Effect of dietary vitamin A on Senegalese sole (Solea senegalensis) skeletogenesis and larval quality

The effects of different levels of vitamin A (VA) in Senegalese sole larval performance and development were evaluated by means of a dietary dose-response experiment using enriched Artemia metanauplii as a carrier of this micronutrient. Larvae were fed from 6 to 27days post hatch (dph) with enriched Artemia containing graded levels of total VA (1.3, 2.1, 4.5 and 12.9µgVAmg⁻¹ DW). The content of VA in live prey directly affected its accumulation in larvae and early juveniles. Retinyl palmitate accumulated during larval ontogeny, whereas retinol showed the opposite trend, decreasing from hatching until 41dph and then remaining constant until the end of the study. In metamorphic larvae (10 and 15dph), VA did not affect the number of thyroid follicles or the intensity of the immunoreactive staining of T₃ and T₄. However, at older stages of development (post-metamorphic larvae: 20, 30, 41 and 48dph), VA decreased the number of thyroid follicles but increased their mean size and enhanced T₃ and T₄ immunoreactive staining. A dietary excess of VA did not affect either larval performance in terms of growth and survival or the maturation of the digestive system. However, the most remarkable impact of this morphogenetic nutrient was detected during skeletal morphogenesis. Dietary VA accelerated the intramembranous ossification of vertebral centrums, which led to the formation of a supranumerary haemal vertebra and a high incidence of fused and compressed vertebrae in fish fed 2.1, 4.5 and 12.9mgVAmg⁻¹ DW. In addition, VA also affected those structures from vertebrae and caudal fin formed by chondral ossification, leading to defects in their shape and fusions with adjacent skeletal elements. In particular, the caudal fin was the region most affected by the dietary treatments. In order of importance, the bones with more developmental anomalies were the modified neural and haemal spines, epural, hypurals and parahypural. The impact of systemic factors such as thyroidal hormones in skeletogenesis should not be neglected since present results revealed that an excess of dietary VA affected the levels of T₃ and T₄, which might have affected bone formation and remodelling, leading to skeletal deformities.