Effect of the supplementation of two capacitive agents in the fertilization medium for the in vitro production of embryos in alpacas (Vicugna pacos)

Abstract

The present study aims to evaluate the effect of heparin and PHE (penicillamine, hypotaurine and epinephrine) in the fertilization medium on the in vitro production of embryos in alpacas. The ovaries were transported in 0.9% physiological saline with gentamicin at 37 ° C for 3 hours. 227 ovaries were used, recovering 1132 oocytes of quality I, II and III; and matured in vitro for 34h. The mature oocytes were randomly distributed in four culture medium treatments, which were supplemented with 2% heparin and PHE according to the following distribution: T1 = Control; T2 = Heparin; T3 = PHE; T4 = PHE + heparin. Then 1x106 sperm/ml selected by the Percoll gradient method (45/90%) and incubated for 18 h. Finally, were cultivated in SOF culture medium for 7 day. The cleavage rate at 24 h in culture were 16.53, 19.92, 21.42 and 23.90 % for T1, T2, T3 and T4, respectively. The group T4 was difference statistically with groups T1, T2 and T3 (P <0.05). The morulae rate at day 7 were 24.23, 29.49, 35.00 and 35.30% for T1, T2, T3 and T4, respectively, with statistical difference between T1 and T2 with respect to T3 and T4. The blastocysts rate at day 7 were 3.60, 3.25, 4.52 and 6.34 % for T1, T2, T3 and T4, respectively, with no statistical difference between treatments. Regarding quality, the embryos (morulae and blastocysts) produced in vitro from the treatments with the use of PHE + heparin and PHE capacitors stood out (p≤0.05) with excellent quality embryos. In conclusion, the use of heparin and PHE in fertilization medium increases the in vitro production of excellent quality embryos.

Keywords

• fertilization
• heparin
• ivf
• alpaca