Carotenoids in diets for salmonids: II. Epimerization studies with astaxanthin in Atlantic salmon

Atlantic salmon (Salmo salar, L.), kept in sea pens were fed diets containing pure carotenoids. No significant preferential utilization of the (3S,3′S), (3R,3′S) or (3R,3′R) optical isomers of astaxanthin was observed. No epimerization occurred in the flesh at the chiral centres at C-3 and C-3′ in astaxanthin.     

The effect of feed pigment type on flesh pigment deposition and colour in farmed Atlantic salmon, Salmo salar L.

The characteristic pink colour of salmonid flesh is a result of deposition of naturally occurring carotenoid pigments. Achieving successful pigmentation in farmed salmonids is a vital aspect of fish farming and commercial feed production. Currently commercial diets for farmed salmonids contain either or both of the synthetic pigments commercially available, astaxanthin and canthaxanthin. Atlantic salmon, Salmo salar L. ( = 220 g initial weight) were given feeds where the pigment source was astaxanthin only, canthaxanthin only or a astaxanthin/canthaxanthin mix. The rearing environment was 12 × 3 m tanks supplied with sea water at the EWOS research farm Lønningdal, near Bergen, Norway. As the proportion of dietary canthaxanthin increased, flesh pigment levels also showed an increase; the pigment content in the muscle of canthaxanthin‐only fed fish was 0.4 mg kg^?1 (or 14%) higher than that of the astaxanthin‐only fed fish, with the mixed pigment fed fish being intermediate between the two extremes. Results of cross‐section assessment for Minolta colorimeter redness (a*) values and Roche Salmofan^TM scores also showed an increase in colour with increasing proportions of canthaxanthin in the feed. The data reported clearly indicates that S. salar ( = 810 g final weight) of this size deposit canthaxanthin more efficiently than they do astaxanthin. These results contrast with those obtained by other authors with rainbow trout, Oncorynchus mykiss (Walbaum), and imply that the absorption or utilization of the pigments differs between species.     

Pharmacokinetics and bioavailabilities of 14C-keto-carotenoids, astaxanthin and canthaxanthin, in rainbow trout, Oncorhynchus mykiss

The pharmacokinetics and bioavailabilities of ¹⁴C‐astaxanthin and ¹⁴C‐canthaxanthin were studied in the blood of rainbow trout following intra‐arterial (i.a.) and oral (p.o.) administration. Sixteen months old 1 kg trout were cannulated in the dorsal aorta. [6,7,6′,7′‐¹⁴C]‐keto‐carotenoids were administered i.a. and p.o. at a dose of 573.5 kBq kg^?1 fish body weight for astaxanthin and 836.2 kBq kg^?1 fish body weight for canthaxanthin. After i.a. distribution, total body clearance (Cl_tot) was 17.30±20.29 mL kg^?1 of fish h^?1 for ¹⁴C‐canthaxanthin and 3.30±1.50 mL kg^?1 of fish h^?1 for ¹⁴C‐astaxanthin. The volume of distribution at steady‐state (V_ss) was 208.32±124.79 mL kg^?1 of fish and 71.84±64.15 mL kg^?1 of fish for ¹⁴C‐canthaxanthin and ¹⁴C‐astaxanthin respectively. Less than 0.4% of the administered radioactivity was recovered in urine. Radioactivity (expressed as percent of the dose) excreted in the bile of fish that received ¹⁴C‐canthaxanthin by i.a. route was 20‐fold higher than that observed for fish treated p.o. This ratio was lower for ¹⁴C‐astaxanthin (7.6‐fold). The mean keto‐carotenoid bioavailabilities calculated were 10–15% for both compounds. Findings suggest one daily astaxanthin application is preferable, while 12‐h time intervals between applications are preferable for canthaxanthin.     

