Pellet Method of Semen Cryopreservation: Effect of Cryoprotectants, Semen Diluents and Chicken Lines

Abstract

The present study evaluated the effect of cryoprotectants, semen diluents and chicken lines during pellet method of semen cryopreservation. Three different experiments were conducted; Experiment 1 -semen was cryopreserved using dimethylformamide (DMF) at 6% and 9% concentrations in two semen diluents (Lake and Ravie diluent and TES/NaCl diluent), Experiment 2 -semen was cryopreserved using dimethylacetamide (DMA) at 6% and 9% with or without sucrose (100mM), Experiment 3-semen from two chicken lines (PD1 and PD6) was cryopreserved using DMA (6% and 9%). Semen was evaluated pre and post cryopreservation for progressive motility, live and abnormal sperm. Semen pellets were stored in cryovials for at least seven days before examination and insemination. Thawed semen was inseminated intravaginaly to study fertility. All the parameters studied were significantly lower (p<0.05) in cryopreserved semen. DMF in Lake and Ravie diluent gave very low fertility and TES/NaCl diluent no fertile eggs. DMA as cryoprotectant gave fertility up to 9.22 %. Addition of sucrose along with DMA produced fertility similar to other cryopreservation treatment groups. No difference in in vitro semen parameters between chicken lines was observed. There is difference in cryopreservation outcome due to semen diluent and type of cryoprotectant. HIGHLIGHTS Cryoprotectants, diluents and chicken lines effect in semen vitrification evaluated  Dimethylformamide gave low fertility in Lake and Ravie diluent and no fertility in TES diluent  Dimethylacetmide gave fertility up to 9.22 % and along with sucrose produced similar fertility  Post thaw in vitro semen parameters similar between chicken lines Shanmugam, M; Mahapatra ,R.

Cryopreservation of Ghagus chicken semen: effect of cryoprotectants diluents and thawing temperature

“…A higher live sperm, mitochondrial activity and lower acrosome damage was obtained in Korean native chicken semen cryopreserved with DMF (Choi et al, 2013). However, semen vitrified using DMF as a cryoprotectant resulted in negligible fertility (Shanmugam and Mahapatra, 2019). In the present report using DMF as cryoprotectant resulted in fertility ranging from 19 to 30%.…”

“…The diluent used during semen cryopreservation should essentially help in maintaining the structure and functional capacity of the sperm during and after cryopreservation. Few studies have compared the effect of diluents/extenders during chicken semen cryopreservation (Nabi et al, 2016; Rakha et al, 2016; Roushdy et al, 2014; Shanmugam and Mahapatra, 2019). In addition to energy and buffering components in diluents used for semen cryopreservation substances such as polyvinylpyrrolidone and glutamic acid are included that supports the sperm during cryopreservation procedure.…”

“…In addition to energy and buffering components in diluents used for semen cryopreservation substances such as polyvinylpyrrolidone and glutamic acid are included that supports the sperm during cryopreservation procedure. A diluent with minimal components that may support sperm during liquid storage may not be of use during cryopreservation (Shanmugam and Mahapatra, 2019).…”

Effect of Permeable Cryoprotectants and Dextran in Cryopreserving Semen of Broiler Breeder Lines

“…Non-permeable cryoprotectants are high molecular weight compounds that do not cross the cell membrane and were used during semen cryopreservation. Disaccharides such as trehalose and sucrose are the reported non permeating sugars used in chicken or turkey semen cryopreservation [6,[8][9][10][11]. The tonicity of extender is increased and cell membrane stabilized by non-permeating disaccharides [12].…”

Optimization of a Protocol for the Cryopreservation of Sperm in Pellets for the Common Pheasant (Phasianus colchicus mongolicus)

“…Finally, the last parts of the process are the thawing method and temperature of thawing [ 1 ]. For all of the factors mentioned above, specific cryopreservation protocols are needed for each species or breed [ 11 ]. In birds, several studies have described different protocols for various domestic and non-domestic birds [ 1 , 4 , 12 , 13 , 14 , 15 , 16 ].…”