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.     

Effect of GnRHa injection on milt volume in recently stripped rainbow trout Oncorhynchus mykiss

In this study, the effect of gonadotropin‐releasing hormone agonist (GnRHa) injection on milt production in spent rainbow trout was investigated. On day 0, 25 newly matured male rainbow trout (Oncorhynchus mykiss) were stripped manually, and sperm quantity (vol: mL fish^?1) and quality, spermatocrit (%), sperm count (cell mL^?1), motile sperm percentage and motility duration (s) were evaluated. After stripping, fish were randomly divided into five groups: intact; sham (injected with propylene glycol as a hormone vehicle); and groups receiving 4, 8 or 16 μg kg^?1 BW of [d ‐Ala⁶ Des‐Gly¹⁰] mGnRHa. On day 7, the fish were stripped again and the same sperm characteristics as on day 0 were measured. At the beginning of the experiment, there were no significant differences in any of the sperm quantity characteristics between groups. On day 7, expressible milt volume was significantly reduced compared with day 0 (P<0.05, t‐test) in the intact and sham groups but milt quality remained the same (P>0.05, t‐test). The present study shows that GnRHa injection with a concentration as low as 4 μg kg^?1 BW after first stripping could prevent a significant reduction in milt quantity collected 7 days later without any adverse effects on sperm quality.     

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.     

Some characteristics and short-term preservation of spermatozoa of Deccan mahseer, Tor khudree (Sykes)

Over the years, the natural stocks of Tor khudree (Cyprinidae) have depleted due to anthropogenic activities and hence it is considered a threatened species in India. Several in situ and ex situ conservation strategies have been suggested for the revival of T. khudree stocks. The total volume of milt obtained from hormone‐injected (gonadotropin‐releasing hormone analogue+domperidone) T. khudree were significantly higher (six to seven times) than that of uninjected fish. The total number of spermatozoa per fish was also higher in injected fish (6.5 × 10⁸–7.6 × 10⁸) than that obtained from uninjected counterparts (1.3 × 10⁸–1.8 × 10⁸). On the contrary, the spermatozoa density and spermatocrit were found to be lower in injected fish than that of the controls. Spermatozoa density and spermatocrit ranged between 4.1 × 10⁸–4.4 × 10⁸ spermatozoa mL^?1 and 38.1–39.4%, respectively, in injected fish, whereas the figures fluctuated between 6.0 × 10⁸–7.8 × 10⁸ spermatozoa mL^?1 and 61.5–63.1%, respectively, in uninjected fish. However, there was no significant difference in the spermatozoa motility rates between experimental and control fish. Different spermatozoa‐activating media revealed no significant difference in spermatozoa motility between hormone‐injected and uninjected mahseers. But motility duration was the longest with NaCl+urea (190–193 s) and the shortest with tap water (50–55 s) in the experimental and control groups. Short‐term preservation of the spermatozoa of T. khudree indicated that spermatozoa stored at 4°C had higher motility rates than those preserved at room temperature either in the presence or absence of oxygen.     

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.     

Sperm density, seminal plasma composition and their physiological relationship in the endangered Caspian brown trout (Salmo trutta caspius)

Sperm density, mineral and organic composition of the seminal plasma and their physiological relationship were investigated in the Caspian brown trout (Salmo trutta caspius). To establish a rapid and accurate method for assessment of sperm density, three different techniques were used: sperm counting, spectrophotometry (at 480 nm) and determination of the spermatocrit. The seminal plasma contained 159.26±8.84 mM sodium (Na), 33.72±2.01 mM potassium (K), 133.04±5.96 mM chlorine (Cl), 1.68±0.2 mM calcium (Ca) and 0.988±0.13 mM magnesium (Mg). The following organic components were found: total protein 0.75±0.14 mg 100 mL⁻¹, cholesterol 2.86±0.58 mg L⁻¹ and glucose 3.81±1.04 mM L⁻¹. The mean sperm density was estimated to be 3.3 × 10⁹ spermatozoa mL⁻¹. The spermatocrit (%) ranged from 25 to 52 in sperm samples. Highly significant linear relationships were found between sperm density and spermatocrit (R²=0.703, P<0.001) and sperm density and optical density (R²=0.909, P<0.001), indicating that optical density can be used as a quick and accurate method of estimating sperm density. Significant relationships were also found between sperm density and Ca, Mg and total protein of seminal plasma. A significant correlation was also observed between the Ca and Mg concentrations (R²=0.774, P<0.01). The following correlations were observed between mineral and organic components: total protein and Ca (R²=0.462, P<0.05), total protein and Mg (R²=0.518, P<0.05) and glucose and Cl (R²=0.374, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of sperm.     

