Application of The UV-Irradiated Homologous and Heterologous Sperm for Activa Tion of the Rainbow Trout (Oncorhynchus Mykiss) Eggs and Production of the Gynogenetic Stocks

Meiotic gynogenetic development of the rainbow trout (Oncorhynchus mykiss, Walbaum 1792) was induced in the course of egg activation performed by the UV-irradiated homologous and heterologous (European grayling Thymallus thymallus Linnaeus, 1758) spermatozoa. To recover diploidy in the gynogenetic zygotes, activated eggs were subjected to the high pressure shock in order to inhibit extrusion of the second polar body. Gynogenetic rainbow trout progeny hatched from the eggs activated by the irradiated rainbow trout and grayling milt with similar hatching rates of 28.19% and 29.22%, respectively. However, gynogenetic rainbow trout produced with grayling semen had shown lower survival than gynogenotes provided with the homologous spermatozoa during two years of rearing. Viable hybrids are not produced between rainbow trout and grayling which ensured that fish obtained in this experiment were true gynogenetic progenies. A Robertsonian polymorphism characteristic for the rainbow trout from the studied strain was also observed among the gynogenetic specimens that exhibited diploid chromosome number ranging from 58 to 62 and stable chromosome arm number (FN= 104). No radiation-induced fragments of the paternal chromosomes were observed in the gynogenetic individuals. Fish produced in both experimental variants were genotypic (XX) and phenotypic (gonads) females. The results confirmed that the gynogenetic protocol used in the present research is an efficient means of producing all-female gynogenetic rainbow trout stocks.

Evaluation of the function of fresh and frozen - thawed sex-sorted and non-sorted stallion spermatozoa using a heterologous oocyte binding assay

The aim of the present study was to evaluate the potential oocyte binding ability and functional integrity of fresh or frozen–thawed, sex-sorted or non-sorted stallion spermatozoa. In the absence of effective IVF procedures in the horse, a heterologous sperm-binding assay was used as an indicator of fertilising capacity to assess differences in the ability of stallion spermatozoa to bind to bovine oocytes. The functional integrity of four treatment groups was assessed: (1) fresh non-sorted spermatozoa; (2) fresh sex-sorted spermatozoa; (3) frozen–thawed non-sorted spermatozoa; and (4) frozen–thawed sex-sorted spermatozoa. Spermatozoa found in association with the zona pellucida of the bovine oocytes were deemed ‘attached’ or ‘bound’ depending on their characterisation as either acrosome intact or acrosome reacted, respectively. Significantly less frozen–thawed spermatozoa were found attached to the oocytes compared with fresh spermatozoa. No significant differences were identified between the number of attached sex-sorted and non-sorted frozen–thawed spermatozoa. However, significantly more sex-sorted than non-sorted fresh spermatozoa were found attached to the oocytes after 1 h coincubation, although after 3 h coincubation this difference was no longer apparent. In conclusion, sex-sorted fresh and frozen–thawed stallion spermatozoa are functionally capable of attaching and binding to bovine oocytes in vitro. Furthermore, fresh sex-sorted spermatozoa attach better than non-sorted spermatozoa, suggesting that they have a more advanced capacitation-like status.