Changes in the flagellum morphology of intact and frozen/thawed Siberian sturgeon Acipenser baerii (Brandt) sperm during motility

Morphological investigations on the changes in flagellar beating was carried out on native (taken from the milt) and thawed sperm of the Siberian sturgeon Acipenser baerii (Brandt). Immediately after activation, the pattern of flagellar wave formation and distribution was the same in native and thawed sperm but, after 27–42 s, depending on the samples, the thawed flagella showed asymmetric and poorly developed waves. The swimming trajectories recorded during 1‐s exposure were much shorter in thawed than in native sperm after 26–28 s motility. In native sperm, the flagella remained in the same axis as the head during the entire motility course, while the head of thawed sperm showed a right angle after 47 s. It is concluded that the freezing/thawing procedure induces some alteration in the dynamics of flagellar beating in many sperm, but these sperm still show progressive displacement. Therefore, the change in morphology of the flagellum during motion is a parameter that should be taken into account in the evaluation of the impact of various treatments on sperm motility.     

Properties and activities of blood‐ or seawater‐contaminated wild‐caught Striped Jewfish (Stereolepis doederleini) sperm

Understanding the effects of environmental factors in sperm qualities will be helpful in the development of optimal artificial reproduction methods and contributes towards the knowledge base of better short‐ and long‐term fish semen preservation conditions The objectives of this study were to determine properties and activities of wild‐caught striped jewfish Stereolepis doederleini sperm contaminated with blood or seawater and compare them with data reported in the literature on other freshwater and marine fish species, for effective short‐ and long‐term storage of fish semen. Overall, we observed that the sodium, chloride, glucose, total protein concentrations of normal sperm were not significantly different from blood‐ or seawater‐contaminated sperm. The salinity and osmolality concentration of sperm contaminated with blood were lower than sperm contaminated with seawater and were not significantly different from normal sperm. In addition, the spermatozoa motility (SM) and duration of spermatozoa motility (DSM) in blood‐contaminated sperm were higher than seawater‐contaminated sperm and also not significantly different from normal sperm. The best condition for SM and DSM in normal sperm was dilution rate of 1:50. Sperm was immotile in distilled water, and cationic factors were shown to stimulate the initiation of spermatozoa activation. The maximum SM and DSM were observed in solution containing 0.4 M NaCl, 0.6 M KCl, 0.6 M CaCl₂ and 0.4 M MgCl₂. This study provides some basic and important knowledge about striped jewfish sperm sensitivity to a cationic condition. In this regard, Na⁺ is the major inhibitory factor of spermatozoa motility in this fish species.     

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.     

A diluent for sperm cryopreservation of gilthead seabream, Sparus aurata

Sperm of gilthead seabream, Sparus aurata, was diluted with solutions of different osmolarities and pH. The effect of the different diluents on sperm motility (intensity and percentage of motile sperm) was studied. Motility was induced as early as 10 s after mixing the sperm with diluents having an osmotic pressure higher than 500 mOsm/l. The intensity of motility decreased when the osmotic pressure was reduced, and was zero or significantly inhibited when the osmotic pressure of the diluent (300–380 mOsm/l) was close to that of the fish's seminal plasma (364.6±3.03 mOsm/l). The pH of the diluent did not have any effect on sperm motility (in a range of 6.8 to 8.9). A diluent which prevents spermatozoa motility (osmotic pressure 375 mOsm/l and pH 7.35) was successfully used to cryopreserve S. aurata sperm at −196°C. This diluent is considered promising for the long-term preservation of gilthead seabream sperm.     

Changes of sperm morphology,volume, density,and motility parameters in northern pike during the spawning period

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.     

Motility Parameters of Rainbow Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Spermatozoa in Relation to Sequential Collection of Milt,Time of <Emphasis Type="Italic">Post-mortem</Emphasis> Storage and Anesthesia

