25Although a relatively high number of sperm quality biomarkers have been reported over 26 the years in several fish species, sperm motility is nowadays considered the best 27 biomarker for fish spermatozoa. The first scientific reports focusing on fish sperm 28 motility date from a century ago, but the objective assessment allowed by computer-29 assisted sperm analysis (CASA) systems was not applied to fish species until the mid 30 1980's. Since this date, a high number of sperm kinetic parameters from more than 170 31 fish species have already been reported in more than 700 scientific articles, covering a 32 wide range of topics such as i) sperm physiology, ii) sperm storage, iii) broodstock 33 management, iv) the phenomenon of sperm competition, v) ecotoxicology studies, and vi) 34 understanding the life cycle of the species. To sum up, the sperm kinetic parameters 35 provided by CASA systems can serve as a powerful and useful tool for aquaculture and 36 ecological purposes, and this review gives an overview of the major research areas in 37 which fish sperm motility assessed by a CASA system have been applied successfully. Sperm motility as a sperm qualitative biomarker in fish 43 Over the years, a relatively high number of sperm parameters have been used to assess 44 sperm quality in fish (reviewed by Fauvel et al. 2010). These sperm biomarkers have so 45 far been documented in scientific articles, and several traits, such as osmolality, plasma 46 composition; enzymatic activity; ATP concentration; sperm density or sperm morphology 47 have been linked to the ability of sperm to fertilize ova (Cabrita et al. 2014). However, 48 sperm motility is currently considered the most useful parameter for assessing sperm 49 quality in fish (Rurangwa et al. 2004), and more than 1500 scientific articles focusing on 50 a large number of topics have been published over the last century. The most commonly 51 used technique for assessing sperm motion in these articles has been subjective 52 evaluation, but some problems have emerged from this method (Verstegen et al. 2002). 53Subjective assessment depends on an experienced observer, and several aspects such as 54 sperm density, sperm velocity, and drift can be over-or underestimated (Rosenthal et al. 55 2010). Therefore, the low reproducibility of motility analyses that use subjective 56 evaluation (which can result in variations of 30-60% in the same sample) often makes it 57 difficult to interpret and compare the results between labs (Verstegen et al. 2002). 58In this sense, the gradual appearance of computer assisted sperm analysis (CASA) 59 systems has made it possible to estimate a higher number of sperm kinetic parameters 60 using objective, sensitive and accurate techniques (Table 1). These systems are the 61 evolution of multiple photomicrograph exposures and videomicrography techniques for 62 sperm tracking, and with the benefits of a computer equipped with imaging software, 63 detailed information on sperm kinetics can be extracted (Cabrita et al. 20...