Excessive rumen-protected choline in the daily diet compromises sperm quality of male dairy goats as a result of aberrant DNA methylation modification

Context Choline is added to the diet of ruminants to improve animal growth, development and reproduction; however, little information is available regarding effects of dietary choline supplementation, in the form of rumen-protected choline (RPC), on fertility of male ruminants. Excess RPC in the diet might damage ram fertility through abnormal alteration of methylation patterns at the imprinting control region (ICR) of imprinted genes H19/IGF2. Aims The present study evaluated the influence of different levels of RPC supplementation on the sperm quality of male Saanen dairy goats. Methods Different proportions of RPC (0%, 0.5%, 1.0% and 2.0% of daily concentrate feed) were added to the diet of Saanen bucks. Sperm quality parameters, subsequent in vitro embryo development potential, and kidding rates post artificial insemination were examined. In addition, differences in methylation status of the global DNA, and at 20 CpG sites in the ICR of imprinted genes H19/IGF2, were compared. Key results Supplementation of the daily diet with 0.5% RPC significantly improved sperm quality, and increased subsequent embryo development and kidding rates. However, the two higher RPC-supplemented groups showed significantly reduced kidding rate. Moreover, methylation levels of both the ICR of H19/IGF2 and the global DNA increased significantly with increasing supplemental RPC, and the expression of IGF2 was significantly inhibited in sperm samples from the 1.0% and 2.0% RPC groups, whereas H19, which should had been silenced, showed high expression. Conclusions Adding excessive RPC (≥1%) to the daily diet of male goats might disturb the process of spermatogenesis and is associated with abnormal methylation modification caused by aberrant expression of DNMT1, DNMT3a and DNMT3b in sperm. Implications This study determines the safe amount of choline to add to the diet during the breeding of male dairy goats. This provides a reference for improving the breeding efficiency and saving the breeding cost of dairy goats.