The present study investigated the effects of sequential collection of milt, time of post-mortem storage and anesthesia on rainbow trout (Oncorhynchus mykiss) sperm motility parameters (using computer-assisted sperm analysis – CASA) as well as seminal plasma osmolality and sperm concentration. The post-mortem storage and time of anesthesia altered motility characteristics of rainbow trout sperm to different extents. The moderate impact of time of anesthesia was manifested in a shortened duration of sperm motility after 10 min exposure of fish to anesthetic. The prolonged post-mortem storage (≥40–60 min), in addition to lowering sperm motility duration, also significantly influenced sperm motility parameters, such as sperm velocities, percentage of motile sperm and sperm trajectory parameters. These results clearly demonstrate that when milt from sacrificed fish is used for sperm motility studies, the time of post-mortem storage significantly alters sperm motility characteristics. Since sperm motility rate and swimming velocity could predict fertilizing ability, detrimental effects of prolonged post-mortem storage may lead to reduced fertilization success. Sperm concentration and seminal plasma osmolality were lower in the first fractions and increased with successive collections of milt. It suggests the presence of urine contamination of the first milt fractions which were collected by stripping. Therefore, testing of sperm concentration and/or seminal plasma osmolality should be mandatory while handling stored milt.     

A study of the dynamics of volume changes during the period of active motility in carp, Cyprinus carpio L., spermatozoa

Carp, Cyprinus carpio L., spermatozoa swelling in NaCl hypotonic solutions (18.8, 37.5, 75, 100 mM) was studied by the methods of photometry and resistance impulses spectroscopy (RIS). The possibility of application of the value of relative optical density of cell suspension, as a qualitative characteristics of the extent of spermatozoa swelling in the period of their movement, was demonstrated. It has been noted, that the moment when swelling starts coincides with the moment of the start of sperm motility. Movement duration, extent and time of swelling depend on the osmotic pressure of the activation solution and moreover the maximum motile period is observed during the activation by 75 mM NaCl solution.     

Effect of water temperature on the physiology of fish spermatozoon function: a brief review

Motility is a key factor in function of the spermatozoon and determines semen quality and fertilizing capacity. Effective motility occurs when sperm is diluted in a swimming solution, the adequacy of which is determined by factors varying according to fish species. Spermatozoon motility rate and velocity, as well as duration of the motility period, are influenced by the temperature of the water in which broodfish are held. Increase in temperature of swimming medium beyond the optimal increases cell metabolism, leading to an increase in velocity with rapid depletion of energy resources, promoting early cessation of movement. The aim of this review was to discuss current information on the influence of temperature on quantitative spermatozoon properties, which could affect sperm function. Our findings provide a greater understanding of fish sperm physiology and a biological foundation for the further development of spermatozoon motility investigations as well as reproduction technologies.     

Motility of sea urchin Paracentrotus lividus spermatozoa in the post‐activation phase

The characterization of sperm motility patterns, particularly post‐activation changes, is the first step in setting up species‐specific protocols involving gamete management and embryo production, for both aquaculture and laboratory research purposes. This study is aimed at the characterization of the sperm motility pattern of the purple sea urchin Paracentrotus lividus. Semen samples were individually diluted in artificial sea water for sperm motility activation. They were then incubated at 18°C for up to 24 hr. Motility was evaluated on dilution, and 1 hr, 3 hr and 24 hr after activation, by computerized analyser. The semen fertilization capacity was also evaluated. Under our experimental conditions (dilution 1:1,000 in artificial sea water plus 0.05% BSA, 18°C, in the dark), P. lividus semen remained viable for up to 24 hr, as the total motile sperm and the fertilization percentages did not change significantly during the incubation time. In contrast, the mean curvilinear velocity and the subpopulation of rapid sperm (those having a curvilinear velocity > 100 µm/s) slightly but significantly decreased after 3 hr, thereafter remaining unchanged for up to 24 hr after activation. In conclusion, our results show that diluted P. lividus semen can be used for a longer period than that of most fish species, with no need for motility inhibition procedures, supporting its wider use in laboratory research. In addition, the development of artificial fertilization protocols for aquaculture production is simplified by long‐lasting sperm motility.     

Age-Related Sperm Quality of Captive Striped Bass Morone saxatilis

Three groups of captive-reared striped bass Morone saxatilis ages 1, 3 and 12 yr, were examined for age-related changes of sperm characteristics including short-term storage. All groups had similar ranges of the following parameters (mean× SEM): expressible milt (5.6× 0.5 mI/kg body weight (BW) to 7.5× 2.1 mL/kg BW), percentage of motile sperm (55× 6% to 60× 2%), duration of sperm motility (69× 3 sec to 72× 5 sec) and percentage of viable sperm (91× 2% to 93× 2%). Compared to the 1 and 12-yr-old fish, the 3-yr-old fish produced the greatest number of spermatozoa (1,190× 370× 10⁹ spermatozoa/kg), sperm concentration (120× 8 × 10⁹ spermatozoa/mL) and spermatocrit (74× 4%). In addition, during short-term storage at 4 C, extender-preserved sperm samples of the 3-yr-old group showed a significantly higher ( P < 0.05) percentage of motile sperm and duration of sperm motility, compared to the other two groups. This suggests that short-term storage may be affected by the age of the male fish. Sperm longevity of the 3-yr-old group was successfully maintained for as long as 15 d, longer than that of the 1-yr-old group (9 d) and 12-yr-old group (7 d). Overall, the 3-yr-old fish appeared to have superior sperm quality than the 1 or 12-yr-old fish based on higher sperm production and increased sperm longevity.     

