Standardization of sperm motility analysis by using CASA-Mot for Atlantic salmon (Salmo salar), European eel (Anguilla anguilla) and Siberian sturgeon (Acipenser baerii)

Abstract

It is essential to define an optimized standard method to assess the fish sperm quality to minimize the differences between the results obtained by different laboratories. Only this optimization and standardization can make them useful from academia to industry. This study presents the validation of sperm motility assessment using a CASA-Mot system for three endangered diadromous fish species: European eel (Anguilla anguilla), Atlantic salmon (Salmo salar) and Siberian sturgeon (Acipenser baerii). To attain this goal, different technical and data processing methods were tested: 1) magnification lens (x10 and x20), 2) Spermtrack® reusable chambers (10 and 20 µm depth) and 3) different frame rates (50 ≥ FR ≤ 250). The results suggested that the sperm motility assessment for eel, salmon and sturgeon should be performed at 200, 250 and 225 frames s -1 , respectively. Moreover, to obtain a high number of analysed spermatozoa in less time and a natural movement of the sperm cells, it is recommended to use x10 objective and 20 µm depth.In conclusion, different technical settings influence sperm kinetic parameters and should be validated for each fish species to allow the comparison of results between laboratories.

Kinematic Sub-Populations in Bull Spermatozoa: A Comparison of Classical and Bayesian Approaches

“…The multivariate approach of PCs and cluster analysis conducted for several species underscores that ejaculates are nevertheless heterogeneous in that they contain spermatozoa with different motility and kinematic patterns [ 17 , 22 , 31 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 ]. In the present study, we identified four different sperm SPs described from three PCs that represented velocity, progressiveness, and cell undulatory movement.…”

“…The intrinsic variability of semen samples, as well as individual variation or differences arising as a result of treatments, can be studied by using computer-assisted sperm analysis (CASA)-Mot systems that allow for the generation of huge datasets consisting of kinematics trajectories from thousands of spermatozoa [ 8 ]. CASA systems have evolved rapidly during the last decade due to major innovations in technology [ 9 ] such as increases in computational memory [ 10 , 11 ], use of 3D technology [ 12 ], considerations of the effect of frame rate capture [ 13 , 14 , 15 , 16 ], improvements in camera acquisition [ 17 ], or new approaches for sperm cell tracking [ 11 , 18 , 19 ]. Current CASA-Mot systems can be used to analyze individual sperm kinematics more accurately and this information can be submitted to a multivariate procedure, such as cluster analysis, for an overview of distinct sperm patterns grouped into SPs or clusters [ 20 ].…”

Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

“…Additionally, in recent studies, while using a high-performance camera and the same study design, depending on each species that the sperm shows non-linear trajectory will need more captures to define the correct track. This explains the differences obtained in the calculation of the OFR in the bull (256 fps) [24], the boar (212 fps) [25], and the salmon (250 fps) [26].…”

The application of computer-assisted semen analysis (CASA) technology to optimise semen evaluation. A review

“…The collection can be performed using an artificial vagina (Barszcz et al, 2012), massage (Valverde et al, 2018) and electro-ejaculation (Palmer et al, 2004). Sperm motility (total and progressive) and concentration are the principal variables assessed at the AI centres or farms to calculate AI doses (Caldeira et al, 2019). For dilution of the sample species, specific extenders must be defined for both refrigeration and cryopreservation of the samples (Foote, 2002;Víquez et al, 2020).…”