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1.
Abstract. The effect of four environmental conditions was investigated upon sperm output in turbot, Scophthalmus maximus (L.), submitted to three different rhythms of stripping. Males kept under a natural light cycle and under a 6-month contracted light programme released a similar sperm output in terms of total volume of semen produced per fish during the experimental period (4·9 ± 0·9ml), mean sperm concentration (29·4 ± 2·8 × 109 spermatozoa/ml) and total sperm number (163·2 ± 40·5 × 109 spermatozoa). Attempts to stimulate spermiation for a second time just after the end of the natural reproduction period resulted in the release of low sperm output (total volume of semen: 1·6 ± 0·4 ml; mean sperm motility: 2 min 36s ± 0 min 47s; mean sperm concentration: 47·6 ± 10·2 × 109 spermatozoa/ml; total sperm number: 84·5 ± 25·3 × 109 spermatozoa). Stripping frequency had no effect on total volume of semen, mean sperm motility and total sperm number. Monthly collection did not modify sperm samples in relation to stripping rank. However, decreasing volume, motility and sperm concentration were observed when males were stripped fortnightly and weekly. During the natural spawning period, the presence of females in the tank enhanced mean sperm motility (from 3 min 27s + 0 min 52s to 6 min 38s ± 1 min 38s).  相似文献   

2.
The yamú Brycon siebenthalae is an endemic fish of the Orinoco river basin, but wild stocks are decreasing because of the disruption of their habitat. We evaluated a protocol for the cryopreservation of yamú sperm to contribute to the preservation of this endangered genetic resource. Milt was mixed with a cryoprotectant medium (5.5% glucose, 12% egg yolk, and 5%, 10%, or 15% dimethyl sulfoxide - DMSO) in a ratio 1:4 (milt:medium), stored in 0.5-mL French straws, frozen in nitrogen liquid vapor (-76 C), then immersed and stored in liquid nitrogen for 10 d or 12 mo. Motility of thawed spermatozoa was higher ( P < 0.001) in 10% DMSO medium than 5% DMSO or 15% DMSO mediums; but lower than the control ( P < 0.001). With sperm cryopreserved, the highest level of fertilization was achieved with 10% DMSO ( P < 0.001) after 10 d or 12 mo of cryopreservation. Fertilization of eggs inseminated with 6.4 × 109 spermatozoa per g of eggs was higher ( P <0.05) than with 1.6 × 109 spermatozoa per g of eggs. There was no difference (P > 0.05) in fertilization between insemination doses of 3.2 × 109 and 6.4 × 109 spermatozoa per g of eggs. Cryopreservation of yamu milt can be performed successfully with a simple medium combined with 10% of DMSO as cryoprotectant. The highest level of fertility was achieved using between 3 × 109 and 6 × 109 spermatozoa per g of fresh eggs.  相似文献   

3.
Abstract.— Captive-reared, 10-mo-old, male striped bass Morone saxatilis were sampled monthly for testicular development between February and June 1994. One of the five males sampled in February showed precocious testicular development and had a gonado-somatic index (GSI) of 1.26%. while the other four fish had immature testes with a mean GSI ± sx, of 0.17 ± 0.03%. Spermiating individuals were present from April to June. In April the average body weight (BW) of spermiating males was 65 ± 4 g and their GSI reached a mean value of 4.75 ± 0.52%. In June, milt collected from ten precocious males contained motile spermatozoa with a mean of 31 ± 7% of the sperm showing forward movement. Mean milt volume and sperm concentration were 1.67 ± 0.41 mL/kg BW and 92.3 ± 1.8 ± 109 spermatozoa/mL. respectively. These data show that male striped bass reared in captivity can reach sexual maturity during their first year. This is one year earlier than previously reported for striped bass in mid-Atlantic regions.  相似文献   

4.
Abstract. The sperm of tench, Tinca tinca L., is characterized by a milky colour and consistency, and is of very low density. After collecting the sperm, motion of spermatozoa was recorded even without water activation. A better motility value (value 4·36 on average) was observed in spermatozoa collected in immobilizing solution (collecting medium) and stored for 3h, when compared with spermatozoa without collecting medium. Average total and relative numbers of spermatozoa were 12·16 × 109 per male and 18·50 × 109 per kg of body weight, respectively. When testing the effect of activating solution in artificial propagation of tench, the highest fertilization rates (81·3 and 85% in two cases) were found for NaCl solution with an osmotic concentration of 34 or 69 mOsmol and for fresh water, respectively. The fertility rate was reduced significantly ( P < 0·01) by any increase above 105 mOsmol in NaCl concentration in the activating solution. In the tests of optimal method of artificial fertilization, the highest hatching rate of sac fry (71·35%) was found in sperm collected into immobilizing solution. The application of immobilizing solution significantly increased the number of sac fry at the levels P < 0·1 and P < 0·01, if compared with intact sperm stored for 3 h and fresh sperm, respectively.  相似文献   