Effects of E/Z Isomers and Coating Materials of Astaxanthin Products on the Pigmentation and Antioxidation of Rainbow Trout,Oncorhynchus mykiss

The objective of this study was to compare the effects of four astaxanthin preparations with different ratio of E/Z (trans/cis) isomers and different coating materials on the pigmentation and antioxidation properties of rainbow trout. Five diets were designed as basal diet (without astaxanthin supplementation) and four astaxanthin diets (100 mg/kg diet) supplemented with four astaxanthin products, BASF (79% all‐E, gelatin coated), Wisdom‐B (85% all‐E, gelatin coated), Wisdom‐C (94% all‐E, carrageenan coated), and Wisdom‐D (94% all‐E, gelatin coated). After 4 wk feeding, the flesh astaxanthin content, redness, and yellowness of astaxanthin‐supplemented groups were higher than those of control group at the second and fourth week (P < 0.05). Among these astaxanthin‐supplemented groups, the Wisdom‐B group had the highest flesh redness and astaxanthin content. The astaxanthin‐supplemented groups also had the lower flesh frozen loss and malondialdehyde level than control group at the fourth week (P < 0.05). The results indicated that dietary 100 mg/kg astaxanthin could improve the flesh redness and antioxidation abilities of rainbow trout. The ration of E/Z isomers and properties of coating materials of astaxanthin preparations influenced the pigmentation and antioxidation properties, and Wisdom‐B had a better pigmentation effect on the flesh than other astaxanthin preparations.     

Effects of dietary astaxanthins on pigmentation of flesh and tissue antioxidation of rainbow trout (Oncorhynchus mykiss)

The present study was conducted to evaluate the effects of three commercial astaxanthin preparations (100 mg kg^?1 diet) with different solubilities in water, from DSM (Dutch State Mines), BASF (Badische Anilin and Soda Fabrik) and Wisdom Company on pigmentation of flesh and antioxidation of flesh, serum and liver in rainbow trout with an initial weight of 52.07 g. After 60 days of feeding, there were no significant differences in growth or flesh proximate composition of rainbow trout among groups (P > 0.05); the Salmo Fan score, redness and astaxanthin content of flesh in rainbow trout fed diets supplemented with astaxanthins were higher than those of the control group (P < 0.05). At 0, 12, 24, 48 and 72 h after thawing, the flesh malondialdehyde (MDA) content of the three astaxanthin groups was lower than that of the control group (P < 0.05). The total antioxidation capacity (T-AOC) of liver in the three astaxanthin groups was significantly higher, but serum catalase (CAT) activities were lower, than that of the control group (P < 0.05). The results indicate that addition of 100 mg kg^?1 astaxanthin from DSM, BASF or Wisdom to the diet could improve flesh redness and liver T-AOC, reduce serum CAT, SOD and flesh MDA and extend the shelf life of flesh, in spite of the different solubilities of the three sources in water.     

Changes of carotenoids contents and analysis of astaxanthin geometrical isomerization in Haematococcus pluvialis under outdoor high light conditions

In this study, carotenoid contents of Haematococcus pluvialis during outdoor high light cultivation were measured, moreover, changes of astaxanthin geometrical isomers and biotic factors which may affect isomerization were investigated. During the incubation, contents of both astaxanthin and its precursors (zeaxanthin and canthaxanthin) increased over time, whereas the relative content of lutein decreased. Contents of all astaxanthin isomers increased, while the proportion of different astaxanthin geometrical isomers fluctuated during the incubation. All‐trans‐astaxanthin of total astaxanthin (T/A) was higher during the astaxanthin accumulation phase than that in the cell transformation phase, which was in contrast to results of 13‐cis‐astaxanthin of total astaxanthin (13C/A) and 9‐cis‐astaxanthin of total astaxanthin (9C/A). Farnesyl diphosphate synthase (trans (2E,6E)‐FPPS, EC2.5.1.10) and geranylgeranyl diphosphate synthase (GGPS, EC2.5.1.29), the key proteins involved in geometrical isomerization, decreased during the astaxanthin accumulation phase. Moreover, the presence of cis (2Z,6E)‐FPPS was firstly confirmed in H. pluvialis by HPLC‐MS/MS shotgun method. The results indicated the biotic factor (trans‐FPPS, cis‐FPPS and GGPS) may play an important role in astaxanthin geometrical isomerization of H. pluvialis, but not a crucial role. This study would help in optimizing the regulation of astaxanthin geometrical isomers in H. pluvialis, with great significance in theory and production.     

Assessment of flesh colour in Atlantic salmon, Salmo salar L.

