Abstract

Computer-assisted sperm class analyser (CASA) analysis of avian semen following cryopreservation indicates that their semen motility and viability parameters become compromised, due in part to oxidative stress. To mimic these observations we have treated cockerel semen with an oxidative stress inducing agent, namely hydrogen peroxide (H2O2) and monitored the motility, kinematic and viability parameters over time. Briefly, five healthy and fertile South African Venda cockerels were selected and their semen was collected using the abdominal massage technique. The semen was then treated with H2O2 at 0 &microM, 5 &microM, 50 &microM and 200 &microM concentrations for 0, 3, 16 and 24 hrs. The semen motility, kinematic and viability parameters were then determined using the CASA system while the viability was determined using the SYBR-14/PI staining. The Pearson’s correlation coefficient was determined to test the relationships between the levels of induced oxidative stress, period of exposure to oxidative stress inducing agent and the motility plus kinematic parameters. Our data revealed that in raw cockerel semen, there was high and positive correlations between total motility (TM), progressive motility (PM), rapid (RAP), curvilinear velocity (VCL), straight line velocity (VSL) and average path velocity (VAP) while the kinematic parameters LIN, STR, WOB, ALH and BCF had low or negative correlations with them. Furthermore, TM, PM, RAP, VCL and VSL remained highly and positively correlated with the induced oxidative stress and also, linearity (LIN), straightness (STR), wobble (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) remained negatively correlated with the induced oxidative stress, after 3 hrs. After 24 hrs, TM, PM, RAP, VCL, VSL, VAP and ALH, became negatively correlated with the induced oxidative stress while LIN, STR, WOB and BCF became positively correlated with the induced oxidative stress. Conversely, when the H2O2 concentration used was correlated with motility and kinematic parameters over time, TM, PM, RAP, VCL, VSL, VAP became negatively correlated with oxidative stress while LIN, STR, WOB, ALH and BCF show negative or low correlations with the induced oxidative stress. This data indicates that LIN, STR, WOB, BCF and to some extend ALH, reveal the least correlations with the induced oxidative stress under persistent oxidative stress conditions in cockerel semen. In conclusion, cockerel semen, like buck semen, does not easily succumb to oxidative stress since the raw semen correlations of CASA analysed parameters are comparable to these observed after 3 hrs of H2O2 treatment. In addition, the oxidative stress levels tolerated by cockerel semen should not 5 &microM H2O2 oxidative stress levels. Lastly, lack of correlation between LIN, STR, WOB, BCF and ALH and induced oxidative stress can be used in cockerel semen to show intolerable cryopreservation conditions.

Highlights

  • Cryopreservation of semen from various species has been shown to undergo oxidative stress [1] [2] [3] [4]

  • Computer-assisted sperm class analyser (CASA) analysis of avian semen following cryopreservation indicates that their semen motility and viability parameters become compromised, due in part to oxidative stress

  • We show that the kinematic parameters beat cross frequency (BCF), WOB, LIN, STR show positive correlations with induced oxidative stress over extended periods in cockerel semen

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Summary

Introduction

Cryopreservation of semen from various species has been shown to undergo oxidative stress [1] [2] [3] [4]. The cryopreservation induced oxidative stress in semen resulted in cellular damage including lipid peroxidation, DNA damage, membrane damage and reduced motility, velocity and viability parameters [2]. Our local Venda chickens have recently received much attention as part of the primary source of protein in poverty alleviation strategies [7] They possess desirable traits enjoyed by resource-poor small-scale farmers, which includes sexual maturity at 20 weeks and excellent fertility rates [8]. Previous cryopreservation of their semen has revealed that their fertility post-thaw have low semen motility, semen velocity and viability parameters, due in part to oxidative stress [8] [9]. CASA technology allows for accurate and automated analysis of motility and kinematic patterns of each spermatozoa analysed showing their TM, PM, RAP VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF values [11]

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