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The mechanisms behind the beneficial effects of dietary peptides on fish skeletal development remain unrevealed. In the present study, we evaluated the effect of 0% (C, Control), 6% (P6) and 12% (P12) levels of small peptide incorporation on European sea bass (Dicentrarchus labrax) early larval skeletal development and post-larval skeletal integrity against a swimming challenge test. Survival was not affected by the peptide diets, whereas P6 presented the lowest growth rate. Larval quality control underlined the advantageous effect of P12 on reducing the frequency of cephalic deformities (e.g., branchiostegal rays, operculum and cross-bite), pre-haemal lordosis and vertebrae bone loss. Simultaneously, individuals from P12 group exhibited an earlier mineralization of the vertebral column and were less prone to develop swimming-induced haemal lordosis (16.0 ± 0.1%) and scoliosis (3.3 ± 0.6%). Expression analysis of genes involved in digestive function, protein transport, muscle ontogeny and bone mineralization revealed a peptide-enhanced larvae development of the P12 group. An early nutritional programming of the post-larval musculoskeletal system is proposed. Limitations induced by the differential free amino-acid profiles are discussed. A potential developmental-stage-specific incorporation of peptide diets in European sea bass rearing is suggested.

Incorporation of dietary peptides has been correlated with decreased presence of skeletal abnormalities in marine larvae. In an attempt to clarify the effect of smaller protein fractions on fish larval and post-larval skeleton, we designed three isoenergetic diets with partial substitution of their protein content with 0% (C), 6% (P6) and 12% (P12) shrimp di- and tripeptides. Experimental diets were tested in zebrafish under two regimes, with inclusion (ADF-Artemia and dry feed) or lack (DF-dry feed only) of live food. Results at the end of metamorphosis highlight the beneficial effect of P12 on growth, survival and early skeletal quality when dry diets are provided from first feeding (DF). Exclusive feeding with P12 also increased the musculoskeletal resistance of the post-larval skeleton against the swimming challenge test (SCT). On the contrary, Artemia inclusion (ADF) overruled any peptide effect in total fish performance. Given the unknown species’ larval nutrient requirements, a 12% dietary peptide incorporation is proposed for successful rearing without live food. A potential nutritional control of the larval and post-larval skeletal development even in aquaculture species is suggested. Limitations of the current molecular analysis are discussed to enable the future identification of the peptide-driven regulatory pathways.

Fish skeletal development has long been correlated with nutritional factors. Lack of zebrafish nutritional standardization, especially during the early stages, decreases the reproducibility of the conducted research. The present study represents an evaluation of four commercial (A, D, zebrafish specific; B, generic for freshwater larvae; C, specific for marine fish larvae) and one experimental (Ctrl) early diets on zebrafish skeletal development. Skeletal abnormalities rates in the different experimental groups were assessed at the end of the larval period (20 days post-fertilization, dpf) and after a swimming challenge test (SCT, 20–24 dpf). At 20 dpf, results revealed a significant effect of diet on the rate of caudal-peduncle scoliosis and gill-cover abnormalities, which were relatively elevated in B and C groups. SCT results focused on swimming-induced lordosis, which was comparatively elevated in diets C and D (83% ± 7% and 75% ± 10%, respectively, vs. 52% ± 18% in diet A). No significant effects of dry diets were observed on the survival and growth rate of zebrafish. Results are discussed with respect to the deferential diet composition between the groups and the species requirements. A potential nutritional control of haemal lordosis in finfish aquaculture is suggested.

Incorporation of dietary peptides has been correlated with decreased presence of skeletal abnormalities in marine larvae. In an attempt to clarify the effect of smaller protein fractions on fish larval and post-larval skeleton, we designed three isoenergetic diets with partial substitution of their protein content with 0% (C), 6% (P6) and 12% (P12) shrimp di- and tripeptides. Experimental diets were tested in zebrafish under two regimes, with inclusion (ADF-Artemia and dry feed) or lack (DF-dry feed only) of live food. Results at the end of metamorphosis highlight the beneficial effect of P12 on growth, survival and early skeletal quality when dry diets are provided from first feeding (DF). Exclusive feeding with P12 also increased the musculoskeletal resistance of the post-larval skeleton against the swimming challenge test (SCT). On the contrary, Artemia inclusion (ADF) overruled any peptide effect in total fish performance. Given the unknown species’ larval nutrient requirements, a 12% dietary peptide incorporation is proposed for successful rearing without live food. A potential nutritional control of the larval and post-larval skeletal development even in aquaculture species is suggested. Limitations of the current molecular analysis are discussed to enable the future identification of the peptide-driven regulatory pathways.

Fish skeletal development has long been correlated with nutritional factors. Lack of zebrafish nutritional standardization, especially during the early stages, decreases the reproducibility of the conducted research. The present study represents an evaluation of four commercial (A, D, zebrafish specific; B, generic for freshwater larvae; C, specific for marine fish larvae) and one experimental (Ctrl) early diets on zebrafish skeletal development. Skeletal abnormalities rates in the different experimental groups were assessed at the end of the larval period (20 days post-fertilization, dpf) and after a swimming challenge test (SCT, 20–24 dpf). At 20 dpf, results revealed a significant effect of diet on the rate of caudal-peduncle scoliosis and gill-cover abnormalities, which were relatively elevated in B and C groups. SCT results focused on swimming-induced lordosis, which was comparatively elevated in diets C and D (83% ± 7% and 75% ± 10%, respectively, vs. 52% ± 18% in diet A). No significant effects of dry diets were observed on the survival and growth rate of zebrafish. Results are discussed with respect to the deferential diet composition between the groups and the species requirements. A potential nutritional control of haemal lordosis in finfish aquaculture is suggested.

