Motility and fertilizing ability of cryopreserved Caspian brown trout (Salmo trutta caspius) sperm: Effect of post-thaw storage time and different sperm-to-egg ratios

doi:10.1016/j.cryobiol.2015.08.005

Cryobiology

Volume 71, Issue 2, October 2015, Pages 360-363

https://doi.org/10.1016/j.cryobiol.2015.08.005 Get rights and content

Abstract

This study was designed to test the effect of post-thaw storage time on sperm motility parameters of Caspian brown trout (n = 7). Furthermore, we investigated the effect of sperm-to-egg ratios of 100,000:1, 300,000:1 and 600,000:1 on fertility of cryopreserved Caspian brown semen. Quality was assessed by measuring sperm motility parameters and fertilization rates at the eyed and hatching stages. The percentage of post-thawed sperm motility, curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) were not affected by 60 min of storage, whereas a decrease in straight line velocity (VSL), average path velocity (VAP) and linearity (LIN) were found in cryopreserved semen. Thus, the cryopreserved sperm of Caspian brown trout could be stored up to 60 min without loss of the percentage of sperm motility. The fertilization rate was not affected by 60 min of post-thaw storage and was over 70% for sperm-to-egg ratios of both 300,000 and 600,000:1. To our knowledge, this study is the first to report the high post-thaw fertilization ability of Caspian brown trout semen at a sperm-to-egg ratio as low as 300,000:1. This procedure after scaling up can be recommended for routine Caspian brown trout sperm cryopreservation.

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Statement of funding

This work was supported by Islamic Azad University, Tehran, Iran.

Conflict of interest

The authors declare there is no conflict of interest.

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Figueroa et al. (2016) found a significant decrease in the motility rate, VCL, VSL and VAP in post-thaw sperm of Atlantic salmon, whereas Dziewulska et al. (2011) only found a significant decrease in the first three parameters in most of the treatments evaluated in the same species. Golshahi et al. (2015) also found a significant decrease in the VSL and VAP of cryopreserved sperm of Caspian brown trout (Salmo trutta caspius). However, some other studies in salmonids have produced conflicting results, with no significant effects on some post-thaw parameters (Ciereszko et al., 2014; Nynca et al., 2016) or even higher values for thawed spermatozoa compared with fresh.
Sperm cryopreservation is an important tool for reproductive management; however, it has not been possible to perform this procedure for coho salmon. This study was designed to assess the effect of cryopreservation on sperm functional parameters, oxidative stress markers and fertilizing capacity of coho salmon. Eight extenders were tested, consisting of two intracellular cryoprotectant agents (CPA-I): dimethylsulfoxide (DMSO) at 8 and 10% and methanol (MeOH) at 9 and 12%, two extracellular cryoprotectants (CPA-E): 0.14 M trehalose and 0.18 M glucose, supplemented in all cases with 10% egg yolk and Storfish®. Milt samples from 18 males were evaluated in three trials. In the first trial, the toxic effect of cryoprotectants on motility was evaluated after 0, 15, 30, and 60 min of equilibration time. The second trial was conducted to select the extender which produced the best post-thaw sperm motility. In both trials, sperm motility was analysed by a subjective method. In the third trial, milt samples were frozen in the three best extenders; post-thawed samples were assessed for motility by computer-assisted sperm analysis CASA, viability, superoxide anion production (O⁠₂⁠^•−), mitochondrial membrane potential (ΔΨm) by flow cytometry, lipid peroxidation (malondialdehyde, MDA) and fertilizing capacity were also analysed. In Trial 1, no negative effect of the extenders tested was observed on sperm motility during the different equilibration times. In Trial 2, the post-thaw sperm motility was significantly affected by CPA-I, CPA-E and their interaction, in other words, the protection capacity was dependent on the type of CPA-I and at the same time on the CPA-E. Thus, the best results were obtained with DMSO combined with glucose, with mean motility of 16.44 ± 12.55% (8% DMSO + glucose) and 11.57 ± 10.23% (10% DMSO + glucose). In Trial 3, all the sperm parameters were significantly affected after freezing. Mean post-thaw sperm motility varied from 20.50 ± 15.83% to 35.80 ± 17.64%, cell viability ranged from 56.23 ± 11.05% to 59.42 ± 12.00% and the ΔΨm varied from 12.50 ± 8.9% to 14.63 ± 10.09%, without significant differences between the three extenders assessed in this trial. On other hand, the percentage of living spermatozoa producing O⁠₂⁠^•− and the MDA levels increased significantly after freezing, which suggests that sperm cryodamage was caused in part by oxidative stress. The combination of either 8% or 10% DMSO with glucose yielded the best fertility rates (30.00 ± 11.73% and 31.00 ± 10.25% respectively). In conclusion, DMSO in combination with glucose and hen egg yolk is the most suitable extender for preserving the sperm quality and fertilizing capacity of coho salmon.
Optimization of conditions for the cryopreservation of yellow catfish (Pelteobagrus fulvidraco) sperm
2017, Cryobiology
Citation Excerpt :
In the present study, yellow catfish sperm could retain its motility following 2 h of post-thaw storage. Corresponding to our result, sperm of Persian sturgeon and Caspian brown trout can maintain post-thaw quality for 30 and 60 min, respectively [3,13]. According to Horváth et al. [14], post-thaw storage for 1 h did not affect sperm fertilizing capacity in Adriatic grayling and brown trout, indicating that immediate use of thawed sperm for fertilization may not mandatory.
Yellow catfish (Pelteobagrus fulvidraco) is a promising aquaculture species in China with an increasing market demand. To serve the growing demand of male broodstock for artificial fertilization and the preservation of valuable strains for selective breeding, we tried to develop a species-specific cryopreservation protocol for yellow catfish sperm in this study. Important factors such as cryoprotectant, freezing height above the liquid nitrogen (LN) surface, dilution ratio, equilibration time, thawing temperature and cool storage before freezing were standardized. Among the cryoprotectants tested here, 10% Me₂SO was the most suitable for sperm cryopreservation. Freezing at 7 cm above the LN surface for 10 min yielded the highest post-thaw motility. Further evaluation showed that dilution ratio of 1:3 and 1:5 produced higher post-thaw motility than semen diluted at 2:1, 1:1, 1:9 or 1:19. Equilibration times from 0 to 30 min did not cause significant differences in both equilibrated and post-thaw motility. Also, cool storage up to 24 h did not affect the suitability of sperm for cryopreservation. After thawing, sperm could be stored at 4 °C for 2 h without a reduction in motility parameters. With the combination of optimized freezing conditions, the fertilization and hatching rate of cryopreserved sperm were 87.1 ± 5.2% and 78.5 ± 7.4%, respectively, which were similar to those of fresh sperm (91.8 ± 3.5% and 83.7 ± 2.5%). In general, the cryopreservation protocol optimized here would facilitate breeding practice and hatchery operation in this economically important fish.
CRYOPRESERVED SEMEN OF Piaractus orinoquensis (Serrasalmidae): POST-THAW STORAGE TIMES AND ACTIVATING SOLUTIONS
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Standardization of sperm management for laboratory assessment of sperm quality and in vitro fertilization in Eurasian perch (Perca fluviatilis)
2021, Aquaculture International
Semen cryopreservation for the Mediterranean brown trout of the Biferno River (Molise-Italy): comparative study on the effects of basic extenders and cryoprotectants
2019, Scientific Reports

