的含量与体长或体重之间的相关性均不显著(P>0.05)。结果显示,长江朱杨江段该8种鱼体均受到重金属Pb和Cd污染;Pb在铜鱼和圆口铜鱼体内的含量表现出了随年龄增长而增加的生物积累作用,Cd仅在圆口铜鱼中表现出了与年龄相关的生物累积作用。     

不同规格圆口铜鱼幼鱼的化学组成和能量密度

2014年11月-12月在金沙江收集体长(L)150~290mm,体重(W)64.1~422.7g的圆口铜鱼23尾,根据体长将其分为4组,分别测定其化学组成,估算其能量密度。结果显示:圆口铜鱼幼鱼含水量(WAT)、蛋白质含量(PRO)、脂肪含量(FAT)、灰分含量(ASH)和能量密度(E)占其湿重的百分比分别为65.87%~69.99%,15.65%~18.17%,9.95%~14.07%,2.69%~3.80%,7.23~10.06kJ/g。经统计分析,圆口铜鱼幼鱼含水量与其蛋白质含量、脂肪含量、灰分含量和能量密度均存在显著的负线性关系。另外,圆口铜鱼幼鱼体长与其含水量存在显著的负线性关系,而与蛋白质含量、脂肪含量、灰分含量和能量密度均存在显著的正线性关系。结果表明,可以用圆口铜鱼幼鱼的含水量和体长估测其化学组成和能量密度,圆口铜鱼高脂肪含量与其长距离繁殖洄游密切相关。     

圆口铜鱼精子超低温冷冻保存

为建立圆口铜鱼(Coreius guichenoti)精子超低温冷冻保存方法,采用计算机辅助精子分析系统(CASA)评价了4种稀释液(D15、D20、L1、D1), 3种抗冻保护剂[二甲基甲酰胺(DMF)、二甲基亚砜(DMSO)、甲醇(METH)]在3种体积浓度(7.5%、10%、12.5%, V/V)下对圆口铜鱼精液的冷冻保存效果,并比较了150 mOsm/kg NaCl与超纯水作为激活剂对精子活力的影响。结果表明,D1组稀释液的精子活率(MOT)最高,为(74.64±13.17)%,与鲜精无显著差异(P0.05);以10%甲醇作为抗冻保护剂的圆口铜鱼精子经超纯水激活后,测得MOT最高,达到(78.11±14.74)%,与鲜精无显著差异(P0.05),精子运动速度达最大,平均曲线运动速度(VCL)、平均直线运动速度(VSL)、VAP(平均路径运动速度)分别达到(50.28±12.46)μm/s、(35.06±10.82)μm/s、(39.44±12.46)μm/s,精子快速运动时间和寿命分别为(8.67±1.15) s、(33.33±5.00) s,显著低于鲜精(P0.05); 7.5%甲醇组和7.5%二甲基甲酰胺组的MOT次之,分别为(77.71±17.74)%、(76.42±12.49)%,抗冻剂二甲基亚砜组的MOT显著低于甲醇组、二甲基甲酰胺组(P0.05)。12.5%的3种抗冻保护剂中,几乎无精子存活;在不同种类不同浓度的抗冻剂保护下,10%甲醇组,相较于使用超纯水激活,用150 mOsm/kg NaCl激活后的精子活率和寿命更高,说明Na~+有延长冻精寿命,提高精子活率的作用。研究表明, D1+10%METH (7.8 g/L NaCl+0.5 g/L KCl+15 g/L葡萄糖+10%METH)可用于圆口铜鱼精液超低温冷冻保存,为圆口铜鱼的繁育工作和种质资源保护奠定了基础。     