The degree of pigmentation in muscle of Atlantic salmon, Salmo salar L., fillets of fish that were fed eight diets fortified with 10, 20, 40, 60, 80.100, 150 and 200 mg astaxanthin kg^?1 and a non-supplemented control diet from 3 to 21 months was assessed using different methods. A tristimulus colorimeter (Minolta Chroma Meter) was used to measure the colour composition of the fillets instrumentally. The colour was also determined using the Roche Colour Card for Salmonids. The concentration of astaxanthin in the muscle was measured by chemical analyses. All measurements for colour were done directly on the epaxial muscle anterior to the dorsal fin. The lightness factor (L *). the red/green chromaticity (a*), the yellow/blue chromaticity (b*) and the saturation C* of the colorimetric readings and the Colour Card scores were compared with the chemical analyses. The astaxanthin concentration in the flesh varied from 1 to 10 mg kg^?1 and the visual appearance of the fillets varied from yellowish-white to red. The relationship between the a*, b* and C* values and the astaxanthin concentration in the muscle was non-linear. Non-linear regression lines were found between the a* value and the astaxanthin concentration in the flesh (r²= 0.974) and the b* value and the astaxanthin concentration in the flesh (r²= 0.984). The instrument was not able to detect differences in astaxanthin concentration at astaxanthin levels above 3-4 mg kg^?1 using the presented method directly on the fillet. The instrument might be useful for rejecting groups of salmon with poor pigmentation. A good linear regression was found between the Colour Card score and the mean astaxanthin concentration in the flesh (r² - 0.992). The Colour Card provided a better prediction of the astaxanthin concentration at higher astaxanthin levels than the Chroma Meter. None of the methods provided a satisfactory prediction of the astaxanthin concentration in the muscle of individual fish using the presented methods.     

The effects of synthetic pigments on pigmentation of Pethia conchonius (Hamilton, 1822)

This study evaluated the effects of diets containing 20, 40, 60, 80 and 100 mg kg^?1 diet astaxanthin or canthaxanthin on Pethia conchonius (Hamilton, 1822) pigmentation. A completely randomized experimental design was developed with ten treatments and three replicates. Three hundred rosy barb with a mean weight of 0.92 ± 0.06 g were assigned to thirty aquaria for period of eight weeks. Carotenoid contents of fish fed canthaxanthin were always lower than those fed astaxanthin. Yellowness (b*) was not affected by pigments. While Luminosity (L*) decreased in fish fed astaxanthin diets, this parameter increased by feeding on canthaxanthin. The most pronounced effect was higher a* values in fish fed astaxanthin. Astaxanthin retention rate was higher than that of canthaxanthin. The present results demonstrate that canthaxanthin cannot be considered as a proper replacement with astaxanthin. Inclusion of 80 and 100 mg astaxanthin kg^?1 diet can be suitable dietary levels to ensure pigmentation and this condition may improve market value of rosy barb.     

Stability of Krill Meal,Astaxanthin, and Canthaxanthin Color in Cultured Rainbow Trout (Oncorthynchus mykiss) Fillets During Frozen Storage and Cooking

Rainbow trout were pigmented with diets containing synthetic astaxanthin, canthaxanthin, or dried krill meal to 6 mg carotenoid/Kg (wwb) flesh. Vacuum packaged frozen fillets were held at -18°C, -28°C or -80°C for 90 d, 180 d, or 90 d, thawed, and refrozen for an additional 90 d. Tristimulus color (L*,a*,b*), carotenoid concentration, fatty acid composition and TBARS were measured for raw and cooked fillets. We observed no change in pigment content or in a* values after 180 d frozen storage or following a thaw/refreeze cycle compared to fresh fish, even though a higher a* values were seen in fillets from fish fed synthetic astaxanthin or canthaxanthin after 90 d frozen storage suggesting that care should be used when interpreting tristimulus color values for grading programs.     

Pigmentation of Artic Char (Salvelinus alpinus) by Dietary Carotenoids

Feeds formulated to contain 75 ppm astaxanthin or canthaxanthin were fed to Artic char (Salvelinus alpinus, Labrador strain) for 15 weeks. After 9-15 weeks of feeding, the level of carotenoids in fillets of fish exceeded 4 mg/kg, which is considered sufficient for visual colour impression on the fillets. Significant correlations were observed between length of time the cartenoid-containing diets were administered and total carotenoid content of both flesh and skin for both the astaxanthin and canthaxanthin-fed fish. The Hunter a, redness, colour values were correlated with total carotenoids content in the flesh for both astaxanthin-fed and canthaxanthin-fed Artic char.     

