Distribution of canthaxanthin in immature rainbow trout Oncorhynchus mykiss serum

• 1.1. The present study was undertaken in order to define the distribution of canthaxanthin between the lipoprotein fractions in serum of immature rainbow trout fed a diet supplemented with synthetic canthaxanthin (80 mg/kg).
• 2.2. Lipoproteins were separated by density-gradient ultracentrifugation.
• 3.3. Canthaxanthin was found in all lipoprotein fractions, in different amounts according to the density of the lipoprotein fraction: VLDL, 13.9%; LDL, 15.2% or LDL, 29.1% since the density of the first fraction was 1.006 g/ml; HDL, 60.4% and VHDL, 10.5%.

     

Antibody structural variation in rainbow trout fluids

Rainbow trout (Oncorhynchus mykiss) were immunized with trinitrophenylated-keyhole limpet hemocyanin (TNP-KLH) and the redox structure of induced anti-TNP antibodies from the serum, mucus, egg and ovarian fluid was examined. In conducting these studies it was determined that all TNP-specific antibody from each source possessed the mAb-specific H chain (1-14) epitopes, which facilitated the direct structural analysis of the induced antibodies. A protocol was developed which ensured complete adsorption of all specific anti-TNP antibody from each fluid. Together these protocols permitted the unbiased compositional analysis of all redox forms of the anti-TNP antibodies from each source. All antibodies, regardless of source, possessed the same molecular mass, characteristic of the trout tetramer (800 kDa). It was found that specific antibody titers were significantly higher in male than female trout, while the degree of disulfide polymerization was relatively invariant in male antibodies, while being highly variable in female antibodies. Within the females, no distinctively different redox ratios were between antibodies isolated from sera, ovarian fluid or eggs: however, mucus antibodies possessed a unique redox structure consisting of halfmeric constituents that were not observed in antibodies from other fluids.     

Genetic (co)variation in skin pigmentation patterns and growth in rainbow trout

From a physiological-behavioral perspective, it has been shown that fish with a higher density of black eumelanin spots are more dominant, less sensitive to stress, have higher feed intake, better feed efficiency and therefore are larger in size. Thus, we hypothesized that genetic (co)variation between skin pigmentation patterns and growth exists and it is advantageous in rainbow trout. The objective of this study was to determine the genetic relationships between skin pigmentation patterns and BW in a breeding population of rainbow trout. We performed a genetic analysis of pigmentation traits including dorsal color (DC), lateral band (LB) intensity, amount of spotting above (SA) and below (SB) the lateral line, and BW at harvest (HW). Variance components were estimated using a multi-trait linear animal model fitted by restricted maximum likelihood. Estimated heritabilities were 0.08±0.02, 0.17±0.03, 0.44±0.04, 0.17±0.04 and 0.23±0.04 for DC, LB, SA, SB and HW, respectively. Genetic correlations between HW and skin color traits were 0.42±0.13, 0.32±0.14 and 0.25±0.11 for LB, SA and SB, respectively. These results indicate positive, but low to moderate genetic relationships between the amount of spotting and BW in rainbow trout. Thus, higher levels of spotting are genetically associated with better growth performance in this population.     

Genetic analysis of androgenetic rainbow trout.

We analyzed a number of genetic characteristics in androgenetic rainbow trout (Oncorhynchus mykiss) and their progeny. The androgenetic progeny of individual androgenetic males appeared genetically identical to each other based on eight enzyme loci. Their viability was no higher than that of androgenetic progeny of outbred males. Homozygous androgenetic female rainbow trout produced very poor quality eggs. When common eggs and sperm from outbred individuals were used to produce androgenetic and gynogenetic progeny, the yield of gynogenetic progeny was higher but some were heterozygous at protein loci, while no androgenetic progeny were heterozygous. Some androgenetic diploid rainbow trout were successfully produced from cryopreserved sperm. The progeny of some androgenetic males crossed to normal females were virtually all males, while the progeny of other males were virtually all females. This suggests that both XX and YY androgenetic individuals may develop as males. Androgenesis is likely to be useful for generating homozygous clones for research and for recovering strains from cryopreserved sperm.     