Gonadotropin releasing hormone‐analogue (GnRHa) treatment improves the milt production and sperm motility of endangered Caspian brown trout,Salmo trutta caspius,over the course of a spawning season

To determine the effect of gonadotropin releasing hormone‐analogue (GnRHa) treatment on the milt quality of endangered Caspian brown trout, Salmo trutta caspius, the sperm motility (percentage and duration of motility), sperm production (sperm density, spermatocrit and milt volume) and milt pH were measured for GnRHa‐treated (the treatment group) and untreated groups (the control group) during the spawning season. For untreated brooders, the values of the motility per cent, sperm density and spermatocrit decreased continuously during the spawning season while the milt volume, duration of motility and milt pH showed only a significant decrease at the end of the season. For GnRHa‐treated males, these parameters increased 14 days after GnRHa treatment (first milt collection) and then decreased continuously towards the end of the season. In addition, the values of milt and sperm density yielded per treated male were higher than that in the untreated group, although these were not statistically different. In any case, the total sum of yielded milt from the treatment group over the spawning season was higher than that in the untreated group. In this experiment, significant positive correlations were found between milt parameters as follows: sperm motility vs. milt pH; sperm density vs. spermatocrit; milt volume vs. spermatocrit; and milt volume vs. sperm density. The results show that the treatment of Caspian brown trout by GnRHa can improve the milt quality in terms of sperm motility and sperm production during a spawning season.     

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.     

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 quality of Brazilian flounder Paralichthys orbignyanus throughout the reproductive season

The aim of this study was to evaluate the sperm quality of Brazilian flounder Paralichthys orbignyanus throughout its reproductive season. Sperm was collected at the beginning, middle and end of the breeding period. Spermatozoa density was maximum at the beginning (12.7 ± 0.92 × 10⁹ cells mL^?1) and at the end (11.8 ± 0.39 × 10⁹ cells mL^?1) of the breeding season (P<0.05). Sperm production and the percentage of spermatozoa moving fast forward increased significantly towards the end of the breeding season (P<0.05). The mean duration of progressive motility of spermatozoa was around 10 min. No difference was observed during the reproductive season in the percentage of motile cells, pH, osmolality and K⁺, Cl^? and Mg²⁺ concentrations in seminal plasma. The concentration of Na⁺ increased throughout the breeding season, reaching 174.62 ± 12.68 mmol L^?1 at the end (P<0.05). There was a decline in the concentration of Ca²⁺ (12.31 ± 3.08 mmol L^?1) in the middle of the breeding season, which coincided with the shortest motility duration of spermatozoa. The information reported in this study should help to improve management and optimize the development of protocols for short‐term storage and cryopreservation of Brazilian flounder semen.     

Induced spermiation in 3-year-old sterlet, Acipenser ruthenus L.

Spermiation in 3‐year‐old sterlet, Acipenser ruthenus (L.), males maintained under warm water conditions was induced by intramuscular injection of either (i) (D‐Ala⁶)‐GnRH‐ProNHEt (Kobarelin); (ii) mammalian GnRH analogue+metoclopramide (Ovopel); or (iii) human chorionic gonadotropin (Biogonadyl). The volume of milt, sperm concentration and motility were measured. A higher percentage of spermiating males was obtained after Kobarelin or Ovopel treatment (81.8% and 77.7% respectively) in comparison with fish treated with Biogonadyl (40.0%). However, only stimulation with Ovopel guaranteed motile spermatozoa in all spermiating males. Moreover, treatment with Ovopel resulted in the highest average milt volume, sperm concentration and motility. The average total number of motile spermatozoa (milt volume×sperm concentration×sperm motility) per individual was 0.99×10⁹, 5.31×10⁹ and 0.02×10⁹ after stimulation with Kobarelin, Ovopel or Biogonadyl respectively. Our data indicate that Ovopel is a good stimulator of spermation in sturgeons. Moreover, the time of sexual maturation could be significantly reduced in sturgeons maintained in cages under warm water conditions.     

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.     

Some properties of bream Abramis brama L. sperm and its cryopreservation

Concentration and motility of spermatozoa, total protein content and its electrophoretic profile, glucose content, activity of aspartate aminotransferase (AspAT) and acid phosphatase (AcP) were assessed in 18 samples of semen from common bream Abramis brama L. males, which were hormonally stimulated to spermiation. Also, milt pooled from four donors was cryopreserved as pellets in vapours of liquid nitrogen (?80 °C) using four extenders (each with or without the addition of hen egg yolk). Mean spermatozoa concentration was 11.68 × 10⁹ mL^?1, and mean spermatozoa motility was about 60%. Protein content in seminal plasma was 2.08 mg mL^?1; both PAGE and SDS–PAGE showed considerable heterogeneity of protein fractions. Mean glucose content was over 11 mg%. AspAT and AcP activities were detected in both seminal plasma and spermatozoa extracts. As calculated to 1 × 10⁹ spermatozoa, AcP and AspAT activities were almost sixfold and 46-fold higher in spermatozoa than in seminal plasma respectively. In the best variant, cryopreservation attempts resulted in 66.6% of eyed embryos (compared with control fertilization) obtained after fertilization of eggs with cryopreserved semen.     

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.     

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.     

