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1.
To improve the efficiency of gynogenetic induction, the effect of UV light on sperm of Heteropneustes fossilis was optimized. The sperm suspension was diluted to 1 × 107 sperm/ml?1 in Hank's Balanced Salt Solution. Sperm suspension was irradiated under UV light for different exposure times ranging from 15 to 360 seconds (7500 ergs/mm?2 for 60 seconds). Sperm motility and egg activation efficiency were assessed for the different exposure durations. Complete inactivation of sperm genetic material was recorded from 120 seconds onwards, and a good motility score was recorded until 240 seconds. A typical “Hertwig effect” in the yield of hatched larvae was observed with doses of UV exposure greater than 75 seconds. Larvae resulting from sperm UV irradiated above 120 seconds were 100% haploid. The genetic inactivation of paternal chromosomes was confirmed by chromosome counting (n = 29) from the resulting embryo, which also had a characteristic haploid syndrome. The resulting embryo (with 29 chromosomes) exhibited haploid syndrome and died after yolk sac absorption.  相似文献   

2.
This study reports the results on induced meiotic diploid gynogenesis and female homogametic nature in the Indian catfish, Heteropneustes fossilis. The eggs of H. fossilis were inseminated with conspecific sperm. The sperm suspension was diluted to 1 × 107 sperm mL−1 in Hanks balanced salt solution. Sperm were irradiated under UV light, with the exposure time ranging from 15 to 360 s (7500 ergs mm−2 for 60 s). The genetic inactivation of paternal chromosomes was confirmed by chromosome counting from the larval cells and the larvae also had a characteristic haploid syndrome. A typical ‘Hertwig effect’ in the yield of hatched larvae was observed with doses of UV exposure >75 s (9375 ergs mm2). Larvae resulting from sperm UV irradiated above 120 s (15 000 ergs mm2) were 100% haploids. Application of heat shock to the activated eggs was effective in suppressing the release of the second polar body (meiotic gynogenesis) and resulted in diploid gynogenetic larvae morphologically identical to those of the control. The best yield of diploid gynogens (49.3% with respect to the control) was found to be at 6 min after egg activation and the heat shock at 41 °C for a 1-min duration, at an ambient water temperature of 27 °C. A total of 113 diploid gynogenetic fry from seven different female fish were reared and subjected to sexing. All gynogenetic fish were female in contrast to the control, which had a mean sex ratio of 56.7% females (which was not significantly different from 50% female). From these results, the sex determination mechanism in H. fossilis was presumed to be female homogamety.  相似文献   

3.
Haploid gynogenesis was induced in Siberian sturgeon (Acipenser baeri Brandt) using UV‐irradiated bester (hybrid of Huso huso L. and Acipenser ruthenus L.) sperm. The inactivated heterologous sperm was used successfully to produce gynogenetic haploids of Siberian sturgeon. All haploid progeny were screened using microsatellite DNA analysis and uniparental transmission in gynogenetic haploids was confirmed. The objective of this study was to obtain haploids of Siberian sturgeon and describe the inheritance of microsatellite loci in haploid embryos of this species. This initial work, based on inheritance in haploid Siberian sturgeon, suggests that three studied microsatellite loci segregate disomically, three tetrasomically and one octasomically.  相似文献   

4.
Large yellow croaker, Pseudosciaena crocea, exhibit sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. We have established a protocol to produce all-female croaker P. crocea through induction of meiotic gynogenesis with homologous sperm. The first set of experiments investigated the ultra-violet (UV) irradiation on sperm motility and duration of sperm activity to determine the optimal UV dosage for genetic inactivation of sperm, yet retaining adequate motility for activation of eggs. Milt from several males was diluted 1:100 with Ringer’s solution and UV irradiated with doses ranging from 0–150 J cm−2. The results indicated that motility and duration of activity generally decreased with increased UV doses. At UV doses greater than 105 J cm−2, after fertilization, motility was <10% and fertilization rates were significantly lower. Highest hatching rate was obtained at 75 J cm−2. A second set of experiments was carried out to determine appropriate conditions of cold shock for retention of the 2nd polar body in P. crocea eggs after fertilization with UV-inactivated sperm by altering the timing, temperature and duration of shock. At 20°C, shock applied at 3 min after fertilization resulted in higher survival rate of larvae at 6 h after hatching. Results of different combinations of three shock temperatures (2°C, 3°C or 4°C) and five shock durations (4 min, 8 min, 12 min, 16 min or 20 min) at 3 min after fertilization demonstrated that shocks of 12 min gave highest production of diploid gynogens. Statistical analysis revealed that maximum production of diploid gynogens (44.55 ± 2.99%) were obtained at 3°C. The results of this study indicate that the use of UV-irradiated homologous sperm for activation of P. crocea eggs and cold shock for polar body retention is an effective method for producing gynogenetic offspring.  相似文献   

