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- Impact of dietary protein hydrolysates on skeleton quality and proteome in Diplodus sargus larvaePublication . Vareilles, Mahaut; Richard, Nadège; Gavaia, Paulo J.; Silva, Tomé S.; Cordeiro, O.; Guerreiro, Inês; Yúfera, M.; Batista, I.; Pires, C.; Pousão-Ferreira, P.; Rodrigues, Pedro; Rønnestad, I.; Fladmark, K. E.; Conceição, L. E. C.In order to investigate the effects of dietary protein hydrolysates (PH) on larval growth performance, skeleton quality and proteome expression, triplicate groups of white seabream (Diplodus sargus) larvae were co-fed from first-feeding with live feed and three microencapsulated diets differing in the molecular weight of their PH fraction (Control – inclusion of CPSP-90; H – inclusion of a high amount in 0.5–30 kDa hydrolysates; L – inclusion of a high amount in <0.5 kDa hydrolysates). At 15 days after hatching (DAH), proteome expression changes were assessed in entire larvae by twodimensional gel electrophoresis and the quality of larval skeleton was analysed at 28 DAH through double staining of cartilage and bone. Dietary PH fractions tested affected growth, the larvae fed diet L being significantly larger than those fed diet H, but it did not affect the incidence of deformed larvae, nor the number of deformities per fish. Two-dimensional analysis of larvae proteome allowed the detection and the comparative quantification of a total of 709 protein spots having a pI between 4 and 7, around half of which had an expression significantly affected by dietary treatment, the main difference being between proteome of Control larvae with those of both groups L and H. From these spots, 52 proteins involved in diverse processes such as cytoskeletal dynamics, energetic, lipoprotein, amino acid (AA), and nucleotide metabolisms, protein chaperoning and degradation, and signal transduction, were identified. This study revealed that the molecular weight of the dietary protein hydrolysate fraction had a minor impact on skeletal deformities in white seabream larvae, but affected growth performance and had a strong impact on larvae whole body proteome.
- Skeletal deformities in larval and juvenile stages of cultured white seabream Diplodus sargus and common two-banded seabream Diplodus vulgarisPublication . Nicolau, L.; Pousão-Ferreira, P.; Gavaia, Paulo J.Species diversification is presently a major concern amongst aquaculture researchers and producers trying to find new profitable species for commercial production and restocking. White seabream Diplodus sargus (Linnaeus, 1758) and common two-banded seabream Diplodus vulgaris (Saint-Hillaire, 1817) are two new promising sparid species for aquaculture production, as an alternative to gilthead seabream Sparus aurata and sea bass Dicenthrarchus labrax, since they present different spawning seasons and good growth ratios.
- Larval and juvenile development of dusky grouper Epinephelus marginatus reared in mesocosmsPublication . Cunha, M. E.; Re, P.; Quental-Ferreira, H.; Gavaia, Paulo J.; Pousão-Ferreira, P.The larval development of the dusky grouper Epinephelus marginatus up to the benthic juvenile stage is described in detail to establish a reference for their larval identification. Development is described in terms of ontogenetic changes in morphology, growth, pigmentation, fin structure and skeletal structure. Larvae were reared in mesocosms at a mean temperature of 24·3◦ C, salinity of 36·5, dissolved oxygen of 6·4mg l−1 and pH of 8·2. Newly hatched larvae had an estimated total length (LT) of 2·3 mm. On the second day post hatching the yolk was almost fully absorbed with traces of the oil globule still present, the eyes were already pigmented and mouth and gut functional. At this stage the cranial skeletal elements for feeding and breathing (mouth and gills) and the pectoral-fin support were already present. About 50% of the observed larvae had food in their guts. Pigmentation was very characteristic, consisting of two large chromatophores visible on the edge of the primordial fin, close to the midpoint of the post-anal region of the body and over the midgut and hindgut and post-anal portion of the body. At 2·9mm LT the emergence of the second dorsal-fin spine, characteristic of the Epinephilinae, was clearly visible. The pre-flexion stage started in larva of 3·2mm LT. At 5·5mm LT the larvae possessed posterior preopercular angle spines, and the dorsal and pelvic spines presented serrated edges and were pigmented. The water surface-tension-related death of the yolk sac and pre-flexion larvae described in the rearing of several other grouper species did not occur during E. marginatus culture. Notochord flexion, with initial ossification of the caudal-fin supporting elements, started at 6·6mm LT. At this stage the major melanophores, preopercular, dorsal and pelvic spines and mandibular teeth were already present. Transformation of larvae into juveniles occurred when larvae averaged 13·8mm LT. Juveniles with a mean LT of 20·1mm started to settle and most of them were benthic with a mean LT of 26·8 mm.
- Development of chondroid tissues in lordotic-kyphotic Senegalese sole (Solea senegalensis)Publication . Cardeira Da Silva, João; Mendes, A. C.; Pousão-Ferreira, P.; Cancela, Leonor; Gavaia, Paulo J.Vertebral deformities are frequent and constitute a major drawback for many aquaculture produced species. Lordosis and kyphosis, which are severe curvatures of the vertebral column, are particularly problematic, compromising fish marketing and profitability. Senegalese sole is a species with increasing interest in this industry and, like many others, presents high frequencies of vertebral deformities under rearing conditions.