Genetic variation in the nine rainbow trout (Oncorhynchus mykiss) populations in South Africa

• 1.1. Twenty-six protein coding loci in nine populations of rainbow trout, from different localities in South Africa, were analyzed by starch and polyacrylamide gel electrophoresis to estimate the amount of genetic diversity present.
• 2.2. Heterozygosity ranged between 0.0445 and 0.0745.
• 3.3. The relatively high heterozygosity levels are probably due to the historical mixing of strains, and the differences in heterozygosity indicate that the populations originate from different sources, each with different genetical characteristics.
• 4.4. Differences in phenotypic characters (i.e. growth) seem to relate to genotypic variation, whilst food conversion rate and survival performance results did not seem to relate to genotypic variation.

     

Response of serum carotenoid levels to dietary astaxanthin and canthaxanthin in immature rainbow trout Oncorhynchus mykiss

Rainbow trout were fed a diet supplemented with astaxanthin (89 mg/kg) or canthaxanthin (116 mg/kg) in two different experiments: experiment 1 was designed to measure the kinetics of the appearance and disappearance of carotenoids in the serum; experiment 2 was undertaken to establish the serum dose-response to synthetic astaxanthin and canthaxanthin for immature rainbow trout. The serum carotenoid concentrations of immature rainbow trout increased when fish were fed carotenoid supplemented feed and then reached a plateau after 1 day of intake for astaxanthin and after 2 days for canthaxanthin. Circulating astaxanthin represented a value 2.3 times that of canthaxanthin. After dietary supplementation was discontinued, the serum carotenoid concentrations decreased within 3 days for both carotenoids. The average decreasing slopes for the two carotenoid pigments were parallel, indicating a similarity in the rate of which astaxanthin and canthaxanthin are utilized by rainbow trout. The serum dose-response of trout that received dietary keto-carotenoids increased with increasing pigment levels. The hypothesis that absorption of dietary carotenoids in 12.5–200 mg/kg range of concentration across the gut wall may be by passive diffusion is proposed.     

Cold storage effects on flesh quality of rainbow trout Oncorhynchus mykiss (Walbaum, 1792) fed diets containing different vegetable oils

The intention of this experiment was to assess the effects of different sources of dietary lipid on the fatty acid composition of the fillet and liver and the flesh quality traits of rainbow trout (Oncorhynchus mykiss) after a 70‐day feeding period. Four iso‐nitrogenous (approx. 51% crude protein) and iso‐lipidic (approx. 14% crude lipid) experimental diets were formulated. The control diet contained only fish oil (FO) as the primary lipid source. In the other three dietary treatments, fish oil was replaced by 100% (LO30/SO35/SFO35) and 70% (FO30/LO35/SO35 or FO30/SO35/SFO35) sesame oil (SO), linseed oil (LO), or sunflower oil (SFO). Triplicate groups of 40 rainbow trout (~46 g) held under similar culture conditions were hand‐fed daily to apparent satiation for 70 days. At the end of the feeding trials, no difference in growth performance among experimental groups was noted (P > 0.05). There were some differences in the proximate composition of fish fillets (P < 0.05): the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels were highest in fish fed the control diet (P < 0.05); and EPA and DHA levels in fish fed the FO30/LO35/SO35 diet were closest to the control diet (P < 0.05). In contrast, fish fed the diet containing 100% plant oils (LO30/SO35/SFO35) had the highest level of total n‐6 fatty acids in the fillet and liver. In a 12‐day refrigerated storage at 1°C the thiobarbituric acid (TBA), trimethylamin nitrogen (TMA‐N) and pH values gradually increased in all dietary groups (P < 0.05). The chemical indicators of spoilage, TBA, TMA‐N, and pH values were within the limit of acceptability for human consumption.     

