Katarzyna Dziewulska

Post-thawed motility and fertility from Atlantic salmon (Salmo salar L.) sperm frozen with four cryodiluents in straws or pellets

The cryopreservation of salmonid sperm is a complex process involving the interplay of many factors. Although cryopreservation protocols can be evaluated through a range of responses at various stages in the process, the number of progeny is the ultimate indicator of success. We compared reproductive success from freezing Atlantic salmon (Salmo salar L.) sperm using the eight combinations of (1) the penetrating cryoprotectants, 10% dimethyl sulfoxide (DMSO) or methanol (MeOH); (2) the nonpenetrating cryoprotectants glucose (0.3 M) or sucrose (0.6 M), and freezing in 0.1 mL pellets or 0.25 mL straws. All cryodiluents were supplemented with 10% (v/v) of hens egg yolk. Response variables were the percentage and degree of motility of thawed and activated sperm using computer assisted sperm analysis (CASA), and rates of eyed embryos, hatch and egg sac larvae. Growth rates of alevins were assessed to two months post hatch. Atlantic salmon milt cryopreserved in straws had higher spermatozoa motility and fertilization success than milt cryopreserved in pellets (P < 0.05). Type of sugar tested did not significantly affect the response variables. In the MeOH treatment, thawed spermatozoa achieved higher speed and a higher fertilization rate evaluated at the eyed embryo stage than spermatozoa subjected to the DMSO treatment. Higher mortality rate (especially before hatching) of MeOH offspring than DMSO offspring led to equal numbers of progeny for the two treatments from the swimming stage to the end of the study. Moreover, during feeding fish from the MeOH group produced significantly lower weight larvae than the DMSO and control groups. Even so, the weight of the MeOH group was satisfactory. Length and the condition factors did not differ significantly among the larvae groups. Significant positive correlations were found between fertilization success (measured in number of eyed eggs) and both motility (rs = 0.81), and velocity (rs = 0.49). Freezing in straws gave betters results than freezing in pellets for cryopreservation of salmon milt; whereas type of sugar tested (glucose vs sucrose) did not have significant effects. Penetrating cryoprotectants DMSO and MeOH differed in their effect on post-thawed sperm velocity, fertilization rate and mortality rate of progeny, suggesting the need for further research on the influence of these cryoprotectants on frozen sperm and and post-fertilization devopmental processes.

Effect of pH and cation concentrations on spermatozoan motility of sea trout (Salmo trutta m. trutta L.).

Environmental conditions during external fertilization in fish have a significant effect on spermatozoan motility (MOT) and fertilization ability. Even in the same family of fish, spermatozoa might differ in sensitivity to ions present in the external medium. Elucidation of such differences within a species would help to understand spermatozoan biology and to determine external conditions that would optimize spermatozoan MOT and successful fertilization. Objectives of the current study were to determine the effects of pH and of sodium, potassium, and calcium ion concentrations in the activation solution on sea trout spermatozoan MOT. Six parameters characterizing MOT (MOT, curvilinear velocity [VCL], linearity, amplitude of lateral head displacement, beat cross frequency, and duration of MOT) in spermatozoa activated in prepared buffers were traced by computer-assisted sperm analysis. Sea trout spermatozoa were motile over a wide range of pH values, and increasing pH did influence MOT, VCL, linearity, amplitude of lateral head displacement, and MOT duration. The optimum pH for sperm MOT was established at approximately 10. Increasing K(+) ion concentration within the observed range caused a decrease in MOT and VCL. Spermatozoan movement ceased at 8 mM KCl concentrations. In Ca(2+) buffers, sperm were motile within the range of 0 to 70 mM CaCl(2) concentration; although beyond 8 mM concentration, VCL and MOT gradually declined. Spermatozoan aggregation was observed at the highest ion concentrations tested. Increasing CaCl(2) concentration affected MOT pattern from initiation to termination of spermatozoan movement in a similar manner as changes associated with increasing pH. At concentrations of CaCl(2) higher than 0.5 mM and in buffers with pH values 10 to 11, movement of spermatozoa was characterized by high initial linearity followed by its gradual reduction. In contrast to the effects of KCl and CaCl(2), increasing NaCl concentration up to 90 mM Na(+) concentration prolonged the duration of spermatozoan movement and, up to 60 mM Na(+) concentration, slightly increased sperm velocity as well. Above the concentration of 90 mM NaCl, these parameters decreased; and at 240 mM of Na(+), spermatozoa did not activate.

ANNUAL DEVELOPMENT OF GONADS OF PUMPKINSEED, LEPOMIS GIBBOSUS (ACTINOPTERYGII: PERCIFORMES: CENTRARCHIDAE) FROM A HEATED-WATER DISCHARGE CANAL OF A POWER PLANT IN THE LOWER STRETCH OF THE ODER RIVER, POLAND

The pumpkinseed, Lepomis gibbosus (Linnaeus, 1758), known also as pumpkinseed sunfish, is native to eastern and central North America (Tomeček et al. 2007). Because of its attractive appearance, the species was introduced to Europe in the 19th century as an ornamental fish for aquaria or park ponds, or a prospective sport fish (Vooren 1972, Witkowski and Grabowska 2012). Natural escapes and intentional introduction of the fish into open waters has resulted in fast spreading of the species over Europe and a successful colonisation. The pumpkinseed has been already found in at least 28 European countries and in Asia Minor (Coop and Fox 2007). The fish are found in shallow, weedy, quiet reservoirs, lakes, river inlets, tributaries, and canals, in cool to moderately warm waters (Miller 1963, Tomeček et al. 2007). The range of the species is limited by the water temperature. The optimum temperature for pumpkinseed growth is 20–30oC (Mischke and Morris 2002). For reproduction, they need the critical temperature range of 14–16.5oC (Burns 1976). ACTA ICHTHYOLOGICA ET PISCATORIA (2014) 44 (2): 131–143 DOI: 10.3750/AIP2014.44.2.07

Survival rate and growth in the wild of sea trout (Salmo trutta L.) fry obtained using frozen semen

Abstract The aim of the study was to assess the growth and development of sea trout, Salmo trutta L., fry obtained from frozen, thawed semen after the fish had been released into the wild and were feeding there. The semen was cryopreserved with either methanol (MeOH) or dimethyl sulfoxide (DMSO). Fresh eggs were collected and fertilized with the thawed semen. The control was eggs fertilized with fresh semen that had been kept on ice. The eggs were incubated and reared in a closed recirculating system. The fertilized eggs were counted for visible eye pigment. After rearing, three groups of fish were tagged and stocked into a stream. The fish aged 6 months were caught from the stream. The fish were measured, weighed, identified to which experimental variant they belonged, and their survival rate was estimated. No significant differences in survival rate, length, weight, or condition factor among the control, DMSO, and MeOH groups were noted during larval development in a closed recirculating system. These parameters were also similar in the control and experimental groups after the fish had grow in a natural stream. The use of frozen semen to fertilize fresh sea trout eggs resulted in a normal incubation process, larval stage, and regular fry growth and survival, both under laboratory conditions, and, most importantly, in the wild. The results indicate it is possible to use frozen semen for fry production and the restoration of wild fish populations.