The need for optimization of technologies of fish sperm cryopreservation is conditioned by instability of fresh sperm quality. Presented protocols of cyprinids sperm cryopreservation have been optimized following the results of experiments carried out on varied-quality biomaterial. These protocols ensure with high effectiveness both freezing big volumes of sperm under laboratory and field conditions. Effectiveness of the protocol with two-stage freezing regime in addition has been confirmed by results of cryopreservation of cyprinids sperm samples for addition to the collection of low-temperature gene bank of VNIIPRKH for the last four years.
The research aimed to determine the effect of different taurine concentrations on the duration of sperm fertility preservation and the results of further use during cryopreservation. The taurine was injected into the sperm of the Siberian sturgeon by various methods under conditions of low positive temperatures. Immediately after preparation, the taurine was injected into native sperm at concentrations of 0.01, 0.05, and 0.1 mmol/ml. Two methods of introducing taurine into native sperm were tested: 1 (dry method) - adding taurine powder directly to native sperm and 2 (wet method) - adding taurine dissolved in saline to native sperm. The studies have shown that injections of taurine in an amount of 0.05 - 0.1 mmol/ml both dry and wet can be recommended as the method of long-term preservation of the native sperm of Siberian sturgeon viability. While storing Siberian sturgeon sperm for further cryopreservation the optimal taurine concentration is 0.01 mmol/ml.
Comparative results of experimental work on the study of stellate sturgeon offspring obtained using cryopreserved (experimental) and native (control) sperm, carried out at the scientific and experimental complex of aquaculture “BIOS” of the Volga-Caspian branch of VNIRO (CaspNIRKH) in this article were presented. Molecular genetic studies of cryopreserved sperm, females and males from the brood stock of the complex were carried out. In order to select the optimal parental pairs, calculations of genetic relationship of fish spawners were made. The scheme of the experiment was developed. The technological process of working with stellate sturgeon spawners was described: obtaining mature reproductive cells, insemination, incubation of caviar. Holding and breeding of the studied groups of stellate sturgeon juveniles up to standard weight was carried out. Each stage of the research work was subjected to comparative analysis. The genetic diversity of juveniles was studied. As a result of the work carried out, it was found that the group of fish obtained using of cryopreserved sperm had higher weight indicators, compared with the juveniles of the control group obtained from males from the brood stock of “BIOS”, by an average of 16%. In addition, the experimental group had a higher homogeneity compared to the control group, as evidenced by the values of the coefficients of variation. The survival rate of juveniles of the experimental group of fish was higher than the control group by an average of 10%. The stellate sturgeon obtained using cryopreserved sperm was characterized by higher values of the total number of alleles and the absence of a deficit of heterozygotes compared to the control group of juveniles, which is especially important for preserving the diversity of this valuable sturgeon species. The results of the research work carried out will help develop proposals for the formation of artificially created sturgeon brood stocks by replenishing them with juveniles obtained using cryopreserved sperm and having higher fish breeding biological and genetic indicators. This will allow the herd to have a stock of working material from different generations of sturgeon species, which is a promising factor for the conservation of aquatic biological resources and selection in commercial aquaculture.
Analysis of information from published sources was carried out with the aim of determining the most perspective tendencies and technological approaches in the field of research of salmoniformes sperm cryopreservation methods. Comparative evaluation of currently used methods, analysis of their effectiveness, and discussion of possibility of cryopreservation protocols standardization for the use in aquaculture are presented in this review.
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