Improved timed-mating, non-invasive method using fewer unproven female rats with pregnancy validation via early body mass increases

Abstract

For studies requiring accurate conception-timing, reliable, efficient methods of detecting oestrus reduce time and costs, whilst improving welfare. Standard methods use vaginal cytology to stage cycle, and breeders are paired-up using approximately five proven females with proven males to achieve at least one conception on a specific day. We describe an alternative, fast, consistent, non-invasive method of timed-mating using detection of lordosis behaviour in Wistar and Lister-Hooded rats that used unproven females with high success rates. Rats under reverse lighting had body masses recorded pre-mating, day (d) 3–4, d8, d10 and d18 of pregnancy. Using only the presence of the oestrus dance to time-mate females for 24 hours, 89% of Wistar and 88% of Lister-Hooded rats successfully conceived. We did not observe behavioural oestrus in Sprague-Dawleys without males being present. Significant body mass increases following mating distinguished pregnant from non-pregnant rats, as early as d4 of pregnancy (10% ± 1.0 increase cf. 3% ± 1.2). The pattern of increases throughout gestation was similar for all pregnant rats until late pregnancy, when there were smaller increases for primi- and multiparous rats (32% ± 2.5; 25% ± 2.4), whereas nulliparous rats had highest gains (38% ± 1.5). This method demonstrated a distinct refinement of the previous timed-mating common practice used, as disturbance of females was minimised. Only the number required of nulli-, primi- or multiparous rats were mated, and body mass increases validated pregnancy status. This new breeding management method is now established practice for two strains of rat and has resulted in a reduction in animal use.