Preliminary semen sample management from paralichthys orbignyanus for sperm motility objective analysis by using a novel open-source suite of algorithms

MICAELA GRECO; JUAN IGNACIO PASTORE; BALLARIN, VIRGINIA L.; FRANCISCO BUCHELLY; MARIELA RADONIC; ANDREINA CESARI; LUCIA ZALAZAR; ANDREA LÓPEZ; EDDIE ARISTIZABAL

Ciudad Autónoma de Buenos Aires

Congreso; Reunion Conjunta de Sociedades de Biociencias; 2017

SAIC-SAIB-SAB-SAFIS-SAFE

A novel suit of open-source algorithms for sperm motility evaluation was previously reported by our group. The proposed method was successfully compared with the Microptic?s Sperm Class Analyzer®. Regarding the precision of the system, number of detected motile trajectories and the percentage of fragmented paths to measure the tracking reliability of ram sperm, our method presented a better performance according to those indicators compared to the commercial system. Knowledge related to marine fish sperm motility is poor. In view to improve gamete cryopreservation in our country, we performed preliminary assays to standardize the sample collection by stripping and evaluation of semen quality from the autochthonous flounder Paralichthys orbignyanus.This species is currently cultured under captivity in the Mariculture Experimental Station of National Institute for Fisheries Research and Development. We performed preliminary motility measurements by videomicroscopy using the recently developed algorithm. Sperm movement was recorded at 30 fr/sec using a videocamera attached to a microscope with a phase-contrast objective. Since fish sperm functionality depend on several factors, firstly, it was necessary to assay sample dilutions in salt water in order to activate the sperm and to find an appropriate sperm concentration that allow detecting individual trajectories. Concentration should not be over 30*106 cells/ml to guarantee the correct tracking. Motility parameters of progressive sperm were within 52.4 to 886.7 µm/s for VAP, 1.7 to 878.1 µm/s for VSL; 201.3 to 885.5 µm/s for VCL; 0 to 4.9 µm for ALH and 0 to 26.6 Hz for BCF, consistent with measurements reported in other marine fishes such as Sparus aurata. Even though there is not much published in fish sperm motility parameters, these result are promising to evaluate the quality of fish semen of interest for aquiculture and eventually to progress with fertility studies as well as cryopreservation protocols