Short-term preservation of Pecari tajacu ovarian preantral follicles using phosphate buffered saline (PBS) or powdered coconut water (ACP®) media

Abstract

We compare protocols for the short-term preservation of collared peccarie's ovarian preantral follicles (PFs) by using phosphate buffered saline-(PBS) or powdered coconut water-(ACP®) based medium. For morphology analysis each pair of ovaries collected from six females was divided into nine fragments. One fragment was destined for morphology analysis (histology and transmission electron microscopy - TEM), constituting the control group and the other fragments were placed in tubes with PBS or ACP®, packed in 5 L Styrofoam boxes, stored for 4h, 12h, 24h, and 36h, and then analyzed. For viability analysis a pair of ovaries from two additional females was divided into nine fragments; one fragment was immediately destined for viability analysis (Trypan blue test) and the other fragments were stored as previously described, until 24h and then analyzed. After 4h storage in ACP® medium, the follicular integrity was similar to control (87.8% vs 94.4%, respectively); however, ultrastructural analyses revealed swollen mitochondria as the first signals of PF degeneration. It was observed that ACP® (66.7%) was more efficient than PBS (49.4%) to preserve the morphological integrity after 36h storage (P<0.05); however, no differences were observed on follicular viability (P>0.05). In conclusion, the use of the ACP® is recommended for the short-term preservation of Pecari tajacu preantral follicles.