In vitro competitive binding index using fluorochrome-labelled spermatozoa for predicting bull fertility.

This work evaluated if an in vitro test, with the combined power of the statistical evaluation of spermatozoa and zona pellucida (ZP) competitive binding ability and a rapid method for accessory sperm counts, could predict the bull fertility. Ten Holstein Friesian bulls of known field fertility (five of high and five of low fertility) were selected. An in vitro heterospermic insemination approach, based on differential staining, was tested on 45 possible pairs of bulls (two batches per bull). Motility and quality (abnormalities and membrane status) seminal characteristics and estimated relative conception rates (ERCR) highlighted only one association between membrane integrity and ERCR (p = 0.007). Differences in ZP binding allowed us to rank bulls into two categories based on low and high binding ability. For eight bulls, this classification reflected the ERCR. Differences between batches were reported for two bulls, in which the effect of heterospermic insemination (the number of sperm binding to ZP from different bulls not in a 1:1 ratio) showed a significant bull-related effect (p < 0.001) in the first batch and no effect (p > 0.05) in the second batch for both bulls. Reduction of the number of oocytes per assay from 25 to 5 had no effect (p > 0.5) on the bulls' ranking. Our results suggest that in vitro competitive binding is a promising approach for estimating bull fertility and support concepts for further implementation, e.g. drastic reduction of oocyte number in a single pair assay and larger scale testing for batches.