圆口铜鱼呼吸代谢的温度响应特征研究

为探讨圆口铜鱼呼吸代谢的温度响应特征,研究了不同水温(10、15、20、25、30℃)下圆口铜鱼(Coreius guichenoti)耗氧率、排氨率、氧氮比以及不同水温范围温度系数(Q₁₀),测定了不同水温下血红蛋白、血糖、乳酸、皮质醇等血液生理生化指标。研究结果显示：圆口铜鱼耗氧率、排氨率和血红蛋白含量均随水温升高呈现先上升后下降趋势。耗氧率、排氨率的Q₁₀值均在25～30℃水温范围时显著下降;10℃组氧氮比值显著低于其他组,除10℃组外其他各组间无显著差异;血糖随水温升高整体呈现下降趋势,除25℃组外,10℃组显著高于其他各组;乳酸含量随水温升高呈现先升高后下降趋势;皮质醇含量呈现先下降后升高的趋势。此外,水温对圆口铜鱼耗氧率昼夜变化具显著影响。15～25℃范围内,圆口铜鱼耗氧率昼夜变化类型为夜间大于白天类型,而10、30℃组属于昼夜差异不明显类型。15～25℃组耗氧率整体均高于10、30℃组。综上所述,水温15～25℃时,圆口铜鱼呼吸代谢水平较稳定。     

长江中上游圆口铜鱼的种群死亡特征及其物种保护

根据葛洲坝(1998～2007年)、重庆(2006～2007年)和合江江段(1998～2005年)的渔业资源调查资料,对圆口铜鱼的种群死亡特征进行了评估,并利用Beverton-Holt模型的单位补充量渔获量方程,分析和探讨了长江中上游圆口铜鱼资源的合理利用.结果表明:1)3个江段各年间的圆口铜鱼开发率和捕捞死亡系数均远远大于相应年份最大允许的开发率和捕捞标准的基准尺度F0.1,3个江段的圆口铜鱼资源均处于严重过度捕捞状况;2)葛洲坝江段圆口铜鱼的最适开捕年龄为4龄(体长330mm),重庆和合江江段圆口铜鱼的开捕年龄应至少为5龄(体长375 mm).为保护圆口铜鱼的资源,建议葛洲坝江段三层流刺网的网日(2a)应大于75 mm为宜,重庆和合江江段的网目(2a)应不小于90 mm且同时需要控制各种渔具的日均作业次数.     

长江干流圆口铜鱼的年龄与生长研究

采用鳞片材料对采自长江干流宜昌和重庆江段圆口铜鱼(Coreius guichenoti)的年龄进行鉴定,并对其生长方程、生长拐点等生物学特征进行了研究。结果表明,调查江段圆口铜鱼的年龄组成以1、2龄个体为主;体长(L)和鳞径(R)呈线性关系,L=15.327R+71.349;体重(W)与体长(L)呈幂指数关系,W=0.00002L2.9942;体长von Bertalanfy生长方程为Lt=730.15[1-e-0.12(t+1.01)];体重生长方程为Wt=7493.05[1-e-0.12(t+1.01)]2.9942;其生长拐点年龄为8.13龄。圆口铜鱼体长生长3龄前为快速期,之后生长减缓。为了保护长江干流圆口铜鱼资源,建议以278mm为最小捕捞个体的体长。     

野生圆口铜鱼人工驯养初探

<正>圆口铜鱼是长江上游特有经济鱼。随着水利枢纽工程的开发，阻隔了圆口铜鱼的生殖洄游通道和长江中下游圆口铜鱼对金沙江繁殖群体的补充通道，这是对其资源量造成不利影响的重要原因。然而圆口铜鱼的人工驯养工作也面临着极大的难题，本文探索了一种圆口铜鱼的人工驯养方法，旨在为圆口铜鱼的人工驯养繁殖工作提供参考。     

