Sex differences in serum steroid hormone levels during embryonic development in hen eggs.

This experiment was conducted to explore the differences of serum steroid hormones with age and sex in hen eggs during incubation periods for identification of males and females. The concentrations of estrone (E1), estradiol (E2), estriol (E3), testosterone (T), androstenedione (A4), and dihydrotestosterone (DHT) in serum were measured by enzyme-linked immunosorbent assays in chicken embryos on 8, 10, 12, 14, and 16 D, respectively. During the development of chicken embryo, egg weight loss was closely related to age but no sex. However, it was found that both age and sex significantly affected hormones and had obviously more effect on androgens levels. Besides E2 and T, other steroids such as E1, A4, and DHT were also significantly correlated with sex (P < 0.05). Notably, the level of T and the ratio of T to E2 were significantly higher in males than females (P < 0.05). The ratio of E1 to E2 displayed different trends in different sexes, which increased in males but decreased in females. The distribution proportions of androgens and estrogens kept stable level during the late hatching periods of 12 to 16 D. The sex differences of steroids were more obvious at the late hatching stage through PCA. These suggested that the serum hormones differences in male and female embryos played a vital role in sexual differentiation. These findings not only provided the theoretical basis for sex determination of fertilized eggs in egg-laying hen strains, but also contributed to develop a non-invasive way to sex determination of fertilized eggs to meet the modern commercial application.