Control of skeletal muscle fibres and adipose cells size in the flesh of rainbow trout

The distributions of the diameters of skeletal muscle fibres and adipocytes were studied in rainbow trout. The cellularity of perivisceral adipose tissues and subcutaneous ventral and dorsal adipose tissues were characterized more specifically. In these tissues, a population of small adipocytes was distinguishable from larger adipocytes. The same was observed in white muscle. The effects of extrinsic factors (dietary lipid in two different thermal conditions) and intrinsic factors (strains in two different saline conditions, growth hormone) on the long-term response of the cellularity of both muscle and adipose tissues were studied. The effects of thermal environment were tested on fish fed the same ration and the effects of saline environment on fish fed ad libitum. The mean size of white muscle fibres was relatively unaffected by the different treatments tested: genetic origin and dietary lipid in different environmental conditions. There were significant differences in growth rate due to genetic origin and saline environment. The possible involvement of hyperplasia in response to these different factors is discussed. Growth hormone supplementation enhanced the percentage of small diameter fibres indicating a role of this hormone in the control of muscle hyperplastic growth. The mean size of adipose cells was affected only slightly by the different treatments tested. An increase in adipose cell size with aging and lipid content was observed. The percentage of small adipocytes also increased with aging. Thus, it is proposed that the development of adipose tissues, and thus fat retention, both result from the recruitment of new adipocytes and from the increase in size of existing adipocytes. The hyperplastic process contributed significantly to the differences in fat retention due to different treatments tested (strains, thermal and saline environments). When partially substituting fish oils for corn oils in the diet, a large increase in the ventral adipose cell size was seen indicating a potential negative effect of n-6 fatty acids on cell proliferation. Growth hormone treatment, on the contrary, induced a decrease in the size of perivisceral adipocytes. Thus, diet and hormonal status affect adipose cells size through two different metabolic pathways: lipogenesis and lipolysis respectively.     

Diel variation in branchial calcium uptake by the rainbow trout

Branchial Ca uptake varied dielly with a nadir at 1000 hours and two peaks at 1600 and 0400 hours in rainbow trout. This variation profile essentially reflected Ca uptake through the arterio-arterial circulation.     

Behavioural variation between piscivore and insectivore rainbow trout Oncorhynchus mykiss

A proactive-reactive continuum integrating multiple (i.e., 3+) dimensions of animal behaviour has been reported as a major axis of behavioural differentiation, but its stability along a biological hierarchy from individuals to populations remains speculative. Piscivore and insectivore rainbow trout (Oncorhynchus mykiss) represent closely related ecotypes with strong ecological divergence driven by selection for a large-bodied piscivorous lifestyle with fast juvenile growth vs. selection for smaller adult body size and lower growth associated with an insectivorous diet. To evaluate whether differences in behaviour between ecotypes are consistent with a proactive-reactive axis and consistent along a biological hierarchy, the authors examined variation in emergence time from a shelter, exploration, activity and predator inspection among individuals, populations and ecotypes of juvenile piscivore and insectivore rainbow trout O. mykiss. As expected, the faster-growing piscivore ecotype was more proactive (i.e., shorter emergence time, exploration and predator inspection) than the more reactive insectivore ecotype. This behavioural contrast was partly maintained across populations, although activity differences were most pronounced among populations, rather than emergence time. Insectivore fry showed substantial variation in behavioural expression among individuals within populations; by contrast, piscivores showed highly similar proactive behaviours with significantly lower inter-individual variation in behavioural expression, suggesting intense selection on behaviour supporting their faster growth. This work suggests that piscivore and insectivore O. mykiss broadly differ in behaviour along a proactive vs. reactive continuum, and highlights the greater multidimensionality of behavioural expression within the insectivore ecotype. Contrasting behaviours between ecotypes may result from differential selection for slow vs. fast juvenile growth and associated metabolism, and may contribute to adult trophic specialization.     

