Enzyme immunosorbent assay

Hair from the forehead of the calves was clipped on d—2, 14 and 28 after castration. Samples were stored in plastic bags at room temperature and handled and analyzed as described by Moya et al. [16 ]. Cortisol was quantified using an enzyme-immunosorbent assay (Salimetrics, State College, PA). The intra-assay and the inter-assay’s CV were 8.8% and 11.0%, respectively.

Salivary samples were collected on d -1, T0, 0.5, 1, 2, 4, 24, 48, 72, 144, 336, 505, 672 h after castration. Salivary samples were collected using a cotton swab that was stored in a plastic tube and frozen at– 20° C for further analysis [33 (link)]. Salivary cortisol concentrations were quantified using an enzyme immunoassay kit (Salimetrics, State College, PA). The inter-assay coefficient of variation (CV) was 10.3% while the intra-assay CV was 9.2%. Hair from the forehead of the calves was clipped on d– 1 and 672 h and stored in plastic bags at room temperature and samples were handled as described by Moya et al. [34 ]. Cortisol was quantified using an enzyme-immunosorbent assay (Salimetrics, State College, PA). The intra-assay and the inter-assay’s CV were 9.7% and 12.1% respectively.