Temperature sex determination in the European sea bass,Dicentrarchus labrax (L., 1758) (Teleostei,Perciformes, Moronidae): critical sensitive ontogenetic phase

The temperature sex determination (TSD) mechanism in the European sea bass (Dicentrarchus labrax L.) was studied in respect to: a) the TSD sensitivity during the different developmental stages; and b) the intrapopulation correlation of sex determination with the growth rate up to the end of the TSD-sensitive period. At the stage of half-epiboly, eggs from the same batch were divided into four groups and subjected to different thermal treatments: a) 15 degrees C (G15 group) and b) 20 degrees C (G20 group) up to the middle of metamorphosis stage; c) 15 degrees C up to the end of yolk-sac larval stage and subsequently to 20 degrees C (G15-5 group); and d) 15 degrees C up to the end of the preflexion stage and then to 20 degrees C (G15-10 group). At the end of the treatments, size grading was applied and four additional populations were established from the upper (L) and lower (S) size portions of the G15 and G20 populations: G15L, G15S, G20L, and G20S. During the following growing phase, all populations were subjected to common rearing conditions. The sex ratios of each population were macroscopically determined at 190-210 mm mean total length. Female incidence was significantly affected (P < 0.05) by the different thermal treatments: 66.1% in the G15, 47.1% in the G15-10, 37.6% in the G15-5, and 18.1% in the G20 group. In addition, sex ratio was correlated with the growth rate of the fish up to the end of the TSD-sensitive period, with the larger fish presenting a significantly higher (P < 0.01) female incidence than the smaller fish in both thermal regimes tested: 73.1% in G15L vs. 57% in G15S, and 36.6% in G20L vs. 22.5% in G20S group. Results provide, for the first time, clear evidence that the sea bass is sensitive to TSD during all different ontogenetic stages up to metamorphosis, and that sex ratio is correlated with the growth rate of the fish well before the differentiation and maturation of the gonads.     

Rearing temperature enhances hepatic glucokinase but not glucose-6-phosphatase activities in European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) juveniles fed with the same level of glucose

The aim of this work was to elucidate if the previous results observed in hepatic glucokinase (GK) and glucose-6-phosphatase (G6Pase) activities in European sea bass and gilthead sea bream are due to temperature per se or to differences in feed intake at different water temperatures. For that purpose triplicate groups of fish (30 g initial body weight) were kept at 18 degrees C or 25 degrees C during two weeks and fed a fixed daily ration of a glucose-free or 20% glucose diet. At the end of the experimental period, plasma glucose levels in both species were not influenced by water temperature but were higher in fish fed the glucose diet. Higher hepatic GK activity was observed in the two fish species fed the glucose diet than the glucose-free diet. In the glucose fed groups, GK activity was higher at 25 degrees C than at 18 degrees C. Glucose-6-phosphatase activities in both species were not influenced by water temperature. In European sea bass and in contrast to gilthead sea bream it was observed an effect of dietary composition on G6Pase activities with surprising higher activities recorded in fish fed the glucose diet than in fish fed the glucose-free diet. Overall, our data strongly suggest that European sea bass and gilthead sea bream are apparently capable to strongly regulate glucose uptake by the liver but not glucose synthesis, which is even enhanced by dietary glucose in European sea bass. Within limits, increasing water temperature enhances liver GK but not G6Pase activities, suggesting that both species are more able to use dietary carbohydrates at higher rearing temperatures.     

Rapid metabolic adaptation in European sea bass (Dicentrarchus labrax) juveniles fed different carbohydrate sources after heat shock stress

A study was conducted to evaluate the effect of two dietary carbohydrate sources (waxy maize starch and glucose) on the metabolic adaptation of sea bass juveniles (initial weight: 24 g) to a heat shock treatment (temperature rise from 18 degrees C to 25 degrees C within 24 h). Two isonitrogenous and isolipidic diets were formulated to contain 20% waxy maize starch (WS diet) or 20% glucose (GLU diet). Triplicate groups of fish were fed to near satiation for 4 weeks at both temperatures (18 degrees C and 25 degrees C). Then, fish previously maintained at 18 degrees C were submitted to a heat shock (18 degrees C to 25 degrees C) and continued to be fed with the same diets during 1 more week. The higher water temperature significantly improved growth performance, feed efficiency, as well as protein efficiency ratio, independently of diet. At 25 degrees C, but not at 18 degrees C, growth of fish fed the WS diet was higher than that of fish fed the GLU diet. Plasma glucose levels were higher in sea bass fed the GLU diet and not influenced by water temperature. Fish fed a glucose diet or reared at high temperatures (25 degrees C) showed enhanced liver glycolytic, lipogenic and gluconeogenic capacities compared to fish fed a starch diet or reared at low temperatures (18 degrees C). For the majority of the enzymes studied, 1 week seemed to be enough time for metabolic adaptation in sea bass submitted to an acute heat shock. Irrespective of carbohydrate source, HSP70 gene expression was similar in both cold water (18 degrees C) and warm water (25 degrees C) acclimated sea bass. A weak down regulation was observed after heat shock only in fish fed the GLU diet. This suggests that HSP70 gene expression is not affected by the rearing temperature per se.     