5.
The sex ratios and sex determination mechanism of gynogenetic diploids of ship sturgeon, Acipenser nudiventris, have been investigated to verify the possibilities of sex control by chromosome manipulation in this species. Meiotic gynogenesis was induced in a female ship sturgeon using cold shock after egg activation with UV-irradiated sperm of a male Siberian sturgeon. Microsatellite DNA analysis was applied for verification of uniparental inheritance in the gynogenetic diploid group of fish. All the analyzed gynogenetic diploids possessed only maternal genotype in the examined experimental group of fish. In this study, a minimum of two distinctly selected diagnostic loci in the offspring was used to confirm exclusively maternal contribution. Also, these fish were analyzed for sex diagnostic. Histological analysis of gonads from gynogenetic diploids, obtained from one family, showed 73.3 % of females and 27.7 % of males. The observed sex ratio has suggested that the ship sturgeon have a female heterogametic sex determination system. Gynogenesis in this species with female heterogametic sex determination system will have important role in breeding program and reclamation of its natural population to produce both female and male progeny, while this species has been introduced in the red list of IUCN (International Union for Conservation of Nature and Natural Resources).  相似文献   

6.
This study reports the results on induced diploid androgenesis in the Indian catfish, Heteropneustes fossilis. Ultraviolet (UV) irradiation was used to inactivate maternal genome of H. fossilis. Complete inactivation of maternal genome was recorded at 12,500 ergs/mm2. These genome‐inactivated eggs of H. fossilis were inseminated with conspecific sperm. The sperm suspension was diluted to 1 × 107 sperm ml/L in Hank's Balanced Salt Solution. Egg viability was assessed for different exposure durations at fertilization, hatching, haploidy, and diploidization. Majority of the larvae derived from irradiated eggs had abnormal appearance. Complete inactivation of maternal genome was detected by haploid syndrome and confirmed by chromosome counting (n = 29). These eggs activated with sperm were subjected to heat shock at 40 and 41 C for different postactivation times and durations. Diploid androgens had a normal appearance as controls and confirmed by chromosome counting (n = 58). A maximum of 21 and 14% of diploidization was recorded at 30 min after activation, at 40 and 41 C, which corresponds to the first cleavage suppression time.  相似文献   

7.
Oxidative stress is a possible source of spermatozoa function deterioration. Seminal fluid (SF) protects spermatozoa against reactive oxygen species (ROS) attack during development in testes and transit through the reproductive tract. Spermatozoa curvilinear velocity and percent of motile cells as well as changes in thiobarbituric acid-reactive substance (TBARS) content, superoxide dismutase, and catalase activity, and uric acid concentration in SF were evaluated in sterlet sperm collected from testes 24 h after hormone induction of spermiation and from Wolffian ducts at 12, 24, 36, and 60 h after hormone injection (HI). While testicular spermatozoa motility was not initiated in activating medium, Wolffian duct sperm showed low motility at 12 h, significant increase at 24 and 36 h, and decrease at 60 h. Testicular SF was characterized by the highest level of TBARS and activity of studied enzymes compared with SF from Wolffian duct sperm at 24 h post-HI. In fluid from Wolffian duct sperm, a significant increase in TBARS content was shown at 36–60 h post-HI. In contrast to testicular SF, in SF from Wolffian duct sperm, this increase was not counterbalanced by changes in the studied variables of antioxidant system. This may be the source of the observed decrease in spermatozoa motility parameters 60 h post-HI. The results may confirm a dual role of ROS in fish sperm physiology. The data with respect to decrease in sturgeon spermatozoa motility parameters at 60 h post-HI should be taken into account in artificial sturgeon propagation.  相似文献   