封闭循环水系统和船体网箱驯养圆口铜鱼幼鱼的效果比较

以长江野生圆口铜鱼(Coreius guichenoti)幼鱼(约150~400 g)为试验对象,开展了封闭循环水系统和船体网箱驯养圆口铜鱼的比较试验。每种驯养条件下的试验鱼均按规格分2组进行驯养。封闭循环水系统中2组试验鱼的放养密度分别为11.48 kg/m3和17.65 kg/m3,驯养时间为279 d,平均水温为(14.5±3.4)℃。船体网箱中2组试验鱼的放养密度分别为4.40 kg/m3和7.06 kg/m3,驯养时间244 d,平均水温为(20.0±3.1)℃。所有试验鱼均采用人工配合饲料进行饱食投喂。结果显示,封闭循环水系统和船体网箱驯养试验鱼的成活率分别为97.75%和83.78%,且大规格试验鱼比小规格试验鱼成活率略高。封闭循环水系统中两组试验鱼的体重特定生长率分别为0.08%/d和0.11%/d,船体网箱中两组试验鱼的体重特定生长率分别为0.17%/d和0.21%/d。船体网箱试验鱼的丰满度总体大于封闭循环水系统的试验鱼。封闭循环水系统中全年未暴发重大疾病,船体网箱中暴发过一次较大规模的小瓜虫、车轮虫等寄生虫病。结果表明,封闭循环水系统驯养圆口铜鱼的成活率高但生长较慢,船体网箱驯养圆口铜鱼的生长优势明显但易遭受疾病侵袭。综合而言,封闭循环水系统和船体网箱养殖均是当前比较可行的圆口铜鱼驯养方式。     

北方铜鱼

<正>北方铜鱼(Coreius septentrionalis)属鲤形目鲤科!亚科铜鱼属,除北方铜鱼外,该属还有铜鱼(Coreius heterodon)和圆口铜鱼(Coreius guichenoti)二种。铜鱼(Coreius heterodon)和圆口铜鱼(Coreius guichenoti)当前在长江水系还有较大的种群量,而北方铜鱼为黄河水系特有鱼,由于数量稀少,已被列入《中国濒危动物红皮书(鱼类)》。     

Quality of testicular semen of the African catfish Clarias gariepinus (Burchell, 1822) and its relationship with fertilization and hatching success

Quality differences of testicular semen of the African catfish, Clarias gariepinus, and their influence on fertilization and hatching success were investigated. In accordance with an earlier study, two semen types of the African catfish were distinguished according to testicular maturity stage. Semen type I derived from males with white mature testes whereas type II semen derived from males with grey, partly mature testes. Semen volume, sperm cell concentration and seminal plasma pH was significantly higher in type I semen than in type II semen, while sperm motility was similar. Similar fertilization percentages were obtained with semen type I and semen type II. However, the hatching percentage was higher and the percentage of deformed hatched larvae was lower for type I semen. There were significant (P<0.01) positive correlations between sperm motility and fertilization percentage, seminal plasma pH and hatching percentage and a negative correlation between seminal plasma pH and percentage of deformed larvae. Therefore seminal plasma pH and sperm motility are useful to predict semen quality of the African catfish.     

Relationship between biochemical constituents of fish semen and fertility: the effect of short-term storage

Spermatozoa and seminal plasma obtained from rainbow trout and whitefish were analyzed in respect to their aspartate aminotransferase (AspAT) and alkaline phosphatase activities. In particular, the experiments characterized AspAT optimum pH, optimization of assay conditions and action of coenzyme, pyridoxal 5-phosphate (vitamin B₆). The effect of short-term semen storage at 0°C on biochemical indicators and fertilization rate was examined in both species. The concentrations of reduced and oxidized ascorbic acid in seminal plasma of both species were several folds higher than in spermatozoa and blood plasma of fish. Highly significant correlations were found for both species between AspAT activity (sperm or seminal plasma) and fertilization rate (% of eyed-stage or hatched embryos). For rainbow trout, highly significant correlations were found between sperm concentration, motility and fertilization rate. These results suggest that several biochemical indicators of seminal plasma can be used as measures of sperm quality of fish. Some common biochemical parameters for fish and mammal's semen provide evidence for using fish sperm as a model in biomedical research.     

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.     