Gonadal sex differentiation and effect of rearing temperature on sex ratio in black rockfish (Sebastes schlegeli)

Temperature-dependent sex determination has been demonstrated in some species of fish, and a high temperature during the period of sex differentiation typically produces a male-dominant population. This research investigated the gonadal sex differentiation and effect of rearing temperature on the sex ratio in larval black rockfish Sebastes schlegeli, which is a viviparous species. Two types of gonads were histologically distinguishable in fish 20 mm in total length (TL). The putative ovary started forming an ovarian cavity, while the putative testis was not clearly differentiated until 51 mm TL. In a temperature-controlled experiment, the proportions of females were 45% at 10°C, 46% at 14°C, 50% at 18°C, 63% at 22°C, and 83% (significantly different from 1:1 sex ratio) at 24°C. These results suggest that morphological sex differentiation in black rockfish occurs at approximately 20 mm in TL, and it is possible that high temperatures (24°C) induce not a male- but a female-dominant population in this species.     

Temperature effects and genotype-temperature interactions on sex determination in the European sea bass (Dicentrarchus labrax L.)

The effect of temperature on sex-ratios in 27 families of sea bass reared in the same tank from the fertilization stage onward was investigated. An excess of males (68%) was found in the groups that were reared at high temperature (mean +/- standard deviation: 20+/-1 degrees C) until they reached the mean size of 8.1 cm (Standard Length, 149 days post-fertilization [p.f.]). Masculinization was higher (89% of males) in the groups maintained at low temperature (13 degrees C), from fertilization to a mean length of 6.5 cm (346 days p.f.). Shifts from high to low temperature at 8.1cm and from low to high temperature at 6.5 cm had no consequence on the sex-ratio. The percentage of males showing intratesticular oocytes was higher at low temperature (63%) than at high temperature (36%), suggesting that these males may be sensitive fish that have been masculinized by environmental factors. Fish sampled in the groups reared at high (2,200 fish) and low (500 fish) temperature were genotyped on three microsatellite loci. This allowed them to be assigned to the breeders used in the crossing design, thus permitting an analysis of parental influence on sex-ratios. In groups reared at high temperature, both parents had a significant additive effect on the percentage of females, and the interaction between sire and dam was not significant. Genotype temperature interactions were also detected and their existence suggests the interesting possibility of selecting nonsensitive genotypes in breeding programs.     

Temperature effects on cranial deformities in European sea bass, Dicentrarchus labrax (L.)

The present study investigates the effect of water temperature on the development of deformities during embryonic and larval stages of European sea bass, Dicentrarchus labrax (L.). Two temperature conditions were examined in duplicate by studying the presence of skeletal deformities in a sample of 45 to 51 fish taken from each population at the end of the rearing trials [20–23 mm total length (TL)]. The results indicated that water temperature during the embryonic and larval phase has significant effects on deformation of the branchiostegal rays (P < 0.001), but not of the mouth and the fins (P > 0.05). At 15°C, 27.2–33.4% of the examined fish had branchiostegal rays of abnormal shape and/or orientation, whereas at 20°C this deformity had a frequency of only 4.0–4.1%. The frequency distribution graph of branchiostegal counts demonstrated a significant deviation in the deformed fish from the normal (seven rays on each side of the body) phenotype at both temperatures tested. This deviation was mainly expressed as a lack of one to four rays (56.7% of deformed fish), or the formation of one extra ray (26.7% of deformed fish) (P < 0.001, G‐test). The results are discussed in respect to the possible mechanisms of temperature effects on the development of skeletal deformities.     