5.
Five hormonal treatments with human chorionic gonadotropin (hCG) were tested for the induction of maturation and spermiation in male farmed eels. The main aim was to optimize previously used hormonal treatments to achieve shorter induction treatments, longer spermiation periods and/or higher sperm quality. Fish treated for just 3 weeks (treatment E) or until the onset of spermiation (treatment C) showed the worst results, while the treatment consisting of weekly administration of 1.5 IU hCG g?1 fish (treatment A) induced the highest percentage of spermiating males, the highest number of sperm samples and sperm volumes and densities similar to the rest of the treatments (B: half hormone dosage, or D: biweekly administration). Evaluation of the sperm quality was performed by computer‐assisted sperm analysis (CASA), considering the percentage of total motile spermatozoa, the percentage of fast and medium‐velocity spermatozoa, as well as different motility parameters. Sperm samples from A‐D groups showed between 44% and 54% motile spermatozoa, and between 10% and 15% fast spermatozoa, while samples from E‐treated males showed 0% motile cells. No significant differences were found in the spermatozoa straight line velocity (VSL), curvilinear velocity (VCL) or the angular velocity (VAP), neither spermatozoa beating cross frequency (BCF) between A–D groups.  相似文献   

6.
This paper reports an initial trial to cryopreserve semen from two freshwater South American fishes, the curimbatá (Prochilodus scrofa) and the dourado (Salminus maxillosus). Motility and duration of motility were observed in curimbatá and dourado fresh sperm. Semen mixed with extender (0.8% NaCl) was frozen using vials (1 ml) with subsequent storage in liquid nitrogen. Samples were thawed in 1% NaHCO3 or in 0.8% NaCl solutions. Post-thawing motility and duration of motility were verified. A simple extender consisting of 0.8% NaCl plus 10% DMSO was able to initiate motility in fresh spermatozoa. The percentage of motile cells and duration of motility were similar in both thawing solutions, but lower than in fresh sperm.  相似文献   

7.
Cryopreservation of Sperm of Farmed European Eel Anguilla anguilla   总被引：1，自引：0，他引：1  
Sexual maturation and sperm release were induced in farmed European eels Anguilla anguilla kept exclusively in fresh water by using two dosages of human chorion gonadotropin (100 International Unit (IU)-Group one and 250 IU/individual per week-Group two). Sperm release took over 13 wk in both groups. The quality of sperm was investigated on the eighth, ninth, and tenth wk. The average cell densities were 0.94 ± 0.4 × 1010 (Group one) and 0.93 to. ± 0.6 × 1010 (Group two) spermatozoa/mL. The estimated motility of eel sperm was 33, 55, and 49% on the eighth, ninth, and tenth wk of treatment, respectively. The estimated average motility of samples selected for cryopreservation was 73 ± 10%, while the post-thaw motility of cryopreserved samples was 36 ± 11%. The extender originally developed for common carp sperm crypreservation together with methanol as cryoprotectant was found suitable for the cryopreservation of European eel sperm.  相似文献   

8.
The fertilization efficiency of cryopreserved sperm was compared with fresh sperm from striped catfish, Pangasius hypophthalmus . Of the two sets of experiments carried out, the first compared four sperm doses using fresh sperm and fresh eggs. The second experiment compared six concentrations of cryopreserved sperm ranging from 6.94 × 107 to 6.94 × 1010 to fertilize 100 eggs per batch. Fertilization, hatch and survival rates were compared between cryopreserved and fresh sperm. The highest fertilization rate (53.75±1.62%) was achieved with a sperm dose of 6.94 × 108. Increasing the sperm dose to 3.47 × 109 did not increase the fertilization rate, indicating that the optimum sperm:egg ratio lies between 6.94 × 106 and 3.47 × 107 sperm per egg. Both highest (6.94 × 1010) and the lowest (6.94 × 107) sperm doses resulted in lower fertilization rates (2.04% and 16.90% respectively). No significant differences were found among four fresh sperm doses compared. Mean hatch and survival rates resulting from fresh and cryopreserved sperm were similar. The experiment shows that while only 1.89 × 106 fresh spermatozoa was required to fertilize a fresh egg, 6.94 × 106 (or 3.67 times more) cryopreserved sperm was required to achieve the same level of fertilization. This provides important information for making decision to cryopreserve sperm for commercial and/or conservation purposes.  相似文献   

