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Microplastics are present in marine habitats worldwide and may be ingested by low trophic organisms such as fish larvae, with uncertain physiological consequences. The present study aims at assessing the impact of polyethylene (PE 10-45 mu M) microbeads ingestion in European sea bass (Dicentrarchus labrax) larvae. Fish were fed an inert diet including 0, 10(4) and 10(5) fluorescent microbeads per gram from 7 until 43 days post-hatching (dph). Microbeads were detected in the gastrointestinal tract in all fish fed diet incorporating PE. Our data revealed an efficient elimination of PE beads from the gut since no fluorescent was observed in the larvae after 48 h depuration. While the mortality rate increased significantly with the amount of microbeads scored per larvae at 14 and 20 dph, only ingestion of the highest concentration slightly impacted mortality rates. Larval growth and inflammatory response through Interleukine-1-beta (IL-1 beta) gene expression were not found to be affected while cytochrome-P450-1A1 (cyp1a1) expression level was significantly positively correlated with the number of microbeads scored per larva at 20 dph. Overall, these results suggest that ingestion of PE microbeads had limited impact on sea bass larvae possibly due to their high potential of egestion.

Ocean warming, eutrophication and consequent decrease in oxygen lead to smaller average fish size. Although such responses are well-known in an evolutionary context, involving multiple generations, it appears to be incompatible with current rapid environmental change. Rather, phenotypic plasticity could provide a means for marine fish to cope with rapid environmental changes. However, little is known about the mechanisms underlying plastic responses to environmental conditions that favour small phenotypes. Our aim was to investigate how and why European sea bass that had experienced a short episode of moderate hypoxia during their larval stage subsequently exhibited a growth depression at the juvenile stage compared to the control group. We examined whether energy was used to cover higher costs for maintenance, digestion or activity metabolisms, as a result of differing metabolic rate. The lower growth was not a consequence of lower feed intake. We measured several respirometry parameters and we only found a higher SDA (Specific Dynamic Action) duration and lower SDA amplitude in a fish phenotype with lower growth; this phenotype was also associated with a lower protein digestive capacity in the intestine. Our results contribute to the understanding of the observed decrease in growth in response to climate change. They demonstrate that the reduced growth of juvenile fishes as a consequence of an early-life hypoxia event was not due to a change of fish aerobic scope, but to a specific change in the efficiency of protein digestive functions. The question remains of whether this effect is epigenetic and could be reversible in the offspring.

Ocean warming, eutrophication and the consequent decrease in oxygen lead to smaller average fish size. Although such responses are well known in an evolutionary context, involving multiple generations, this appears to be incompatible with current rapid environmental change. Instead, phenotypic plasticity could provide a means for marine fish to cope with rapid environmental changes. However, little is known about the mechanisms underlying plastic responses to environmental conditions that favour small phenotypes. Our aim was to investigate how and why European sea bass that had experienced a short episode of moderate hypoxia during their larval stage subsequently exhibited a growth depression at the juvenile stage compared with the control group. We examined whether energy was used to cover higher costs for maintenance, digestion or activity metabolisms, as a result of differing metabolic rate. The lower growth was not a consequence of lower food intake. We measured several respirometry parameters and we only found a higher specific dynamic action (SDA) duration and lower SDA amplitude in a fish phenotype with lower growth; this phenotype was also associated with a lower protein digestive capacity in the intestine. Our results contribute to the understanding of the observed decrease in growth in response to climate change. They demonstrate that the reduced growth of juvenile fishes as a consequence of an early life hypoxia event was not due to a change of fish aerobic scope but to a specific change in the efficiency of protein digestive functions. The question remains of whether this effect is epigenetic and could be reversible in the offspring.