Hepatic glucokinase and glucose-6-phosphatase responses to dietary glucose and starch in gilthead sea bream (Sparus aurata) juveniles reared at two temperatures

The effects of carbohydrate sources/complexity and rearing temperature on hepatic glucokinase (GK) and glucose-6-phosphatase (G6Pase) activities and gene expression were studied in gilthead sea bream juveniles. Two isonitrogenous (50% crude protein) and isolipidic (19% crude lipids) diets were formulated to contain 20% waxy maize starch or 20% glucose. Triplicate groups of fish (63.5 g initial body weight) were fed each diet to near satiation during four weeks at 18 degrees C or 25 degrees C. Growth, feed intake, feed efficiency and protein efficiency ratio, were higher at the higher water temperature. At each water temperatures fish growth and feed efficiency were higher with the glucose diet. Plasma glucose levels were not influenced by water temperature but were higher in fish fed the glucose diet. Hepatosomatic index and liver glycogen were higher at the lower water temperature and within each water temperature in fish fed the glucose diet. No effect of water temperature on enzymes activities was observed, except for hexokinase and GK which were higher at 25 degrees C. Hepatic hexokinase and pyruvate kinase activities were not influenced by diet composition, whereas glucose-6-phosphate dehydrogenase activity was higher in fish fed the glucose diet. Higher GK activity was observed in fish fed the glucose diet. GK gene expression was higher at 25 degrees C in fish fed the waxy maize starch diet while in fish fed the glucose diet, no temperature effect on GK gene expression was observed. Hepatic G6Pase activities and gene expression were neither influenced by dietary carbohydrates nor water temperature. Overall, our data suggest that in gilthead sea bream juveniles hepatocytes dietary carbohydrate source and temperature affect more intensively GK, the enzyme responsible for the first step of glucose uptake, than G6Pase the enzyme involved in the last step of glucose hepatic release.     

Hepatic glucokinase and glucose-6-phosphatase responses to dietary glucose and starch in gilthead sea bream (Sparus aurata) juveniles reared at two temperatures.

The effects of carbohydrate sources/complexity and rearing temperature on hepatic glucokinase (GK) and glucose-6-phosphatase (G6Pase) activities and gene expression were studied in gilthead sea bream juveniles. Two isonitrogenous (50% crude protein) and isolipidic (19% crude lipids) diets were formulated to contain 20% waxy maize starch or 20% glucose. Triplicate groups of fish (63.5 g initial body weight) were fed each diet to near satiation during four weeks at 18 degrees C or 25 degrees C. Growth, feed intake, feed efficiency and protein efficiency ratio, were higher at the higher water temperature. At each water temperatures fish growth and feed efficiency were higher with the glucose diet. Plasma glucose levels were not influenced by water temperature but were higher in fish fed the glucose diet. Hepatosomatic index and liver glycogen were higher at the lower water temperature and within each water temperature in fish fed the glucose diet. No effect of water temperature on enzymes activities was observed, except for hexokinase and GK which were higher at 25 degrees C. Hepatic hexokinase and pyruvate kinase activities were not influenced by diet composition, whereas glucose-6-phosphate dehydrogenase activity was higher in fish fed the glucose diet. Higher GK activity was observed in fish fed the glucose diet. GK gene expression was higher at 25 degrees C in fish fed the waxy maize starch diet while in fish fed the glucose diet, no temperature effect on GK gene expression was observed. Hepatic G6Pase activities and gene expression were neither influenced by dietary carbohydrates nor water temperature. Overall, our data suggest that in gilthead sea bream juveniles hepatocytes dietary carbohydrate source and temperature affect more intensively GK, the enzyme responsible for the first step of glucose uptake, than G6Pase the enzyme involved in the last step of glucose hepatic release.     

Toxicity and pathological effects of orally and intraperitoneally administered ivermectin on sea bass Dicentrarchus labrax

The toxicity and histopathology of ivermectin was studied in 3 and 35 g sea bass Dicentrarchus labrax L. following in-feed, oral intubation and injection administration at dose rates ranging from 0.5 to 3.5 mg kg(-1). Estimated LD50 values for 3 g fish were 0.335 and 0.106 mg kg(-1) following oral intubation and injection administration respectively, for fish reared at 11 degrees C; and 0.839 and 1.023 mg kg(-1) following oral intubation and injection administration, respectively for fish reared at 20 degrees C. For 35 g fish reared at 11 degrees C, the estimated LD50 was 0.523 and 0.361 mg kg(-1) following oral intubation and injection administration respectively. No signs of toxicity were observed when the compound was administered via the feed at 0.5 and 0.7 mg kg(-1). However, toxicity (> 10%) was observed at dose rates of 0.2 mg kg(-1) and higher when the compound was administered via oral intubation and at 0.5 mg kg(-1) when administered via injection. The compound was significantly more toxic to fish reared at 11 degrees C than at 20 degrees C. Further, ivermectin was more toxic to 3 g than to 35 g sea bass when administered via injection. Histopathological examination of the major organs revealed pathology was largely restricted to gills and intestinal tissue. In 3 g sea bass, lesions were also found in the kidneys.     

