Cryopreservation of the Mediterranean mussel (Mytilus galloprovincialis) spermatozoa.
The effects of cryoprotectants, cooling rate and freezing on the mussel Mytilus galloprovincialis sperm were evaluated. At the end of each step of the experimental protocol, motility and fertilization ability of sperm were analyzed, compared to fresh semen. Five cryoprotectants were tested in their toxicity level: dimethylsulfoxide, ethylene glycol, 1-2 propylene glycol at 5%, 7%, 10%, 15% and 20% concentration; glycerol and methanol at concentration of 5%, 7% and 10%. The incubation times were 10, 20 and 30 min at 20+/-1 degrees C. Only dimethylsulfoxide, ethylene glycol and 1-2 propylene glycol at 5%, 7% and 10% were chosen for the following pre-freezing step. Five adaptation/chilling rates were analyzed: 10 min at 20+/-1, -2, -1, -0.5 and -0.25 degrees C/min and the last one was used for testing the best freezing procedure among seven gradients. Particularly, two rapid rates, three slow rates and two double step rates were conducted. Thawing results showed that M. galloprovincialis sperm are very sensitive to rapid pre-freezing and freezing protocols and only a slow procedure assured good motility and fertilization percentages.
Di Matteo
O
O
Dipartimento delle Scienze Biologiche, Laboratorio di Criobiologia, University of Naples Federico II, Via Mezzocannone 16, 80134 Napoli, Italy.
Langellotti
A L
AL
Masullo
P
P
Sansone
G
G
D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry / Q000494:pharmacology* / Q000633:toxicity; D005307:Fertilization in Vitro / Q000187:drug effects; D008297:Male; D049878:Mytilus; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology
D003451:Cryoprotective Agents