Fertilization by refrigerator stored sperm of the Indian major carp, Labeo calbasu (Hamilton, 1822)

This study reports on the spermatological properties, and on the development of a protocol for refrigerator storage (4°C) of Labeo calbasu (Hamilton, 1822) sperm for artificial breeding. Volume, motility, concentration and pH of the freshly collected sperm were 2.21 ± 0.53 (μL g^?1 of fish weight) (mean ± SD), 95 ± 1 (%), 1.93 ± 0.44 × 10⁹ (cells mL^?1) and 7.56 ± 0.17 respectively. Sperm activation was evaluated at different osmolalities of NaCl solution, and motility ceased completely when osmolality of the extender was ≥287 mOsmol kg^?1. Sperm retained motility for 24, 72 and 108 h, after refrigerator storage when sperm were undiluted, suspended in Alsever's solution and suspended in Alsever's solution containing 5% methanol respectively. Fertilization rate of fresh eggs with sperm stored at 4°C in Alsever's solution and Alsever's solution containing 5% methanol was 77% and 60% with a hatching rate of 60% and 43% respectively. The fertilization and hatching success of the stored sperm suggests potential to use refrigeration for transporting genetic material to hatcheries for artificial breeding of L. calbasu in Bangladesh.     

Oxygen consumption and ingestion rate of Penaeus setiferus larvae fed Chaetoceros ceratosporum, Tetraselmis chuii and Artemia nauplii

The effects of the density and type of food on oxygen consumption and ingestion rate of larvae of the white shrimp Penaeus setiferus fed diatoms Chaetoceros ceratosporum, flagellates Tetraselmis chuii and Artemia franciscana nauplii were analysed. Diatoms, flagellates and Artemia nauplii were fed at five densities from 10 to 5 × 10³ cells mL^?1, 0 to 4 × 10³ cells mL^?1, and 0.1, 0.5, 1.0, 1.5 and 2 nauplii mL^?1, respectively. In three experiments, two of three types of food were maintained constant at concentrations of 30-40 × 10³ cells mL^?1 (diatoms), 2 × 10³ cells mL^?1 (flagellates) and 1 Artemia nauplii mL^?1. The oxygen consumption in three experiments increased with larval stage, reaching maximum values in Mill except at lower feed concentrations. A maximum ingestion peak in MI was recorded in larvae fed diatoms, whereas that peak was observed in Mil in larvae fed flagellates. The maximum ingestion rate of Artemia nauplii was observed in Mill. Feed concentrations that produced an optimum metabolic rate as a consequence of equilibrium between ingested food and larval stages were obtained with 20 and 30 × 10³ cells mL^?1 of C. ceratosporum, 2 and 3 × 10³ cells mL^?1 of T. chuii, and 1.0 Artemia nauplii mL^?1. These concentrations would be the most suitable for producing P. setiferus postlarvae.     

Standardization of photometric measurement of sperm concentration from diploid and tetraploid Pacific oysters, Crassostrea gigas (Thunberg)

To provide necessary standardization of procedures for cryopreservation of sperm, a spectrophotomeric method was developed to determine the sperm concentration of diploid and tetraploid Pacific oysters, Crassostrea gigas. Wavelengths of 380, 550, 581 and 780 nm were compared, and 550 and 581 nm were found to be the most sensitive and reliable. A linear relationship between sperm concentration and photometric absorbance was observed for sperm concentrations between 2 × 10⁷ and 2 × 10⁹ cells mL^?1. The regression equation for the standard curve at 550 nm for sperm of diploid oysters was Y=?8.528+1.165 log X. The equation for sperm of tetraploid oysters was Y=?8.844+1.236 log X. The equation at 581 nm for sperm of diploid oysters was Y=?8.07+1.104 log X. The equation at 581 nm for sperm of tetraploid oysters was Y=?8.331+1.167 log X. Comparisons derived from the standard curves at 581 nm between observed values and the predicted values indicated good agreement for sperm from diploid (coefficient of determination, r²=0.983) and tetraploid (r²=0.980) oysters.     

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.     

Hormone-induced ovulation, natural spawning and larviculture of Brazilian flounder Paralichthys orbignyanus (Valenciennes, 1839)

Mature Brazilian flounders Paralichthys orbignyanus were captured in coastal southern Brazil and their reproduction in captivity was studied. Brazilian flounder will spawn naturally in captivity when the water temperature is around 23 °C and 14 h of light is provided daily. Females were induced for ovulation and hand stripping using human chorionic gonadotropin, luteinizing hormone‐releasing hormone analogue or carp pituitary extract. There was no need to inject males, as running milt was observed during the spawning season. Fertilization and hatching rates were above 80% independent of the hormone used. Notochord length at hatching was 2.18±0.07 mm for larvae hatching from naturally spawned eggs. Larvae were reared in salt water (30–35 g L^?1) at 24 °C and under continuous illumination. Larviculture was with green water (Tetraselmis tetrathele 50 × 10⁴ cells mL^?1). Rotifers (10–20 ind mL^?1) were offered as first food 3 days after hatching and gradually replaced by Artemia nauplii (0.5–10 ind mL^?1). Larvae settled to the bottom 20 days after hatching and completed metamorphosis within a week after that. The total length for newly metamorphosed juveniles was 12.9±2.2 mm and the mean survival was 44.8%. The results demonstrate the feasibility of producing Brazilian flounder fingerlings for stock enhancement or grow‐out purposes.