Organic Zinc and Copper Supplementation-Associated Changes in Gene Expression and Protein Profiles in Buck Spermatozoa.

Mineral supplementation has greater impact on male reproduction; however, the mechanism of action has not been studied in detail. The present study was aimed to deal with the lacuna in mechanism of action of mineral supplementation on improvement in sperm characteristics. A group of 40 bucks (aged 5 months) were assigned to 10 groups (4 in each group) based on their body weight and fed with concentrate mixture: basal roughage (minimal diet) in equal proportion to all the bucks. Among the 10 groups, one was considered as control, without any additional mineral supplementation, and the remaining 9 were treatment groups (3 groups each in Zn, Cu, and Zn + Cu). In treatment groups, organic Zn was fed in three different doses as 20, 40, and 60 mg/kg DM; organic Cu was fed in three different doses as 12.5, 25, and 37.5 mg/kg DM; and organic Zn + Cu was combinedly supplied as 20 + 12.5, 40 + 25, and 60 + 37.5 based on their mg/kg DM for 8 months period. The neat semen samples were processed for spermatozoal gene (stress- NOS3, HSP70, HIF1A; fertility- MTF1, MTA1, TIMP2, TNFa, and EGFR) expression studies through qRT-PCR and protein profile changes through single- and two-dimensional gel electrophoresis. Significantly, the stress-responsive genes were downregulated, and fertility-related genes were upregulated in treatment groups. A significant correlation had been noticed among the genes studied: HIF1A with MTA1 (P < 0.05) and MTF1 with EGFR, TIMP2, TNFa, and NOS3 (P < 0.01) respectively. The organic Zn and Cu feeding modulated the expression of stress- and fertility-related genes and protein abundance, thereby improved the sperm characteristics.

Organic Zn and Cu supplementation imprints on seminal plasma mineral, biochemical/ antioxidant activities and its relationship to spermatozoal characteristics in bucks.

This experiment was conducted to study the effect of mineral supplementation on seminal plasma minerals level, biochemical constituents and total antioxidant capacity of Osmanabadi bucks. The study comprised of forty healthy bucks, aged five months were randomly assigned to ten groups (n = 4 per group). The control group was fed with a basal diet without any additional mineral supplementation. In addition to basal diet, treatment bucks were supplemented with three graded doses of organic Zinc (Zn) as 20, 40 and 60 mg/kg dry matter (DM); organic Copper (Cu) as 12.5, 25, 37.5 mg/ kg DM and combination of Zn + Cu as Zn20+Cu12.5, Zn40+Cu25, Zn60+Cu37.5 mg /kg DM basis respectively. Minerals were supplemented for 8 months and the separated seminal plasma used for analysis of minerals, biochemical profile, total antioxidant capacity (TAC), lipid peroxidation (LPO), and protein carbonylation (PC). In treatment groups, significantly lower LPO and PC were observed, except Zn60 and Zn60+Cu37.5, where higher malondialdehyde (MDA) (P < 0.05) formed. The TAC was relatively higher (P < 0.05) in Zn20, Zn40, Cu12.5 and Zn60+Cu37.5 than control. The minerals and biochemical parameters were significantly altered and positive relationship was observed among them. From this study, it was concluded that supplemented minerals changed the seminal plasma minerals profile (Zn- 7-13; Cu- 0.5-1.9 mg/L), reduced the stress (LPO and PC of control Vs treatment as 0.3 Vs 0.1 nmol/ml and 25.7 Vs 4.3 nmol protein carbonyl/mg protein), which improved the sperm quality in Zn40, all Cu treatments and Zn60+Cu37.5 groups respectively.