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In our study, a traditionally used (Grayling, already used in cyprinid species) and a newly tested (Pike) extender was tested to avoid sperm agglutination phenomenon following thawing during carp sperm cryopreservation. A large‐scale (elevated volume of sperm) freezing method in a controlled‐rate freezer using 5 ml straw and 10 ml cryotube was also systematically established. In all experiments, the sperm cryopreserved in using Grayling extender (except only one sample) showed an agglutination phenomenon (damaged and intact cells adhered to each other) after thawing where Pike extender resulted the regular cell suspension. No significant difference was observed between the two cryopreserved groups (Pike and Grayling extender) in all motility parameters using the 0.5 ml straw and the polystyrene box. Similarly, motility parameters did not show a significant difference in the two frozen groups with the 5 ml straw, also in the polystyrene box. A significantly higher progressive motility (pMOT, Grayling: 54% ± 8%, Pike: 37% ± 5%), straight line velocity (VSL, Grayling: 50 ± 5 µm/s, Pike: 39 ± 4 µm/s) and beat cross frequency (BCF, Grayling: 20 ± 1 Hz, Pike: 17 ± 1 Hz) was observed in the case of the grayling extender by the 5 ml straw cryopreserved in a controlled‐rate freezer (CRF) compare to the pike extender. A significantly higher VSL (Grayling: 45 ± 3 µm/s, Pike: 38 ± 4 µm/s) was observed by the grayling extender using the 10 ml cryotube than with the pike extender. Despite the randomly occurring differences in a few parameters, our new controlled freezing method using the newly tested Pike extender, the 5 ml straw or the 10 ml cryotube can be a good solution for the preservation of elevated volume of carp sperm.
In our study, the comparison of 5 ml straw and 10 ml cryotube during sperm cryopreservation in a Hungarian carp landrace (Cyprinus carpio carpio morpha accuminatus) was carried out. Three different dilution ratios (1:1, 1:4 and 1:9) were also tested using the cryotube. A significantly higher pMOT was recorded using the cryotube in comparison with the straw. VCL and STR were similar in both groups. Cryopreservation had a negative effect on pMOT and VCl using the cryotube and also the straw where, STR was not reduced significantly. An increasing tendency was observed using higher dilution of sperm during cryopreservation; however, significant difference was not recorded between the three groups. VCL and STR were similar in all groups. Cryotube was applicable for the sperm cryopreservation of the Hungarian carp landrace. However, the standardization for the freezing method specified for this volume is recommended. The different dilution ratios needed be tested also during fertilization.
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