Ovarian fluid pH enhances motility parameters of rainbow trout (Oncorhynchus mykiss) spermatozoa

All evidence to date suggest that sperm motility is the primary determinant of fertilization success in externally fertilizing fish species. Ovarian fluid, which comprises 10–30% of the total egg volume in salmonids, enhances sperm motility with respect to swimming speed, trajectory and the duration of movement. It was recently demonstrated that there is individual variability in sperm motility enhancing potential of ovarian fluid of particular females. In the present study we examined the effect of particular ovarian fluids collected from 31 females on the sperm motility parameters of one male of rainbow trout (Oncorhynchus mykiss) using computer-assisted sperm analysis (CASA). During our experiment we also monitored the pH of ovarian fluid. We found that particular fluids differed in the ability to activate spermatozoa; sperm remained immotile in four fluids and exhibited 50–100% motility in 27 samples. The percentage of motile sperm, velocity and duration of movement positively correlated with ovarian fluid pH (r² = 0.34–0.62). These data strongly suggest that the pH of the ovarian fluid is the primary determinant of sperm motility in rainbow trout under natural conditions of fertilization.     

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.     

鱼类精子质量评价研究进展

在鱼类人工繁育中,研究者主要关心的是卵子质量,长期以来对精子质量未引起足够重视.而精子质量同样会影响繁育效果的重要因素.鱼类精子质量的评价指标有多种,如精子活力、运动时间、密度、形态、受精率和生理功能等.其中最传统的评价指标是精子活力,其测定方便,能较准确地预测受精率.将精子运动时间和活力综合考虑可更好地反映精子的运动能力.而精子受精率则是精子质量的直接反映,但会受到卵质等因素的影响.质膜完整性、线粒体功能、染色质结构完整性等可体现精子的质量,但测定方法较繁琐.近年来,鱼类精子质量检测技术迅速发展,计算机辅助精子分析(CASA)、流式细胞术(FCM)分析、低渗肿胀(HOS)、单细胞凝胶单泳(SCGE)等技术的建立,使得测定指标更多样、客观、准确.本文逐一介绍了评价精子质量的各种指标,并对各指标的测定方法、测定原理、国内外研究情况进行详细叙述,旨为我国鱼类精子质量评价研究提供背景资料.     

Factors affecting the activation, motility and cryopreservation of the spermatozoa of the yellowfin bream, Acanthopagrus australis (Günther)

Abstract. The effects of varying temperature, salinity and pH on the activation and subsequent motility of sperm of the yellowfin bream, Acanthopagrus australis (Günther), were assessed using a linear scale based on the overall activity of the sperm over time. Motility half-life was calculated using log transformation.
Conditions reflecting the natural habitat of the fish, oceanic salinity (1200 mOsM) and slight alkalinity (pH 8-8-5), were shown to produce both maximum activation and subsequent motility duration. The half-life of activated sperm was shown to be greater at 4°C than at 20-23°C (5·13 min:22·8 min). Storage of freshly stripped semen was shown to be most successful at 4°C with a half-life of 98·9 min.
The cryopreservation of semen was tested using the cryoprotectants glycerol and dimethyl sulphoxide at concentrations ranging from 0·5M to 2·0M, and pre-freezing equilibration times of 5 and 15 min. Glycerol at 20 M was shown to give significantly superior results. There was no significant difference between sperm activation or sperm half-life for fresh-stripped semen and frozen semen, using glycerol at 2·0M as the cryoprotectant.     

