The use of antifreeze protein type III for vitrification of in vitro matured bovine oocytes.

The aim of this study was to evaluate the use of antifreeze protein type III (AFP III) into vitrification medium on meiotic spindle morphology of in vitro matured bovine oocytes as well as the fertilization and blastocyst rates. Mature cumulus-oocyte complexes (COC) were distributed in four groups: control (untreated), vitrified without supplementation (AFP0) or supplemented with 500 (AFP500) or 1000 ng/mL (AFP1000) into vitrification solutions. Samples from each group were used to analyze the organization of meiotic spindle by confocal microscopy and the remaining COC were submitted to in vitro fertilization and culture for eight days. Control group exhibited only 15% of abnormal spindle. However, the spindle morphology was affected in all vitrified groups regardless to AFP concentration: 75.8%, 76.1% and 69.2% (P > 0.05) for AFP0, AFP500 and AFP1000, respectively. Similar cleavage rate was obtained among the vitrified groups (AFP0 = 17.9%, AFP500 = 16.9% and AFP1000 = 17.8%), but lower (P < 0.05) compared with control group (68.7%). At Day 5 of culture, embryo production rate of AFP500 (30.8%) and AFP1000 (25.0%) were similar to control group (49.4%). However, at Day 8 of culture, AFP0, AFP500 and AFP1000 groups exhibited lower (P < 0.05) blastocyst rates (10.0%, 3.8% and 9.4%, respectively) when compared to control (41.1%). In conclusion, AFP III did not preserve meiotic spindle organization against the cryoinjuries. However, the use of AFP III improved embryo development at Day 5 of culture, although this effect was not maintained up to the blastocyst formation.

Evaluation of hematologic, blood gas, and select biochemical variables in ovine whole blood stored in CPDA-1 bags.

BACKGROUND: The economic consequences from mortality of sheep after blood loss can be considerable. To date there are no reports addressing hematologic, blood gas, and biochemical changes in ovine blood stored in CPDA-1 bags. OBJECTIVES: The aim of this study was to investigate hematologic, blood gas, and biochemical alterations resulting from storage of ovine blood in CPDA-1 bags to establish transfusion protocols in sheep. METHODS: From each of 7 healthy 8-month-old sheep 450 mL of blood were collected into CPDA-1 bags and stored for 35 days in at 3-6°C. Samples were taken from the bags at days 0, 7, 14, 21, and 35. Whole blood was used to assess PCV, MCV, RBC count, pH, pO2 , pCO2 , and concentrations of bicarbonate, sodium, and lactate. Plasma was used to measure potassium, hemoglobin, and glucose concentrations. RESULTS: The PCV remained stable throughout the storage period, while plasma hemoglobin and MCV began to increase on days 7 and 21, respectively. The RBC count began to decrease on day 21. Blood pH decreased and pCO2 increased steadily throughout the storage period. Potassium concentration increased from 3.8 to 18.3 mmol/L on day 7 and remained high thereafter. In contrast, sodium concentration began to decrease on day 7. CONCLUSIONS: The results show that ovine blood undergoes hematologic, blood gas, and biochemical changes during storage. Further studies are required to establish RBC viability in CPDA-1 bags after a storage period of 35 days.