Saline preferendum for the European sea bass, Dicentrarchus labrax, larvae and juveniles: effect of salinity on early development and sex determination

The sea bass, Dicentrarchus labrax, is an eurhyaline marine fish. Juveniles of this species are known to frequent estuaries and lagoons where salinity is lower than in the open sea. Sex determination occurs during this phase of fish life and has been shown labile and sensitive to environmental factors. In this work, the effect of rearing salinity on sex determination and early development of the sea bass was investigated. An excess of males (87%) was found in all groups and the salinity level [(natural sea water salinity, mean: 37‰) vs. (15‰)], when maintained constant, had no effect on the sex-ratio. The transfer from low to high salinity at 93 days post-fertilization (p.f.) increased the percentage of males (93%) suggesting that sexual differentiation in this species may be influenced by such an osmotic stress. Growth was improved by a 15‰ salinity at the beginning of larval rearing (14 days p.f.) and at the end of pre-growing (234-458 days p.f.), periods during which low temperatures were applied. Survival during larval rearing and nursery were also improved in the groups reared at low salinity and so was swimbladder inflation. These results show that sea bass juveniles have a low saline preferendum, a finding that corresponds to the conditions they actually encounter in the wild during their juvenile ecophase; sex determination is not directly modulated by the salinity level but seems to be subjected to complex environmental regulations.     

Spontaneous haemolytic activity of Atlantic halibut (Hippoglossus hippoglossus L.) and sea bass (Dicentrarchus labrax) serum

The spontaneous haemolytic (SH) activity of sera was compared in groups of cultured halibut and sea bass. The optimum assay temperature was determined for each species and different red blood cell donors were tested. The effects of heat inactivation, storage temperature and of different agents like EDTA, EGTA, yeast cell components and bacterial LPS were compared. Halibut sera gave optimum lysis with sheep red blood cells (RBC) at 16 degrees C whereas sea bass sera showed optimum lysis with rabbit RBC at 37 degrees C. The haemolytic activity of halibut sera was inactivated at 45 degrees C while sea bass sera were inactivated at 56 degrees C. The haemolytic activity of halibut sera was significantly reduced during short-term storage at -80 degrees C, whereas the sea bass sera maintained fairly good activity after 1-year storage at -80 degrees C. EGTA and EDTA inhibited the spontaneous haemolytic activity of sera from both the species. Zymosan and MacroGard from yeast cells also inhibited the haemolytic activity of the sera of both species, whereas LPS had a very slight effect. Considerable variation in haemolytic activity was observed within both the halibut and sea bass groups studied.     

Temperature-dependency of Betanodavirus infection in SSN-1 cell line

This study examined the in vitro effects of temperature on Betanodavirus infection in the SSN-1 cell line. A Betanodavirus isolated from moribund sea bass fry Dicentrarchus labrax farmed in the Adriatic Sea and characterised as a RGNNV (Redspotted Grouper Nervous Necrosis Virus) genotype was used. Virus-infected SSN-1 cells were incubated at temperatures between 10 and 30 degrees C and observed for cytopathic effects daily for 15 d. Cell-free and cell-associated viral growth were evaluated by 50% tissue culture infectious dose (TCID50) titration at 0, 24, 48, 72, 96, 144, 192, 240, 312 and 360 h post-infection. Virus replication was observed at all temperatures from 15 to 30 degrees C. The optimal temperature for virus growth was 25 degrees C. A temperature of 10 degrees C was detrimental to the growth of the SSN-1 cells and cell death interfered with interpretations of viral growth. The isolate of Betanodavirus from Italian sea bass in this study demonstrates a different temperature range for growth compared to previous reports for related Betanodavirus strains, most likely due to an adaptation to the normal environmental temperatures of the host fish species of origin.     