8.
Turbot Scophthalmus maximus exhibits sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. In this paper, gynogenetic diploids of turbot were induced by activating egg development with ultraviolet (UV)-irradiated left-eyed flounder Paralichthys olivaceus sperm combined with cold shock to prevent extrusion of the second polar body. The results of UV irradiation experiments showed that survival, motility, and duration of activity of P. olivaceus sperm generally decreased with increase in UV dose. The typical Hertwig’s effect was observed after fertilized turbot eggs with UV-irradiated P. olivaceus sperm and the optimal UV dose for gynogenetic haploid production was 36,000 erg mm−2. At 15°C, appropriate timing of cold shock for retention of the second polar body in turbot eggs was at 6 min after fertilization. Results of different combinations of two shock temperatures (1 or 3°C) and four shock durations (15, 25, 35 or 45 min) at 6 min after fertilization demonstrated that shock of 25 min at 1°C gave the highest production of diploid gynogens (39.58% relative to its diploid control). The results of this study reveal that the use of UV-irradiated P. olivaceus sperm for activation of turbot eggs and cold shock for polar body retention is an effective method to produce gynogenetic offspring.  相似文献   

9.
The aims of this study were to evaluate the efficiency of simple and complex extenders in prolonging the cold storage of sperm (Experiments 1 and 2) and to test the diluted‐cooled sperm in the best extender with regard to sperm quality parameters (Experiment 3) in the streaked prochilod, Prochilodus lineatus. In all the experiments, aliquots of 0.3 mL of sperm were diluted 1:10 in extenders and stored at 4–6 °C. Sperm diluted in simple extenders (NaCl and glucose solutions) yielded 0–26% sperm motility, whereas sperm diluted in complex extenders (BTS?, M III? and Androstar?) yielded 62–81% sperm motility on day 4 after cold storage. When Androstar? was further investigated, the following was observed on day 4: 53% motility with 94 s of duration; 47% live spermatozoa; 26–61% fertility rate; and 22–60% hatching rate. The use of Androstar? improves the sperm fertility of the streaked prochilod during a 4‐day storage period and can therefore be used to facilitate artificial reproduction.  相似文献   

10.
In this study, the effects of spermiation time are investigated on milt quality of Russian sturgeon over the course of the spawning season. The milt samples were collected from three broodstock batches at three time points including: the beginning, middle and at the end of the spawning season. According to the results, the milt quality parameters including pH, sperm density, spermatocrit, duration of sperm motility and percentage of sperm motility were significantly low in the beginning and end of season than middle of season. The values of milt quality parameters in the middle of season were as follows: (motility percentage: 69.6 ± 3.5, motility duration: 460.3 ± 37.2 s, sperm density: 8.7 ± 0.4 × 109, milt volume: 86.3 ± 8.1 and milt pH: 8.3 ± 0.15). Significant positive correlations were also found between milt pH and sperm motility as well as between sperm density and spermatocrit. In conclusion, our study showed that the middle of season is the best time for collection of milt with appropriate quality in Russian sturgeon. Selection of milt with good quality is necessary aim to cryopreservation of spermatozoa in endangered fish species including Russian sturgeon.  相似文献   

