view - Department of Reproduction, Obstetrics and Herd Health
view - Department of Reproduction, Obstetrics and Herd Health
view - Department of Reproduction, Obstetrics and Herd Health
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3.3.1. Abstract<br />
CHAPTER 3.3<br />
Routine semen analysis includes evaluation <strong>of</strong> concentration combined with seminal volume,<br />
morphology <strong>and</strong> motility. Subjective analysis <strong>of</strong> these parameters is known to be inaccurate,<br />
imprecise <strong>and</strong> subject to variability. Automated semen analysis could lead to an increased<br />
st<strong>and</strong>ardization in <strong>and</strong> between laboratories but for that to happen automated devices need to be<br />
validated. A new device, the sperm quality analyzer V equine (SQA-Ve) version 1.00.43, was<br />
evaluated for its repeatability <strong>and</strong> agreement with light microscopy (LM), for raw <strong>and</strong> extended<br />
equine semen. Results were compared with computer assisted sperm analysis (CASA), which was<br />
also tested for its repeatability <strong>and</strong> agreement with LM. The SQA-Ve showed a good repeatability<br />
<strong>and</strong> fine agreement for assessing sperm concentration <strong>of</strong> raw semen based on scatter <strong>and</strong> Bl<strong>and</strong>-<br />
Altman plots. This was in contrast with the motility parameters, which had a low repeatability.<br />
Morphology assessment with SQA-Ve was poorly repeatable as well as in poor agreement with LM.<br />
For extended semen, the findings were comparable. The SQA-Ve did well for concentration, whereas<br />
for the motility parameters repeatability was only just acceptable, with however, no agreement with<br />
LM. This sharply contrasted the CASA findings that were highly repeatable <strong>and</strong> almost in perfect<br />
agreement with LM. Based on these findings, the tested version <strong>of</strong> the SQA-Ve is insufficiently<br />
accurate to be used for analyzing raw or extended equine semen.<br />
3.3.2. Introduction<br />
Although a plethora <strong>of</strong> specialized tests is available for analyzing semen samples (Varner,<br />
2008), a routine semen analysis includes the evaluation <strong>of</strong> concentration, seminal volume (Perreault,<br />
2009), morphology <strong>and</strong> motility (Varner, 2008). Microscopical measurements <strong>of</strong> sperm numbers,<br />
motility <strong>and</strong> morphology have been shown to be inaccurate <strong>and</strong> imprecise (Davis <strong>and</strong> Katz, 1993)<br />
<strong>and</strong> subject to high subjectivity (Matson, 1995). In order to overcome these inaccuracies, automated<br />
systems have been developed. In 1981, the Sperm Motility Analyzer was described by Bartoov et al.<br />
(1981) as a practical tool to evaluate overall sperm quality. This device, in later articles called the<br />
Sperm Quality Analyzer (SQA), registers fluctuations in optical density <strong>of</strong> light passing through a<br />
capillary which contains the semen sample. These fluctuations are registered by a photometric cell<br />
<strong>and</strong> converted to a numerical output. As such, the SQA does not recognize the actual sperm cells<br />
during the analysis.<br />
Computer-assisted semen analysis (CASA) was introduced both for humans <strong>and</strong> animal<br />
species about two decades ago (Verstegen et al., 2002). Using the combination <strong>of</strong> a microscope <strong>and</strong><br />
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