Nathalie R. Le François

The Salmonids (Family : Salmonidae)

As a taxon, the salmonids comprise 11 genera, with 65-70 species. All species occur naturally in the northern hemisphere but several salmonid species have been introduced to the southern hemisphere, where they often form the basis of sport fisheries or aquaculture enterprises. There are both freshwater and anadromous species, but some of the anadromous species also have populations that are strictly confined to fresh waters.

Protein synthesis is lowered while 20S proteasome activity is maintained following acclimation to low temperature in juvenile spotted wolffish (Anarhichas minor Olafsen).

SUMMARY The effects of temperature on protein metabolism have been studied mostly with respect to protein synthesis. Temperature generally has a parabolic effect on protein synthesis with a maximum rate being observed at optimal growth temperature. The effect of temperature on protein degradation is poorly understood. The 20S proteasome is mainly responsible for the degradation of short-lived and oxidatively modified proteins and has been recently identified as a potentially good proxy for protein degradation in fish. The aim of this experiment was to examine the relationships between the rate of protein synthesis, activity of the 20S proteasome, oxidative stress markers and antioxidant capacity in white muscle of juvenile spotted wolffish (Anarhichas minor) acclimated at three temperatures (4, 8 and 12°C). The rate of protein synthesis was lower at 4°C than at 8°C while it was intermediate at 12°C. Despite the decrease of protein synthesis at low temperature, the activity of 20S proteasome activity was maintained high in fish acclimated at lower temperature (4°C), reaching levels 130% of that of fish acclimated at 8°C when measured at a common temperature. The oxidative stress markers TBARS and protein-carbonyl content did not change among temperature groups, but reduced glutathione concentration was higher in cold-acclimated fish, suggesting a higher antioxidant capacity in this group. Our data suggest that lower growth rate in cold temperature results from both high 20S proteasome activity and a reduced rate of protein synthesis.

Ontogenesis of catabolic and energy metabolism capacities during the embryonic development of spotted wolffish (Anarhichas minor).

The catabolic and energy metabolism capacities during spotted wolffish (Anarhichas minor) embryogenesis were investigated. We assessed the embryos ability to catabolize proteins (trypsin-like proteases) and lipids (triglyceride lipase) and examined the development of metabolic capacities using enzymatic assays: ability to use carbohydrates (pyruvate kinase), amino acids (aspartate aminotransferase) and fatty acids (hydroxyacyl-CoA dehydrogenase) for energy production, and aerobic (citrate synthase) and anaerobic (lactate dehydrogenase) energy production. Functional enzymatic systems were detected from the eyed stage (350 degree-days), except for fatty acids, which was detected from 540 degree-days. To compare the development of 1) aerobic and anaerobic pathways and 2) the capacity to mobilize the different energy substrates, enzymatic ratios were calculated. Anaerobic capacity appeared to increase at a significantly higher rate than the aerobic capacity. Ratios revealing the relative capacity to use specific energy substrates showed a significantly slower increase during development in the capacity to use carbohydrates than amino acids and fatty acids. The end of embryogenesis was characterized by a significant decrease in the use of carbohydrates for aerobic energy production but an increasing capacity to use amino acids. Egg survival as affected by the variability in metabolic parameters is discussed.

Reproductive events and associated reduction in the seawater adaptability of brook charr (Salvelinus fontinalis): evaluation of gill metabolic adjustments

Abstract We tested the hypothesis that along with gill Na+K+ATPase activity reduction, branchial energetic metabolism of brook charr (Salvelinus fontinalis) is altered during sexual maturation. Maturing and sterile fish were transferred from freshwater (FW) to seawater (SW). The activity of gill pyruvate kinase (PK), cytochrome C oxidase (COX), citrate synthase (CS) and lactate dehydrogenase (LDH) was measured at different intervals. Following 1 month in SW, twofold increases in Na+K+ATPase activity were recorded in all groups, while mean metabolic enzyme activities were sharply reduced (COX: 0.051 ± 0.029 to 0.036 ± 0.018; PK: 1.670 ± 0565 to 1.210 ± 0.340 LDH: 2.245 ± 0.690 to 1.642 ± 0.381 μmol mg–1 protein min–1). Interestingly, during this period, no mortality occurred. After 5 months, comparison of Na+K+ATPase:CS and Na+K+ATPase:LDH ratios of mature and sterile fish held either in FW or SW, indicated that the response of Na+K+ATPase largely exceeds the response of the metabolic enzyme apparatus in hyperosmotic conditions. Hence, the reduced iono-osmoregulatory capacity and higher mortality observed in SW-maturing fish during the reproductive season appears to be mainly attributable to Na+K+ATPase activity reduction rather than the alteration of gill metabolic capacity, since no concurrent increase of metabolic enzyme activity with Na+K+ATPase activity occurred.

