Use of laparoscopic intrauterine insemination associated with a simplified superovulation treatment for in vivo embryo production in sheep: a preliminary report

Abstract

This study evaluated the reproductive performance of sheep that were subjected to superovulatory treatment and intrauterine insemination with two different doses of semen. The main objective was to determine whether intrauterine insemination is associated with higher rates of embryo recovery than existing procedures. In the first experiment 30 Ojalada de Soria ewes were used at the end of their productive life and nine younger ewes of the same breed were used in Experiment 2. Synchronisation of oestrus was conducted with intravaginal sponges and the superovulation treatment consisted of an intramuscular injection of 210 IU pFSH and 500 IU eCG administered 24 h before sponge removal. After 48 h of sponge withdrawal, ewes were inseminated by laparoscopy. They were divided into two groups: low dose group (LD, 25 × 106 sperm; n = 14 Expt 1 and n = 5 Expt 2) and high dose group (HD, 100 × 106 sperm; n = 14 Expt 1 and n = 4 Expt 2). Embryos were recovered 7 days after the onset of oestrus by laparotomy and uterine flushing. Two blood samples were collected in order to analyse cortisol, at sponge insertion and the day the embryos were recovered. The same protocol was repeated twice in Experiment 1, with an interval of 2 months between two consecutive treatments. Results show a significant effect of the number of recovery on ovulation rate (21.7 ± 2.2 v. 11.8 ± 1.1 corpora lutea for first and second recoveries, respectively), and number of structures recovered (11.4 ± 1.6 v. 5.2 ± 1.1, P < 0.01). Differences of embryo performance in the second recovery compared with the first one were more evident in the LD group compared with the HD group. In Experiment 2, results reflected a significant superiority of the HD group compared with the LD group, especially rate of fertilisation, which was 25% higher. Sheep producing non-fertilised embryos had higher cortisol levels at the time of flushing than those with embryos (P < 0.01). It is concluded that the use of simplified and consecutive superovulatory treatments associated with intrauterine insemination is a potential means of obtaining embryos, although the response seems to depend on donor sheep age. The use of a higher number of sperm cells per millilitre results in better outcomes in terms of embryos produced.