11.
Three experiments were performed to develop protocols for cryopreservation of Persian sturgeon Acipenser persicus, sperm. In the first experiment, sperm from six males was individually split in three subsamples and cryopreserved using Modified Tsvetkova's extender (mT) supplemented with dimethyl sulfoxide (DMSO), methanol (MeOH), glycerol (Gly) and ethylene glycol (EG) at concentration of 5%, 10%, 15% and 20%. In the second set of experiments, the effects of six equilibration times (0, 5, 10, 20, 40 and 60 min) and dilution ratios (volume sperm: volume extender 1:0.5, 1:1, 1:2, 1:3, 1:5 and 1:10) and the additive advantage of bovine serum albumin (BSA; 0, 2.5, 5 and 10 mg mL?1) and ascorbic acid (0, 2.5, 5 and 10 U mL?1), on the post‐thaw survival of sperm (triplicate set of six fish) were evaluated. Then, sperm was diluted in 1:1 mT extender with 10 mg mL?1 BSA with selected cryoprotectants (15% MeOH and 10% DMSO) for 5 min. After a month of storage in liquid nitrogen, post‐thawed sperm motility; fertilization and hatching rate and viability of derived larvae were measured (Exp.3). Evaluation of cryoprotectants efficiency showed that MeOH 15% and DMSO 10% were suitable for cryopreservation of Persian sturgeon sperm. Gly and EG resulted in very low post‐thaw motility rates even at lowest concentration. No significant difference was observed among the four different equilibration times (0, 5, 10, 20 min) (P > 0.05) although higher equilibration times than 20 min resulted low post‐thaw motility (P < 0.05). The motility of frozen–thawed sperm did not significantly change when dilution ratio was increased from 1:0.5 to 1:3 (P > 0.05). However, higher dilution ratios (1:5 and 1:10) reduced the percentage of motile sperm. Supplementation of the cryoprotectant solution with 10 mg mL?1 BSA significantly improved post‐thaw motility (P < 0.05), but ascorbic acid did not improve post‐thaw motility (P > 0.05). The results of experiment 3 showed that the highest fertilization (30.2 ± 5.75) and hatching rates (28.2 ± 5.25) were observed when samples were frozen with 15% MeOH (P > 0.05). Our study indicates that the use of mT extender consisting of 10 mg mL?1 BSA in 15% MeOH diluted with sperm at 1:1 ratio for 5 min can be recommended cryopreservation method for Persian sturgeon sperm.  相似文献   

12.
In this study, the efficiency of a novel droplet vitrification technique along with different doses of fish antifreeze protein (AFP) type III on Persian sturgeon thawed spermatozoa quality (motility duration and motility percentage) was investigated. Semen of seven male individuals was pooled in equal volumes and diluted with 4°C Tris‐Hcl (100 mM), pH = 8 extenders containing 0, 5, 10, 15 μM of AFP type III in a ratio of 1:1 (semen/extenders). Treated semen was dropped into liquid nitrogen. Solidified droplets were stored for 2, 60 and 120 days and thawed by plunging them into a tube containing 5 mL Tris‐Hcl (100 mM), pH=8 with 1% BSA at 37°C. Motility duration in all treatments had no significant difference comparing to fresh sperm (P > 0.05), but their motility percentage was significantly lower. Treatment with 10 μM of AFP had significantly higher motility percentage (16.11 ± 0.5%) comparing to other treatments (P < 0.05). There was no significant difference between 0, 5, 15 μM of antifreeze protein treatments (P > 0.05), suggesting that antifreeze protein effectiveness are highly dose dependent, and dose of 10 μM is appropriate in Persian sturgeon spermatozoa droplet vitrification. Besides, the present technique obtained higher quality of spermatozoa comparing to its analogue techniques.  相似文献   

13.
The motility and fertilizing ability of the Persian sturgeon, Acipenser persicus, spermatozoa were investigated. Optimum ionic content (Na+, K+, Ca2+ and Mg2+) and pH of activation solution as well as the optimum dilution rate were determined. The results show optimum motility characteristics of spermatozoa in buffered solutions containing 25, 0.2, 3 and 10 mM L?1 Na+, K+, Ca2+ and Mg2+, respectively, at dilution rate 1:50 and pH 8.0. To test the fertilizing ability of sperm, two buffered saline solutions were used as activation solution of sperm motility. The present study indicated (1) spermatozoa motility is one of key factors that influence on fertilizing ability of sperm, (2) a high fertilizing ability of sperm is obtained after dilution in saline solutions rather than in freshwater and (3) a maximum fertilization rate occurs in buffered saline solution containing 0.2 mM L?1 K+. There is also a good correlation between biochemical characteristics of seminal plasma and fertilizing ability of sperm.  相似文献   

14.
The objective of this study was to investigate the effects of extenders and storage time on motility, viability and fertilization of preserved black sharkminnow, Labeo chrysophekadion spermatozoa. Sperm were diluted 1:3 in one of five extenders: modified Cortland solution (MC); Hanks' balanced salt solution (HBSS); 0.9% sodium chloride (NaCl); Kurokura solution (KU); and modified extender, and undiluted sperm samples were used as control and stored at 4°C for 5 days. Motility, viability and fertilization rates were evaluated every day. After a storage time of three days, the highest motility, viability and fertilization rates (61.27 ± 2.26%, 58.60 ± 2.29% and 40.58 ± 0.57, respectively) were achieved with sperm diluted with modified extender. Motility, viability and fertilization rates decreased significantly (P < 0.05) with increasing storage time in all treatments. In addition, this study found that motility, viability and fertilization had a positive significant correlation (P < 0.01). The results indicate that isotonic extender is suitable for the short‐term preservation of black sharkminnow spermatozoa.  相似文献   

