EFFECT OF DIFFERENT CONCENTRATIONS OF SEMINAL PLASMA ON SPERM VIABILITY AND ACROSOMAL STATUS IN LLAMA

FUMUSO FG; CARRETERO MI; CETICA P; NEILD DM; MIRAGAYA MH; GIULIANO SM

Tours

Congreso; 18th International Congress on Animal Reproduction; 2016

In various species it has been reported that dilution of seminal plasma (SP) induces a process similar to capacitation [1]. It is currently unknown if this occurs in South American Camelids sperm. The objective of this study was to evaluate the effect of different concentrations of SP on llama sperm viability and acrosomal status in raw semen. A total of eighteen ejaculates were collected from 6 llamas (n=6, r=3), using electroejaculation. Each ejaculate was diluted 4:1 in 0.1% collagenase in Hepes-TALP (HT) medium and incubated 4 minutes at 37 ºC with the objective of separating spermatozoa from SP. Immediately after incubation each ejaculate was divided in four and centrifuged for 8 min at 800g. The pellets were resuspended with different concentrations of SP in HT: Controls (0 and 100% SP) and Treatments (10 and 50% SP). All samples were maintained at 37 ºC until evaluation at 0, 1.5 and 3 hours. Acrosomal status and viability were evaluated with FITC-PNA/PI (Fluorescein isothiocyanate - Arachis hypogaea agglutinin/Propidum iodide). A Kruskal Wallis test was carried out to compare the FITC-PNA/PI patterns: live acrosome intact (LI), live acrosome reacted (LR), dead acrosome intact (DI) and dead acrosome reacted (DR). When comparing acrosomal status, we observed a greater percentage of intact acrosomes (LI and DI) in samples with high concentrations of SP (50% and 100%) compared to those with low concentrations (0% and 10%) at all incubation times, indicating a possible protective effect of SP on sperm acrosomes. In addition, the percentages of LR sperm were significantly greater (P