Interaction between Dietary Vitamin D3 and Vitamin K3 in Gilthead Seabream Larvae (Sparus aurata) in Relation to Growth and Expression of Bone Development-Related Genes

Vitamins D and K are essential fat-soluble nutrients that intervene in bone development processes among other biological functions. The present study is aimed at investigating the potential combined effect of dietary supplementation with vitamin D3 (cholecalciferol) and vitamin K3 (menadione) in gilthead seabream (Sparus aurata) larvae. For that purpose, seabream diets were supplemented with different combinations of vitamin D3/vitamin K3 (mg/kg diet) as follows: 0.00/0, 0.06/70, 0.06/170, 0.13/70, 0.13/170, 0.40/70, and 0.40/170. Feeding gilthead seabream larvae (22 days post hatch) for 21 days with the diets supplemented with 0.06-0.13 mg/kg vitamin D3 and 70 mg/kg vitamin K3 (diets 0.06/70 and 0.13/70) led to the highest larval growth and survival and the highest expression of important biomarkers of both bone development and health, such as bmp2, osx, and mgp, and calcium homeostasis, such as pthrp and casr. However, the increased supplementation with both vitamins at 0.40 mg/kg vitamin D3 and 170 mg/kg vitamin K3 (diet 0.40/170) reduced larval growth and survival, downregulated bmp2 and pthrp expressions, and upregulated osx and mgp, causing an unbalance in the relative expression of these genes. The results of the present study have shown the interaction between vitamin D3 supplementation and vitamin K3 supplementation in larval performance and gene expression related to bone development and calcium homeostasis, denoting the significance of a correct balance between both vitamins in larval diets.publishedVersio

Effects of the dietary supplementation of copper on growth, oxidative stress, fatty acid profile and skeletal development in gilthead seabream (Sparus aurata) larvae

The effect of dietary copper (Cu) levels on growth, oxidative stress, fatty acid profile and bone health of gilthead seabream larvae (Sparus aurata) was studied in 47 days-post-hatching fish fed 4 experimental diets containing 17, 18, 19 and 25 mg Cu/kg diet. After 21 days of feeding, larval growth and survival were not affected by the dietary Cu levels. However, larvae fed the non-supplemented diet (17 mg Cu/kg) showed clear symptoms of Cu deficiency, including increased lipid peroxidation and reduced bone mineralization. Increased peroxidation risk caused a reduction in essential fatty acids and n-3 PUFA (polyunsaturated fatty acids) contents in whole body lipids, whereas the poor vertebral body mineralization in seabream larvae was related to a marked increase in abdominal vertebrae anomalies. On the contrary, dietary Cu supplementation raised whole-body contents in DHA, EPA and n-3 PUFA and reduced MUFA, TBARS and the incidence of severe, cranial, branchiostegal rays and abdominal vertebral anomalies. Polynomial regression models applied to all these parameters suggested optimum dietary Cu levels to be 21.5–22.6 mg/kg in gilthead seabream larvae microdiets supplemented with CuSO4.5H2O. These levels (21.5–22.6 mg/kg) are in the range of the Cu contents in copepods (12–38 mg/kg) and on the verge of the dietary levels authorized by EU Commission in feeds for cultured fish species (25 mg Cu/kg diet).publishedVersio

Interaction between Dietary Vitamin D3 and Vitamin K3 in Gilthead Seabream Larvae (Sparus aurata) in Relation to Growth and Expression of Bone Development-Related Genes

Vitamins D and K are essential fat-soluble nutrients that intervene in bone development processes among other biological functions. The present study is aimed at investigating the potential combined effect of dietary supplementation with vitamin D3 (cholecalciferol) and vitamin K3 (menadione) in gilthead seabream (Sparus aurata) larvae. For that purpose, seabream diets were supplemented with different combinations of vitamin D3/vitamin K3 (mg/kg diet) as follows: 0.00/0, 0.06/70, 0.06/170, 0.13/70, 0.13/170, 0.40/70, and 0.40/170. Feeding gilthead seabream larvae (22 days post hatch) for 21 days with the diets supplemented with 0.06-0.13 mg/kg vitamin D3 and 70 mg/kg vitamin K3 (diets 0.06/70 and 0.13/70) led to the highest larval growth and survival and the highest expression of important biomarkers of both bone development and health, such as bmp2, osx, and mgp, and calcium homeostasis, such as pthrp and casr. However, the increased supplementation with both vitamins at 0.40 mg/kg vitamin D3 and 170 mg/kg vitamin K3 (diet 0.40/170) reduced larval growth and survival, downregulated bmp2 and pthrp expressions, and upregulated osx and mgp, causing an unbalance in the relative expression of these genes. The results of the present study have shown the interaction between vitamin D3 supplementation and vitamin K3 supplementation in larval performance and gene expression related to bone development and calcium homeostasis, denoting the significance of a correct balance between both vitamins in larval diets

Effects of dietary vitamin D3 levels on survival, mineralization, and skeletal development of gilthead seabream (Sparus aurata) larvae

Vitamin D is an essential fat soluble micronutrient that helps in growth, bone development, calcium homeostasis and other metabolic process. The study on effect of vitamin D3 in marine fish larvae were very scarce irrespective of species. The present study determines the impacts of dietary vitamin D3 on growth performance, calcium absorption, mineralization, and skeletal anomalies during the development of gilthead seabream (Sparus aurata) larvae was assessed until 47 days post hatching. Diets containing four levels of vitamin D3 (0, 25, 30, 384 μg kg−1 or 11.6, 1000, 1200, 15,360 IU kg−1) were formulated to determine the effect of vitamin D3 at deficient, excess, and optimum levels. The gilthead seabream larvae in the present study fed with this wide range of vitamin D3 presented a constant growth with all the diets but presented signs of toxicity in excess level, affecting the survival, calcium uptake, and bone biomarker mechanism in larvae, which resulted in increased skeletal anomalies and mortality. An increase of dietary vitamin D3 up to 384 μg kg−1 significantly raised the whole body vitamin D3 content, calcium, and phosphorus intake and increased the incidence of skeletal anomalies, particularly cranial anomalies. The appearance of skeletal anomalies in larvae fed 384 μg kg−1 vitamin D3 was in association with the upregulation of bmp2, alp, and oc gene expression. However, larvae fed 0, 25, 30 μg kg−1 vitamin D3 showed higher survival than the group fed 384 μg kg−1 vitamin D3. Meanwhile vitamin D3 deficient diet 0 μg kg−1 presented with lower mineralization rate and increase incidence of maxillary anomaly. Thus, the current study revealed the evidence of vitamin D3 deficiency as well as toxicity in gilthead seabream larvae during the developmental process and conclude that the recommended dietary vitamin D3 level for gilthead seabream larvae may range between 25 and 30 μg kg−1 which improves larval survival, calcium and phosphate level and vertebral mineralization with reduced incidence of skeletal anomalies in gilthead seabream larva