The investigation of post-thaw chilled storage and the applicability of large-scale cryopreservation in chub (Squalius cephalus) sperm

G Bernáth; B Csorbai; B Nagy; E Csókás; J Molnár; T Bartucz; Z L Láng; M Gyurcsák; Á Hegyi; J Kobolák; J D Griffitts; Á Ferincz; B Urbányi; Z Bokor

doi:10.1016/j.cryobiol.2023.104588

The investigation of post-thaw chilled storage and the applicability of large-scale cryopreservation in chub (Squalius cephalus) sperm

Cryobiology. 2023 Dec:113:104588. doi: 10.1016/j.cryobiol.2023.104588. Epub 2023 Oct 7.

Authors

G Bernáth¹, B Csorbai², B Nagy², E Csókás², J Molnár², T Bartucz², Z L Láng², M Gyurcsák², Á Hegyi², J Kobolák², J D Griffitts³, Á Ferincz⁴, B Urbányi², Z Bokor²

Affiliations

¹ Department of Aquaculture, Institute for Aquaculture and Environmental Safety, Hungarian University of Agriculture and Life Sciences, H-2100, Gödöllő, Hungary. Electronic address: Bernath.Gergely@uni-mate.hu.
² Department of Aquaculture, Institute for Aquaculture and Environmental Safety, Hungarian University of Agriculture and Life Sciences, H-2100, Gödöllő, Hungary.
³ Department of Environmental Toxicology, Institute for Aquaculture and Environmental Safety, Hungarian University of Agriculture and Life Sciences, H-2100, Gödöllő, Hungary.
⁴ Department of Freshwater Fish Ecology, Institute for Aquaculture and Environmental Safety, Hungarian University of Agriculture and Life Sciences, H- 2484, Agárd, Hungary.

PMID: 37813176
DOI: 10.1016/j.cryobiol.2023.104588

Abstract

Chub (reophillic cyprinids) is one of the most sensitive bioindicator fish of environmental changes following anthropogenic activities. The improvement of different biotechnological procedures could help support its conservation and strengthen the natural populations. The aim of this study was to compare the effects of two different hormonal agents (carp pituitary extract and Ovopel™) on various motility parameters (pMOT-%, DAP-μm, VCL μm s^-1, VSL-μm s^-1, LIN-%, ALH-μm, BCF-Hz) of fresh and cryopreserved/thawed sperm (stored at 4 °C for 6 h). Additionally, we sought to develop a novel, large-scale cryopreservation method for chub sperm, assessing freezing methods (Styrofoam box and a controlled-rate freezer) and different containers (0.5, 5 mL straw and 4 mL cryotube) for sperm cryopreservation. The results of this study indicated no difference between the carp pituitary extract and Ovopel treated groups in either the fresh or frozen/thawed sperm (at 0, 3, 6, hour post thawing, P = 0.4351). In contrast, the quality of the thawed chub sperm was negatively affected after 3 h chilled storage in both hormonal treatments (P = 0.0036, P < 0.0001). When assessing the motility parameters of the sperm between the 5 mL straw and 4 mL cryotube groups cryopreserved in a Styrofoam Box, no difference was observed (P = 0.103). Additionally, sperm loaded in 4 mL cryotubes showed no difference in motility when cryopreserved with either the Styrofoam box or controlled-rate freezer methods (P = 0.109). A similar hatching rate was observed in sperm preserved using the Styrofoam box (35 ± 7 %) and controlled rate freezer (25 ± 9 %) methods (P = 0.300). In a second fertilization trial, hatching rate was similar between control (72 ± 19 %) and cryopreserved (4 mL cryotube and Styrofoam box, 61 ± 5 %) groups. (P = 0.257). Based on our findings and its standard features (less species specific, precise dose calculation), Ovopel can be a good candidate for the stimulation of spermiation in chub sperm prior to cryopreservation. Furthermore, our study presents a novel and applicable method for the large-scale cryopreservation of chub sperm.

Keywords: Chub sperm motility; Hatching rate; Hormonal treatment; Large-scale sperm cryopreservation; Post-thaw storage.

MeSH terms

Animals
Carps*
Cryopreservation / methods
Cryoprotective Agents / pharmacology
Cyprinidae*
Male
Semen
Semen Preservation* / methods
Semen Preservation* / veterinary
Sperm Motility
Spermatozoa

Substances

styrofoam
Cryoprotective Agents