15.
Sexually mature males (BW?=?1600?±?150 g and TL?=?235?±?30 mm) of northern pike (Esox lucius L.) were randomly selected from a pond to record changes in their sperm quality parameters (spermatozoa morphology, sperm volume, density, and motility parameters) during the spawning season. The morphological and motility parameters changed significantly during the reproductive season with following trends. Only, head width was not changed during the spawning season. The longest spermatozoa and its flagellar length were found at the middle of spawning period (TL?=?38.24?±?0.37 μm and 35.14?±?0.26 μm) and shortest at the beginning of spawning period (TL?=?34.81?±?0.29 μm and 32.53?±?0.18 μm). Other morphological characters were always the lowest at the beginning of spawning period. Sperm volume was changed from 0.33?±?0.3 ml in February, 0.43?±?0.2 ml in March to 0.24?±?0.1 ml in April, and density from 16.2?±?0.2?×?109 spermatozoa ml?1 in February, 19.4?±?0.2?×?109 spermatozoa ml?1 in March to 4.8?±?0.2?×?109 spermatozoa ml?1 in April. Same sperm velocity was observed in all spawning terms at 10 and 20 s after activation. Higher velocity was found at 30 and 40 s after activation in sperm collected at the middle and the end of spawning period. Significantly, higher percentage of motile sperm was observed at 20, 30, and 40 s after activation in sperm sampled at the end of spawning period. This study supports the hypothesis that longer spermatozoa swim faster.  相似文献   

16.
This paper provides data concerning biochemical composition of milt of two sturgeon species, Siberian sturgeon bred in aquaculture facility of Inland Fisheries Institute in North Poland and sterlet (from two different populations from Danube and Odra). Milt plasma of Siberian sturgeon and sterlet, when compared to teleost fish, is characterized by much lower osmolality (up 70 to 96 mOsm kg−1) and protein concentration (0.24–0.58 g l−1). In spite of the presence of an acrosome and acrosin the antiproteinase activity of seminal plasma was low (12.79–25.40 U l−1). Activities of arylsulfatase and β-N-acetylglucosaminidase were found in spermatozoa. This agrees with the presence of an acrosome in sturgeons sperm. Similarly to mammals, these enzymes are also present in milt plasma. We determined a range of enzymatic activities from the minimal (seminal plasma) to the maximal damage (supernatants obtained after freezing-thawing without cryoprotectant). Activities of arylsulfatase, β-N-acetylglucosaminidase, lactic dehydrogenase and acid phosphatase were released from spermatozoa after freezing-thawing. For this reason they are good potential candidates as a markers of cryoinjury to sperm acrosome and midpiece. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

17.
人工养殖西伯利亚鲟精子超低温冷冻保存研究   总被引：7，自引：6，他引：7  
刘鹏  庄平  章龙珍  王斌  闫文罡 《海洋渔业》2007,29(2):120-127
研究了人工养殖西伯利亚鲟精子的生物学特征及超低温冷冻保存方法。西伯利亚鲟的产精量为113.67±39.86 ml,精子密度为(6.49±3.10)×108/ml,精子活力为(85.4±9.5)%,精子寿命为353±23 s。精子密度与精子快速运动时间、精子寿命之间均存在线性相关,用方程分别表示为:y=1.0384x+1.5089(R2=0.7325);y=2.9069x+74.289(R2=0.6967)。结果表明精子密度可作为一项精子质量评价的标准。通过比较西伯利亚鲟精子在不同稀释液、不同抗冻剂和抗冻剂浓度、降温速率、解冻温度下的保存效果,结果表明:配方2作为稀释液,18%甲醇作为抗冻剂,二步法超低温(-196℃)冷冻保存精子,40℃水浴解冻取得最好的冻后活力,解冻后活力为(51.8±5.8)%。西伯利亚鲟授精的最佳精卵比为106∶1。在此精卵比下用冻精授精分别得到了(72.3±3)%的受精率和(52.9±4.1)%的孵化率,其中受精率与鲜精没有显著性差异,孵化率与鲜精有显著差异(P<0.05)。  相似文献   