中华倒刺鲃、白甲鱼和岩原鲤精子的生理特性比较

对中华倒刺鲃(Spinibarbussinensis)、白甲鱼(Onychostomasimus)和岩原鲤(Procyprisrabaudi)3种鱼的鲜精在不同水溶液和不同浓度梯度的NaCl溶液中的活力以及精子形态、密度和精液pH值、浓度等进行了比较研究。结果表明:3种鱼的精液pH值都偏弱碱性,位于7·2~7·7之间;精液浓度依次为71·1%,76·1%和71%;精子头长依次为(3·89±0·53)μm,(2·98±0·08)μm和(6·42±0·59)μm,全长依次为(41·63±3·66)μm,(65·67±2·97)μm和(58·89±5·25)μm;精子密度依次为1·527×1010尾/mL,1·336×1010尾/mL和1·362×1010尾/mL。分析表明,3种鱼的精子大小与密度无相关性。在不同水溶液中,3种鱼的精子活力均以池塘水中最高;在不同浓度NaCl溶液中,3种鱼精子的最适浓度位于0·45%~0·55%之间,有效运动时间以中华倒刺鲃最高,为(85·23±12·02)s,其次是岩原鲤,为(50·45±6·89)s,白甲鱼最短,为(31·44±5·53)s。3种鱼的精子全长与精子活力存在负相关性,即精子越长,活力越低,精子越短,活力越高。     

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.     

Sperm quality and selected biochemical markers of seminal plasma at the beginning of the reproductive period of common carp, Cyprinus carpio L.

Changes in semen quality and selected biochemical markers were analyzed during a week of spawning season of common carp Cyprinus carpio L. Semen was obtained twice, on May 30 and on June 7, and each time it was collected 24 h after hormonal stimulation using Ovopel [(d-Ala⁶, Pro⁹ NEt)-mGnRH + metoclopramide] in 1 pellet kg^?1. The total volume of semen (ml), volume of semen per kg of body weight (ml kg^?1 b.w.), sperm concentration (×10⁹ ml^?1), total number of sperm per kg of body weight (×10⁹ kg^?1 b.w.), pH of semen, pH of seminal plasma, seminal plasma osmotic pressure (mOsm kg^?1) and the total protein content in seminal plasma (mg ml^?1) were determined. A 10 mM Tris buffer containing 100 mM NaCl with 0.5 % BSA (pH 9.0, osmolality 200 mOsm kg^?1) was used to activate sperm. The following computer-assisted sperm analysis (CASA) parameters were determined: percentage of motile sperm (MOT, %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, μm s^?1), straight-line velocity (VSL, μm s^?1), movement linearity (LIN, %), wobbling index (WOB, %), amplitude of lateral head displacement (ALH, μm) and beat cross-frequency (BCF, Hz). The volume of semen per kg of BW, total number of sperm per kg of BW and semen pH were significantly lower at the second semen sampling compared to the first semen sampling. Volume of semen at the second sampling correlated positively with CASA parameters. A lack of differences among CASA parameters between both collection periods indicates good quality of carp sperm hormonally stimulated with Ovopel twice at a 1-week interval.     

Low seminal plasma buffering capacity corresponds to high pH sensitivity of sperm motility in salmonids

The buffering capacity of air-equilibrated seminal plasma from the chinook salmon Oncorhynchus tshawytscha and steelhead Oncorhynchus mykiss were determined. Buffering capacities were significantly less at pH values at and above 7.3 than over the range 6.0 to 7.0 and were particularly low between pH values of approximately 7.5 to 8.5. The potential of chinook salmon sperm to become motile upon addition to water was highly sensitive to pH. Only very low levels of motility were demonstrable when sperm had been maintained at pH values at or below 7.5 prior to water exposure while maximum motility was noted when sperm had been maintained at pH values of about 8.0 and above. Thus, the potential for motility appeared especially sensitive to that pH range over which seminal plasma exhibited its lowest buffering capacity. Buffering capacity, protein content and inorganic phosphate content of chinook salmon and steelhead seminal plasmas were less than those of corresponding blood plasma or ovarian fluid suggesting that seminal plasma is unique among salmonid body fluids in its composition and low buffering capacity. This low buffering capacity is likely to facilitate the role of the sperm duct epithelium in semen pH regulation and thus, in the sperm's acquisition of the potential for motility upon exposure to water. This revised version was published online in August 2006 with corrections to the Cover Date.     