Phaffia rhodozyma as an astaxanthin source in salmonid diets

The red yeast Phaffia rhodozyma has possible application as a component of diets for use in aquaculture. Its primary value lies in its content of astaxanthin, which is much higher (5–50 times) than that found in crustacean meals. When fed to rainbow trout, the deposition of astaxanthin in the fish flesh was dependent on the proper preparation of yeast cells before their inclusion into the feed. No astaxanthin was nutritionally available from intact yeast. If P. rhodozyma was mechanically ruptured its pigments were transferred to the flesh of rainbow trout, coloring it salmon-pink. The most efficient deposition of astaxanthin in trout occurred when the cell wall of P. rhodozyma was partially removed by enzymatic digestion.The proxiamte composition, amino acid content, fatty acid profile, and astaxanthin content were determined for P. rhodozyma. When grown under the conditions used in this study, P. rhodozyma has a low protein content (～25%) and a high total lipid content (～17%) compared with most other microorganisms. Its amino acid profile is well balanced but is deficient in methionine. The predominant fatty acids present in the yeast are oleic, linoleic and palmitic acids.     

Reconstitution of muscle F‐actin from Atlantic salmon (Salmo salar L.) with carotenoids—binding characteristics of astaxanthin and canthaxanthin

The binding of carotenoids to the myofibrillar protein F‐actin purified from the white muscle of Atlantic salmon (Salmo salar L.) was studied using in vitro reconstitution. The binding of astaxanthin and canthaxanthin was saturable, and analysis revealed the presence of a single carotenoid‐binding site. The dissociation constants (K_d) for actin prepared from 2.5 kg FW (Fresh Weight) fish were 1.04 ± 0.13 μg carotenoid per milligram of actin and 0.54 ± 0.11 μg/mg for astaxanthin and canthaxanthin, respectively. The saturation binding level (B_max) for astaxanthin was 1.39 ± 0.07 μg/mg and 1.04 ± 0.08 μg/mg for canthaxanthin. These values were higher for F‐actin prepared from organic and small (~0.5 kg FW) salmon than for non‐organic and larger, mature fish. The structural specificity of carotenoid binding revealed a preference for carotenoids that possess a keto group at C‐4 on the β end group of the molecule, but the presence of hydroxyl groups at C‐3 or C‐4 reduced overall binding efficiency. The study suggests that the ability of myofibrillar proteins to bind carotenoids is not a limiting factor governing the deposition of carotenoids in the muscle of salmonids.     

Astaxanthin deposition in the flesh of Atlantic Salmon, Salmo salar L., in relation to dietary astaxanthin concentration and feeding period

Atlantic salmon, Salmo salar L., were fed nine experimental diets containing from 0 to 200 mg astaxanthin per kg^?1 for six time periods, ranging from 3 to 21 months, in sea cages at Matre Aquaculture Research Station, Matredal, Norway. The sampled fish had an initial mean weight of 115 g and reached a weight of 3.2 kg at the termination of the experiment. Every third month, 10 fish from each dose and time group were sampled and the astaxanthin concentration in the flesh determined. The amount of astaxanthin in the flesh ranged from 0.7 to 8.9 mg kg^?1 at the termination of the experiment. This paper discusses deposition of astaxanthin in the flesh of Atlantic salmon in relation to dietary carotenoid levels in the 0–200 mg kg^?1 range and feeding times of 3–21 months. Under the conditions of this experiment, no significant effect on astaxanthin deposition rate could be achieved by increasing the astaxanthin level above 60 mg kg dry feed^?1. Atlantic salmon should be fed astaxanthin-supplemented diets during the whole seawater stage in order to obtain maximal astaxanthin level in the flesh.     

