Morphological and molecular characterization of developing vertebral fusions using a teleost model

<p>Abstract</p> <p>Background</p> <p>Spinal disorders are a major cause of disability for humans and an important health problem for intensively farmed animals. Experiments have shown that vertebral deformities present a complex but comparable etiology across species. However, the underlying molecular mechanisms involved in bone deformities are still far from understood. To further explicate the mechanisms involved, we have examined the fundamental aspects of bone metabolism and pathogenesis of vertebral fusions in Atlantic salmon (<it>Salmo salar</it>).</p> <p>Results</p> <p>Experimentally, juvenile salmon were subjected to hyperthermic conditions where more than 28% developed fused vertebral bodies. To characterize the fusion process we analyzed an intermediate and a terminal stage of the pathology by using x-ray, histology, immunohistochemistry, real-time quantitative PCR and <it>in situ </it>hybridization. At early stage in the fusion process, disorganized and proliferating osteoblasts were prominent at the growth zones of the vertebral body endplates. PCNA positive cells further extended along the rims of fusing vertebral bodies. During the developing pathology, the marked border between the osteoblast growth zones and the chondrocytic areas connected to the arches became less distinct, as proliferating cells and chondrocytes blended through an intermediate zone. This cell proliferation appeared to be closely linked to fusion of opposing arch centra. During the fusion process a metaplastic shift appeared in the arch centra where cells in the intermediate zone between osteoblasts and chondrocytes co-expressed mixed signals of chondrogenic and osteogenic markers. A similar shift also occurred in the notochord where proliferating chordoblasts changed transcription profile from chondrogenic to also include osteogenic marker genes. In progressed fusions, arch centra and intervertebral space mineralized.</p> <p>Conclusion</p> <p>Loss of cell integrity through cell proliferation and metaplastic shifts seem to be key events in the fusion process. The fusion process involves molecular regulation and cellular changes similar to those found in mammalian deformities, indicating that salmon is suitable for studying general bone development and to be a comparative model for spinal deformities.</p

Molecular pathology of vertebral deformities in hyperthermic Atlantic salmon (Salmo salar)

<p>Abstract</p> <p>Background</p> <p>Hyperthermia has been shown in a number of organisms to induce developmental defects as a result of changes in cell proliferation, differentiation and gene expression. In spite of this, salmon aquaculture commonly uses high water temperature to speed up developmental rate in intensive production systems, resulting in an increased frequency of skeletal deformities. In order to study the molecular pathology of vertebral deformities, Atlantic salmon was subjected to hyperthermic conditions from fertilization until after the juvenile stage.</p> <p>Results</p> <p>Fish exposed to the high temperature regime showed a markedly higher growth rate and a significant higher percentage of deformities in the spinal column than fish reared at low temperatures. By analyzing phenotypically normal spinal columns from the two temperature regimes, we found that the increased risk of developing vertebral deformities was linked to an altered gene transcription. In particular, down-regulation of extracellular matrix (ECM) genes such as <it>col1a1</it>, <it>osteocalcin</it>, <it>osteonectin </it>and <it>decorin</it>, indicated that maturation and mineralization of osteoblasts were restrained. Moreover, histological staining and <it>in situ </it>hybridization visualized areas with distorted chondrocytes and an increased population of hypertrophic cells. These findings were further confirmed by an up-regulation of <it>mef2c </it>and <it>col10a</it>, genes involved in chondrocyte hypertrophy.</p> <p>Conclusion</p> <p>The presented data strongly indicates that temperature induced fast growth is severely affecting gene transcription in osteoblasts and chondrocytes; hence change in the vertebral tissue structure and composition. A disrupted bone and cartilage production was detected, which most likely is involved in the higher rate of deformities developed in the high intensive group. Our results are of basic interest for bone metabolism and contribute to the understanding of the mechanisms involved in development of temperature induced vertebral pathology. The findings may further conduce to future molecular tools for assessing fish welfare in practical farming.</p

Growth, sensory and chemical characterization of Mediterranean yellowtail (Seriola dumerili) fed diets with partial replacement of fish meal by other protein sources

[EN] An 84-day trial was performed to assess the use of alternative protein sources in Seriola dumerili. Three diets were used, FM100 diet, as a control diet without fishmeal substitution, and FM66 and FM33 diets with a fishmeal replacement of 330 g/kg and 660 g/kg, respectively. At the end of experiment, fish fed the FM66 diet showed the no differences in growth, nutritional parameters and fatty acid composition. Heavy metals present some differences but are always lower than risk levels. In sensory analysis, differences between diets appeared in pH and color, and also in some texture parameters between FM33 and the other two diets. No differences appeared between diets related to flavor. In summary, long periods of feeding with high fish meal substitution diets, affects Seriola dumerili growth; despite this the quality of the fillet was not affected even with a 66 % of substitution.This project was financed by "Generalitat Valenciana. Ayudas para grupos de investigacion consolidables."Monge-Ortiz, R.; Martínez-Llorens, S.; Lemos-Neto, M.; Falco, S.; Pagán Moreno, MJ.; Godoy-Olmos, S.; Jover Cerda, M.... (2020). Growth, sensory and chemical characterization of Mediterranean yellowtail (Seriola dumerili) fed diets with partial replacement of fish meal by other protein sources. Aquaculture Reports. 18:1-10. https://doi.org/10.1016/j.aqrep.2020.100466S11018Abbas, K. A., Mohamed, A., Jamilah, B., & Ebrahimian, M. (2008). A Review on Correlations between Fish Freshness and pH during Cold Storage. American Journal of Biochemistry and Biotechnology, 4(4), 416-421. doi:10.3844/ajbbsp.2008.416.421Álvarez, A., García García, B., Garrido, M. D., & Hernández, M. D. (2008). The influence of starvation time prior to slaughter on the quality of commercial-sized gilthead seabream (Sparus aurata) during ice storage. 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Thermal imprinting modifies bone homeostasis in cold-challenged sea bream (Sparus aurata)

Fish are ectotherms and temperature plays a determinant role in their physiology, biology and ecology, and is a driver of seasonal responses. The present study assessed how thermal imprinting during embryonic and larval stages modified the response of adult fish to low water temperature. We targeted the gilthead sea bream, which develops a condition known as winter syndrome when it is exposed to low water temperatures. Eggs and larvae of sea bream were exposed to four different thermal regimes and then the response of the resulting adults to a low temperature challenge was assessed. Sea bream exposed to a high-low thermal regime as eggs and larvae (HLT; 22 degrees C until hatch and then 18 degrees C until larvae-juvenile transition) had increased plasma cortisol and lower sodium and potassium in response to a cold challenge compared with the other thermal history groups. Plasma glucose and osmolality were increased in cold-challenged HLT fish relative to the unchallenged HLT fish. Cold challenge modified bone homeostasis/responsiveness in the low-high thermal regime group (LHT) relative to other groups, and ocn, ogn1/2, igf1, gr and tr alpha/beta transcripts were all downregulated. In the low temperature group (LT) and HLT group challenged with a low temperature, alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) activities were decreased relative to unchallenged groups, and bone calcium content also decreased in the LT group. Overall, the results indicate that thermal imprinting during early development of sea bream causes a change in the physiological response of adults to a cold challenge.Seventh Framework Programme project Lifecycle [EU-FP7 222719