Seasonal changes in sperm production and quality in the red porgy Pagrus pagrus (L.)

Cultured red porgy Pagrus pagrus (L.) males (n=6) were sampled every 2 weeks for milt, in order to monitor changes in sperm quality parameters during a whole spawning period. On 11 January 2001, 60% of the fish were spermiating, increasing to 100% in mid‐February and dropping to 30% by mid‐April. Sperm density showed a slight increasing trend, with mean values ranging between 8.6 and 23.7×10⁹ spermatozoa mL^?1. Sperm motility percentage exhibited a significant improvement during the spawning season (analysis of variance (anova ) P=0.0001). The duration of forward motility for the major part of the monitoring period ranged between 2 and 4 min. Red porgy spermatozoa maintained their viability for many days after whole storage of milt at 4°C. During the monitoring period there were significant changes in the mean duration of sperm survival after cold storage, ranging from 5 to 12 days. The total volume of expressible milt was maximal on 28 March, increasing from a mean value of 1.7 mL to 5.3 mL kg^?1. Milt production of captive‐reared red porgy does not appear to be limiting, when compared with the volume of expressible milt produced by other cultured marine fishes.     

Sperm characteristics and androgens in Acipenser ruthenus after induction of spermiation by carp pituitary extract or GnRHa implants

Spermiation and changes in androgen (testosterone, T and 11-ketotestosterone, 11-KT) levels were studied in sterlet (Acipenser ruthenus) treated with GnRH agonist implants (dAla⁶-Pro⁹-LHRHa) at 25 and 75?μg?kg^?1 b.w. and compared with those males treated with 4?mg?kg^?1 b.w. of carp pituitary extract (CPE) and 3 pellets of Ovopel kg^?1 b.w., which contains dAla⁶-Pro⁹NEt-mGnRH and metoclopramide. Sperm quality (sperm mass, spermatozoa concentration and sperm motility and velocity) was evaluated 24, 48 and 72?h after hormonal treatments. Males did not release sperm in the control group injected with physiological solution, while sperm could not be collected 7?days after treatments in all hormonally treated groups. Spermiation rates were 100?% in the CPE and Ovopel groups and 25–50?% in the GnRHa-treated groups. Sperm production was significantly lower in the GnRHa-treated groups than in the CPE and Ovopel groups and decreased 72?h after hormonal treatment. Sperm motility and velocity were higher in the Ovopel and GnRHa (75?μg) groups compared to the CPE and GnRHa (25?μg) groups and decreased 72?h after hormonal treatment. Androgens were only affected in spermiating males and changed in the Ovopel and GnRHa (75?μg) after hormonal treatment. Significant correlations were observed between sperm production, sperm motility and sperm velocity, but not androgens. The present study suggests involvement of dopamine in sturgeon spawning. Additionally, better sperm quality observed in the Ovopel group and particularly sperm motility in the GnRHa (75?μg) suggests enhancement of sperm quality in sturgeon treated with GnRHa. Therefore, further study is needed to induce fully spermiation using GnRHa implants in combination with a dopamine inhibitor.     

Effect of two commercial preparations containing different GnRH analogues with dopamine antagonists on barbel Barbus barbus (L.) sperm quantity and quality

The effectiveness of applying Ovaprim [(D-Arg⁶, Pro⁹NEt)-sGnRH + domperidone] and Ovopel [(D-Ala⁶, Pro⁹NEt)-mGnRH + metoclopramide] to male barbel Barbus barbus (L.) 6, 12 and 24 h after hormonal stimulation was analyzed. The control group (Control) during each time interval was stimulated with 0.9 % NaCl. Milt was collected from seven fish only once (n = 7) for Ovopel, Ovaprim and Control group in order to determine total volume of milt, volume of milt per kg of body weight, sperm concentration, total sperm production, seminal plasma osmotic pressure, pH of milt and pH of seminal plasma. Woynarovich’s solution (68 mM NaCl + 50 mM urea) with the addition of 0.5 % BSA (pH 7.7; 181 mOsm kg^?1) was used as the activating liquid. Selected parameters of sperm motility (MOT %) and progressively motile sperm (%), curvilinear velocity (VCL, μm s^?1), straight-line velocity (μm s^?1), movement linearity (%), wobbling index (%), amplitude of lateral head displacement (μm) and beat cross frequency (Hz) were determined using the Computer-assisted sperm analysis system. A time of 6 h proved to be too short to obtain milt from barbel following hormonal stimulation with Ovaprim and Ovopel. Extending the time to 12 h, however, resulted in 100 % spermiation in males, regardless of hormonal preparation used for stimulation. The stimulation of spermiation in barbel is best performed using Ovopel 12 h upon application. Extending the latency period to 24 h following the application of this preparation results in a significant decrease in the volume of milt obtained, sperm count and motility parameters, including MOT and VCL, which may influence sperm fertilization ability.     

