Denis Coves

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.

Cloning and retinal expression of melatonin receptors in the European sea bass, Dicentrarchus labrax

Melatonin contributes to synchronizing behaviors and physiological functions to daily and seasonal rhythm in fish. However, no coherent vision emerges because the effects vary with the species, sex, age, moment of the year or sexual cycle. And, scarce information is available concerning the melatonin receptors, which is crucial to our understanding of the role melatonin plays. We report here the full length cloning of three different melatonin receptor subtypes in the sea bass Dicentrarchus labrax, belonging, respectively, to the MT1, MT2 and Mel1c subtypes. MT1, the most abundantly expressed, was detected in the central nervous system, retina, and gills. MT2 was detected in the pituitary gland, blood cells and, to a lesser extend, in the optic tectum, diencephalon, liver and retina. Mel1c was mainly expressed in the skin; traces were found in the retina. The cellular sites of MT1 and MT2 expressions were investigated by in situ hybridization in the retina of pigmented and albino fish. The strongest signals were obtained with the MT1 riboprobes. Expression was seen in cells also known to express the enzymes of the melatonin biosynthesis, i.e., in the photoreceptor, inner nuclear and ganglion cell layers. MT1 receptor mRNAs were also abundant in the retinal pigment epithelium. The results are consistent with the idea that melatonin is an autocrine (neural retina) and paracrine (retinal pigment epithelium) regulator of retinal function. The molecular tools provided here will be of valuable interest to further investigate the targets and role of melatonin in nervous and peripheral tissues of fish.

Tagging juvenile seabass and sole with telemetry transmitters: medium-term effects on growth

The effects of tagging with acoustic transmitters on the growth of juvenile seabass, Dicentrarchus labrax (L.) (initial mean mass±SD, 173 g±23.4) in a 47 d tank experiment, and sole, Solea solea (L.) (103.2 g±14.8) in a 72 d tank experiment and (104.0 g±18.4) in a 58 d salt marsh mesocosm experiment were examined. Twenty externally tagged seabass grew more slowly than the 20 with surgically implanted tags, which reached the same mass as nine control fish. Movements of the external transmitters harness caused abrasions of the skin and loss of the tag in 60% of the cases. We thus recommend implanting transmitters for telemetry studies of juvenile seabass weighing between 120 and 214 g and carrying a tag that represents 2.2–2.5% of body mass. Both tank and mesocosm experiments conducted on juvenile sole concluded that the externally attached tag retention rate was good, but at the expense of the fish growth rate.

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.

Typology of individual growth in sea bass (Dicentrarchus labrax)

The individual growth variability of passive integrated transponder tagged sea bass was studied using data sets from two different experiments. In experiment 1 (n = 485), fish submitted to different photoperiod regimes were held in fourteen groups of individual weight of 88 ± 13 g (mean ± SD). In experiment 2 (n = 748, initial weight 243 ± 30 g) fish were held in fifteen groups and had either free or restricted access to diets with three lipid levels. After adjustment for treatment and tank effects, individual growth curves were analysed using multivariate analysis (principal component analysis and clustering) and were modelled using the summary statistics technique. Different growth profiles where characterized. All of them appeared to be curvilinear. They differed in their level (initial and final weight), slope (slope, specific growth weight, gain) and especially the ratio of males, which showed sexual growth dimorphism. The fish with similar initial weight proved to have very different growth performances, regardless of the treatment effect. Within the same sex, part of the variability between the growth profiles could be explained by differences in the social interactions and in the genetic potential of growth among individuals.

Development of the posterior lateral line system in Thunnus thynnus, the atlantic blue-fin tuna, and in its close relative Sarda sarda

The lateral line system of amphibians and fish comprises a large number of individual mechanosensory organs, the neuromasts, and their sensory neurons. The pattern of neuromasts varies markedly between species, yet the embryonic pattern is highly conserved from the relatively basal zebrafish, Danio rerio, to more derived species. Here we examine in more detail the development of the posterior lateral line (PLL) in embryos and early larvae of one of the most derived fish species, the blue-fin tuna Thunnus thynnus, and of its close relative, the Atlantic bonito Sarda sarda. We show that the basic features of embryonic PLL development, including the migratory properties of the PLL primordium, the patterning of neuromasts and their innervation, are largely conserved between zebrafish and tuna. However, Thunnus and Sarda embryos differ from Danio in three respects: the larger size of the neuromast cupula, the capability of mature neuromasts to migrate dorsally, and the presence of a single, precisely located terminal neuromast.

Developmental origin of a major difference in sensory patterning between zebrafish and bluefin tuna

The posterior lateral line system (PLL) of teleost fish comprises a number of mechanosensory organs arranged in defined patterns on the body surface. Embryonic patterns are largely conserved among teleosts, yet adult patterns are highly diverse. Although changes in pattern modify the perceptual abilities of the system, their developmental origin remains unknown. Here we compare the processes that underlie the formation of the juvenile PLL pattern in Thunnus thynnus, the bluefin tuna, to the processes that were elucidated in Danio rerio, the zebrafish. In both cases, the embryonic PLL comprises five neuromasts regularly spaced along the horizontal myoseptum, but the juvenile PLL comprises four roughly parallel anteroposterior lines in zebrafish, whereas it is a simple dorsally arched line in tuna fish. We examined whether this difference involves evolutionary novelties, and show that the same mechanisms mediate the transition from embryonic to juvenile patterns in both species. We conclude that the marked difference in juveniles depends on a single change (dorsal vs. ventral migration of neuromasts) in the first days of larval life.

Somatotropic axis genes are expressed before pituitary onset during zebrafish and sea bass development.

The somatotropic axis, or growth hormone-insulin-like growth factor-1 (GH-IGF-1) axis, of fish is involved in numerous physiological process including regulation of ionic and osmotic balance, lipid, carbohydrate and protein metabolism, growth, reproduction, immune function and behavior. It is thought that GH plays a role in fish development but conflicting results have been obtained concerning the ontogeny of the somatotropic axis. Here we investigated the developmental expression of GH, GH-receptor (GHR) and IGF-1 genes and of a GH-like protein from fertilization until early stages of larval development in two Teleosts species, Danio rerio and Dicentrarchus labrax, by PCR, in situ hybridization and Western blotting. GH, GHR and IGF-1 mRNA were present in unfertilized eggs and at all stages of embryonic development, all three displaying a similar distribution in the two species. First located in the whole embryo (until 12 hpf in zebrafish and 76 hpf in sea bass), the mRNAs appeared then distributed in the head and tail, from where they disappeared progressively to concentrate in the forming pituitary gland. Proteins immunoreactive with a specific sea bass anti-GH antibody were also detected at all stages in this species. Differences in intensity and number of bands suggest that protein processing varies from early to later stages of development. The data show that all actors of the somatotropic axis are present from fertilization in these two species, suggesting they plays a role in early development, perhaps in an autocrine/paracrine mode as all three elements displayed a similar distribution at each stage investigated.