Gelatinolytic and anti-trypsin activities in seminal plasma of common carp: relationship to blood, skin mucus and spermatozoa

Proteases and protease inhibitors were detected in the seminal plasma, blood plasma, skin mucus and spermatozoa. Their molecular weights were estimated using SDS-PAGE under non-reducing conditions. The results demonstrate that the two main bands of anti-proteinase activity (APA) detected earlier in common carp seminal plasma with molecular weight of approximately 47 and 58 kDa are also present in other fluids. The intensity of staining was highest in blood and seminal plasma. The intensity in skin mucus was visible, but in a sperm extract it was faint. An additional fast migrating band (30 kDa) was observed only in seminal plasma. Highest APA was found in blood and seminal plasma followed by skin mucus. The activity in sperm extracts was low. The two serine-like proteases with molecular weight of 79 and 189 kDa in seminal and blood plasma have also been found. In skin mucus, protease of 79 kDa was also present. Metalloproteinases with molecular weights of 61 and 69 kDa were found in seminal and blood plasma but metalloproteinases of 44 and 38 kDa were observed only in seminal plasma. Although metalloproteinases, of molecular weight ranging between 61 and 75 kDa, were also visible in a sperm extract, the experimental approach used in this study did not allow unequivocal identification of unique proteases of spermatozoa. Blood plasma contains a serine protease and protease inhibitor not present in seminal plasma. Skin mucus also showed the profile of three unique proteases (two EDTA stimulated and one metalloproteinase). These results indicate that the analysis of proteases and their inhibitors makes it possible to distinguish contamination of milt with either blood or skin mucus. The physiological role of the detected protease-inhibitory system in fish seminal plasma is still unknown. It is possible that the protease inhibitor and proteases, unique for seminal plasma, are involved in a specific function of milt or testis (e.g. the control of spermatogenesis)

Characterization of protease inhibitors of seminal plasma of cyprinids

Anti-proteinase activity was demonstrated in the seminal plasma of cyprinid fish species (bream, chub, ide, dace, asp, goldfish, roach, common carp) using electrophoretic techniques combined with a detection method based on inhibition of bovine trypsin. We found species-specific protease inhibitors in the seminal plasma of cyprinids. At least three bands of protease inhibitors with different migration rates could be identified by native PAGE. Higher variability was characterized for bands with slower migration rates. Visualization of inhibitors after SDS-PAGE under non-reducing conditions allowed estimation of their molecular weights. Apparent molecular weights were within the range of 51–59 and 47–54 kDa for the bands with slower and moderate migration rates, respectively. The molecular weight of fast migration bands for roach and common carp were estimated to 23 and 30 kDa, respectively. Inhibitors of common carp seminal plasma differed in their affinity toward serine proteases. Three inhibitors in common carp seminal plasma could be visualized using cod and bovine trypsin, but only two inhibitors (of high molecular weight) were recognized with chymotrypsin. There were differences in anti-proteinase activity and seminal plasma protein concentration in relation to the origin of common carp seminal plasma (breeding lines) and time of milt collection (spawning vs. post-spawning season)

Gynogenesis in northern pike: UV-inactivation of spermatozoa and the heat shock inducing meiotic diploidization

ABSTRACT Gynogenetic northern pike (Esox lucius L.) were produced using UV irradiated sperm and heat shock applied to inseminated eggs shortly after gamete activation. Milt was diluted in immobilizing solution (1:9) and UV irradiated (6.4 W·m -2 ) for 2-20 min, with dosage in the range of 768-7680 J·m -2 . Genetic inactivation of spermatozoa was most efficient when milt was irradiated for 8 min (3072 J·m -2 ). Insemination of eggs with irradiated milt yielded 100% haploid larvae with hatching rate at 72.1±0.8% (mean±SD), expressed as a percentage of inseminated eggs. Haploid embryo developed and most of them hatched (showing "haploid syndrome") but all haploid larvae died within 48 hours after hatching. After insemination with irradiated sperm the eggs were exposed to a thermal shock of 34°C or 34.5°C, lasting 3 or 5 min, applied 11-16 min after gamete activation. The efficiency of heat shock and survival in experimental groups significantly depended on the source (individual female effect) and quality of eggs. The time of application and duration of the heat shock also affected the survival of embryo and percentage of gynogenetic larvae. Gynogenetic larvae were obtained in all experimental groups with hatching rate as high as 24.2±3.6%

Artificial reproduction of wild and cultured barbel (Barbus barbus, Cyprinidae) under controlled conditions

The aim of this work was to compare the effects of controlled reproduction of cultured and wild common barbel, Barbus barbus (L.). Preparations containing different GnRH analogues and dopamine receptor antagonists (Ovopel, Ovaprim) as well as human chorionic gonadotropin (hCG) (in the case of cultured fish) were applied and their influence on ovulation, spermiation and quality of gametes obtained was determined. No differences in the qualitative or quantitative parameters of semen were found between fish stimulated with different hormonal preparations and those not receiving hormonal stimulation. The high suitability of Ovaprim for ovulation induction in (cultured and wild) barbel was confirmed. The highest synchronisation of ovulation was obtained after the application of Ovopel (18 ± 3 h), but the best results of controlled reproduction (expressed as the percentage of ovulations and survival of embryos) were obtained by applying Ovaprim (83.2 ± 4.1). A significantly higher percentage of ovulation was obtained in cultured fish (80–90%) than in wild fish (< 25%)

SUMMARY

The following article is a summary of research on the influence of season on the reproductive processes in undomesticated animals. The results presented below show: a / an annual hormonal profile of domestic pig and wild boar crossbreed and the antioxidant blood system in the different seasons, b / the possibility of gonadptropic hormone stimulation in chinchillas which are in diestrus or infertile, c / the possibility of using bison’s semen (collected

Influences of heavy metals and 4nonylphenol on reproductive function in fish Xeno

Many industrial and agricultural chemicals (including heavy metals and alkylphenols) present in the environment have adverse effects on the reproductive function in fish. Three studies were conducted to assess toxicity of these chemicals towards reproduction of freshwater fish. It was shown that heavy metals added to the diets accumulate in brain tissue of carp, and this accumulation results in inhibition of the secretion of noradrenaline and stimulation of the secretion of dopamine in the hypothalamus. These processes results in a disturbance of hormonal equilibrium of the hypothalamo-pituitary system, which can unfavorably influence the efficiency of artificial spawning in fish. Quality of salmonid and sturgeon sperm was impaired after in vitro exposure to heavy metals. The degree of this toxic effect was species-specific.