Effect of different methods for the induction of spermiation on semen quality in European eel

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.     

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.     

The parameters of energetic status of the wild Atlantic salmon (Salmo salar L.) spermatozoa

The energetic status of fresh sperm of wild Atlantic salmon (Salmo salar L.) was presented in selected males (n=10) with motility rate ≥80%. Purine and pyridine nucleotides: adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), guanosine triphosphate, guanosine diphosphate, guanosine monophosphate, inosine monophosphate, nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate; nucleosides: adenosine, guanosine and inosine; bases: hypoxanthine, xanthine and uric acid were determined using the high‐performance liquid chromatography method. The average value of adenylate energy charge in the group was 0.77 ± 0.10, and the mean total adenine nucleotide content (TAN) was 65.1 ± 18.3 pmol × 10^?6 spermatozoa. The mean concentrations of ATP, ADP and AMP were 43.5, 11.8 and 9.9 pmol × 10^?6 spermatozoa respectively. The concentrations of the other energetic nucleotides studied were lower. In all males studied (n=23) with a motility rate from 0% to 95%, no statistically significant correlation between the per cent of motile sperm and ATP concentration was found (R_s=0.35), whereas the correlation between the per cent of motile sperm and ADP was statistically significant (R_s=0.60). A negative correlation was found between hypoxanthine and the per cent of motile spermatozoa (R_s=?0.44). Our results suggest that hypoxanthine is the final product of salmon spermatozoa of adenine nucleotide catabolism.     

Comparative analysis of reproductive performance between spring and autumn forms of Persian sturgeon,Acipenser persicus,Bordin, 1897: with an emphasis on sex steroids and milt quality

To compare the reproductive performance of spring and autumn forms of Persian sturgeon males, Acipenser persicus, the relative abundance of spermiating and non‐spermiating males, milt quality, sex steroids and cortisol were investigated after pituitary preparation (PP) treatment and at the time of spermiation. Our results showed that higher numbers of spring males are spermiated after PP treatment than autumn individuals. The autumn spermiating males had higher and lower values of sperm density and duration of motility than spring males respectively. The milt volume was higher in spring males than in autumn individuals. The percentage of motility was statistically equal in both the groups examined. In both forms of brooders, the serum levels of testosterone (T), 11‐ketotestosterone (11‐KT), progesterone (P), 17α, 20β, 21‐trihydroxy‐4‐pregnen‐3‐one (20βS) and cortisol (C) increased after PP treatment, although such increases did not occur in non‐spermiating males. Our results showed the probable involvement of 20βs, P, T, 11‐KT and C in the final maturation of Persian sturgeon. In addition, it was demonstrated that the spring form of Persian sturgeon shows better reproductive performance than the autumn form in terms of a response to artificial induction and milt quality.     

Exposure to 17α,20β-dihydroxy-4-pregnen-3-one changes seminal characteristics in Nile tilapia, Oreochromis niloticus

This experiment was conducted to evaluate the seminal characteristics of Nile tilapia males exposed to water‐borne 17α,20β‐dihydroxy‐4‐pregnen‐3‐one (17,20βP). Male Nile tilapia (Oreochromis niloticus L.) were exposed to the steroidal pre‐ovulatory pheromone 17,20βP, added to water at a concentration of 5×10^?9 M. The pheromone‐exposed males had higher sperm volume and concentration. In addition, the spermatozoa contained in the sperm had higher motility and the motility duration was longer than ethanol‐exposed males (control group). The percentage of live spermatozoa was not affected by the treatments. Our results suggest that this pheromone can improve sperm quality characteristics and could become a non‐invasive method for enhancing spawning in Nile tilapia.     

Spermiation time affect the milt quality indices of the Russian sturgeon,Acipenser gueldenstaedtii,Brandt & Ratzeburg, 1833

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.     