Low temperature has opposite effects on sex determination in a marine fish at the larval/postlarval and juvenile stages

Temperature‐dependent sex determination (TSD) can be observed in multiple reptile and fish species. It is adaptive when varying environmental conditions advantage either males or females. A good knowledge of the thermosensitive period is key to understand how environmental changes may lead to changes in population sex ratio. Here, by manipulating temperature during development, we confirm that cold temperature (16°C) increases the proportion of fish that develop as females in European sea bass (Dicentrarchus labrax) until 56 days posthatching, but show that it has an opposite effect at later stages, with the proportion of males reaching ~90% after 230 days at 16°C. This is the first observation of opposite effects of temperature at different time periods on the sex ratio of a vertebrate. Our results highlight the potential complexity of environmental effects on sex determination.     

Effect of temperature on the development and viability of Gryon gallardoi (Brethes) (Hymenoptera: Scelionidae) parasitizing Spartocera dentiventris (Berg) (Hemiptera: Coreidae) eggs.

The development and viability of Gryon gallardoi (Brethes) (Hym.: Scelionidae) in Spartocera dentiventris (Berg) (Hem.: Coreidae) eggs were studied under four temperatures: 15, 20, 25, and 30 +/- 1 degree C, with a 12-h photophase. No parasitoid developed at 15 degrees C. Otherwise, viability reached 98.8% without varying significantly over the temperature range tested. The duration of development for males and females was inversely proportional to the temperature increase, varying respectively from 46.2 +/- 0.13 and 47.1 +/- 0.11 days (20 degrees C) to 13.3 +/- 0.07 and 13.4 +/- 0.06 days (30 degrees C). Males developed faster than females. The values estimated for the lowest thermic thresholds of development and the thermic constants were 15.5 degrees C and 185.19 DD for males and 15.6 degrees C and 192.31 DD for females, respectively. Given the average weather conditions in Porto Alegre, RS (30 degrees 01' S and 51 degrees 13' W), Brazil, G. gallardoi could annually produce 8.54 and 8.07 generations of males and females, respectively. The low rates of parasitism observed in the field during the first generation of its host may be due to the small number of G. gallardoi generations in this period.     

Induced sex change in black sea bass

Experiments were conducted to identify factors involved in sex change in the protogynous black sea bass Centropristis striata . Black sea bass maintained in the ratio of 8 females (F):0 males (M) for 9 months reversed sex while those kept at the ratios of 6F:2M or 4F:4M did not. Female black sea bass implanted with 1·0 mg 11-ketotestosterone (11-KT) or 10 mg fadrozole (FAD) changed sex and began spermiating while those implanted with 0·1 mg 11-KT or 1·0 mg FAD underwent incomplete sex reversal. One fish implanted with 1·0 mg FAD initiated sex change but was not spermiating at the end of the study. One fish in the control group, the largest fish in the study, initiated sex change. These results suggest that the presence of males may restrict sex reversal in black sea bass and that high 17β-oestradiol:11-KT is required for maintaining ovarian function.     

Sex ratios in offspring of sex-reversed sea bass and the relationship between growth and phenotypic sex differentiation

Groups of sexually undifferentiated sea bass Dicentrarchus labrax were fed with the androgen 17α-methyltestosterone (MT) during sex differentiation. MT treatment increased males from 79±3% in the controls (the usual 3:1 male:female sex ratio of cultured sea bass) to 100±0%, implying that in the treated groups one out of each five resulting males was a masculinized female (neomale). Thirteen males from the MT treated groups were taken as the parental generation and their sperm used to individually fertilize a pool of eggs from unrelated females. The probability of having at least one neomale was 95% and most probably two or three of the males used were neomales. The offspring from each family were reared separately under the same environmental conditions. Samples were taken at 11 and 15 months of age, during and after sex differentiation, respectively. Results showed that females predominated among the larger fish whereas males and undifferentiated fish predominated among the smaller ones. Intersexes exhibited an intermediate size. All fish with a body length smaller than 12 cm were undifferentiated. These results suggest that sex differentiation is more dependent on length than on age. At 15 months, sex ratios were male-biased in all families, except one (females ranged from 5 to 50%) and only two families had sex ratios not significantly different from 1:1, suggesting that the mechanism of sex determination in the sea bass is not of a XX/XY or ZW/ZZ type since no family exhibited a female-biased progeny, as would be expected from both types. Results support the hypothesis that factors other than genetic, i.e., environmental, may act epigenetically on the sex determination mechanisms of sea bass, as has been demonstrated in other fishes.     

