Effect of cryopreservation on semen biochemical parameters including lipid profile in beetal and Assam hill goat
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Date
2021-01
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Abstract
A total of 72 ejaculates comprising six ejaculates from each of 12 bucks (six Beetal
and six Assam Hill Goat) were used to study the effect of cryopreservation on the physical
and biochemical characteristics of semen including sperm ultrastructure. The physical
characteristics of fresh semen viz., ejaculate volume, mass activity, initial sperm motility,
live sperm, sperm concentration, cold shock resistance index, acrosomal integrity, HOSTreacted
sperm and sperm abnormalities were studied by conventional methods. The
biochemical characteristics viz., sodium, potassium, calcium, total cholesterol, total lipid,
AST, ALT and lipid profile were studied in seminal plasma, and total cholesterol, total
lipid and lipid profile were also studied in spermatozoa of fresh and frozen semen. Semen
was extended in Optixcell extender and frozen in 0.25 ml straws using liquid nitrogen
vapour and stored in liquid nitrogen. Each sample was evaluated on the following day for
sperm motility, live sperm, acrosomal integrity and HOST-reacted sperm. The
biochemical characteristics studied after freezing in extracellular medium were the same
as in case of seminal plasma. Sperm ultrastructure was studied both in fresh and frozenthawed
spermatozoa.
Physical and biochemical characteristics of semen of Beetal and Assam Hill Goats
were within normal range and differed significantly between breed for ejaculate volume
(P<0.001), sperm concentration (P<0.001), cold shock resistance (P<0.05), Host-reacted
sperm (P<0.01), sodium (P<0.001), potassium (P<0.001) and total lipid (P<0.001). Postthaw
sperm parameters viz. sperm motility, live sperm, acrosomal integrity and HOST–
reacted sperm, and content of biochemical constituents viz. sodium, potassium, calcium,
total cholesterol and total lipid decreased after freezing in semen of both the breeds, while
AST and ALT increased. Thirteen and fourteen fatty acids were identified in the seminal
plasma of Beetal and Assam Hill Goat bucks, respectively. Pentanedioic acid was
identified only in seminal plasma of Assam Hill Goat. In the extracellular medium of
frozen Beetal semen, all the fatty acids of seminal plasma of fresh semen were present,
however, in Assam Hill Goat bucks both Dodecadienoic acid and Pentanedioic acid which
were present in seminal plasma of fresh semen were found to be absent in frozen semen.
Breed variation was observed in respect of Pentanedioic acid of fresh semen. Freezing had
significant effect in alteration of membrane stabilizing fatty acids viz. Erucic acid,
carboceric acid, linoleic acid, tricontanoic acid, arachidic acid, eicosadienoic acid,
margaric acid, mentanic acid, tetradecadienoate, tridecanoic acid, heneicosylic acid,
cerotic acid and tricosanoic acid. Major ultrastructural changes in spermatozoa after
freezing were separating and ruptured plasma membrane, fusion of plasma membrane with
outer acrosomal membrane, swelling of acrosome and loss of acrosomal content.
Based on the physical and biochemical parameters studied it could be concluded
that cryopreservation of goat semen has deleterious effects on fatty acid profile of seminal
plasma and sperm plasma membrane, and also on sperm ultrastructure in both Beetal and
Assam Hill Goat. The proportion of loss of plasma membrane fatty acids after freezing
was lower in Assam Hill Goat as compared to that of Beetal goat. Post-thaw semen quality
in Assam Hill Goat was superior to that of Beetal goat.