Passive liquefaction and colloidal centrifugation demonstrate improved semen quality following liquefaction of semen collected in the bonobo (Pan paniscus).

Passive liquefaction and colloidal centrifugation best preserve bonobo sperm motility and normal morphology post-collection.

This experimental cross-sectional study evaluated four semen processing methods for liquefying the coagulum in ejaculates voluntarily collected from three male bonobos (Pan paniscus). A total of 22 ejaculates were divided among passive liquefaction (PL; n=6), HEPES-balanced salt solution (n=4), trypsin enzymatic digestion (n=6), and colloidal centrifugation (CC; n=6) groups. Outcome measures included subjective total and progressive motility, total and progressively motile sperm counts per ejaculate, and morphology. Statistical analysis employed Shapiro-Wilk normality testing followed by Kruskal-Wallis testing with significance at P<0.05. Both PL and CC groups demonstrated the highest mean total motility (43.3±31.25% and 51.7±32.81%, respectively) and progressive motility (35.0±28.28% and 45.8±30.07%, respectively), both significantly exceeding the HEPES group, which yielded 0% motility. Normal morphology was significantly higher in PL (60.67±9.65%) and CC (58.67±13.97%) groups compared to the trypsin group (30.5±8.87%), suggesting enzymatic digestion adversely affects sperm morphology. No significant differences were detected among groups for total or progressively motile sperm counts per ejaculate. These findings indicate that passive liquefaction and colloidal centrifugation are the most effective methods for preserving bonobo sperm motility and morphology, with important implications for assisted reproductive technologies in this endangered great ape species.

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