9.
A high percentage (98.3%, N = 60) of the marbled grouper Epinephelus microdon individuals captured from spawning aggregations during July and August 1993 in the waters surrounding the island of Koror, Republic of Palau, Micronesia, were in the stage of maturity at which final maturation and spawning could be hormonally induced. The sex ratio of the captured fish was highly skewed towards males (4 male:1 female). Sexually immature females comprised the smallest size class, (<0.6 kg body weight (BW) or 33.0 cm total length (TL)), while sexually mature females were restricted to the 0.6–1.5 kg BW (33.0–46.4 cm TL) groups. Males predominated in size classes >0.6 kg BW, and individuals >1.5 kg BW (46.4 cm TL) were exclusively male. All females with oocytes that averaged ( N = 50) >400 μm in diameter were successfully induced to spawn by a two-injection protocol using human chorionic gonadotropin (HCG) at total dosages of 2,100–3,200 IU/kg fish. All males used in the spawning trials were administered a single injection of HCG at dosages of 500 or 1,000 IU/kg fish. Fecundity ranged between 7.96 × 105−1.24 × 106kg BW, average spawned egg diameters ranged between 769–832 μm, percent fertilization ranged between 32.6%–99.9%, and hatching percentages were >90.0%. Total fat content of eggs obtained from a pooled spawning event was 14.1 mg/100 mg dry weight. The data indicate that HCG is a suitable treatment for the induction of spawning in marbled grouper females that possess a mean oocyte diameter of 400 μm or greater.  相似文献   

10.
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   

11.
Abstract.— Two experiments were designed to improve upon existing methods for cryopreserving striped bass Morone saxatilis , semen. In the first experiment, two extenders, two cryoprotectant concentrations, and two freezing rates were evaluated on the basis of post-thaw semen motility after 1, 7, and 30 d of storage at −196 C. Semen samples cryopreserved at a freezing rate of −40 C/m resulted in a significantly higher percentage of motile sperm ( P < 0.001) and longer duration of spermatozoa motility ( P < 0.001) than samples cryopreserved at a freezing rate of -30 Chin. Also, the cryoprotectant dimethyl-sulfoxide yielded a significantly higher percentage of motile sperm ( P < 0.001) and longer duration of spermatozoa motility ( P < 0.001) when a 5% concentration was used instead of 7.5%. In the second experiment, the two extenders from Experiment I were re-evaluated and a new extender, which was a modified version of Extender 1, was tested. The samples were cryopreserved at -40 C/min with 5% DMSO and thawed in a 25 C water bath. Spermatozoa motility and fertilization ability were evaluated, and semen cryopreserved in Extender 2 yielded the longest duration of spermatozoa motility ( P < 0.001). the highest percentage of motile sperm ( P < 0.001). and the highest percentage of fertilized eggs ( P < 0.002) in comparison to Extenders I and 3.  相似文献   

12.
The common carp, Cyprinus carpio L., sperm motility parameters were analyzed by using computer‐assisted sperm analysis system. The percentage of motile sperm (MOT, %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, µm/sec), average path velocity (VAP, µm/sec), the wobbling index (WOB, %), movement linearity (LIN, %), beat cross frequency (BCF, Hz), and amplitude of lateral head displacement (ALH, µm) were determined. Five activation solutions (As) were used to activate sperm movement. As 1 solution: 68 mM NaCl, 50 mM urea, 0.5% bovine serum albumin (BSA), pH: 7.7, 181 mOsm/kg; As 2 buffer: 100 mM NaCl, 10 mM Tris, 0.5% BSA, pH: 9.0, 199 mOsm/kg; As 3 solution: 86 mM NaCl, 0.5% BSA, pH: 7.4, 167 mOsm/kg; As 4 buffer: 5 mM KCl, 45 mM NaCl, 30 mM Tris, 0.5%, pH: 8.0, 160 mOsm/kg; and As 5 solution: distilled water with the addition of 0.5% BSA, pH: 7.3, <3 mOsm/kg. Among five tested solutions, a buffer with a pH of 9.0 and osmolality of approximately 200 mOsm/kg (As 2) was the most suitable. After its activation, a significant increase in MOT and ALH values was observed, which can be of importance to the effectiveness of egg fertilization .  相似文献   

