The comparison of two different extenders for the improvement of large‐scale sperm cryopreservation in common carp (Cyprinus carpio)

Abstract

Contents 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.

Optimizing aquaculture-scale common carp artificial reproduction: a novel approach to sperm cryopreservation using large-volume containers and elevated thawing temperatures

“…Consistent with numerous prior investigations on common carp motility parameters, our results align with reported trends in motility percentage, VCL, and LIN (Boryshpolets et al, 2009;Dzyuba et al, 2010;Dzyuba et al, 2013;Bernath et al, 2016;Boryshpolets et al, 2017;Marinovićet al, 2021). However, the observed disparity in BCF outcomes contradicts data presented in several studies (Vaŕkonyi et al, 2019;Marinovićet al, 2021) and aligns with research conducted in non-cyprinid species (Gallego et al, 2017). Although the underlying reasons for these discrepancies remain challenging to elucidate due to the limited number of comparable examples, it is supposed that both cryopreservation and post-activation time may contribute to increased dribbling of spermatozoa during their motility, as suggested by the observed rise in BCF (Wilson-Leedy and Ingermann, 2007).…”

“…This critical task was, among others, solved in the current study, which provides, for the first time, the proper combination of cryoprotective medium and freezing and thawing rates appropriate for the cryopreservation of carp sperm in containers of large volume and low dilution rate. This combination distinguishes the described method from the methods already described for big sperm volume cryopreservation, which often involves high dilution rates (Horvath et al, 2007;Vaŕkonyi et al, 2019;Bokor et al, 2023) or low dilution rates; the fertilizing ability was not assessed (Vaŕkonyi et al, 2018).…”

Crioconservación seminal en peces de agua dulce: aspectos biotecnológicos, celulares y bioquímicos

“…Las pajillas son el sistema de empaque más empleado para la crioconservación seminal puesto que su alta relación/superficie volumen facilita la congelación y descongelación a tasas uniformes (Maria et al, 2015). Se han evaluado pajillas de diferentes volúmenes, siendo las de 0.25 ml y 0.5 ml las más utilizadas (Bernáth et al, 2016;Martínez-Páramo et al, 2017) aunque las pajillas de mayor volumen (1.2 -5.0 ml), también llamadas macrotubos, han sido probadas exitosamente (Medina-Robles et al, 2007;Ding et al, 2011;Várkonyi et al, 2019). Para el caso de peces que producen poco semen, como es el caso del pez cebra (Danio rerio), se han diseñado pajillas de doble chaqueta con una capacidad de tan sólo 0.1 ml, con un peso incorporado en cada extremo para evitar la flotabilidad sobre el agente criogénico (Grevle y Ruud, 2019).…”

“…Además, su compatibilidad con las placas de pozos facilita su procesamiento automatizado. El efecto de los criotubos sobre la calidad seminal es independiente de su volumen de empaque (Maria et al, 2015;Várkonyi et al, 2019).…”

Effect of fish antifreeze protein type iii supplementation to semen extender on cryopreserved dog spermatozoa

“…Composition of CPA in the extender can affect the freezing behaviour of the specific cell, which effect on the rate of transmembrane water transport, ice nucleation and ice crystal growth. By the mean, differences in composition of extender have to be followed by specific optimum freezing protocols (Le et al, 2019;Pabon et al, 2019;Varkonyi et al, 2019). The adverse result in our study can be related with those factors, which are related with the ice crystal formation in freezing or thawing processes.…”