Effects of ambient colour on colour preference and growth of juvenile rainbow trout Oncorhynchus mykiss (Walbaum)

Colour preference of individual juvenile rainbow trout Oncorhynchus mykiss was tested at 1 and 12° C, and also at 12° C after a 42 day growth experiment under white, blue, green, yellow or red ambient colour. All experiments were carried out under controlled laboratory conditions and the preference was assessed by the location of the fish in a preference tank with four chambers. Rainbow trout showed a preference for blue and green at 1° C and for green at 12° C. After the growth experiment the fish reared in blue tanks preferred blue and green but green was the most preferred colour for the fish reared in green, yellow and red tanks. Yellow and especially red chambers were avoided, irrespective of the ambient colour during the growth trial. The final mass of fish reared in the red aquaria was significantly smaller than that of the fish in green tanks. In addition, when the data of the preference tests were correlated with the data of the growth experiment using mean values of the four tested colours, a very good linear relationship was observed between the preference ( i.e. visit frequency in coloured compartments) and growth rate as well as food intake. When considering the results both from the preference and growth trials it is suggested that green is the best environmental colour for rearing juvenile rainbow trout while rearing in a red environment cannot be recommended.     

Alterations in the synthesis of proteins in hepatocytes of rainbow trout fed cyclopropenoid fatty acids

Hepatocytes from rainbow trout reared on a diet containing cyclopropenoid fatty acids were analyzed for alterations in protein composition and synthesis by double label experiments. Both cytosolic and microsomal hepatocyte fractions were investigated. In the cytosolic fraction, the synthesis of proteins in the range of 68,000 to 74,000 daltons were significantly decreased. The identity of these proteins remains uncertain. A pronounced depression in both the mass and apparent synthesis of a 200,000 to 240,000 dalton microsomal protein was also observed. Immunoblotting with antibodies raised against goose acetyl-CoA carboxylase and avidin-peroxidase staining suggest that this protein is acetyl-CoA carboxylase. Moreover, synthesis of this protein as well as mass of the protein in cyclopropenoid fatty acid-fed fish were less than 20% of that found in control fish.     

Tissue astaxanthin and canthaxanthin distribution in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar)

A comparative investigation of tissue carotenoid distribution between rainbow trout, Oncorhynchus mykiss, and Atlantic salmon, Salmo salar, was undertaken to identify the relative efficiency of utilization of astaxanthin and canthaxanthin. Higher apparent digestibility coefficients (ADCs) (96% in trout vs. 28-31% in salmon; P<0.05), and pigment retention efficiencies (11.5-12.5% in trout vs. 5.5% in salmon; P<0.05), for both astaxanthin and canthaxanthin, were observed for rainbow trout. Astaxanthin deposition was higher than canthaxanthin in rainbow trout, while the reverse was true for Atlantic salmon, suggesting species-specificity in carotenoid utilization. The white muscle (95% in trout vs. 93% in salmon) and kidneys (0.5% in trout vs. 0.2% in salmon) represented higher proportions of the total body carotenoid pool in rainbow trout than in Atlantic salmon (P<0.05), whereas the liver was a more important storage organ in Atlantic salmon (2-6% in salmon vs. 0.2% in trout; P<0.05). The liver and kidney appeared to be important sites of carotenoid catabolism based on the relative proportion of the peak chromatogram of the fed carotenoid in both species, with the pyloric caecae and hind gut being more important in Atlantic salmon than in the rainbow trout. Liver catabolism is suspected to be a critical determinant in carotenoid clearance, with higher catabolism expected in Atlantic salmon than in rainbow trout.     

Genetic variability and components of fitness in hatchery strains of rainbow trout

Two fitness components, development rate and egg size, were examined in six hatchery strains of rainbow trout, Oncorhynchus mykiss (syn. Salmo gairdneri Richardson), with different amounts of enzyme heterozygosity. The average expected heterozygosities per strain ranged from 4 to 8%, based upon an electrophoretic analysis of the protein products of 42 loci. Strains with higher heterozygosities had faster development rates, as measured by hatching time, than strains with lower heterozygosity. Concordance between hatching time and another measure of development rate, degree of yolk-sac resorption, suggests that hatching time is a valid measure of embryonic development rate in salmonid fishes. Earlier-hatching embryos were longer and heavier at the time of yolk-sac resorption than later-hatching fish. Females from more-heterozygous strains also had a tendency to have larger eggs. These data suggest that genetic variation is an important biological resource to be conserved in hatchery stocks.     

