1. Spermatozoa DNA and plasma membrane integrity after pellet optimized processing for cryopreservation in meat type chicken breeders.
- Author
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Gliozzi, T. M., Zaniboni, L., Iaffaldano, N., and Cerolini, S.
- Subjects
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CHICKENS , *POULTRY breeding , *SPERMATOZOA , *CELL membranes , *CRYOPRESERVATION of organs, tissues, etc. , *STRUCTURAL optimization - Abstract
1. Aim of this study was the development of an optimised cryopreservation pellet procedure for chicken semen and the assessment of DNA and membrane integrity in frozen/thawed spermatozoa in a Hubbard F15 meat type selected strain. 2. The following semen processing conditions were studied: spermatozoa working concentration (SWC), 1.5vs2 × 109cells/ml in pre-freezing extender; equilibration of diluted semen at 5°C, 20vs40 min; dimethylacetamide concentration, 6%vs9%; dimethylacetamide equilibration time at 5°C, 1vs30 min; thawing at 60°C for 10vs50°C for 30 sec. Spermatozoa viability (EtBr exclusion procedure โ stress test), mobility (Accudenz® swim-down test) and subjective motility were assessed in fresh and frozen-thawed semen. 3. The lower SWC (1.5 × 109cells/ml) and the higher dimethylacetamide concentration (9%) had positive significant effects on the recovery rate of motile (22%vs16%) and viable spermatozoa (39vs34%), respectively. 4.Membrane (SYBR14-PI staining) and DNA integrity (comet assay) were assessed before and after freezing/thawing according to the optimised protocol. 5. Recovery rates of spermatozoa with undamaged plasma membrane and DNA were 41% and 76%, respectively. The distribution of spermatozoa in classes of DNA damage was also analysed and discussed. 6. It was concluded that pellet cryopreservation was a damaging process mainly for plasma membrane rather than nuclear DNA in chicken spermatozoa. [ABSTRACT FROM PUBLISHER]
- Published
- 2017
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