Sperm characteristics of chub Leuciscus cephalus (L.) collected in artificial condition after Ovopel and Ovaprim treatment

The effect of two commercial preparations containing different GnRH analogues with dopamine antagonists on quantitative and qualitative parameters of semen from chub Leuciscus cephalus L. collected in artificial conditions were examined. Semen was collected after the application of [(D‐Ala⁶, Pro⁹ NEt)‐mGnRH + metoclopramide] (Ovopel, n = 9), [(D‐Arg⁶, Pro⁹ Net)‐sGnR + domperidone] (Ovaprim, n = 9) and from the control group (0.9% NaCl, n = 9). Afterwards, semen volume, sperm concentration, total sperm production and semen pH were determined. Osmolality and pH of seminal plasma were also determined. Using the Computer‐assisted sperm analysis system (CASA), selected sperm parameters such as sperm motility (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) were analysed. While Ovopel can also be used to stimulate chub spermation, the application of Ovaprim was much more effective for obtaining higher amounts of semen.     

Milt quality and spermatozoa morphology of captive Brycon siebenthalae (Eigenmann) broodstock

In order to develop artificial reproduction in freshwater fish for potential species to be developed in South American aquaculture, milt quality and sperm morphology were studied in yamú (Brycon siebenthalae) under captive conditions during the natural middle spermiation period. The volume of milt collected for each male was 1.8±1.2 mL and the sperm concentration was 13.9±4.0 × 10⁹ spermatozoa mL^?1. Spermatocrit (41.5±10.8%) was positively associated (r²=0.30) with sperm density calculated using a corpuscle counting chamber. Sperm motility was 88±9% and the average duration of forward motility was 41±7 s. Fertilization rate was 84±8% and there was no association between this trait and sperm motility (r²=0.009) or with sperm density (r²=0.073). These results suggest that captive B. siebenthalae broodstock can be reproduced successfully.     

Spectrophotometric determination of sperm concentration and short‐term cold‐storage of sperm in Atlantic croaker Micropogonias undulatus L. broodstock

The objective of this study was to optimize the methodology for spectrophotometric determination of sperm concentration in Atlantic croaker Micropogonias undulatus L. milt and to estimate its potential for short‐term cold‐storage. The spectrophotometric determination of sperm concentration was evaluated using milt samples from six males serially diluted in Hank's balanced salt solution at 200 mOsm kg^?1 (HBSS). The predictive power of regression models between sperm concentration and absorbance was determined from 200 to 500 nm and found to be highest within the visible spectrum despite a peak of milt absorbance at 288 nm. Absorbance reading at 400 nm was selected for further analysis to maximize the absorbance of the sample hence the sensitivity of the method while minimizing the impact of potential sample contamination with blood. The standard‐curve of correlation between sperm absorbance at 400 nm and concentration was validated and held an accuracy ranging between ?7.40% and +4.56% across males. Total sperm motility duration and the proportion of motile spermatozoa were significantly higher in milt samples diluted 1:3 in HBSS than in the undiluted control during up to 30 h of cold‐storage.The methodologies investigated in this study can be applied to optimize sperm usage and achieve predictable artificial fertilization protocols in Atlantic croaker.     

Sperm Characteristics of Precocious 1-year-old Male Striped Bass Morone saxatilis

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 ± s^x, 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.     

Retention of sperm motility in turbot, Scophthalmus maximus L.: the effects of time from activation, thermal shock and adenosine triphosphate levels

Abstract Four parameters were examined in order to define sperm quality in turbot Scophthalmus maximus L., sperm: (1) sperm motility, measured by direct counts of the number of active spermatozoa, expressed as % of total spermatozoa; (2) retention of motility after activation, measured by direct counts, 0–60min after activation, expressed as a % of the initial level of activity; (3) resistance to thermal stress, measured as change in retention of motility, and (4) adenosine phosphate (ATP) concentration, determined for samples of non-activated sperm. The proportion of motile spermatozoa at activation ranged from 34·8% to 97·6% (mean 76·3%) for the individual males tested. Turbot sperm retained on average 52% (range 27–90%) of its initial activity one hour after activation. Sperm samples which were stressed by cooling to –27°C retained only 8·6% (range 0–25%) of initial activity, compared to control samples which retained 49% (range 38–63%) of initial activity. The retention of motility after activation was not significantly related to the initial motility or the levels of ATP. Concentrations of ATP in turbot sperm (mean 0·46mg ATP/10⁶ spermatozoa, equivalent to 9·2nmol ATP/10⁸ spermatozoa) were comparable to those measured in mammals.     

