It was hypothesized that dimethyleacetamide (DMA) can be used as an alternate to glycerol for cryopreservation of Indian red jungle fowl semen. Four concentrations of DMA (4%, 6%, 8% and 10%) in extender were compared with previously optimized cryopreservation protocol based on 20% glycerol (control) for Indian red jungle fowl. Sperm motility, plasma membrane integrity, viability, and acrosome integrity were assessed at the stage of post-dilution, cooling, equilibration, and freeze-thawing. The whole experiment was repeated/replicated for five times independently. Sperm motility, plasma membrane integrity, viability and acrosome integrity were recorded highest (P < 0.05) at post-dilution, cooling, equilibration, and freeze-thawing in extender having 6% DMA compared to control and other experimental extenders. The highest (P < 0.05) recovery rates of all aforementioned parameters were also recorded in extender having 6% DMA; thus, 6% DMA was further compared with control (20% glycerol) for fertility after artificial insemination. Eggs were collected for five days after artificial insemination with semen cryopreserved in extender containing 6% DMA and control. The higher no. of fertilized eggs, fertility, no. of hatched eggs, hatch (%) and hatchability were recorded with semen cryopreserved in extender having 6% DMA compared to control. It is concluded that 6% DMA maintained higher post-thaw quality and fertility of Indian red jungle fowl semen and is a better replacement of glycerol.

Aim: The study was designed to elucidate the effects of quercetin in an extender on oxidative stress, mitochondrial activity and quality of Indian red jungle fowl ( Gallus gallus murghi ) sperm during cryopreservation. Materials and Methods: Semen was collected from seven adult males through abdominal massage and evaluated for semen volume, concentration, and motility. The qualifying semen ejaculates having >80% motility were diluted in red fowl extenders with 0 (control), 5, 10, 15, and 20 mM quercetin. Diluted semen was frozen following a glycerol-based protocol. Semen quality (motility, plasma membrane integrity, viability, acrosome integrity, and chromatin condensation status) and biochemical parameters (mitochondrial activity, ferric reducing antioxidant power, and malondialdehyde [MDA]) were determined at various stages of cryopreservation. Results: Sperm motility, plasma membrane integrity, viability, acrosome integrity, and chromatin condensation were recorded highest ( p < 0.05) with 15 mM quercetin compared with 5, 10, and 20 mM quercetin and control at post-dilution, cooling, equilibration, and freeze–thawing. Nevertheless, mitochondrial activity and antioxidant potential were recorded highest with 15 mM quercetin compared with all experimental extenders at post-equilibration and freeze–thawing. MDA concentration in sperm and seminal plasma were recorded lowest ( p < 0.05) in the extender having 15 mM quercetin at post-equilibration and freeze–thawing. Cryopreservation stages showed negative effects ( p < 0.05) on semen quality parameters, irrespective of experimental extenders. Conclusions: It is concluded that quercetin (15 mM) supplementation in red fowl extender improves sperm motility, plasma membrane integrity, viability, acrosome integrity, chromatin condensation, and mitochondrial activity by elevating the total antioxidant potential and ameliorating lipid peroxidation during cryopreservation.