Non-invasive measurement of cortisol in saliva is of prime importance as it represents a bioavailable neuroendocrine marker for stress. Therefore, in this study, we developed an enzyme immune assay that was suitable for salivary cortisol measurements. For that purpose, rabbit polyclonal antibody was raised against cortisol-3-CMO:BSA conjugate. The test was based on competition of liquid phase cortisol with conjugated cortisol on the solid phase. Primary antibody was used to bind available sites on the conjugate, which was proportional to numbers of cortisol in liquid phase. Biotinylated secondary anti-rabbit antibody was used to detect primary antibodies by addition of streptavidin peroxidase and substrate, respectively. Color formation was stopped and yellow color was read by a plate-reader spectrophotometer. Additionally, validated test was used to met all validation criteria including. Test developed was used to establish cortisol awakening response (CAR) in saliva samples collected in the morning after awakening (0, 15, 30, and 60^th min) from women (n = 4) and men (n = 4) at 8 or 4 different days, respectively. Diurnal cortisol levels were assessed (n = 8) at after awaking 60 min at morning, 12:00, 19:00, and 22:00 hr. In conclusion, an enzyme immunoassay test was successfully produced, validated and used for cortisol measurement in saliva samples.

The genetic relevance of Turkish sheep has recently been revealed by mtDNA analysis, which has led to the identification of a third maternal lineage in that species. We present here a study of the genetic variation of these breeds at a nuclear level from the analysis of 30 microsatellite loci. Parameters of variability indicated a high level of variation in the breeds analysed. Mean number of alleles per locus ranged from 7.8 to 10.4 and gene diversity varied between 0.69 and 0.74. Considerable differences were detected between breeds regarding the pattern of within-breed variability. Breed-specific alleles were detected at most loci and all breeds analysed, a total of 75 private variants being identified. The Hemsin sheep showed noticeably lower genetic variation in accordance with the present status of this breed. Percentages of shared alleles (from 54.8% to 69.5%) and gene flow estimates (from 5.98 to 28.32) gave information about the relative genetic differentiation of breeds. The results presented here for nuclear DNA complement those reported for mtDNA and bear out the need to preserve this invaluable genetic material.