The quality of development in intensive or mesocosm hatchery-reared Dicentrarchus labrax larvae was investigated using physiological indicators assessing ontogeny. Larvae were reared in intensive (120 L tanks) and in mesocosm systems (20 m(3) enclosures) with the same feeding sequence, excluding the wild zooplankton naturally available in mesocosms. Faster growth was recorded since early development [16 day after hatching (DAH)] in the mesocosm. Maturation of the digestive system also occurred earlier as indicated by the higher amylase secretion ratios, the intestinal maturation index (alkaline phosphatase/leucine-alanine peptidase and aminopeptidase-N/leucine-alanine peptidase ratios) and the more developed intestinal epithelium at 23 DAH. Nevertheless, the delay in digestive maturation in the intensive system seemed retrieved within few days. In both the groups, the number of vertebrae ranged between 24 and 26, with the dominant class being 25 vertebrae. However, the distributions differed between treatments for meristic characteristics, ossification stages and incidence of malformation types. Loss of a vertebra was more frequent in the intensive system, while the appearance of an additional vertebra was more frequent in the mesocosm. Ossification at 37 DAH was also more advanced in the mesocosm in addition to a lesser rate and severity of skeletal malformations. It is suggested that the early nutritional contribution of mesocosm wild zooplankton, yet at densities of 0.2-0.7 prey mL(-1), had key effects on larvae development since the early stages.

International audience; The influence of dietary vitamins on growth, survival, and morphogenesis was evaluated until day 38 of posthatching life in European sea bass larvae (Dicentrarchus labrax). A standard vitamin mix (VM), at double the concentration of the U. S. National Research Council's recommendations, was incorporated into larval feeds at 0.5%, 1.5%, 2.5%, 4.0%, and 8.0% to give treatments VM 0.5, VM 1.5, VM 2.5, VM 4.0, and VM 8.0, respectively. The group fed the VM 0.5 diet all died before day 30. At day 38, the larvae group fed VM 1.5 had 33% survival, while the other groups, with higher vitamin levels, showed at least 50% survival. The higher the percentage VM in the diet, the lower the percentage of column deformities. High dietary vitamin levels positively influenced the formation of mineralized bone in larvae: the higher the dietary vitamin level, the higher the ossification status. In the larvae group fed at the highest vitamin levels, we observed a temporal sequence of coordinated growth factor expression, in which the expression of bone morphometric protein (BMP-4) preceded the expression of IGF-1, which stimulated the maturation of osteoblasts (revealed by high osteocalcin expression levels). In groups fed lower proportions of vitamins, elevated proliferator peroxisome-activated receptors (PPAR-gamma) expression coincided with low BMP-4 expression. Our results suggest that high levels of PPAR-gamma transcripts in larvae-fed diets with a low VM content converted some osteoblasts into adipocytes during the first two weeks of life. This loss of osteoblasts is likely to have caused skeletal deformities.

A study was designed to evaluate the maximum level of substitution of fish meal in diets for turbot when a mixture of feedstuffs of plant origin was used and when the amino acid profile was adjusted to reflect amino acid profile of a reference fish meal based diet. Five diets were formulated: a reference diet (FM40) containing 40% fish meal and four other diets (FM30, FM20, FM10 and FM0) in which protein from fish meal was substituted by protein from lupin, corn gluten and wheat gluten meal. Amino acids profile of each diet was balanced by addition of crystalline amino acids. Turbot, initially weighing 26 g were fed these diets for 12 weeks and growth parameters were recorded. Protein (96%), energy (83%) and dry matter (73%) digestibility were very high in all diets tested. Feed intake was significantly decreased by the incorporation of plant protein in diets (from 10.2 g/kg ABW/day to 8.2 g/kg ABW/day in fish fed diets FM40 and FM0, respectively), growth rate was only affected in fish fed diets FM10 and FM0 (respectively, 2.2 and 1.9%/day against an average of 2.4%/day for the other treatments) and no effect on feed utilisation was observed. The whole body composition of fish and the nitrogen retention were significantly negatively affected when dietary fish meal incorporation was equal or less than 20% diet. Activities of alanine amino transferase (ALAT) were significantly increased by the dietary incorporation of plant protein. These results suggest that a sizeable amount of fish meal can be replaced by plant feedstuffs in diets of turbot when an adequate mixture of plant feedstuffs along with amino acid supplementation are used.