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Cryobiology

Abstract

Section snippets

Statement of funding

Conflict of interest

References (12)

Reproduction in rainbow trout: sex differentiation, dynamics of gametogenesis, biology and preservation of gametes

Aquaculture

Cryopreservation of rainbow trout semen using a glucose-methanol extender

Aquaculture

Spermatozoa concentration influences cryopreservation success in sea trout (Salmo trutta m. trutta L.)

Theriogenology

Determination of semen quality of the rainbow trout, Oncorhynchus mykiss, by sperm motility, seminal plasma parameters, and spermatozoal metabolism

Aquaculture

Cryobanking as tool for conservation of biodiversity: effect of brown trout sperm cryopreservation on the male genetic potential

Theriogenology

Effect of cryopreservation on sperm motility parameters and fertilizing ability of brown trout semen

Aquaculture

Cited by (5)

Cryopreservation of coho salmon sperm (Oncorhynchus kisutch): Effect on sperm function, oxidative stress and fertilizing capacity

Optimization of conditions for the cryopreservation of yellow catfish (Pelteobagrus fulvidraco) sperm

CRYOPRESERVED SEMEN OF Piaractus orinoquensis (Serrasalmidae): POST-THAW STORAGE TIMES AND ACTIVATING SOLUTIONS

Standardization of sperm management for laboratory assessment of sperm quality and in vitro fertilization in Eurasian perch (Perca fluviatilis)

Semen cryopreservation for the Mediterranean brown trout of the Biferno River (Molise-Italy): comparative study on the effects of basic extenders and cryoprotectants

Brief CommunicationMotility and fertilizing ability of cryopreserved Caspian brown trout (Salmo trutta caspius) sperm: Effect of post-thaw storage time and different sperm-to-egg ratios

Abstract

Section snippets

Statement of funding

Conflict of interest

Aquaculture

Aquaculture

Theriogenology

Aquaculture

Theriogenology

Aquaculture

Brief Communication
Motility and fertilizing ability of cryopreserved Caspian brown trout (Salmo trutta caspius) sperm: Effect of post-thaw storage time and different sperm-to-egg ratios