Changes in Sperm Production,Sperm Motility,and Composition of Seminal Fluid in Caspian Brown Trout,Salmo trutta caspius,Over the Course of a Spawning Season

This study was carried out to evaluate milt quality in male Caspian brown trout (Salmo trutta caspius) over the course of the winter spawning season. Milt samples were collected biweekly during December and January. Chemical composition of seminal fluid, sperm production (milt volume, sperm density, spermatocrit,) and sperm motility characteristics (percentage and duration of motility) were measured. Milt volume, sperm density, osmolality, seminal minerals (Ca²⁺, Mg²⁺, K⁺, Na⁺, Cl^?), and total protein gradually decreased over the spawning season. Glucose and triglyceride content of milt did not show significant changes over the spawning season. Milt pH and the percentage and duration of motility were comparatively stable, declining only at the end of the season. Significant positive correlations were found between sperm density and seminal minerals, total protein and spermatocrit; percentage of motile spermatozoa and seminal minerals, total protein; and duration of motility and K⁺, Cl^?, total protein, and pH. Results show that season has a significant influence on milt quality in male Caspian brown trout, with the best milt being available at the beginning of spawning season.     

Influence of the length of time after hormonal stimulation on selected parameters of milt of ide Leuciscus idus L.

Milt of the Leuciscus idus L. was collected from five experimental groups, and selected parameters of its quality were analysed for 36 h (group II), 60 h (group III), 84 h (group IV) and 108 h (group V), respectively, after hormonal stimulation with Ovopel (1 granule kg^?1 of body weight). The control (group I) fish were not subjected to hormonal stimulation. The highest milt volume was obtained from the fish in group IV (0.70 ± 0.55 mL), where the largest volume of milt expressed per kilogram was also obtained (3.03 ± 1.94 mL kg^?1). Significant differences were also found in milt volumes obtained between group I and groups III (P<0.01) and IV (P<0.05). The highest percentage of motile spermatozoa was found in the milt of group IV (59%); significant differences were found between group I and groups II (P<0.01) and III (P<0.001). The value of osmotic pressure of seminal plasma was the highest in group IV (203.19 ± 37.63 mOsm kg^?1), and the lowest in group I (118.31 ± 41.13 mOsm kg^?1). Parameters determining milt quality and quantity indicate that the period of 60–84 h after hormonal stimulation with Ovopel is optimal for obtaining milt from ide.     

Sperm motility and seminal plasma characteristics in Barbus sharpeyi (Günther, 1874)

Spermatozoa concentration, ionic composition, osmolality, glucose and total protein contents of seminal plasma and sperm motility were determined in Barbus sharpeyi (Cyprinidae, Teleosotei). Spermatozoa concentration ranged from 9.77 to 20.20 × 10⁹ spermatozoa mL^?1. Osmolality (mOsmol kg^?1) and ionic contents (mM L^?1) of the seminal plasma were 274.5±9.0, 70.0±3.4 Na⁺, 28.8±0.9 K⁺, 101.7±3.1 Cl^?, 0.9±0.1 Mg²⁺ and 2.1±0.1 Ca²⁺ respectively. Total protein and glucose were 5.3±0.2 g L^?1 and 76.7±4.3 mM L^?1 respectively. Sperm motility was initiated in a hypo‐osmotic condition, composed of either an ionic (KCl or NaCl) or a non‐ionic (sucrose) activation medium. Duration of sperm motility was very short: <2 min after activation in distilled water. Percentage of motile spermatozoa was significantly higher in an activation medium containing NaCl compared with that of distilled water. An activating medium containing NaCl or KCl higher than 150 mM or sucrose higher than 275 mM totally inhibited the activation of sperm motility. Immediately after sperm activation, wave(s) propagated along the flagellum, but waves were restricted to the proximal part of the flagellum (close to the head) at 1 min post activation. Studied characteristics in the present study were compared with those of other cyprinids for understanding inter‐species differences.     