Antibiotics,Penicillin and Sreptomycin improve semen quality indices of endangered Caspian brown trout,Salmo trutta caspius (Kessler, 1870) during in vitro short‐term storage

In this study, we examined the effects of 500 IU mL^?1 penicillin + 500 μg mL^?1 streptomycin sulphate on semen quality indices of endangered caspian brown during 12 days short‐term storage at 4°C. Twenty‐four millilitre semen samples with good quality were considered for the experiment. The semen samples were then stored in the presence and absence of 500 IU mL^?1 penicillin + 500 μg mL^?1 streptomycin sulphate. The semen quality parameters including percentage and duration of sperm motility were measured 0, 3, 6, 9 and 12 days after storage. In the antibiotic receiving group, the values of percentage and duration of sperm motility reduced 3 and 6 days after storage respectively and reduced to lowest levels at day 12. In the antibiotic‐free group, the duration and percentage of sperm motility decreased significantly after 3 days of storage and reached to lowest values at day 12. Also, percentage and duration of sperm motility in each storage time were significantly higher in the antibiotic receiving group than in the antibiotic‐free group. The overall values of percentage and duration of sperm motility for all storage periods were higher in the antibiotic receiving group than in the antibiotic‐free group. In conclusion, our results demonstrated that 500 IU mL^?1 penicillin + 500 μg mL^?1 streptomycin sulphate improves the viability of caspian brown trout during short‐term storage.     

Relationship between transmembrane potential and activation of motility in rainbow trout (Salmo gairdneri)

A hypothesis is developed that activation of motility in rainbow trout spermatozoa is a result of membrane hyperpolarization. This hypothesis was developed to explain experimental observations of a relationship between membrane potential and motility as revealed by the use of voltage sensitive fluorescent dyes. The results lead to the following conclusions: a) Transmembrane potential hyperpolarizes with decreasing KCl concentration in 100 mM NaCl. b) Transmembrane potential hyperpolarizes with decreasing NaCl concentration. c) NaCl is three time less effective in changing transmembrane potential and two orders of magnitude less effective in inhibiting activation of motility than KCl. d) Chloride ions have little effect on transmembrane potential or motility. e) Increases in osmotic pressure with the non-ionic molecule sucrose increased the amount of KCl required to inhibit activation. f) The major effect of Na⁺ on K⁺ inhibition may be osmotic.It is suggested that while sperm cells are in the seminal plasma in the reproductive tract of the male rainbow trout their transmembrane potential is maintained above an activation threshold, probably through Na/K pumps which are found in almost all animal cells. Since K⁺ is the most important ion in determining the transmembrane potential, hyperpolarization of the plasma membrane below an activation threshold occurs when the sperm cells are diluted, during spawning, into the low K⁺ environment of freshwater.     

Protective role of antifreeze proteins on sterlet (<Emphasis Type="Italic">Acipenser ruthenus</Emphasis>) sperm during cryopreservation

The loss of sperm quality in sterlet (Acipenser ruthenus) due to freeze-thaw process in cryopreservation was investigated in the present study. Two antifreeze proteins (AFPI or AFPIII) were used at different concentrations of 0.1, 1, 10, and 100 μg/mL. We compared motility, curvilinear velocity, and plasma membrane integrity of fresh, cryopreserved sperm, and sperm cryopreserved in the presence of antifreeze proteins. Fresh sperm (control) had 85?±?4% motility and 160?±?2 μm/s curvilinear velocity, respectively. After cryopreservation, the motility of frozen-thawed sperm without addition of antifreeze proteins significantly decreased (44?±?9%), compared to the control. The highest motility of frozen-thawed sperm was obtained in cryopreserved sperm with addition of 1 μg/mL of AFPIII (58?±?14%). No significant differences were observed in curvilinear velocity between fresh sperm and cryopreserved sperm with/without addition of AFPI or AFPIII. The flow cytometry analysis revealed that fresh sperm contained 94.5?±?6% live cells, while the cryopreserved sperm only contained 26.6?±?14% live cells. Supplementation of antifreeze proteins has significantly improved the percentage of live cells in frozen-thawed sperm, except 0.1 μg/ml of AFPI group. No significant difference in percentage of live cells was detected in the sperm cryopreserved with 10 μg/mL of AFPI or AFPIII, compared to fresh sperm. Thus, addition of antifreeze proteins to cryopreservation medium could be considered to improve the post-thawed sperm quality of sterlet.     

