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Rezultaty
Measurements of sperm motility in mammals using Computer Assisted Sperm Analysis (CASA) is well established but the method has not been extensively used for fish sperm due to the technical difficulties arising from the differences between mammalian and fish sperm. We have developed a methodology which will rapidly and easily measure the motility of fish sperm using CASA and its morphology using Automated Sperm Morphology Analysis (ASMA). The methodology for motility analysis has been validated by the demonstration of a significant correlation between sperm motility (curvilinear velocity and straight line velocity) and fertilisation rate in several species.
The CASA methodology has been applied to assessing the toxicity of pollutants to fish sperm, monitoring sperm quality of wild fish in polluted waters and developing new methods for sperm cryopreservation. ASMA is able to rapidly measure both the dimensions of the head and the length of the flagellum of fish sperm. It has been applied to demonstrate damage to flagella of sperm exposed to mercury, but is also applicable to assessing changes in sperm morphology resulting from cryopreservation or exposure of fish to environmental pollution. Detailed standard operating protocols are provided for measuring fish sperm motility (FiSMot), assessing the effects of pollutants on sperm quality by in vitro and in vivo exposure (FiSMoTox), testing cryopreservation methods (FiSCryo) and examining sperm morphology (FiSMorph). Supporting validation and description of the application are included in the final report and will be published in scientific peer reviewed publications.
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