Effects of glucose concentration in semen extender and storage temperature on stallion sperm quality following long-term cooled storage.

In Experiment 1, the effects of glucose concentration in extender (0 mM, 67 mM, 147 mM, 270 mM; G0, G67, G147, and G270, respectively) and storage temperature of extended semen (5, 10, 15 and 20 °C) were evaluated after storage for up to 5 days (T0h to T120h). For all time points tested, mean total (TMOT) and progressive (PMOT) sperm motility were lower in G0 than all other treatment groups (P < 0.05). Mean curvilinear velocity (VCL) was lower in G0 than other treatment groups at all time points tested except T0h (P < 0.05). Mean percentage of plasma membrane/acrosome intact sperm (VAI) was similar among treatments at T0h, T72h, and T120h (P > 0.05). Mean TMOT and PMOT, were lower for semen stored at 20 °C than all lower storage temperatures (P < 0.05) at all time points. In Experiment 2, semen was stored at 10 °C in extender containing no added glucose (G0) or 147 mM glucose (G147). Following storage, semen was centrifuged and resuspended in extender containing no added glucose (G0 – G0 or G147 – G0, respectively) or 147 mM of glucose (G0 – G147 or G147 – G147, respectively). Mean TMOT, PMOT, and VCL were higher in G147 than G0 at all time periods tested (P < 0.05), whereas mean VAI was similar between these treatment groups throughout the experiment (P > 0.05). Mean TMOT and PMOT were higher in G0 – G147 than G0 – G0 at T72h and T120h (P < 0.05) and mean VCL was higher in G0 – G147 than G0 – G0 for all time periods. Mean TMOT, PMOT, and VCL were higher in G147 – G147 than G147 – G0 at all time points tested (P < 0.05), whereas mean VAI was similar between these two treatment groups for each of the time points (P > 0.05). In Experiment 3, the minimum concentration of glucose required to maintain sperm quality following long-term cooled storage (T120 h) was evaluated (G0, G5, G10, G20, G40, G67, G147 mM). At T120 h, mean TMOT was lowest in G0, G5, G10, and G20 (P < 0.05), whereas mean PMOT and VCL were lower in G0, G5, G10, and G20 than in G40, G67, and G147 (P < 0.05). Mean VAI was higher in G10 than G67, but similar among G10 and other treatment groups (P > 0.05). In conclusion, the absence of added glucose in extender reduced the motion characteristics of stallion sperm during long-term storage (5 days), but VAI was not affected. The use of temperatures between 5 and 15 °C for long-term storage (5 days) best maintained sperm motility and VAI. The threshold concentration of added glucose in extender required to optimize sperm motion characteristics was 40 mM.