Influence of Osteopontin in Bovine Uterine Tube Fluid on Sperm Binding and Fertilization in RCA-1 Lectin-treated Oocytes     

RF Gonçalves  CG Wolinetz  VH Barnabe  GJ Killian 《Reproduction in domestic animals》2009,44(1):152-155

This study was conducted to determine the effect of pre‐exposure of oocytes to Ricinus communis (RCA‐1) lectin and osteopontin (OPN) in uterine tube fluid (UTF) on in vitro sperm–egg binding and fertilization. In vitro‐matured bovine oocytes were incubated (39°C, 5% CO₂ in air) for 2 h in the following treatments: (i) 500 μl of fertilization medium (FM); (ii) 250 μl of FM with 0.25 ml of non‐luteal ampullary uterine tube fluid (NLAUTF); (iii) 250 μl of FM with 250 μl of NLAUTF and 4 μl of RCA‐1 lectin; (iv) 250 μl of FM with 250 μl of NLAUTF, a rabbit polyclonal antibody (1:200) against purified bovine milk OPN, and RCA‐1 lectin; (v) 500 μl of FM and RCA‐1 lectin. Following incubation, oocytes were washed, placed in FM with 2 μg heparin, and incubated with 1 × 10⁵ frozen–thawed spermatozoa per 10 oocytes. Oocytes used to assess sperm binding were stained with Hoescht 33342, and the number of sperm bound per zona pellucida counted. The remaining oocytes were fixed in acid alcohol, stained with 1% acetate‐orcein and observed to determine the presence of pronuclei. More sperm bound to the zona pellucida (mean ± SEM) when oocytes were incubated in treatment 3 (59.0 ± 5.5) than in treatments 2 (46.4 ± 5.6), 4 (18.1 ± 5.4), 5 (33.4 ± 5.6) or 1 (32.5 ± 5.6). More oocytes were fertilized when incubated in treatment 3 (91% ± 3.0) than in 2 (84% ± 3.0), 4 (40% ± 3.0), 5 (77% ± 3.0) or 1 (76% ± 3.0). As in previous studies, this study suggests that RCA‐1 lectin enhances binding of UTF‐derived OPN to bovine oocytes, resulting in increased sperm–egg binding and fertilization in vitro and a possible role in fertilization.  相似文献   

Phenotyping of Urochloa humidicola grass hybrids for agronomic and environmental performance in the Piedmont region of the Orinoquian savannas of Colombia     

Daniel M. Villegas  Ashly Arévalo  Mauricio Sotelo  Jonathan Nuñez  Danilo Moreta  Idupulapati Rao  Manabu Ishitani  Guntur V. Subbarao  Jacobo Arango 《Grass and Forage Science》2023,78(1):119-128

In the low fertility acid soils of the Orinoquian savannas of Colombia, Urochloa humidicola cv. Tully or Humidicola is one of the most widely planted tropical forage grasses for improving livestock productivity. Low nutritional quality of this grass limits sustainable livestock production in this region. In this study, we conducted a phenotypic evaluation under field and greenhouse conditions of one of the first hybrid populations of U. humidicola generated from the forage breeding program of CIAT. Our objective was to identify a set of new hybrids of U. humidicola that combine improved productivity and nutritional quality plus the biological nitrification inhibition (BNI) trait/ability to reduce nitrogen (N) losses via leaching and nitrous oxide (N₂O) emissions. To this end, we tested 118 hybrids (planted in pots) in the greenhouse for over 6 months and measured potential nitrification rates (NR) using soil microcosm incubation. NR values observed ranged from 0.27 to 5.75 mg N-NO₃⁻ kg soil⁻¹ day⁻¹. Later, 12 hybrids with different levels of NR were selected and field-tested in the Orinoquia region over a 4 years period (2013–2017) for dry matter production, nutrition quality (crude protein, in vitro digestibility and fibres content) and NR in each year. In the rainy season of 2018, two hybrids with superior agronomic performance and contrasting field level NR (Uh08/1149 and 0450) were subjected to analysis of soil-borne N₂O emissions after fertilization during 13 days. The NR values recorded were not directly correlated with the forage quality parameters evaluated, however, the two grasses with the lowest NR values were among those with the highest biomass production, crude protein content, and N uptake. The grass hybrid Uh08/1149 and the germplasm accession CIAT 16888 were found as materials with superior forage value, with production of 14.1 and 14.6 tons dry matter ha⁻¹ year⁻¹ (up to 8% higher than the cv. Tully), crude protein of 11.5 and 9.1% per cut (up to 20% higher than the cv. Tully), and N uptake of 31.6 and 25.7 kg N ha⁻¹ cut⁻¹ (up to 30% higher than the cv. Tully). Additionally, these two grasses are likely to exhibit high-BNI ability, with potential to improve N use efficiency in managed pastures.  相似文献