title: Nitrogen metabolism and fertility in cattle: II. Development of oocytes recovered from heifers offered diets differing in their rate of nitrogen release in the rumen.
abstract: 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.
authors.affiliation: Scottish Agricultural College, Craibstone Estate, Bucksburn, Aberdeen, UK. k.sinclair@ab.sac.ac.uk
authors.forename: K D
authors.initials: KD
authors.lastname: Sinclair
authors.forename: M
authors.initials: M
authors.lastname: Kuran
authors.forename: F E
authors.initials: FE
authors.lastname: Gebbie
authors.forename: R
authors.initials: R
authors.lastname: Webb
authors.forename: T G
authors.initials: TG
authors.lastname: McEvoy
mesh_terms: D000641:Ammonia / Q000097:blood; D000818:Animals; D001826:Body Fluids / Q000737:chemistry; D002417:Cattle / Q000378:metabolism / Q000502:physiology*; D004032:Diet; D004971:Estrus; D005260:Female; D005298:Fertility / Q000502:physiology*; D005306:Fertilization; D005307:Fertilization in Vitro / Q000662:veterinary; D009584:Nitrogen / Q000378:metabolism*; D009865:Oocytes / Q000254:growth & development*; D010053:Ovary / Q000000981:diagnostic imaging; D012417:Rumen / Q000378:metabolism*; D014463:Ultrasonography
chemical_list: D000641:Ammonia; D009584:Nitrogen