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Three Fluorescence Methods for Assessing Boar Sperm Viability
Author(s) -
Juonala T.,
Salonen E.,
Nurttila T.,
Andersson M.
Publication year - 1999
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.1999.tb01388.x
Subject(s) - boar , sperm , fluorescence , biology , andrology , medicine , botany , physics , quantum mechanics
Contents This study compares three fluorescence methods for the evaluation of boar sperm viability and examines the interrelationships between fluorescence viability and motility of liquid semen, stored for 7 days, and fertility. One artificial insemination dose from each of 102 boars was stored for 7 days at 20°C. Plasma membrane integrity was evaluated in three different ways. First, a combination of two fluorophore probes, calcein acetylmethyl ester and ethidium homodimer‐1 (EH), was used to stain samples for fluorescence microscopic analysis by making semen smears. Secondly, semen samples were stained with EH with and without the addition of a detergent (ATP‐releasing reagent), and fluorescence intensities were measured with a fluorometer. Thirdly, fluorometric evaluation was carried out in the same pattern as with EH, by means of a more permeable, less specific DNA fluorochrome (more interference with RNA), Hoechst 33258. Computer‐assisted motility assessment gave the values for total, rapid and progressive motility and path velocity in semen stored for 7 days. Fertility of the boars was determined by the nonreturn (NR%) rate within 60 days of first inseminations and litter size of multiparous farrowings. The results showed that all three fluorescence methods were strongly intercorrelated. All plasma membrane integrity parameters correlated significantly with motility parameters and with fertility parameters (NR% and litter size of multiparous farrowings), but motility parameters did not correlate with NR%. It seems that fluorometric measurement could prove useful for plasma membrane integrity studies in liquid boar semen. Use of the objective and fast fluorometer‐based viability assay is thus suitable for several applications in sperm studies.