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Title: THE EFFECT OF SEMINAL PLASMA AND DILUTION RATE ON THE MOTILITY AND VIABLITYOF FROZEN-THAWED BOAR

Author
item MAXWELL, W.M.C. - UNIVERSITY OF SYDNEY
item Guthrie, Howard
item JOHNSON, L - COLLABORATOR, ARS

Submitted to: International Congress on Animal Reproduction
Publication Type: Abstract Only
Publication Acceptance Date: 4/3/2000
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: This experiment tested the hypothesis that seminal plasma (SP) would improve the survival and acrosome integrity of highly diluted boar sperm after freeze-thawing. SP was prepared from the sperm rich fractions of three boar ejaculates. Sperm from the same boars were diluted in BTS+0.1% (w/v) BSA (BTS) (with or without 10%, v/v, seminal plasma; +/-SP) to provide sperm concentrations of 5, 50 or 500 million/ml in 1 ml. The sperm suspensions were held at RT for 1 hr, centrifuged, and resuspended to 1 ml with Test-yolk (20%, v/v) + 3% (v/v) glycerol (TY) (+/-SP). The samples were held at 17C for 1.5 h, cooled from 17C to 5C in 3.5 h, pellet-frozen (0.2 ml) on dry ice and stored in liquid N2. Pellets were thawed in dry tubes (38C), centrifuged, the supernatant discarded down to .1 ml and the sperm resuspended to 1 ml with BTS (+/-SP). Sperm were incubated at RT and assessed 0, 3 and 6 h after thawing for percentage motile (light microscopy), live (flow cytometry after staining with SYBR-14 & PI; 2) and acrosome reacted (AR, CTC fluorescence assay). SP had no effect in the initial diluent or freezing extender. %motile and %live sperm decreased, while AR increased, during post-thaw incubation. Motility and viability were higher and %AR lower in the presence of SP than in its absence. Motility was higher but viability lower for sperm frozen at a concentration of 500 than at 5 or 50 million/ml. Post-thaw motility was low regardless of freezing concentration but 50% of sperm remained live at RT for 6 h after freezing at concentrations of 5 and 50 million/ml. SP improved motility, viability and acrosome integrity of sperm frozen at low and high dilution rates and might be a useful post-thaw additive for extending the fertilizing life of sex sorted cryopreserved boar sperm.