Temperature affects physiological stress responses to acute confinement in sunshine bass (Morone chrysops x Morone saxatilis)

Sunshine bass (Morone chrysopsxMorone saxatilis) were subjected to a 15-min low-water confinement stressor at temperatures ranging from 5 to 30 degrees C. Physiological responses were evaluated by measuring hematocrit, and plasma chloride, glucose and cortisol. Fish acclimated to 30 degrees C had initial glucose concentrations of 3.13 mM (564 mg/L) which were significantly lower than in fish acclimated to 5 and 10 degrees C (4.32 and 4.82 mM or 779 and 868 mg/l, respectively). Fish survived the conditions imposed at every temperature except 30 degrees C, where 15 out of 42 fish died during the stress and recovery protocol. The general pattern was an initial increase in hematocrit, followed by a delayed decrease in hematocrit and chloride, and an increase in plasma glucose and cortisol. In general, fish stressed at temperatures below 20 degrees C had lower and more delayed changes in plasma glucose and cortisol than fish tested at 20, 25 and 30 degrees C. Initial cortisol concentrations were 65 ng/ml and increased to above 200 ng/ml in fish held at 20 degrees C and above. At the higher temperatures, glucose concentrations were twice the initial concentration after stress and cortisol changes were four to five times the initial concentration after the stress. Quantitative responses for glucose and cortisol were moderate and recovery rapid in fish stressed at 10 and 15 degrees C; therefore, this range of water temperature is recommended when handling sunshine bass.     

Exposure of larvae to daily thermocycles affects gonad development, sex ratio, and sexual steroids in Solea senegalensis, kaup

The effect of water temperature during the development of fish larvae on sex differentiation is well known, but not so well known is the impact of the daily thermocycles. Our aim was to investigate the effect of early exposure of Senegal sole larvae to different temperature cycles on gonad development, sex ratio, and sex steroid (11-ketotestosterone (11-KT); estradiol (E(2) ); and testosterone, (T)) content in muscle extracts of juveniles. From 1 to 97 days posthatching (DPH) fish larvae and post-larvae were subjected to three temperature regimes: Thermophase-Cryophase (TC), Cryophase-Thermophase (CT), and constant temperature. In fish exposed to TC, sex determination occurred earlier, because 90% of soles were males/females at 110 DPH, whereas 45% of fish under CT were undifferentiated at that time. Fish under TC showed the highest growth rates, followed by fish under constant temperature and by fish under CT, the differences being statistically significant between the TC and CT groups. Regarding sex ratio, juveniles exposed to TC showed a higher proportion of females than fish under CT or constant temperature. Under TC, fish showed the highest concentration of E(2) , whereas 11-KT concentration was highest in fish under CT and constant temperature. Fish under constant temperature and CT showed higher T levels than those under TC. These results provide the first insights into the effect of daily thermocycles on sex differentiation in fish, and underline the key role of natural environmental cycles on the control of sex ratios during larval development, which may be applied to the manipulation of sex ratio in aquaculture.     

A retrospective approach to fractionize variation in body mass of Atlantic cod Gadus morhua

Eggs of a single spawning batch from wild-caught Norwegian Atlantic cod Gadus morhua were hatched and first fed on either natural zooplankton or enriched rotifers Brachionus plicatilis during the larval period. Juvenile G. morhua (initial mass 14·2 g) from the two first-feeding groups were then reared for 3 months under a variety of temperature (10 and 14° C) and salinity (15 and 32) combinations. All fish were individually tagged and microsatellite markers were used in a multiplex to trace the pedigree of all fish and body mass variation analysed according to different environmental and genetic sources. After the termination of the laboratory trial, the fish were transferred to land-based tanks and later to sea pens and reared at ambient conditions for 26 months until they were harvested in March 2009. Growth gain from the larval and juvenile periods was persistent during the 26 months of sea pen ongrowing. The final mass of the zooplankton group was 12% higher compared to the B. plicatilis group. Similarly, rearing under a temperature of 14° C and salinity of 15 during the initial 3 month period during the early juvenile stage resulted in 7-13% larger size at harvesting compared to the other three temperature and salinity combinations. The study indicates that the first-feeding method and temperature and salinity manipulation explain nearly 90% of the body mass variation explained by the model. The genetic effect (measured as body mass variation within the families studied) only accounted for c. 2% during the initial rearing period, whereas it has a large effect on growth variation (30%) during the long-term rearing at ambient conditions. Sex proportion and final maturation did not differ between family groups, and no interaction between sex and family group was seen.