- Lordotic-kyphotic vertebrae develop ectopic cartilage-like tissue in Senegalese sole (Solea senegalensis)Publication . Cardeira Da Silva, João; Bensimon-Brito, A.; Pousão-Ferreira, P.; Cancela, Leonor; Gavaia, Paulo J.Lordosis and kyphosis are curvatures of the vertebral column that commonly affect Senegalese sole (Solea senegalensis) under intensive rearing conditions, constituting a problem for the aquaculture industry by increasing labor and operation costs and consequently reducing profits. However, no studies have yet addressed microanatomical changes in vertebral column curvature in this species. We have conducted histological analyses in order to determine the alterations at tissue and cellular levels in these deformities. Development of an ectopic cartilage-like tissue within bone matrix at the growth regions (contact area between two vertebrae) of affected vertebrae was observed in lordotic-kyphotic fish. As previously suggested, the increase in strain supported by deformed vertebrae may trigger the onset of metaplastic tissue formation through an osteochondroprogenitor precursor from the condensation of mesenchymal cells or by trans-differentiation of (pre-)osteoblastic cells to a chondrocytic lineage. The possibility of a notochord contribution, as well as the molecular events related to bone stress and bone types in terms of cellularity, are also discussed.
- Early axial and appendicular skeletal development of the meagre (Argyrosomus regius). A comparison of two rearing systemsPublication . Valles, E.; Cardeira Da Silva, João; Dionísio, Gisela; Estévez, A.; Pousão-Ferreira, P.; Cancela, Leonor; Gavaia, Paulo J.The meagre (Argyrosomus regius) is a new species for aquaculture with high economical value, presenting growth rates of 1 kg/year. Skeletal deformities generally appear in cultured larvae at early stages, reducing the value of the fish. The present study describes the osteological development of the vertebral column and fins in meagre larvae. Meagre larvae were obtained from wild broodstocks, kept at IRTA and IPIMAR, and induced to spawn using GnRHa injection.
- Lack of essential fatty acids in live feed during larval and post-larval rearing: effect on the performance of juvenile Solea senegalensisPublication . Dâmaso-Rodrigues, M. L.; Pousão-Ferreira, P.; Ribeiro, Laura; Coutinho, J.; Bandarra, N.; Gavaia, Paulo J.; Narciso, L.; Morais, S.Despite the large progress obtained in recent years, Senegalese sole (Solea senegalensis) production of high quality juveniles is still a bottleneck. This paper examines the effect of larval and post-larval lipid nutrition on juvenile performance and quality. Four dietary treatments were tested: A—enriched Artemia spp. (EA); B—non-enriched Artemia spp. (NEA); C—EA during the pelagic larval period and NEA after larval settlement; D—50% EA and 50% NEA. Juvenile fatty acid profile at 60 days after hatching (DAH) clearly reflected the larval and post-larval diet composition. Feeding sole larvae on NEA (poor in lipids and essential fatty acids-EFA) had a negative effect, reducing growth (total length and dry weight) after 30 DAH and decreasing digestive enzyme activity at the end of the rearing period (60 DAH). However, relatively good performance compared to the EFA-richest treatment (A) was obtained when larvae were fed 50% EA and 50% NEA (D) or even EA only during the pelagic larval period followed by NEA after larval settlement (C). Malpigmentation was not affected by the dietary regimes and its incidence was very low. However, skeletal deformities were prevalent, particularly in the caudal complex, independently of diet. The results confirm that Senegalese sole appear to have lower larval EFA requirements than most cultured marine species and potentially even lower requirements during the post-larval stage. The importance of studying the impact of early nutrition on later juvenile stages was clearly highlighted in this study.
- Rearing fish larvae for extensive and semi-intensive aquaculture - the “natural” mesocosms systemPublication . Cunha, M. E.; Quental-Ferreira, H.; Boglione, C.; Palamara, E.; Gavaia, Paulo J.; Pousão-Ferreira, P.Non-intensive aquaculture systems are rapidly generating interest due to increased awareness of consumers about environmental issues, including those associated with coastal areas; eutrophication as a result of intensive aquaculture production is seen as a problem, there is concern about animal welfare issues in fish rearing, and food safety is of course a major concern.
- Fish larvae quality descriptors: an appraisal of methods for red porgy Pagrus pagrus and grouper Epinephelus marginatus produced under different rearing conditionsPublication . Andrade, Carlos A. P.; Nogueira, N.; Silva, Paula; Cordeiro, Nereida; Cunha, M. E.; Pousão-Ferreira, P.; Ribeira, Laura; Bandarra, N.; Gavaia, Paulo J.Quality control in fish hatcheries is of paramount importance to achieve the desired characteristics of larvae and fry, either for the market or release to the wild. Quality programmes are applied in a day to day basis having direct implications for production management and final costs of product. Most common criteria used for larval quality assessment are morphometrics, condition factor, histometrical indices, lipid analysis, nucleic acid ratios, enzyme activity and stress tests.
- Skeletal evaluation and technical improvements to decrease the incidence of skeletal deformities in Solea senegalensis, KaupPublication . Gavaia, Paulo J.; Richard, Nadège; Dâmaso, L.; Dinis, Maria Teresa; Pousão-Ferreira, P.; Engrola, S.; Conceição, L. E. C.; Cancela, LeonorThe appearance of skeletal deformities is a major problem associated to intensive, large-scale culture of fish species, affecting growth, development and ultimately, survival of produced fish. Although the mechanisms working behind it are still unclear, it is known that environmental, genetic and nutritional factors play fundamental roles. Given its importance as a vertebrate model to study skeleton-related diseases, the selection of an appropriate dietary protocol is a critical step in order to diminish the nutritional effect on bone and cartilage development. To better understand the effect of diet on skeletal development and deformities, as well as on larvae and juvenile global performance, we have tested four distinct dietary regimes, composed of rotifers Brachionus sp., Artemia nauplii and dry, formulated food.