18.
通过紫外线(UV)对冷冻的鲈(Lateolabrax japonicus)精子进行灭活,利用冷休克和压力休克方法诱导星斑川鲽(Platichthys stellatus)雌核发育二倍体,同时利用灭活鲈精子制备单倍体胚胎,未灭活鲈精子受精制备杂交胚胎,星斑川鲽精子受精制备正常发育胚胎。对以上几种胚胎发育时序、发育生物学特征进行了观察比较。结果表明,卵裂期单倍体、杂交二倍体和雌核发育二倍体胚胎发育速度与普通二倍体胚胎没有明显差异,从低囊胚期开始各实验组胚胎发育速度均慢于普通二倍体胚胎;杂交胚胎在胚体形成期基本死亡,单倍体胚胎在尾芽期停止发育死亡,均不能正常孵出。雌核发育二倍体与普通二倍体具有相似的发育时序,普通二倍体100 h 10 min孵化出膜,而雌核发育二倍体104 h 50 min孵化出膜。雌核发育胚体畸形率(53.59±0.36)%,孵化率(0.11±0.01)%;普通二倍体胚体畸形率(35.11±6.19)%,孵化率(58.01±5.30)%;与普通二倍体相比,雌核发育二倍体胚体畸形率高,孵化率低,但孵化鱼苗能够正常发育,获得了雌核发育群体。本研究为星斑川鲽雌核发育提供了技术方法,同时为单倍体、杂交和雌核发育胚胎的发育生物学研究提供了细胞生物学证据。  相似文献   

19.
Fin ray biopsies are commonly used to age sturgeon, but the effects of fin ray biopsy on Siberian sturgeon Acipenser baerii swimming performance are unknown. Therefore, the effects of two fin ray biopsy methods on swimming performance of hatchery-reared, juvenile Siberian sturgeon were evaluated in this study. Treatment fish were subjected to one of two biopsy methods: removal of a 2- to 4-cm section from a marginal pectoral fin ray, or full removal of a marginal pectoral fin ray. Control fish were only subjected to a sham operation. A modified 2,936-l Brett-type swim tunnel was used to evaluate 10-min critical station-holding speeds (C SHS) and behavioral swimming characteristics of sturgeon immediately after fin ray biopsies were carried out. Fish sizes (range 65- to 84-cm fork length) were comparable among treatments. Mean 10-min C SHS (mean ± SE) were 113 ± 3.4, 109 ± 2.5, and 111 ± 2.8 cm s?1 for the segment removal treatment, full removal treatment, and control treatment, respectively. ANOVA indicated that the two methods had no significant effect on the 10-min C SHS of Siberian sturgeon compared to the control treatment. Results indicate that fin ray removal has no effect on Siberian sturgeon swimming performance.  相似文献   

20.
In a natural environment, seminal plasma provides spermatozoa with protection against reactive oxygen species. Storing semen in cooling conditions requires diluting it with various buffer solutions. Therefore, the protective role of seminal plasma is not sufficient enough. Semen obtained from five male specimens was diluted with the Kobayashi buffer solution at a 1:9 ratio. To determine the influence of antioxidants on semen storage, a buffer solution was used, as before, with the addition of 1 % albumin, 1 mM vitamin C, 1.5 mg ml?1 vitamin E, 5 mM sodium citrate, 5 mM glutathione and 5 mM cysteine. After the preparation of such tests, the parameters of spermatozoa motility were measured every 3–5 days, using the CASA system (Image House CRISMAS Company Ltd.). Among all used antioxidants, the best effects were observed after the addition of glutathione to semen. After 17 days of storage, the percentage of motile spermatozoa in the samples preserved with glutathione addition was 57 %, while without antioxidant addition, it was 44 %. Furthermore, the addition of cysteine and albumin also resulted in the lengthening of the life span of perch sperm cells. The presence of the remaining antioxidants (vitamins C and E, and sodium citrate) did not have any positive influence on spermatozoa viability, and in these samples, no motile spermatozoa were observed after 12 days of storage. Our data show that dilution of perch sperm with buffered solution might be a promising method for short-term storage.  相似文献   

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