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At hatch, the chick skeleton is a miniature of that of the adult bird. The hen deposits calcium, phosphorus, and trace minerals (copper, zinc, and manganese) along with vitamin D into the egg to allow development of the embryonic skeleton. The main source of calcium is the eggshell, whereas phosphorus, trace minerals, and vitamin D are mainly derived from the yolk. Calcium is absorbed from the eggshell and transferred to the embryo and yolk through the chorioallantoic membrane, whereas phosphorus and trace minerals are simultaneously mobilized by the yolk sac membrane. These processes start at day 12 of incubation and peak at around day 17. While the eggshell provides a steady supply of calcium until 19 d of incubation, phosphorus and trace mineral reserves decrease considerably and minimal skeletal development occurs in the last 3 d of incubation. Whether the low levels of phosphorus and trace minerals at late incubation prevent further bone growth, or some other biological control exists preventing further mineralization towards hatching is unknown. Maternal transfer of minerals and the influence of trace mineral form in the hen diet to advance the state of skeletal development at hatch have received increased research attention. Minimal effects on yolk mineral composition and bone growth were observed in the offspring of hens fed different forms of trace minerals. Embryos from young hens had inferior bone development towards the end of incubation and at hatch relative to chicks from older hens. This effect is likely a consequence of limited egg nutrient resources in eggs from young hens. The influence of maternal nutrient transfer on embryonic bone development has been clearly established. However, attempts to increase the state of skeletal development at hatch through increasing egg mineral content have met with limited success. The focus of this paper is the relationship between skeletal mineralization of the chicken embryo throughout incubation and egg mineral supply.

Broiler strain, maternal age, and incubation temperature influence embryo metabolism. Hatching eggs were obtained from young (Y; 28 to 34 wk, $\bar{\rm x}$ = 31.2 wk), mid (M; 36 to 45 wk, $\bar{\rm x}$ = 40.5 wk) and old (O; 49 to 54 wk, $\bar{\rm x}$ = 51.4 wk) Ross 708 (n = 88; Experiment 1) and Ross 308 [(n = 45; Experiment 2: (Y; 25 to 34 wk, $\bar{\rm x}$ = 30.5 wk), (M; 35 to 44 wk, $\bar{\rm x}$ = 40.2 wk), and (O; 49 to 54 wk, $\bar{\rm x}$ = 51.6 wk)] breeders. Eggs were stored for 2 to 4 d (18°C, 73% RH), and incubated for 14 d at 37.5°C and 56% RH. At 15 d (E15), 8 fertile eggs per flock age were incubated in individual metabolic chambers at 36.0, 36.5, 37.0, or 37.5°C until E21.5. Each temperature was repeated one additional time. O2 consumption and CO2 production were used to calculate embryonic heat production (EHP). Embryo temperature was measured as eggshell temperature (EST). Initial egg weight was used as a covariate; significance was assessed at P < 0.05. In Ross 708, daily EHP tended to be higher in M and O than Y treatments at E16; EHP of M was higher than Y and O eggs at E18; M and O were higher than O eggs at E19. Incubation at 37.0°C resulted in the highest EHP from E15 to E21, except at E17. Embryos at 37.5°C had reduced EHP beyond E17. Daily EST from E15 to E21 was higher at 37.5 and 37.0°C than at 36.0 and 36.5°C. In Ross 308, daily EST was highest at 37.5°C except at E20. Incubation temperature and EST were highly correlated (R2 = 0.90 to 0.89; P < 0.001). Ross 708 chicks were longer and hatched earlier at 37.0°C than at 36.0 and 37.5°C. EST and EHP increased with incubation temperature in Ross 708. In Ross 308, maternal flock age and incubation temperature did not impact EHP. However, EST was highest at 37.5°C except at E20. Ross 708 was more sensitive to incubation temperature than Ross 308.

Broiler chickens have an extreme physiology (rapid growth rates) that challenges the correct bone mineralization, being an interesting animal model for studying the development of bone pathologies. This work studies in detail how the mineralization, chemistry, and structural organization of tibiae bone in broiler chickens change with age during the first 5 wk (37 D) from hatching until acquiring the final weight for slaughter. During the early growth phase (first 2 wk), the rapid addition of bone tissue does not allow for bone organic matrix to fully mineralize and mature, and seems to be a critical period for bone development at which bone mineralization cannot keep pace with the rapid growth of bones. The low degree of bone mineralization and large porosity of cortical bone at this period might be responsible of leg deformation and/or other skeletal abnormalities commonly observed in these birds. Later, cortical bone porosity gradually decreases and the cortical bone became fully mineralized (65%) at 37 D of age. At the same time, bone mineral acquires the composition of mature bone tissue (decreased amount of carbonate, higher crystallinity, Ca/P = 1.68). However, the mineral part was still poorly organized even at 37 D. The oriented fraction was about 0.45 which means that more than half of apatite crystals within the mineral are randomly oriented. Mineral organization (crystal orientation) had an important contribution to bone-breaking strength. Nevertheless, locally determined (at tibia mid-shaft) bone properties (i.e., cortical thickness, crystal orientation) has only a moderate correlation (R2 = 0.33) with bone breaking strength probably due to large and highly heterogeneous porosity of bone that acts as structural defects. On the other hand, the total amount of mineral (a global property) measured by total ash content was the best predictor for breaking strength (R2 = 0.49). Knowledge acquired in this study could help in designing strategies to improve bone quality and reduce the incidence of skeletal problems in broiler chickens that have important welfare and economic implications.