Dynamics of sperm DNA fragmentation in the swine: Ejaculate and temperature effects

Sperm DNA fragmentation is one of the major causes of infertility; the sperm chromatin dispersion test (SCDt) evaluates this parameter and offers the advantage of species-specific validated protocol and ease of use under field conditions. The main purpose of this study was to evaluate sperm DNA fragmentation dynamics in both fresh and post-thaw bottlenose dolphin sperm using the SCDt following different cryopreservation protocols to gain new information about the post-thaw differential sperm DNA longevity in this species. Fresh and cryopreserved semen samples from five bottlenose dolphins were examined for sperm DNA fragmentation dynamics using the SCDt (Halomax(®)). Sperm DNA fragmentation was assessed immediately at collection and following cryopreservation (T0) and then after 0.5, 1, 4, 8, 24, 48 and 72 h incubation at 37°C. Serially collected ejaculates from four dolphins were frozen using different cryopreservation protocols in a TES-TRIS-fructose buffer (TTF), an egg-yolk-free vegetable lipid LP1 buffer (LP1) and human sperm preservation medium (HSPM). Fresh ejaculated spermatozoa initially showed low levels of DNA fragmentation for up to 48 h. Lower Sperm DNA fragmentation (SDF) was found in the second fresh ejaculate compared to the first when more than one sample was collected on the same day (p < 0.05); this difference was not apparent in any other seminal characteristic. While there was no difference observed in SDF between fresh and frozen-thawed sperm using the different cryopreservation protocols immediately after thawing (T0), frozen-thawed spermatozoa incubated at 37°C showed an increase in the rate of SDF after 24 h. Sperm frozen in the LP1(℗) buffer had higher levels (p < 0.05) of DNA fragmentation after 24- and 48-h incubation than those frozen in TTF or HSPM. No correlation was found between any seminal characteristic and DNA fragmentation in either fresh and/or frozen-thawed samples.

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“…These results are similar to the DNA fragmentation rate described in the boar (Pérez‐Llano et al. ; Alkmin et al. ) and koala (Johnston et al.…”

Section: Discussionsupporting

confidence: 89%

Effect of Cryopreservation on the Sperm DNA Fragmentation Dynamics of the Bottlenose Dolphin (Tursiops truncatus)

Sánchez‐Calabuig

López‐Fernández

Johnston

et al. 2015

Reprod Domestic Animals

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“…These two parameters are also positively correlated (De Iuliis et al ., ; Zribi et al ., ). Our results show that DNA damage (fragmentation and oxidation) remained similar through incubation time (18 h) and are according with kinetic DNA fragmentation described in bovine (González‐Marín et al ., ) and boar spermatozoa, which only shows a significant increase in DNA fragmentation after 2‐day incubation (Perez‐Llano et al ., ).…”

Section: Discussionmentioning

confidence: 97%

Supplementation of IVF medium with melatonin: effect on sperm functionality andin vitroproduced bovine embryos

et al. 2014

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Gamete co-incubation generates high free radical levels surrounding growing zygotes which may impair subsequent embryo viability. Melatonin eliminates a wide variety of free radicals; hence, we tried to improve in vitro embryo production by adding melatonin to in vitro fertilisation (IVF) media in high (Exp. 1) and low concentrations (Exp. 2), and we evaluated its effect on bull sperm function during IVF co-incubation time (Exp. 3). In Experiment 1, we supplemented IVF media culture with 0.01, 0.1 and 1 mmol of melatonin, along with a no melatonin control group. In Experiment 2, melatonin levels were reduced to 10, 100 and 1000 nmol, with a no melatonin control group. In Experiment 3, spermatozoa were incubated in IVF media with melatonin (as Exp. 2) and functional parameters were analysed at 0, 4 and 18 h. In Experiment 1, only 1 mmol melatonin showed lesser blastocyst rates than control (C: 23.2 ± 6.7% versus 1 mmol: 2.0 ± 1.7%). In Experiment 2, no statistical differences were found in cleavage percentage, blastocyst percentage and total cell count for any melatonin treatment. In Experiment 3, sperm samples with 1000 nmol melatonin had a significantly higher wobbler (WOB) coefficient, a lower percentage of intact acrosomes, a lower percentage of viable spermatozoa with ROS, greater DNA fragmentation and higher DNA oxidation than controls. Total fluorescence intensity for ROS at 10 nmol melatonin was significantly greater than controls (P < 0.05). IVF media with 1 mmol melatonin is deleterious for embryo development, and in lower concentrations, it modulated sperm functionality, but had no effects on embryo production.

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“…Enciso et al [3] mentioned differences in the shape of the halo around the boar spermatozoa head when SCD was performed on fresh semen samples. In addition, Pérez-Llano et al [20] defined two new sperm morphologies after performing a dynamic analysis of sperm DNA damage by SCD test on fresh semen samples: one including a large halo of chromatin dispersion with a highly degraded sperm core and the other showing a compact core lacking the small halo detected around the sperm heads at baseline. However, they only considered relevant the sperm population that showed nucleoids with a large and spotty halo of chromatin dispersion, indicative of sperm with nuclear fragmented DNA [3].…”

Section: Discussionmentioning

confidence: 99%

Freeze-dried dog sperm: Dynamics of DNA integrity

et al. 2015

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Dynamics of sperm DNA fragmentation in the swine: Ejaculate and temperature effects

Cited by 34 publications

References 32 publications

Effect of Cryopreservation on the Sperm DNA Fragmentation Dynamics of the Bottlenose Dolphin (Tursiops truncatus)

Effect of Cryopreservation on the Sperm DNA Fragmentation Dynamics of the Bottlenose Dolphin (Tursiops truncatus)

Supplementation of IVF medium with melatonin: effect on sperm functionality andin vitroproduced bovine embryos

Freeze-dried dog sperm: Dynamics of DNA integrity

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