Ontogenetic effects of diet during early development on growth performance, myosin mRNA expression and metabolic enzyme activity in Atlantic cod juveniles reared at different salinities.

This study investigates the effect of diet during early development on growth and metabolic capacity in the juvenile stage of Atlantic cod. Growth in three groups of Atlantic cod juveniles (10-70 g) was measured at two salinities (15 per thousand or 32 per thousand) in combination with two temperatures (10 degrees C or 14 degrees C). Groups of cod from a single egg batch differed by having been fed with rotifers (R) or natural zooplankton (Z) during the first 36 days post hatch. A third group was fed zooplankton from 1 to 22 dph, after which diet changed to rotifers from 22 to 36 dph (ZRZ). All fish were weaned at 36 dph. Juveniles from the Z and ZRZ groups performed equally well under all experimental conditions, but fish that had received rotifers as a larval diet showed overall significantly lower growth rates. Growth was significantly enhanced by reduced salinity. Metabolic enzyme activity and relative myosin mRNA expression levels were not affected by larval diet. Muscle AAT and MDH were affected by salinity while these enzymes in liver tissue were affected by the interaction between salinity and temperature. Metabolic enzymes were stronger correlated with fish size than growth rates. Our results indicate that larval diet has a pronounced effect on juvenile growth rates under varying environmental conditions as optimal larval diet (zooplankton) increased juvenile growth rates significantly. Metabolic enzyme activity and relative myosin mRNA expression were not affected by larval history, which suggests that the persisting juvenile growth difference is not a result of differing metabolic capacity.

Trypsin-Like Enzyme from Atlantic Wolffish (Anarhichas Lupus) Viscera: Purification and Characterization

ABSTRACT A trypsin-like enzyme was recovered from Atlantic wolffish (Anarhichas lupus) viscera and its properties were compared with those of commercially available bovine trypsin. Trypsin-like purification was carried out by a combination of sulphate ammonium fractionation, acetone precipitation, and ultra-filtration on 100 kDa and 50 kDa NMW membranes. Assays reveal that both sources of enzymes displayed similar pH range optima (pH 7–10) and similar activity profiles with increasing temperature. The wolffish trypsin-like enzyme mainly differed from bovine trypsin by its thermal and alkaline stability. Atlantic wolffish appear to be an interesting source of digestive proteases with unique properties that could be used in food biotechnology.

Preliminary investigations of the physiological adjustments associated with compensatory growth in juvenile brook charr (Salvelinus fontinalis)

ABSTRACT Various physiological factors suspected to govern or be influenced by growth or feed intake of brook charr (80–120 g) were examined under long and short fasting periods during the year. A control (fed to satiation) was included. During winter, fasting (14 and 28 days) followed by refeeding led to a complete compensation. In summer, longer fasting (39 and 61 days) was applied to exacerbate the physiological response. Compensatory growth was observed in the restricted groups, and overall, adjustments/variations in growth and physiological indicators (feed conversion ratio, organosomatic index, enzymatic activities [CS, TRY, CHY]) were more pronounced. Degree of compensation reached was positively related to the deprivation duration. The summer experiment clearly suggests that compensatory growth takes place in two stages after a long starvation period (61 days): (1) restoration of the digestive system (PSI) followed by enzyme activities (trypsin and chymotrypsin) and (2) rebuilding of somatic tissues.