Astaxanthin and canthaxanthin do not induce liver or kidney xenobiotic-metabolizing enzymes in rainbow trout (Oncorhynchus mykiss Walbaum)

This study was designed to assess the effects of dietary carotenoid supplementation on liver and kidney xenobiotic-metabolizing enzymes in the rainbow trout. Twelve rainbow trout (mean weight 266+/-10 g) were assigned to each of three replicate tanks for each of four dietary treatments; astaxanthin, canthaxanthin, negative control and positive control using beta-naphthoflavone, at a target dietary inclusion of 100 mg kg(-1) for each additive. Fish were fed for 3 weeks at a level of 1.2% body wt. day(-1). Serum carotenoid levels were used as indicators of exposure and were not significantly different (P>0.05) between carotenoid-fed trout. Livers and kidney were frozen separately in liquid N(2) by immersion and microsomal fractions from pooled samples (n=3) assayed for xenobiotic-metabolizing enzyme (cytochrome P450 monoxygenase) activities including ethoxyresorufin O-deethylase; methoxyresorufin O-demethylase; pentoxyresorufin O-dealkylase; benzoxyresorufin O-dearylase; and the conjugating enzymes glucuronosyl transferase; and glutathione-s-transferase. Results revealed that carotenoid treatment did not significantly (P>0.05) induce any enzyme system examined. Results are discussed in the context of metabolism of absorbed carotenoids.     

Growth, carcass composition and plasma growth hormone levels in cyclically fed rainbow trout

Growth, body composition and plasma growth hormone levels were recorded weekly for 24 weeks in rainbow trout Oncorhynchus mykiss . Underyearling rainbow trout were individually identified using coded tags and placed on either a cyclic feeding regime of 3 weeks of deprivation followed by 3 weeks of feeding or a daily feeding regime. No significant difference was found in standard length and mass among the cyclically fed and daily fed fish at the end of the experiment. For cyclically fed fish, the absolute specific growth rate and condition factor reached a maximum during the last week of refeeding. Cyclically fed fish had a significantly higher moisture and protein content and lower lipid levels relative to fish fed daily. Absolute mass and fat loss in the deprivation phase of the feeding cycle decreased in intensity with subsequent feeding cycles, indicating that the fish were acclimatizing to the feeding regime. It was proposed that this response was an adaptation against possible adverse effects in the adults ( e.g. locomotor performance, bone ossification rates, fat deposition rate, growth rate and age at sexual maturity). Plasma growth hormone concentrations were not affected by cyclic feeding indicating that variations in plasma growth hormone concentration are not the cause of compensatory growth in rainbow trout.     

Absorption and fate of labelled canthaxanthin 15, 15'-3H2 in rainbow trout (Salmo gairdneri Rich.)

1. Using one force fed meal, absorption of labelled canthaxanthin 15,15'-3H2 was studied by collecting blood via caudal punction at regular intervals and measuring radioactivity in various tissues and organs of rainbow trout. 2. Canthaxanthin absorption showed a large variability between individuals irrespective of their sex. A rapid increase in the radioactivity linked to canthaxanthin 15,15'-3H2 is observed in the blood followed by a slow decrease. 3. Twenty-four and 72 hours after ingestion of labelled canthaxanthin by rainbow trout, the radioactivity was widely distributed. From the total radioactivity given, most was found 24 hours after the meal in the pyloric caeca, ovary and skin. Seventy-two hours after diet, the radioactivity increased in the muscle, liver and kidney. On the other hand, the percentage of radioactivity of intestine and caeca declined. 4. Data from absorption curve indicates that 0.86% of radioactivity ingested is present in the blood, suggesting that canthaxanthin is not readily transferred from the digestive tract. 5. The metabolic clearance of canthaxanthin (lambda = 0.0246/hr +/- 0.005) which would correspond to a half-life of 28 hours gives a preliminary idea of the digestive and metabolic utilization of canthaxanthin by the rainbow trout.