High‐throughput Cryopreservation of Sperm from Sex‐reversed Southern Flounder,Paralichthys lethostigma

The Southern flounder, Paralichthys lethostigma, is a valuable aquaculture fish with established markets in the USA. All‐female production in this species is an important technology for aquaculture because the females usually have body sizes twice those of males at the same age, and sex‐reversed males (genotypic XX neomales) are used for all‐female production by crossing with genetically normal females. However, sperm volume from the neomales is usually small (<0.5 mL) and limits their application for all‐female fish production. Cryopreservation of sperm from these sex‐reversed neomales will provide access on demand with increased efficiency to extend the application of neomales. The goal of this study was to develop a protocol for cryopreservation of sperm from the Southern flounder by using an automated high‐throughput processing system. The objectives were to: (1) determine the effect of osmolality on activation of sperm motility; (2) evaluate the effect of extender solutions on sperm motility capacity; (3) evaluate the acute toxicity of cryoprotectants (dimethyl sulfoxide [DMSO], propylene glycol, and polyethylene glycol) on sperm motility, and (4) estimate the effect of cooling rate on sperm cryopreservation and post‐thaw fertilization. Sperm motility was activated when osmolality was 400 mOsmol/kg or higher. Of the three extender buffers tested, HEPES4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES) at 300 mOsmol/kg resulted in better protection for sperm motility than did Hanks' balanced salt solution and Mounib solution at 300 mOsmol/kg during 7 d of refrigerated storage. After 30 min equilibration with the cryoprotectant of 15% DMSO, sperm motility was 24 ± 21% (fresh sperm motility without any cryoprotectants was 42%). After cooling at a rate of 20 C/min, post‐thaw sperm motility was 8 ± 5% and fertilization was 63 ± 40% evaluated at the 32–64 cell stage (5 × 10⁵ sperm per egg). Overall, a protocol was developed for sperm cryopreservation in the Southern flounder with high‐throughput processing, which provides a tool to preserve the valuable genetic resources from neomale flounders, and enables germplasm repository development for the Southern flounder.     

Subjective and objective assessment of fish sperm motility: when the technique and technicians matter

Fish sperm motility is nowadays considered the best sperm quality biomarker in fish, and can be evaluated both by subjective and computerized methods. With the aim to compare the precision and accuracy of both techniques, fish sperm samples were assessed by subjective methods and by a computer-assisted sperm analysis (CASA-Mot) system, and simultaneously by three different technicians with different degrees of expertise on the sperm quality analysis. Statistical dispersion parameters (CV, coefficient of variation; and RG, range) were estimated in order to determine the precision and accuracy of the techniques and the influence of laboratory staff on sperm motion assessments. Concerning precision, there were not much significant differences between the technical support staff (high, medium, and low experimented technician), and statistical dispersion parameters were quite similar between them independent of the technique used and the sperm motility class analyzed. However, concerning accuracy, experimented technician reported subjective motility values very closed to the values provided by the CASA-Mot system, only 10 percentage points away from the data provided by a CASA-Mot system. However, medium and low experimented technicians often overestimate the CASA-Mot values, and amplitudes up to 30 percentage points were detected in several sperm assessments. To sum up, both the technique (subjective or objective) and the technician (degree of expertise) became key factors in order to reach accurate motility estimations, so the use of both qualified staff and novel CASA-Mot systems seems to be a critical requirement for obtaining satisfying results in fish species with similar motility patterns.     

Changes in Atlantic cod (Gadus morhua L.) sperm quality during the spawning season

The biology of cod reproduction is well described in the scientific literature. However, sperm biology and spermatozoa management are poorly studied in this species. Because of its recent farming expansion, a better knowledge of cod gametes is becoming especially useful. This work aimed at establishing tools to study sperm biology in cod, and also investigated the existence of changes in cod sperm quality during the spawning period. We showed that sperm concentration could be assessed using spectrophotometry at 260 nm. Sperm motility significantly decreased after a 168‐h storage at 4 °C. A 1:9 dilution of sperm in a non‐activating medium (1/3 seawater and 2/3 freshwater, osmotic pressure: 360 mOsm kg^?1) improved sperm storage. Sperm concentration, sperm velocity and storage capacity at 4 °C peaked during the medium period of the spawning season and then decreased to values close to those observed at the beginning of the reproductive period. The measured values of osmotic pressure, pH, protein, Na⁺, Cl^? and Ca²⁺ concentrations of the seminal fluid were modified along the spawning period. Cell damage was noted at the end of the spawning period: local blebs were observed on the flagellum but also loops at its distal part. On the other hand, spermatocrit did not vary with the sampling date. In conclusion, cod sperm quality is modified during the spawning period, the highest‐quality samples being collected during the medium part of this season.