Induced spermiation of Pimelodus britskii (Teleostei: Pimelodidae) during the reproductive period

For the first time was characterized the semen of Pimelodus britskii, hormonally induced and non‐induced induced, during the reproductive period. The experiment 1 was conducted with 12 fish per month, divided into: (i) induced with Carp Pituitary Extract (CPE), and (ii) without hormonal induction (testes macerated). The experiment 2 was conducted, with 30 fish, divided into groups for comparison of different doses of CPE and human Chorionic Gonadotrophin (hCG; T1: control; T2: 3.0 mg kg^?1 CPE; T3: 0.5 mg kg + 3.0 mg kg^?1 CPE; T4: 0.5 mg kg^?1 + 7.0 mg kg^?1 CPE; T5: 200 UI kg^?1 hCG), the same fish were used every month, and the semen was collected by abdominal massage. In both, experiments were assessed the sperm motility and velocity by means of the CASA software. In experiment 1, the concentration of spermatozoids was significantly increase by application of CPE compared untreated males. The volume and motility decreased gradually until the end of the experiment, the highest values were recorded for treatment induced (0.49 ± 0.25 mL and 62.18, respectively). The same occurred to Gonadosomatic Index, showing the smallest value at the end of the reproductive period. The fish from experiment 2 released reduced volume of watery semen (0.1 mL). The values of sperm concentration, motility velocity decreased gradually throughout the months. For P. britskii, the reproductive period influenced the production and sperm quality. Despite small seminal volume released the dose of 7.5 mg kg^?1 of CPE proved effective.     

黑尾近红鲌精子低温保存方法研究与应用

在4℃条件下,以精子活力为指标,研究了不同浓度的Na~+、K~+、Ca~(2+)、Mg~(2+)、葡萄糖、氨基酸等组成的5种(A、B、C、D、E)精子保存液及其适量添加青霉素对黑尾近红鲌(Ancherythroculter nigrocauda)精子活力的影响。结果显示:1)温度为4℃时,精子活力达80%的保存液A、B、C、D、E的保存时间分别为48、24、60、48、48 h,保存液C的保存时间明显高于其它各组;2)在各保存液中分别添加浓度为2.0×104IU/m L的青霉素,精子活力达80%的保存液A、B、C、D、E的保存时间分别为60、36、156、48、144 h,保存时间延长了12~96 h,保存液C和E的延长时间最长,均为96 h;3)人工授精试验证明,经保存的黑尾近红鲌精子能正常用于人工繁殖,受精率达(90.6±0.8)%~(91.8±0.9)%,与对照组精子受精率(92.4±0.8)%无显著差异。添加青霉素,保存液C的保存效果最好,其次是保存液E。     

Identification of calcium-binding proteins in fish seminal plasma

Calcium ions play an important role in the activation of fish sperm movement. The mechanism of their binding in semen is still unknown. The goal of this study was the development of a method for identifying calcium-binding proteins in fish seminal plasma. Two methods of calcium-binding proteins detection were tested with the use of Quin2 and Stains-all dyes. The first method was useful for the identification of calcium-binding proteins of fish seminal plasma. It consisted of proteins separation using SDS–PAGE, transfer on PVDF membrane, incubation with CaCl₂, staining with Quin2 and illumination with UV light to reveal calcium-binding protein bands. Using Quin2 allowed the detection of calcium-binding proteins with low and high molecular weight. Electrophoretic species-specific profiles of calcium-binding proteins were identified in the seminal plasma of carp, whitefish, roach, brook trout, brown trout and rainbow trout. Staining of calcium-binding proteins with Quin2 is a quick and safe method, allowing the identification of calcium-binding proteins in fish semen.