Cortisol and Behavioral Response to Handling (Acute) and Confinement (Chronic) Stressors in Juvenile Spotted Wolffish, Anarhichas minor

The purpose of this study was to determine the cortisol response of spotted wolffish (Anarhichas minor) after exposure to air immersion and netting (acute stress) under a range of increasing densities (chronic stressor). In addition, the presence of a cumulative negative impact from chronic stress was assessed by the application of an additional acute stressor following rearing at various densities, and proxies of behavior were evaluated (feeding and aggression data). The stress response to air immersion and netting led to increasing plasma cortisol values at 0.5, 13, and 37 h post-stress, with the highest value achieved after 13 h. At 168 h, cortisol concentrations returned to similar pre-stress value. Changes in cortisol concentrations were relatively low after acute stress (five to six fold increases) and recovery time long lasting (>37 h). Prolonged rearing at 20, 30, and 40 kg m−2 showed a non-significant trend of increasing cortisol values with increasing density, and the cortisol response after the application of an additional acute stressor was significantly altered, i.e., higher cortisol levels were measured in unstressed (chronic stress only) compared to stressed fish (chronic stress + acute stress). Our results suggest that (1) spotted wolffish is a low responder that additionally displays a long-lasting cortisol elevation following an acute stressor; (2) fixed densities did not lead to significant differences of plasma cortisol concentration or aggression levels, although involved marked changes in daily feed intake and food conversion efficiency were observed; and (3) cortisol levels reached acute stress values after prolonged chronic stress. Although this species displays indications of being relatively resistant to handling disturbances during aquaculture practices, it is nevertheless responsive to chronic stress factors that could alter the physiological response to subsequent acute stressor.

Population Genetic Structure of the Spotted Wolffish, Anarhichas minor, in the North Atlantic

The genetic population structure of spotted wolffish Anarhichas minor (Olafsen) in the North Atlantic was in- vestigated by allozyme electrophoresis and by PCR based RFLP variation in mtDNA. Samples were collected from five sites distributed across the North Atlantic and from second generation Norwegian broodstock. The present data demon- strates that significant differences exist between almost all sampling sites, indicating biologically different groups of indi- viduals, and thus the existence of different management populations of spotted wolffish in the North Atlantic. Overall, the data indicated that isolation by distance is weak among spotted wolffish at the geographic scale investigated in this study, suggesting that gene flow between neighbouring spotted wolffish populations is low. The present study demonstrates a high level of geographic population structure between the western Atlantic, middle and eastern Atlantic and Baltic Sea, and thus, the findings should be useful in devising overall management and conservation strategies for the species.

Thermal tolerance and thermal sensitivity of heart mitochondria: Mitochondrial integrity and ROS production

Cardiac mitochondrial metabolism provides 90% of the ATP necessary for the contractile exertion of the heart muscle. Mitochondria are therefore assumed to play a pivotal role in heart failure (HF), cardiovascular disease and ageing. Heat stress increases energy metabolism and oxygen demand in tissues throughout the body and imposes a major challenge on the heart, which is suspected of being the first organ to fail during heat stress. The underlying mechanisms inducing heart failure are still unclear. To pinpoint the processes implicated in HF during heat stress, we measured mitochondrial respiration rates and hydrogen peroxide production of isolated Arctic char (Salvelinus alpinus) heart mitochondria at 4 temperatures: 10°C (acclimation), 15°C, 20°C and 25°C (just over critical maximum). We found that at temperature ranges causing the loss of an organisms general homeostasis (between 20°C and 25°C) and with a substrate combination close to physiological conditions, the heat-induced increase in mitochondrial oxygen consumption levels off. More importantly, at the same state, hydrogen peroxide efflux increased by almost 50%. In addition, we found that individuals with low mitochondrial respiration rates produced more hydrogen peroxide at 10°C, 15°C and 20°C. This could indicate that individuals with cardiac mitochondria having a low respiratory capacity, have a more fragile heart and will be more prone to oxidative stress and HF, and less tolerant to temperature changes and other stressors. Our results show that, at temperatures close to the thermal limit, mitochondrial capacity is compromised and ROS production rates increase. This could potentially alter the performance of the cardiac muscle and lead to heat-induced HF underlining the important role that mitochondria play in setting thermal tolerance limits.