13.
Basic characteristics of the European smelt (Osmerus eperlanus) sperm are reported here for the first time. Smelt spermatozoa had a bullet-shaped head (1.42 μm length), a short midpiece and a long flagellum (27.72 μm). Two mitochondria were located along the flagella. The volume of smelt sperm was small (30-60 μl) and the duration of sperm motility was short (22 s in distilled water and 41 s in 20 mM sodium bicarbonate solution). Sodium chloride at concentrations ranging from 0-120 mM did not influence the percentage of motile spermatozoa but caused a steady increase in the duration of sperm movement. Potassium ions clearly reduced the percentage of motile sperm at a concentration of 10 mM. Spermatozoa were motile through a broad range of pH with an optimum from 7.5 to 8.5. Testicular spermatozoa had a different motility pattern compared to stripped spermatozoa (the latter exhibiting a reduction of motile spermatozoa by 30%, lower ALH and VCL and higher LIN and VSL). These results indicate that maturation of smelt spermatozoa occurring in sperm ducts is related not only to an increase of the percentage of motile spermatozoa but also to changes in the sperm motility pattern. Maintaining males with females resulted in stimulation of milt production. Our results indicate that European smelt sperm characteristics are similar to those of ayu (Osmeridae).  相似文献   

14.
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.  相似文献   

15.
Sperm physiology, in vivo artificial insemination and spawning of the ocean pout (Macrozoarces americanus L.), a marine bottom fish, were studied. Milt was collected from the reproductive tract of mature males by suction using a catheter. The uncontaminated milt, having a very low sperm concentration, contains highly motile spermatozoa and sperm motility was retained in vitro at 4 °C for at least 24 h in both seminal plasma and ovarian slime collected from the oviduct of pre-spawning females. Instead of activating sperm, dilution in sea water instantly immobilized the spermatozoa of ocean pout. Osmolarity and pH of ocean pout seminal plasma were in the ranges 365–406 mOsM and 7.2–7.5, respectively. A study of the ionic composition of ocean pout seminal plasma demonstrated the presence of various ions including Na+, K+, Ca2+, Mg2+, and Cl−, with a remarkably lower K+ concentration compared to that from other fish species. Since injections of milt containing motile sperm into the ovaries of pre-spawning females, which spawned in the absence of males, yielded fertilized ocean pout eggs, it is concluded that the ocean pout exhibits internal fertilization. The larvae hatched after 3 months of egg incubation in ambient sea water (9–10 °C). With proper timing of in vivo artificial insemination of mature females, fertilized ocean pout eggs can be obtained from fish reared in captivity.  相似文献   

16.
The effects of cryopreservation on the viability, morphology and capability of spermatozoa in Atlantic cod, Gadus morhua L., were studied. The sperm was cryopreserved in straws using Hanks' balanced salt solution, hens' egg yolks and glycerol in the vapor of liquid nitrogen. Straws of cryopreserved sperm were stored in liquid nitrogen and thawed in seawater (35 C) for 8 sec before use. The motility of cryopreserved sperm was low (range 8–19%) compared to motility before freezing (range 69–76%). The fertilization rate (range 94–95%) in control groups using fresh sperm was significantly higher (P < 0.05) than in test groups (range 48–72%). In cryopreserved sperm, a relatively high percentage (range 82–93%) of the spermatozoa had changes in morphology. Many spermatozoa had no mitochondria; when mitochondria were present, the observed number varied from one and five in cryopreserved spermatozoa, and from two and seven in noncryopreserved spermatozoa. In groups where cryopreserved sperm was used, the hatching rate was lower (range 18–38%) than in control groups (range 41–63%), indicating higher mortality during embryonic development. Paternal effects on progeny performance were noted in the proportion of abnormalities but no negative effects were identified in newly hatched larvae produced using cryopreserved sperm.  相似文献   

17.
The role of the seminal fluid antioxidant system in protection against damage to spermatozoa during in vitro sperm storage is unclear. This study investigated the effect of in vitro storage of sterlet Acipenser ruthenus spermatozoa together with seminal fluid for 36 h at 4 °C on spermatozoon motility rate and curvilinear velocity, thiobarbituric acid reactive substance level, and components of enzyme and non-enzyme antioxidant system (superoxide dismutase and catalase activity and uric acid concentration) in seminal fluid. Spermatozoon motility parameters after sperm storage were significantly decreased, while the level of thiobarbituric acid reactive substances, activity of superoxide dismutase and catalase, and uric acid concentration did not change. Our findings suggest that the antioxidant system of sterlet seminal fluid is effective in preventing oxidative stress during short-term sperm storage and prompt future investigations of changes in spermatozoon homeostasis and in spermatozoon plasma membrane structure which are other possible reasons of spermatozoon motility deterioration upon sperm storage.  相似文献   