Effects of bicarbonate ions and pH on acquisition and maintenance of potential for motility in ayu, Plecoglossus altivelis Temminck et Schlegel (Osmeridae), spermatozoa

The effects of extracellular ions on the acquisition and maintenance of the potential for motility in the spermatozoa of the ayu, Plecoglossus altivelis Temminck et Schlegel (Osmeridae), were examined. Testicular spermatozoa and milt spermatozoa were obtained from fully matured males and diluted with buffered solution (BS, 20 mM HEPES–NaOH, pH 7.5). Testicular spermatozoa showed a significantly low rate of motility (0.8 ± 0.4%), whereas milt spermatozoa showed a high rate (89.4 ± 2.1%). The spermatozoa were incubated with various isotonic media for 2 h, diluted with BS, and changes in the rates of motility were then compared. When incubated for 2 h with artificial seminal plasma (ASP), corresponding in terms of ionic constituents to seminal plasma buffered at pH 8.0, both spermatozoa showed a high rate of motility. Testicular spermatozoa acquired and milt spermatozoa maintained the potential for motility in response to the HCO₃^– ion concentrations (between 0 and 20 mM) in the ASP. The differences in the pH of the ASP had a significant effect on the acquisition and maintenance of the potential for motility, and spermatozoa showed the highest rate of motility with the ASP at pH 8.0 and 8.5. These results suggest that the quality of milt in the ayu can be regulated by controlling the concentration of bicarbonate and the pH of the incubating media.     

Production and storage of sperm from the black sea bass Centropristis striata L

Black sea bass Centropristis striata L. are protogynous hermaphrodites that develop and spawn as females before changing sex to male. Since all fish eventually become males, determining the relationship between sperm production, sperm quality and seasonal changes in plasma levels of testosterone (T) and 11‐ketotestosterone (11‐KT) could be useful for identifying appropriate males to maintain as broodstock. Milt and blood samples were collected three times during an 8‐week spawning season. Milt volume (3.5±0.76 mL kg^?1), sperm density (3.2 × 10⁸± 0.31 cells mL^?1), sperm production [11 × 10⁸±3.4 cells kg^?1 body weight (BW)] and sperm motility (80±0.6%) were at their highest during the first sampling interval and coincided with the highest 11‐KT levels (1.0± 0.11 ng mL^?1). All of the sperm indices decreased to their lowest levels during the final 3 weeks of the study. Sperm viability was highly correlated (adjusted R²=0.84) with sperm motility. Sperm cryopreserved in modified Mounib's extender (MME) had the highest post‐thaw motility compared with two other extenders. Post‐thaw motility of sperm cryopreserved in MME was not different from fresh after 90 days of storage. There was no difference in fertilization rates between fresh (69±2.4%) and post‐thaw (67±4.1%) sperm samples taken from the same male or among males. These results demonstrate that the quality of black sea bass spermatozoa is higher earlier in the spawning season and that acceptable post‐thaw fertilization rates can be obtained from cryopreserved sperm.     

Spermatozoon ultrastructure and sperm cryopreservation of the Brazilian dry season spawner fish pirapitinga,Brycon nattereri

The aims of this study were to describe the fresh spermatozoon ultrastructure using scanning and transmission electron microscopy and to improve the sperm cryopreservation methodology for the freshwater fish pirapitinga Brycon nattereri. Extenders (BTS^? and NaCl), straw volumes (0.5 and 4.0 mL), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO₃) were tested. Methylglycol was used as a cryoprotectant agent and sperm was frozen in nitrogen vapour (dry‐shipper). Post‐thawed sperm motility rate, motility quality (score 0=no movement; 5=rapidly swimming spermatozoa), duration of motility and spermatozoon morphology were evaluated. Fresh spermatozoon was 35.06 μm long, the head was ovoid (2.00 × 1.22 μm) with no acrosome, the midpiece was 2.15 μm long and the flagellum was 30.90 μm long with the typical 9+2 axoneme arrangement. Post‐thawed sperm motility rate (70–79% motile sperm), motility quality (score 3.1–3.7) and morphology (9.3–11.6% abnormal spermatozoa) were not affected by any of the parameters tested. The duration of sperm motility was longer when triggered in 1% NaHCO₃ (392–1031 s) compared with 0.29% NaCl (144–338 s). Brycon nattereri sperm cryopreserved under the conditions described above yields over 70% motility and should last long enough to fertilize oocytes, even after 2 years of freezing.