Effect of follicle size on hormonal status of follicular fluid, oocyte ATP content, and in vitro embryo production in prepubertal sheep.

The aim of this study was to assess the effect of follicular size on estradiol (E2) and progesterone (P4) levels in intrafollicular fluid, ATP content in oocytes, and the embryo development rate in prepubertal sheep. Slaughterhouse ovaries were dissected to recover the follicles, which were classified according to the follicle diameter as <3 mm (n = 20) and ≥3 mm (n = 17). Then, follicular fluid was obtained and analyzed by radioimmunoassay to determine the E2 and P4 concentrations. Another group of ovaries was used to recover cumulus-oocyte complexes according to follicle size. In vitro maturation (IVM), in vitro fertilization (IVF), and embryo culture were performed using standard procedures, and ATP level was assessed at 0 and 24 h of IVM. Intrafollicular concentrations of E2 and P4 and E2:P4 ratio were higher in ≥3 mm (18.7 ± 5.9 ng/mL, 7.8 ± 1.2 ng/mL, and 3.6 ± 1.3, respectively) than <3 mm (1.8 ± 0.4 ng/mL, 2.6 ± 0.3 ng/mL and 0.9 ± 0.3, respectively) follicles. The rate of ATP increased during IVM and was higher in oocytes from ≥3 mm than <3 mm (22.4 ± 0.7 and 8.6 ± 2.2-fold change; respectively) follicles. After IVF, the blastocyst development was higher in oocytes recovered from ≥3 mm (11.1 ± 0.9%) than from <3 mm (6.5 ± 0.7%) follicles. These results indicate an improvement in the competence and development of oocytes from ≥3 mm follicles with a higher E2:P4 ratio. Thus, this ratio could be used as reference to design IVM medium and to enhance the in vitro embryo production in lambs.

Pestivirus in alpine wild ruminants and sympatric livestock from the Cantabrian Mountains, Spain.

Bovine viral diarrhoea virus (BVDV) and Border disease virus (BDV) were investigated at the wildlife-livestock interface in the distribution area of chamois in the Cantabrian Mountains, North-Western Spain. From 2010 to 2014, sera from sympatric wild (n=167) and domestic (n=272) ruminants were analysed for pestivirus antibodies by cELISA, virus neutralisation test (VNT) and for the presence of pestiviral RNA using a reverse transcription (RT)-PCR. Results showed a higher seroprevalence in cattle (59.4 per cent, 13/13 of herds) than in domestic small ruminants (5.9 per cent sheep, 2/8 of flocks; 0 per cent goats of 4 flocks) and wildlife (10.8 per cent in red deer, 0 per cent in roe deer and 0 per cent in Cantabrian chamois). High VNT titres were detected in two cattle herds, suggesting the circulation of BVDV-1 and BVDV-2 strains. BVDV-1 RNA was detected in one cattle calf by RT-PCR and sequencing. Conversely to other similar grazing systems, sheep flocks did not play a relevant role in the pestivirus epidemiology in this region. Pestivirus infections in wild ruminants were sporadic and most probably dependent on a domestic source.