Christine Huelvan

Comparison of nitrogenous losses in five teleost fish species

Abstract 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.

Excess dietary arginine affects urea excretion but does not improve N utilisation in rainbow trout Oncorhynchus mykiss and turbot Psetta maxima

Abstract 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.

Influence of dietary bulk agents (silica, cellulose and a natural zeolite) on protein digestibility, growth, feed intake and feed transit time in European seabass (Dicentrarchus labrax) juveniles

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.

Evaluation of the impact of polyethylene microbeads ingestion in European sea bass (Dicentrarchus labrax) larvae

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 μM) 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-1β) 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.

Imbalanced dietary ascorbic acid alters molecular pathways involved in skeletogenesis of developing European sea bass (Dicentrarchus labrax).

The influence of dietary ascorbic acid (AA) on growth and morphogenesis during the larval development of European sea bass (Dicentrarchus labrax) was evaluated until 45days post hatching. Diets incorporated 0, 5, 15, 30, 50 or 400mg AA per kg diet to give AA-0, AA-5, AA-15, AA-30, AA-50 and AA-400 dietary treatments, respectively. Dietary AA levels lower than 15mg/kg reduced larval growth and survival was affected in specimens fed diets devoid of AA. Globally, disruption of the expression of genes involved in AA and calcium absorption in the intestine (SVCT-1, TRPV-6), skeletogenesis (BMP-4, IGF-1, RARγ) and bone mineralization (VDRβ, osteocalcin) were observed in groups fed doses lower and higher than 50mg AA/kg diet. Such disturbances detected at molecular level were associated with disruptions of the ossification process and the appearance of skeletal abnormalities.

Hypoxia tolerance of common sole juveniles depends on dietary regime and temperature at the larval stage: evidence for environmental conditioning

An individuals environmental history may have delayed effects on its physiology and life history at later stages in life because of irreversible plastic responses of early ontogenesis to environmental conditions. We chose a marine fish, the common sole, as a model species to study these effects, because it inhabits shallow marine areas highly exposed to environmental changes. We tested whether temperature and trophic conditions experienced during the larval stage had delayed effects on life-history traits and resistance to hypoxia at the juvenile stage. We thus examined the combined effect of global warming and hypoxia in coastal waters, which are potential stressors to many estuarine and coastal marine fishes. Elevated temperature and better trophic conditions had a positive effect on larval growth and developmental rates; warmer larval temperature had a delayed positive effect on body mass and resistance to hypoxia at the juvenile stage. The latter suggests a lower oxygen demand of individuals that had experienced elevated temperatures during larval stages. We hypothesize that an irreversible plastic response to temperature occurred during early ontogeny that allowed adaptive regulation of metabolic rates and/or oxygen demand with long-lasting effects. These results could deeply affect predictions about impacts of global warming and eutrophication on marine organisms.

Comparison of brown trout (Salmo trutta) reared in fresh water and sea water to freshwater rainbow trout (Oncorhynchus mykiss): II. Phosphorus balance

Brown trout and rainbow trout (average weight 100 g) were reared in fresh water at 12 °C under the same conditions before transferring brown trout to sea water, in order to compare phosphorus utilisation in both species. Apparent phosphorus availability, orthophosphate excretion and phosphorus accretion in the fish were directly determined. Thus, actual phosphorus mass balance was built. Rainbow trout raised in fresh water had a higher phosphorus retention coefficient (maximum 50 %) than brown trout reared in fresh water (maximum 45 %). Transferring brown trout to sea water induced a reduction in phosphorus retention (maximum 39 %). Orthophosphate excretion, ranging 7–20 mg phosphorus per kg wet weight per day, represented 10–20 % of ingested phosphorus. Phosphorus availability was lower in brown trout raised in sea water (65 %) than brown trout raised in fresh water (76 %). Phosphorus balance measurements showed that 90 to 98 % of phosphorus flow could be accounted for.

An early life hypoxia event has a long-term impact on protein digestion and growth in juvenile European sea bass

ABSTRACT 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. Summary: The growth reduction in juvenile sea bass as a consequence of an early life hypoxia event is due to a specific change in the efficiency of protein digestive functions.