Gilbert Dutto

Effect of chronic ammonia exposure on growth of European seabass (Dicentrarchus labrax) juveniles

Chronic effects of ammonia were studied in juvenile seabass, Dicentrarchus labrax (mean WEIGHT=11 g), exposed for 63 days to eight stable ammonia concentrations, ranging from 0.24 to 0.90 mg l−1 unionised ammonia nitrogen (UIA-N), respectively, from 6.1 to 22.3 mg l−1 total ammonia nitrogen (TA-N). Temperature (21.8 °C), pH (8.0), salinity (37.0 ppt), and oxygen concentration (over 80% saturation at the outlet) were maintained constant. Fish were fed using a self-feeder device, and they were starved during the last 8 days. Mortality of 28.9 and 42.6% occurred within the first 8 days at the two highest UIA-N concentrations, respectively, 0.90 and 0.88 mg l−1. From days 0 to 55, a 1.8- fold increase in weight gain was observed under the 0.90-mg l−1 UIA-N condition, compared to a 3.4- fold increase in the control. Weight gains were negatively correlated to ambient ammonia concentrations. Weight loss, or a transient period of growth stagnation, was observed from the onset of ammonia exposure to day 13 in seabass exposed to concentrations above 0.43 mg l−1 UIA-N. After day 13, weight gains were observed in all groups, indicating that the fish were able to adapt to increased ambient ammonia concentrations over time. By the end of the experiment, plasma ammonia levels were positively related to ambient ammonia concentrations, and oxygen consumption recorded in fasting fish was significantly dependent on ammonia concentrations. In seabass juveniles, the 0.26- mg l−1 UIA-N concentration, under an average pH of 8.0, can be considered as a safe long-term limit conditions in seawater.

Nitrogenous and phosphorous waste production in a flow-through land-based farm of European seabass (Dicentrarchus labrax)

A study on nitrogenous and phosphorous waste production in European seabass (Dicentrarchus labrax) was carried out in a flow-through land-based farm. The main objectives were (i) to calculate nitrogenous (N) and phosphorous (P) waste production (particulate and dissolved) from in situ measurements for different rearing ponds with specific fish biomasses, fish sizes and flow rates, (ii) to establish nitrogenous and phosphorous waste production budgets, ratios and equations, and (iii) to compare, for the whole farm, in situ measurements to estimate N and P waste production from waste equations. Waste production was manifest by an increase in concentrations of total ammonia nitrogen, total Kjeldhal nitrogen, particulate nitrogen, dissolved phosphorus and total phosphorus concentrations in the outlet water compared to the inlet water for both batches and whole farm. However, no production of nitrite or nitrate was observed. In our budgets, the N and P amounts from biomass gains and wastes were explained by the N and P derived from feed over the range 83.9–105.2 % and 66.5–104.6 % respectively, depending on the fish batch. Values were respectively 103.4 and 87.5 % for the same calculations in the farm. When the whole-farm waste production was calculated from previous equations derived from batches, and then compared with the measured data, the percentages of recovery (estimated by the ratio predicted data/measured data) were 88 and 94 % for total-N and total-P respectively.

Behavioral and neurophysiological responses of European sea bass groups reared under food constraint

The individual food-demand behavior of juvenile European sea bass (Dicentrarchus labrax, L.) reared in groups under self-feeding conditions was investigated. The triggering activity on self-feeder, i.e. index of the food-demand activity, agonistic interactions and territorial behavior were monitored for periods of 42 to 68 days in six groups of 50 fish. The specific growth rate was calculated and the brain serotonergic activity was used as a stable index of social stress. Inter-individual differences appeared in triggering activity and three groups were distinguished: 3-5 high-triggering fish, 17-30 low-triggering fish and the remaining individuals were null-triggering fish. There were no significant differences in specific growth rates calculated at the end of the experiment (day 42 or day 68) between individuals with high, low, and null food-demand (ANOVA, p>0.05). No territorial or agonistic behaviors were observed, however, there were significant differences in brain serotonergic activity between the three triggering groups (ANOVA, p=0.050 in telencephalon and p=0.004 in cerebellum). Specifically, high-triggering fish had lower serotonergic turnover than low or null-triggering fish. We put forth the hypothesis that fish with low or null-triggering activity could be stressed by the high activity of high-triggering individuals.

Effect of dietary lipid content on circadian rhythm of feeding activity in European sea bass.

In fish, dietary digestible energy (DE) content is a major factor controlling feed intake. It was therefore of interest to determine how circadian rhythm of feeding activity is influenced by the dietary DE levels. To that end, groups of European sea bass were fed on demand by means of self feeders, under light-dark and constant light conditions, with a fixed or an unlimited amount of feed with variable lipid contents. Daily total feed intake, but not the feeding rhythm, was adjusted in relation to the DE content of the diet regardless of the lighting conditions. We conclude that a satiation mechanism was likely responsible for the regulation of feed intake in relation to the dietary fat content but was not acting in itself on the mechanisms that drive the free-running rhythms of feeding activity. These results are giving additional evidence that a true endogenous clock is driving feeding activity rhythms in fish.

Effect of chronic exposure to ammonia on alterations of proteins and immunoglobulins in sea bass (Dicentrarchus labrax) serum

The relationship between serum protein, immunoglobulin concentrations and protein molecular weight profile (PMWP) alterations of sea bass (131.3 ± 4.3 g) reared in sea water with sublethal concentrations of ammonia was studied over two periods totalling 116 days. During the exposure period (62 days) the first group (group 1) lived in sea water with 0.204 mg·L -1 unionized ammonia nitrogen equivalent to 12 % of the lethal concentration for 50 % of a population exposed for 96 h to ammonia (96-h LC50), whilst the second group (group 2) lived in sea water with 0.340 mg-L -1 UIA-N equivalent to 20 % of the 96-h LC50 of ammonia. Then, the two groups were left for a recovery period (54 days) in the same water as the control group (group 0). The determination of the total immunoglobulin (Ig) concentration was carried out by enzyme-linked immunosorbant assay (ELISA). Gel filtration columns were used for the serum PMWP. Serum Ig concentration of the exposed fish (group 2: 1.76 ± 0.43 mg·mL- 1 ; group 1: 1.19 ± 0.33 mg·mL- 1 ) was lower than the control group (3.39 ± 1.01 mg·mL- 1 ) after 21 days of exposure period but this difference was reduced at the end of this period (day 62) and treated group Ig concentrations switched to higher than the control group after the recovery period (day 116, group 2: 9.75 ± 1.84 mg·mL-1; group 1: 7.50 ± 1.22 mg·mL- 1 group 0: 6.38 ± 1.13 mg·mL- 1 ). In fact, at the end of the experiment, the cumulative Ig production difference between fish exposed to ammonia and the control was less than 10 %. Although a similar evolution of the Ig serum occurred with protein concentration, the serum protein concentration deficit of group 1 was restored at the end of the exposure period (group 2: 46.49 ± 2.34 mg·mL -1; group 0: 46.74 ± 1.97 mg·mL- 1 ) and the cumulative production during the experiment was not significantly different between group 1 and group O. However, this remained lower for group 2. During the exposure period, the PMWP of treated fish moved towards smaller molecular weight proteins. This alteration of the PMWP showed that the 0.2-kDa fraction increased, and another fraction appeared at the end of the exposure period. However, at the end of the recovery period, no difference was found between the PMWPs of control and treated fish. The alterations of the serum protein and Ig of fish reared, for a Iimited period (2 months), in water with sublethal concentrations of ammonia (20 % 96-h LC50 or less) should disappear completely after a few months in normal rearing conditions.