18.
A two-factor experiment was carried out to investigate the change in skin colour and plasma cortisol response of cultured Australian snapper Pagrus auratus to a change in background colour. Snapper (mean weight=437 g) were held in black or white tanks and fed diets containing 39 mg unesterified astaxanthin kg−1 for 49 days before being transferred from white tanks to black cages (WB) or black tanks to white cages (BW). Skin colour values [ L * (lightness), a * (redness) and b * (yellowness)] of all snapper were measured at stocking ( t =0 days) and from cages of fish randomly assigned to each sampling time at 0.25, 0.5, 1, 2, 3, 5 and 7 days. Plasma cortisol was measured in anaesthetized snapper following colour measurements at 0, 1 and 7 days. Fish from additional black-to-black (BB) and white-to-white (WW) control treatments were also sampled for colour and cortisol at those times. Rapid changes occurred in skin lightness ( L * values) after altering background colour with maximum change in L * values for BW and WB treatments occurring within 1 day. Skin redness ( a *) of BW snapper continued to steadily decrease over the 7 days ( a *=7.93 × e−0.051 × time). Plasma cortisol concentrations were highest at stocking when fish were held at greater densities and were not affected by cage colour. The results of this study suggest that transferring dark coloured snapper to white cages for 1 day is sufficient to affect the greatest benefit in terms of producing light coloured fish while minimizing the reduction in favourable red skin colouration.  相似文献   

19.
The tench Tinca tinca is an interesting fish from the viewpoint of polyploidy and related atypical reproduction aspects. Triploid tench were produced artificially. Studies of spermiation as well as of sperm motility and structure were performed on several triploid and diploid males simultaneously with individual experimental crosses with diploid females to define their reproductive capacities. The testes of triploids visually looked less developed in the most of cases with lower sperm production (0.05 cm3 sperm per male), GSI and weight of testes compared to diploids (0.58 cm3 sperm per male). Analysis of variance showed significant influence of ploidy level on the percentage of motile spermatozoa. Triploidy did not change percentage of live spermatozoa and velocity of spermatozoa at the first time of sperm movement. The study of sperm structure by scanning electron microscopy revealed that most sperm cells were of normal structure with some anomalies. Sperm heads of triploid and diploid males were mostly round-shaped, 1.86±0.2 and 1.6±0.18 μm in diameter. The midpiece of triploid spermatozoa was slightly narrower than that of diploid ones with typical cylindrical shape. Flow cytometry revealed sperm cells of triploids to be largely aneuploid (1.47 n) with high mosaic DNA, oscillating from haploid DNA content (1.0 n) to diploid DNA content (1.9 n). Experimental crosses between triploid males and diploid females revealed that these males were capable to stimulate effective development with relatively high level of fertilization and hatching rates from 0 to 70%. In conclusion, triploidization does not seem to guarantee sterility of tench.  相似文献   

20.
The spermiation of tench males was stimulated with Supergestran containing mammalian LHRHa lecireline at the following doses: 5, 10, 20 and 40 g kg−1 b.w.; then with carp pituitary suspension (CPS) at a dose of 2 mg kg−1 b.w. and with a control of saline physiological solution. The following days, meaning 24, 48 and 72 h after injection, sperm was collected to evaluate volume and the number of sperm per male per kg body weight (B.W.) The percentage of motile sperm and velocity of spermatozoa were measured 48 h after hormonal injection, and 72 h after hormonal injection the sperm was evaluated for fertilization and hatching ability. All 42 males in experimental groups were diploid. Live weight did not differ significantly among experimental groups. The strongest stimulation of spermiation was achieved with LHRHa in dosage of 20 and 40 g kg−1 b.w. and CPS compared to males of the control group and lower dosage of LHRHa. Analysis of variance showed no significant influence of the treatment on the velocity and percentage of motile spermatozoa. The effect of different treatment on the fertilization capacity (the number of spermatozoa per egg was equilibrated) was significant. Significantly the highest quality of sperm collected 72 h after injection expressed by percentage of fertilization and hatching (62–65% fertilization and 61–64% hatching rates, respectively) was found for LHRHa in dosage of 20 and 40 g kg−1 b.w. Significantly the lowest parameters of fertilization and hatching were found for the control group, on the 12% level.  相似文献   

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