Morphometric characterization of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa using computer-assisted spermatozoa analysis (ASMA)

Abstract

As part of a larger study on sperm quality and cryopreservation methods, the present study characterized the head morphometry of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa, using both scanning electron microscopy (SEM) and computer-assisted morphology analysis (ASMA). The latter method has been used rarely in fish and this is its first application on sharpsnout sea bream and gilthead sea bream spermatozoa. Results obtained using SEM are expensive and time-consuming, while ASMA provides a faster and automated evaluation of morphometric parameters of spermatozoa head. For sharpsnout sea bream spermatozoa, similar head measurement values were obtained using both ASMA and SEM, having a mean standard error length, of 2.57 +/- 0.01 mu m vs 2.54 +/- 0.02 mu m, width of 2.22 +/- 0.02 mu m vs 2.26 +/- 0.04 mu m, surface area of 4.44 +/- 0.02 mu m(2) vs 4.50 +/- 0.04 mu m(2) and perimeter of 7.70 +/- 0.02 mu m vs 7.73 +/- 0.04 mu m using ASMA and SEM, respectively. Although gilthead sea bream spermatozoa, were found to be smaller than those of sharpsnout sea bream,,spermatozoal head morphometry parameters were also found to be similar regardless of evaluation method, having a mean head length of 1.97 +/- 0.01 mu m vs 1.94 +/- 0.02 mu m, head width of 1.80 +/- 0.01 mu m vs 1.78 +/- 0.02 mu m, surface area of 3.16 +/- 0.03 mu m(2) vs 3.18 +/- 0.06 mu m(2) and perimeter of 6.52 +/- 0.04 mu m vs 6.56 0.08 mu m using ASMA and SEM, respectively. The results demonstrate that ASMA can be considered as a reliable technique for spermatozoal morphology analysis, and can be a useful tool for studies on fish spermatozoa, providing quick and objective results.