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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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Temperatures experienced during early ontogeny significantly influence fish phenotypes, with clear consequences for the wild and reared stocks. We examined the effect of temperature (17, 20, or 23 °C) during the short embryonic and yolk-sac larval period, on the swimming performance and skeleton of metamorphosing Gilthead seabream larvae. In the following ontogenetic period, all fish were subjected to common temperature (20 °C). The critical swimming speed of metamorphosing larvae was significantly decreased from 9.7 ± 0.6 TL/s (total length per second) at 17 °C developmental temperature (DT) to 8.7 ± 0.6 and 8.8 ± 0.7 TL/s at 20 and 23 °C DT respectively (p < 0.05). Swimming performance was significantly correlated with fish body shape (p < 0.05). Compared with the rest groups, fish of 17 °C DT presented a slender body shape, longer caudal peduncle, terminal mouth and ventrally transposed pectoral fins. Moreover, DT significantly affected the relative depth of heart ventricle (VD/TL,p < 0.05), which was comparatively increased at 17 °C DT. Finally, the incidence of caudal-fin abnormalities significantly decreased (p < 0.05) with the increase of DT. To our knowledge, this is the first evidence for the significant effect of DT during the short embryonic and yolk-sac larval period on the swimming performance of the later stages. [ABSTRACT FROM AUTHOR]

The anabolic effect of exercise on muscles and bones is well documented. In teleost fish, exercise has been shown to accelerate skeletogenesis, to increase bone volume, and to change the shape of vertebral bodies. Still, increased swimming has also been reported to induce malformations of the teleost vertebral column, particularly lordosis. This study examines whether zebrafish (Danio rerio) develops lordosis as a result of continuous physical exercise. Zebrafish were subjected, for 1 week, to an increased swimming exercise of 5.0, 6.5 or 8.0 total body lengths (TL) per second. Control and exercise group zebrafish were examined for the presence of vertebral abnormalities, by in vivo examination, whole mount staining for bone and cartilage and histology and micro‐computed tomography (CT) scanning. Exercise zebrafish developed a significantly higher rate of lordosis in the haemal part of the vertebral column. At the end of the experiment, the frequency of lordosis in the control groups was 0.5 ± 1.3% and that in the exercise groups was 7.5 ± 10.6%, 47.5 ± 10.6% and 92.5 ± 6.0% of 5.0, 6.5 and 8.0 TL∙s−1, respectively. Histological analysis and CT scanning revealed abnormal vertebrae with dorsal folding of the vertebral body end plates. Possible mechanisms that trigger lordotic spine malformations are discussed. This is the first study to report a quick, reliable and welfare‐compatible method of inducing skeletal abnormalities in a vertebrate model during the post‐embryonic period. [ABSTRACT FROM AUTHOR]

Haemal lordosis is a frequent abnormality of the vertebral column. It has been recorded to develop in different finfish species, during the hatchery rearing phase. Under certain conditions, this abnormality reaches a high prevalence and severity degree, with significant effects on the external morphology of the fish. We show that haemal lordosis recovers during the on-growing of Gilthead seabream in sea cages. At the end of the hatchery phase, 1700 seabream juveniles were tagged electronically and examined for the presence of haemal lordosis. Subsequently, their morphology was examined periodically up to the end of the on-growing period. We found that the prevalence of fish with a lordotic external morphology decreased during the studied period by approximately 50%. Interestingly, 27% of the recovered fish presented a completely normal vertebral column. Geometric morphometric analysis showed no significant differences in the body shape between the fish with a recovered normal phenotype and the fish that were normal since the beginning of the on-growing period. Our results provide the first evidence for the recovery of lordosis during the growth of fish. A mechanism with multiple levels of remodeling of abnormal bones is suggested. [ABSTRACT FROM AUTHOR]