In vitro blastocyst production was determined for oocytes recovered postmortem from 48 beef x dairy heifers offered low (Low NH3) or high (High NH3) plasma ammonia-generating diets during the period of late antral follicle development. Following the establishment of a reference estrus (d 0), the experimental diets were offered for an 18-d period starting on d 3 and during which a second estrus was induced (d 16) 4 d before the animals were slaughtered. Blood samples collected at varying intervals were analyzed for ammonia, urea, progesterone, and LH. Ovarian folliculogenesis was monitored daily by transrectal ultrasonography. Ovaries were collected at slaughter and cumulus-oocyte complexes were aspirated from small (1 to 4 mm) and medium-sized (> 4 to 8 mm) sized follicles. In vitro-matured and -fertilized putative d-1 zygotes were cultured for a further 7 d in vitro and embryo development and metabolism were assessed. Relative to the low-NH3-generating diet, the high-NH3-generating diet increased peak postprandial levels of plasma ammonia (326.1 +/- 43.3 vs 52.1 +/- 7.4 micromol/L; P < .001), mean levels of plasma urea (7.0 vs 5.7 mmol/L; SED = .2; P < .001), peak levels of plasma progesterone prior to induced luteolysis (8.9 +/- .4 vs 6.8 +/- .3 microg/L; P < .001), and follicular fluid levels of ammonia (267 +/- 18 vs 205 +/- 20 nmol/mL; P < .05) and progesterone (351 +/- 69 vs 199 +/- 26 ng/mL; P < .05). The timing and level of the preovulatory LH surge was not affected by dietary treatment. Of oocytes cultured, cleavage (47.4 vs 62.4%; P = .02) and blastocyst production (10.9 vs 20.6%; P = .06) rates were reduced when the oocytes were derived from heifers offered the high- rather than the low-NH3-generating diets. There were interactions between dietary treatment and follicle size class, which indicated that fewer blastocysts were produced from cleaved oocytes derived from medium-sized follicles of heifers offered the high-NH3 treatment but that de novo protein synthesis was increased in such embryos. In conclusion, exposure to high levels of ammonia and(or) urea in vivo can significantly compromise the subsequent capacity of oocytes to develop to blastocysts in vitro, and oocytes recovered from medium-sized follicles are particularly sensitive to this effect.

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The roe deer blastocyst is in diapause between August and December, after which time it expands and elongates rapidly before implantation. Blood samples were taken from 30 animals to define temporal changes in reproductively important hormones to investigate the physiological cues present at embryo reactivation. In 15 of these animals, changes in uterine and conceptus protein synthesis and secretion, and luteal progesterone release during diapause and reactivation, were assessed after culture of these tissues in vitro. Oestradiol concentrations remained low during diapause (1.07 +/- 0.4 pg ml(-1)) and expansion (1.2 +/- 0.4 pg ml(-1)) but increased by 30 times at trophoblast elongation (49.17 +/- 0.37 pg ml(-1)). Prolactin remained at basal concentrations (4.69 +/- 0.86 ng ml(-1)) and increased after implantation (12.34 +/- 2.71 ng ml(-1)). Peripheral progesterone concentrations and luteal progesterone release remained constant throughout diapause, reactivation and implantation (peripheral progesterone: 3.82 +/- 1.97 ng ml(-1); luteal progesterone: 6.72 +/- 0.81 ng mg(-1) protein). Incorporation of a radiolabel into conceptus secretory proteins increased by four times at expansion compared with diapause, whereas incorporation into endometrial secretions remained constant. At elongation, incorporation into endometrial secretions increased two times and conceptus secretions increased 32 times. Two-dimensional electrophoresis and fluorography showed that the profile of endometrial secretory proteins was constant until implantation when qualitative changes were evident. Although a role for an endocrine maternal trigger of reactivation from diapause cannot be dismissed, these data provide no supporting evidence and indicate that the conceptus itself may drive reactivation.