The aim of the study was to evaluate the effects of dietary arginine (Arg) supplementation to either a plant protein-based diet or a fish meal-based diet on nitrogen (N) utilisation and ureogenesis in turbot and rainbow trout. Juvenile turbot (7.4 g) and rainbow trout (9.3 g) were fed for 12 weeks two types of diets (plant protein-based (PM) and fish meal-based (FM)) containing different levels of Arg (1.6%, 3% and 4% diet for PM diets and, 3% and 4% for FM diets). Besides, measurement of growth parameters and protein utilisation, ammonia and urea excretion rates were monitored. Plasma ammonia and urea concentrations and activities of selected enzymes of ornithine urea cycle (carbamoyl phosphate synthetase III, ornithine carbamoyltransferase and arginase) were measured. Both species fed PM-based diets had reduced growth performance compared to fish fed FM-based diets. Amino acid imbalance of the PM-based diets had a significant effect on plasma ammonia levels and ammonia excretion. Dietary Arg supplementation did not lead to any improvement in N utilisation or on ammoniogenesis. In both species, there was a linear relationship between dietary Arg and urea excretion. Our data confirm that ornithine urea cycle is incomplete in the liver and that argininolysis plays a major role in ureogenesis in both species.

Recent studies indicate that urea excretion is responsive to protein intake and that turbot, Psetta maxima, appear to differ from other species by their urea excretion pattern and levels. This study was undertaken to evaluate the influence of dietary nitrogen and arginine on ureagenesis and excretion in turbot. Juvenile turbot (29 g) were fed semi-purified diets containing graded levels of nitrogen (0–8% dry matter) and arginine (0–3% dry matter) for 6 weeks. Growth data showed that turbot have high dietary nitrogen (123 mg/kg metabolic body weight/day) and very low dietary arginine (9.3 mg/kg metabolic body weight/day) requirements for maintenance. Requirements for unit body protein accretion were 0.31 g and 0.15 g for nitrogen and arginine respectively. Post-prandial plasma urea levels and urea excretion rates showed that urea production was significantly (P

We evaluated the influence of dietary nucleic acid (NA) supplied either as an RNA extract (RNA) or as brewer's yeast (BY) on nitrogen (N) utilisation, N excretion and ureogenesis in a freshwater teleost, the rainbow trout (Oncorhynchus mykiss) and a marine teleost, the turbot (Psetta maxima). Five diets containing two levels of NA or BY were formulated for each species, and fed to juvenile rainbow trout (initial body weight (IBW): 21.7 g) and turbot (IBW: 11.8 g) over 8–10 weeks. Besides growth and N utilisation, we monitored total ammonia-N and urea-N excretion rates and measured the activities of selected enzymes (glutamate dehydrogenase, ornithine carbamoyltransferase, arginase and uricase). There was no clear N sparing effect of dietary NA or BY supplementations in either species. Feeding diets containing the RNA extract led to an almost two-fold increase in postprandial plasma urea-N and uric acid concentrations and in urea-N excretion rates. Glutamate dehydrogenase and arginase activities were decreased in fish fed NA supplemented diets. Uricase activities were inversely related to dietary NA levels. Data on N excretion and enzyme activities clearly suggest that ureogenesis is influenced by dietary NA in both species.

The incorporation of various bulk agents by substitution of an equivalent amount of the basal mixture was studied in terms of protein digestibility, growth performance and body composition in European seabass (Dicentrarchus labrax) juveniles. During the growth trial, triplicate groups of 40 seabass (mean initial weight: 7.0 g) were grown in seawater (salinity: 35; temperature: 18 degrees C) over 60 days. Fish were hand-fed, three times a day, one of seven experimental diets. Bulk agents tested at 10 or 20 % level of incorporation were silica, cellulose and a natural zeolite (chabamin). Feeding rates were adjusted in proportion to the percent dilution of the control diet without bulk incorporation. The incorporation of the bulk agents, at a 10 and 20 % level, did not affect protein digestibility or growth performance. Dietary bulk incorporation reduced feed efficiency values, particularly at the 20 % incorporation level. However, this reduction was mostly caused by the dietary nutrient dilution of the bulk-incorporated diets, rather than by a negative effect of the bulk agents as dietary ingredients. In comparison to the control treatment, bulk incorporation at 10 and 20 % level did not affect protein retention values. When compared with the control diet, 20 % bulk agent incorporation changed the evacuation profile of faeces and increased faecal egestion time.

