Fertilizing capacity and motility of tenchTinca tinca(L., 1758) sperm following cryopreservation

Abstract

Experiments were carried out to develop an optimal cryopreservation protocol for tench sperm by testing the fertilizing capacity and motility parameters including progressive motility, curvilinear velocity (VCL) and linearity (LIN) of cryopreserved sperm. Three experiments were designed to this aim: first experiment where we tested the effects of two extenders (sugar-based Grayling and ion-based Kurokura 180) and two cryoprotectants (DMSO and methanol) on fertilization and hatching success; second where we tested the effect of cryoprotectant type (methanol or DMSO) in different concentrations (5%, 10% and 15%) on fertilization and hatching success; and third where we tested the effect of two cryoprotectants (methanol and DMSO) on sperm motility parameters (progressive motility, VCL and LIN) after 4 h post-thaw storage (4°C). Sperm prepared with the sugar-based Grayling extender displayed better fertilization and hatching rates independently of the applied cryoprotectant most likely due to glucose present which acted as an external cryoprotectant. Concerning cryoprotectant concentrations, the use of 10% methanol yielded the highest fertilization (85 AE 15%) and hatching (80 AE 13%) rates, which were significantly higher than in all other groups. During the post-thaw storage time, 5% methanol, 10% methanol and 5% DMSO groups had significantly higher motility parameters than other groups and we observed no significant decline in any of the parameters during the storage time. Overall, we found that a sugar-based extender in combination with methanol as cryoprotectant is suitable for the cryopreservation of tench sperm and allows storage of cryopreserved sperm for up to 4 h post thaw.

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

“…Further measurement of the changes in cell density during the agglutination process can clarify the mentioned effect. In the last 15 years, several publications proved that the composition of the extender is one of the key factors in cyprinid (carp) sperm agglutination but the level of the agglutination is species specific (Horváth et al, 2003;Irawan et al, 2010;Lujić et al, 2017;Marinović et al, 2017). The sugar content is necessary in the extender designed for common carp sperm cryopreservation (Horváth et al, 2003;Irawan et al, 2010).…”

Cryopreservation of Danube barbel Barbus balcanicus sperm and its effects on sperm subpopulation structure

“…On the other hand, Horváth et al [22] reported that sperm of carp cryopreserved with MetOH exhibited significantly higher post-thaw motility than sperm cryopreserved with DMSO. MetOH has been used successfully for sperm cryopreservation in other cyprinid species such as tench [12] and zebrafish [23]. The appropriate cryoprotectant varies among fish species and interactions between extender and cooling rate [24].…”

“…Immediately before stripping, the genital papilla was wiped dry to prevent contamination by water or mucus, and sperm was collected into 2-ml syringes by applying abdominal pressure. To prevent fast agglutination occurring in barbel sperm, stripped sperm was diluted in a glucose-based extender (GBE; 40 mM KCl, 30 mM Tris, 200 mM glucose [11,12]), supplemented with 20 µg/mL DNase I (PanReac AppliChem, Spain, Germany, Italy). Samples were kept on ice until further analysis (maximum duration 20 min).…”

“…In light of reports on the beneficial effects of sugars on sperm motility parameters [12], we tested the additional supplementation of three sugars: glucose (Glu), sucrose (Suc) and trehalose (Tre) at two concentrations, 0.1 M and 0.3 M (v/v). The control group contained no additional sugar supplementation.…”

Outcomes of in vitro fertilization with frozen‐thawed sperm: An analysis of post‐thaw recovery of sperm, embryogenesis, offspring morphology, and skeletogenesis for a cyprinid fish

“…Although post-thaw sperm longevity was not different between the tested CPAs, motility in METH-5% treatment was~63%, which is higher than the other treatments, suggesting it efficiently reduced cryoinjuries as demonstrated for other fishes. [50][51][52][53][54][55] In vitro fertilization trials are a necessary step for validating the effectiveness of sperm cryopreservation protocols due to close correlations between sperm motility and fertility. 56,57 Damage to sperm structure during freezing-thawing may impact fertility capability of the spermatozoon.…”