Haemal lordosis, a frequent skeletal deformity in teleost fish, has long been correlated with increased mechanical loads induced by swimming activity. In the present study, we examine whether juvenile zebrafish can recover from haemal lordosis and explore the musculoskeletal mechanisms involved. Juveniles were subjected to a swimming challenge test (SCT) that induced severe haemal lordosis in 49% of the animals and then immediately transferred them to 0.0 total body lengths (TL) per second of water velocity for a week. The recovery from lordosis was examined by means of whole mount staining, histology and gene expression analysis. Results demonstrate that 80% of the lordotic zebrafish are capable of internal and external recovery within a week after the SCT. Recovered individuals presented normal shape of the vertebral centra, maintaining though distorted internal tissue organization. Through the transcriptomic analysis of the affected haemal regions, several processes related to chromosome organization, DNA replication, circadian clock and transcription regulation were enriched within genes significantly regulated behind this musculoskeletal recovery procedure. Genes especially involved in adipogenesis, bone remodeling and muscular regeneration were regulated. A remodeling tissue-repair hypothesis behind haemal lordosis recovery is raised. Limitations and future possibilities for zebrafish as a model organism to clarify mechanically driven musculoskeletal changes are discussed.

Haemal lordosis, a frequent skeletal deformity in teleost fish, has long been correlated with increased mechanical loads induced by swimming activity. In the present study, we examine whether juvenile zebrafish can recover from haemal lordosis and explore the musculoskeletal mechanisms involved. Juveniles were subjected to a swimming challenge test (SCT) that induced severe haemal lordosis in 49% of the animals and then immediately transferred them to 0.0 total body lengths (TL) per second of water velocity for a week. The recovery from lordosis was examined by means of whole mount staining, histology and gene expression analysis. Results demonstrate that 80% of the lordotic zebrafish are capable of internal and external recovery within a week after the SCT. Recovered individuals presented normal shape of the vertebral centra, maintaining though distorted internal tissue organization. Through the transcriptomic analysis of the affected haemal regions, several processes related to chromosome organization, DNA replication, circadian clock and transcription regulation were enriched within genes significantly regulated behind this musculoskeletal recovery procedure. Genes especially involved in adipogenesis, bone remodeling and muscular regeneration were regulated. A remodeling tissue-repair hypothesis behind haemal lordosis recovery is raised. Limitations and future possibilities for zebrafish as a model organism to clarify mechanically driven musculoskeletal changes are discussed.

Targeting in zebrafish fast growth, high survival rates and improved reproductive performance has led over the last years in variable feeding regimes between different facilities. Despite its significance on fish function and welfare, normal skeletal development has rarely been evaluated in establishing the best feeding practices for zebrafish. The aim of this study was to establish a protocol for normal skeletal development, growth and survival of zebrafish larvae through live feed-to-microdiet transition at an appropriate rate. Four feeding regimes including feeding exclusively on Artemia nauplii (A) or dry microdiet (D), and feeding on both Artemia and microdiet at two different transition rates (slow (B) or fast (C)) were applied from 5 to 24 dpf (days post-fertilization). Results demonstrated a significant effect of feeding regimes on the incidence of skeletal abnormalities (gill cover, fins and vertebral column, p < .05) in zebrafish larvae. The A and B experimental groups presented the highest (88 ± 3 and 84 ± 17%, respectively), but the C and D the lowest (18 ± 14 and 11 ± 2%, respectively), rates of normal fish (fish without any abnormality). Similarly, growth rate was comparatively elevated in A and B groups. No significant differences were observed in fish survival between A, B and C groups. However, D group presented a significantly lower survival rate. To our knowledge, this is the first study to show that the live feed-to-microdiet transition rate influences larval growth, survival and abnormality rates in a non-homogenous pattern.

Targeting in zebrafish fast growth, high survival rates and improved reproductive performance has led over the last years in variable feeding regimes between different facilities. Despite its significance on fish function and welfare, normal skeletal development has rarely been evaluated in establishing the best feeding practices for zebrafish. The aim of this study was to establish a protocol for normal skeletal development, growth and survival of zebrafish larvae through live feed-to-microdiet transition at an appropriate rate. Four feeding regimes including feeding exclusively on Artemia nauplii (A) or dry microdiet (D), and feeding on both Artemia and microdiet at two different transition rates (slow (B) or fast (C)) were applied from 5 to 24 dpf (days post-fertilization). Results demonstrated a significant effect of feeding regimes on the incidence of skeletal abnormalities (gill cover, fins and vertebral column, p < .05) in zebrafish larvae. The A and B experimental groups presented the highest (88 ± 3 and 84 ± 17%, respectively), but the C and D the lowest (18 ± 14 and 11 ± 2%, respectively), rates of normal fish (fish without any abnormality). Similarly, growth rate was comparatively elevated in A and B groups. No significant differences were observed in fish survival between A, B and C groups. However, D group presented a significantly lower survival rate. To our knowledge, this is the first study to show that the live feed-to-microdiet transition rate influences larval growth, survival and abnormality rates in a non-homogenous pattern.