RANKING/GROUPING OF SANTA INÊS RAM FROZEN SEMEN ANALYZED BY CASA AND FLUORESCENT PROBES LEAD THROUGH THE MULTIVARIATE STATISTICS ANALYSIS
Keywords:
sperm subpopulation, cryopreservation of ram semen, computer-assisted semen motility analysis (CASA), mitochondrial activity, fluorescent probesAbstract
The cryopreservation process causes several alterations in semen quality, reducing fertility.
New evaluations of sperm motility have been searched, such as the sperm subpopulations
analysis by computer-assisted motility assessments (CASA). Fluorescent probes have been
studied to evaluate spermatozoon integrity, supplying functional information of the spermatozoa. The aim of this study was to optimize the CASA system in combination with
fluorescent probes as a new method of evaluation of the qualitative parameters of the frozen
semen of rams. Twenty-six frozen semen samples were evaluated by CASA and MITO Test.
The cluster analysis was used with the VARIMAX rotation method and the grouping analysis
was performed with Ward’s method. The individual kinetics of each spermatozoon was
considered for the analysis, extending samples from 26 averages to 5834 spermatozoa. For the
MITO test, the samples were grouped into eight categories. The multivariate exploring
technique pointed out reduction of the dimensionality, leading to three factors: F1 factor
interpreted as a factor related to the forward movement; F2 factor represents a factor of sperm
displacement without considering the movement direction; F3 factor means the energy
available for the sperm displacement. The determination of score coefficients of the factors
was determined by grouping the samples into 9 subpopulations, with distinct qualitative
characteristics. With the use of the multivariate statistics analysis of the frozen semen of the
rams, it was possible to determine physiological and kinematics differences of the
subpopulations associated to the fluorescent probes in the CASA system, with correlations
between the CASA system and the MITO test.
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