Larval performance and skeletal deformities in farmed gilthead sea bream (Sparus aurata) fed with graded levels of Vitamin A enriched rotifers (Brachionus plicatilis)

Several nutritional studies have found a direct effect of several vitamins in chondrogenic and osteogenic development during early life stages of marine fish species. In the present study, the effect of vitamin A (VA) in gilthead sea bream skeletogenesis was evaluated by means of four different dietary regimes (enriched rotifers) containing increasing levels of total VA (75, 109, 188 and 723 ng total VA mg⁻ ¹ DW). Dietary treatments were offered to larvae during the rotifer-feeding phase (4-20 days after hatching), while later all groups were fed with Artemia nauplii and weaned onto the same inert diet. Different dietary doses of VA affected gilthead sea bream larval growth, survival, performance (maturation of the digestive system) and quality (incidence of skeletal deformities). Higher levels of dietary VA than those included in the commercial emulsion for rotifer enrichment led to different levels and typologies of skeletal deformities, indicating that gilthead sea bream larvae were very sensitive to small increases in dietary VA. The degree of ossification was affected by the level of VA in enriched rotifers: the higher amount of VA in the diet, the higher number of skeletal pieces ossified (R =0.585, P =0.04). Dietary VA affected the normal process of bone formation and skeletogenesis, the skeletal structures mostly affected by high amounts of dietary VA were those from the cranial skeleton (splanchnocranium), vertebral centrums and caudal fin complex. The premaxilla, maxilla and dentary bones were the cranial structures affected by dietary VA levels, resulting in a large incidence of animals with compressed snout. Dietary VA also affected the normal development of the opercular complex, and a dose-response dependant effect was observed in relation to the incidence of specimens with incomplete operculum. Body shape was also affected by the level of dietary VA, increasing the incidence of specimens with lordosis, kyphosis and/or scoliosis with the dose of VA, being the prehaemal and caudal vertebrae the most affected regions of the vertebral column with this kind of abnormalities. The caudal fin complex was the most affected region of the skeleton affected by dietary treatments as seen by the high incidence of skeletal deformities in fish fed different doses of dietary VA. Deformities affected all skeletal elements composing the caudal fin, although the most affected ones were, in order of importance, the epurals, hypurals, parahypural, neural arch and uroneurals. Differences in sensitivity to dietary VA amongst caudal fin skeletal elements might be due to their differential ontogenetic development and differences in the exposure time to VA. An excess of dietary VA also accelerated the intramembranous ossification process of vertebral centrums leading to one or two supranumerary vertebrae, and a high incidence of fused and compressed vertebral centrums. The sensibility of the developing skeletal structures to dietary VA levels should incline us to test lower doses of VA in live preys enrichments during early larval stages and higher doses afterwards.