Abstract

Despite passing all quality control checks at animal breeding centres, bulls with apparently normal semen quality can yield unacceptably low field fertility rates. This study took an ex-vivo approach to assess if bulls of divergent field fertility differ in the ability of their spermatozoa to interact with the female reproductive tract and its secretions. Six high and six low fertility Holstein Friesian bulls (+4.0 ± 0.2 and −15.7 ± 3.13, respectively; adjusted mean fertility ± s.e.m. mean of the bull population was 0) were selected from a population of 840 bulls with >500 field inseminations per bull. Thawed spermatozoa from each bull were analysed across a range of in vitro assays to assess their ability to transverse the female reproductive tract including; motility and kinematic parameters using computer-assisted sperm analysis, viability, membrane fluidity and acrosomal integrity using flow cytometry as well as mucus penetration tests, rheotactic behaviour and sperm binding ability to the oviductal epithelium. While there was no significant difference between high and low fertility bulls in most of the sperm motility, kinematic and sperm functional parameters (namely, motility, average path velocity, linearity, straightness, amplitude of lateral head movement), viability, membrane fluidity or acrosome intactness, high fertility bulls had higher curvilinear velocity compared to the low fertility group (P < 0.05) and a higher straight-line velocity was observed although it did not reach statistical significance (P = 0.08). There was no difference between treatment groups in the ability of spermatozoa to penetrate periovulatory cervical mucus or in their rheotactic response (P > 0.05). Interestingly, there was a positive correlation between the straight-line velocity of spermatozoa and their rheotactic response (r = 0.45, P < 0.001) and further linear regression analysis indicated 18.9% of the variance in sperm rheotaxis was accounted for by straight line velocity. A higher number of spermatozoa from the high fertility group compared to the low fertility group bound to oviductal explants (15.1 ± 0.98 and 12.5 ± 0.76, respectively; mean ± s.e.m; P < 0.05). In conclusion, the differences in the kinematics of sperm motility and ability to bind to oviductal explants between high and low fertility bulls were modest and are unlikely to explain the inherent differences in fertility between these cohorts of bulls.

Highlights

  • Artificial Insemination (AI) centres internationally typically use microscopy-based sperm assessments pre- and post-thawing to assess sperm motility, viability and morphology

  • There was no difference in the viability, membrane fluidity or acrosome intactness of spermatozoa from high and low fertility bulls (Table 2; P > 0.05)

  • Through our broad analysis of sperm functional attributes, including kinematic parameters, viability, acrosomal integrity, membrane fluidity, mucus penetration, rheotactic response and sperm binding ability to oviductal epithelium, we identified differences in VCL and in the ability of spermatozoa to bind to the oviductal epithelium which is a pre-requisite for fertilisation

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Summary

Introduction

Artificial Insemination (AI) centres internationally typically use microscopy-based sperm assessments pre- and post-thawing to assess sperm motility, viability and morphology Despite these stringent Quality Control (QC) checks, bulls that have passed, often up to 20e30%, despite the fact that their semen had passed all QC checks in the AI centre. Our group used a linear model of combined data from in vitro assessments to explain 47% of the variation in bull field fertility [9] These studies do not directly inform us of the etiology underpinning sire subfertility and it is unclear if bulls differ in the ability of their spermatozoa to get to the site of fertilisation, interact with the oocyte, complete fertilisation or to sustain embryo and foetal development

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