The aim of this study was to determine whether mortality observed during the larval development of reared bluefin tuna (Thunnus thynnus) could be related to improper expression profiles of key genes involved in digestive or anti-oxidant response capabilities. Tuna larvae were sampled at hatching, 2, 5, 10, 15 and 20 dph (days post hatching) for the relative quantification of transcripts encoded by genes involved in digestive (trypsinogen 1 [TRYP1], alpha-amylase [AMY], aminopeptidase N [ANPEP]) and antioxidant (catalase 32 [CAT]) functions. The levels of expression of ANPEP related to the development and maturation of intestinal function increased from 5 to 20 dph. Furthermore, AMY and TRYP1 genes which are pancreatic enzymes implicated in carbohydrate and peptide digestions exhibit a typical peak of expression at 5 and 15 dph respectively. The antioxidant enzyme, CAT, exhibited higher mRNA levels during the first stage of larval development. In conclusion, our investigation indicates that the expression of genes involved in digestive and antioxidant physiological processes followed typical patterns which could not explain high mortality rate observed during the first stage of larval development, SI

Resume Brown trout and rainbow trout (average weight 100 g) were reared in fresh water at 12OC under the same conditions beforc transfer of brown trout to sea water, in order to compare nitrogen utilisation in the two species. Apparent protein digestibility (ADC), nitrogen (ammonia and urea) excretion, protein productive value (PPV) and actual observed nitrogen mass balance were determined. Rainbow trout raised in fresh water had a higher growth rate (1.1 vs 0.8Yb.d-'), better food conversion ratio (0.7 vs 1.0), better ADC (91 vs 85%) and PPV (45 vs 35%) and lower ammonia excretion rates than brown trout reared in fresh water. Transfemng brown trout to sea water induced lower PPV (30%) and ammonia and urea excretion. Salinity did not rnodify metabolic efficiency in brown trout. Fat content was higher in brown trout (7.7-8.9% ww) than in rainbow trout (5.7-7.6% ww). Nitrogen mass balance indicated that compounds other than arnmonia and urea were produced in higher quantities by seawater brown trout. Behaviour, less domestication and specific ability to utilise protein could explain the differences between the two species.

Three species of marine fish, sea bass, sea bream and turbot, and two salmonids, rainbow trout and brown trout, were raised under similar feeding and environmental conditions in order to compare their nitrogen utilisation. Apparent digestibility of protein, ammonia and urea excretion patterns, plasma ammonia and urea concentrations and liver arginase activities were measured. No differences in protein digestibility were noticed among the five species. Ammonia and urea losses were quantitatively similar in all species, except in turbot, in which ammonia production was significantly lower. Ammonia excretion patterns were linked to ingested nitrogen and showed no inter-species differences. Conversely, urea excretion patterns were specific in turbot and sea bream. Plasma urea levels were higher in marine fish than in salmonids, with the highest values being reached in turbot. Some specificity with regard to liver arginase was also detected in the turbot. Turbot demonstrated some metabolic characteristics that could be connected with its elevated position in the phylogeny of fish.

Laboratory experiments on juvenile turbot. Scophthalmus maximus (L.), were carried out under controlled temperature (12°C) and feeding regimes in a flow-through system. Monitoring of total ammonia nitrogen (TAN) and urea nitrogen (Urea-N) was performed through continuous sampling of the effluent sea water. The effects of ingested nitrogen levels on TAN and Urea-N daily and hourly excretion rates were studied. Strong relationships were found between ingested nitrogen, and both TAN and Urea-N excretion for both daily and hourly maximum excretion rates. Turbot showed a low metabolic activity, confirmed by low excretion levels. Daily patterns for TAN and Urea-N production were different, suggesting specific physiological phenomenon for urea excretion mechanisms. The question of whether turbot are partly ureotelic or ureogenic is underlined.