Given the same dietary carotenoid inclusion, Atlantic salmon, Salmo salar (L.) display higher blood levels of canthaxanthin than astaxanthin

Atlantic salmon, Salmo salar, fitted with permanent dorsal aorta cannulae were fed diets containing either 0, 30, 60 mg kg^?1 or combinations of astaxanthin and canthaxanthin, with the aim of comparing the uptake efficiencies to blood of the two pigments and evaluating possible interactions during absorption when formulated in the same diet. Given either astaxanthin or canthaxanthin in separate diets, at dietary levels of <30 mg kg^?1, an identical linear relationship (R² = 0.97) between dietary levels and blood concentrations was observed for both carotenoids. At dietary astaxanthin inclusions above 30 mg kg^?1, blood astaxanthin concentration approached saturation at an average level of 1.2 ± 0.04 μg mL^?1 (arithmetic mean ± SD), whereas blood levels of canthaxanthin continued to increase linearly throughout the inclusion range tested (0–60 mg kg^?1). When both carotenoids were presented in the same diet, a reduction in the absorption efficiency of both pigments was observed (P < 0.05). This manifested itself as a lower level in blood than the level observed when each carotenoid was administered separately. The negative interaction was most prominent for astaxanthin, the maximum average blood saturation level of which fell (P < 0.05) to 0.73 ± 0.03 μg mL^?1 (arithmetic mean ± SD). Our data support the conclusion that at higher dietary inclusions, canthaxanthin is more efficiently absorbed from the digestive tract into the blood of S. salar than astaxanthin.     

虾青素和角黄素对虹鳟肌肉着色和肝脏总抗氧化能力的影响

分别在基础饲料(对照组)中添加100 mg/kg的虾青素、角黄素,混合色素(50 mg/kg虾青素+50 mg/kg角黄素)饲喂初始体重为(56.60±0.63) g的虹鳟60 d,考察虾青素和角黄素对虹鳟肌肉着色和肝脏总抗氧化能力的影响。结果显示,饲料中添加了虾青素、角黄素和混合色素后对虹鳟增重率、饲料系数及肌肉常规成分、肌肉失水率、含肉率均无显著影响(P> 0.05)。虾青素组、角黄素组和混合色素组虹鳟肌肉的比色卡得分、红度、虾青素含量和血清总类胡萝卜素含量均比对照组有显著提高(P< 0.05);虾青素组虹鳟肌肉比色卡得分(26.25)和红度值(18.40)显著高于角黄素组(22.38, 14.13)和混合色素组(24.00, 15.70)(P< 0.05);虾青素组虹鳟肌肉虾青素含量为4.75 mg/kg (30 d)和6.45 mg/kg (60 d),均显著高于混合色素组的3.87 mg/kg (30 d)和5.48 mg/kg (60 d)(P< 0.05);在虹鳟血清总类胡萝卜素含量方面,虾青素组 > 混合色素组 > 角黄素组;虾青素组、角黄素组、混合色素组虹鳟肝脏的总抗氧化能力之间无显著差异(P> 0.05),分别为2.39 U/mg,2.25 U/mg,2.39 U/mg,均较对照组(2.03 U/mg)显著提高(P< 0.05)。上述结果表明：饲料中添加100 mg/kg虾青素、角黄素及虾青素+角黄素混合(1∶1)均能有效改善虹鳟肌肉颜色,提高肝脏总抗氧化能力,虾青素、虾青素+角黄素混合(1∶1)对虹鳟肌肉的着色效果优于角黄素。     

Utilization of carotenoids from various sources by rainbow trout: muscle colour,carotenoid digestibility and retention

Rainbow trout (Oncorhynchus mykiss) with a mean (sd) weight of 120 (2) g were fed diets supplemented with astaxanthin extracted from the yeast Phaffia rhodozyma (OY1 = 50 mg carotenoids kg^–1 feed, OY2 = 100 mg carotenoids kg^–1 feed), astaxanthin (AX = 100 mg astaxanthin kg^–1 feed) and canthaxanthin (CX = 100 mg canthaxanthin kg^–1 feed) for 4 weeks. Muscle analyses at the end of the experiment indicated a significantly higher carotenoid concentration in the AX group, while CX and OY1 groups were similar in spite of the differences in dietary concentration. The measure of total muscle colour difference (E^* _ab) between initial samples and 4 week ones was higher for the AX fish group but showed no significant difference between OY1, OY2, and CX. The hue and the reflectance ratio (R₆₅₀:R₅₁₀) of fish muscle increased in proportion to carotenoid intake. Digestibility (ADC) of yeast astaxanthin in OY1 and OY2 groups was significantly higher than that in the AX group. Canthaxanthin ADC was about one sixth of that of astaxanthin (AX group). Carotenoid retention in the muscle, expressed as a percentage of carotenoid intake, was higher for the AX group than that recorded for OY1 and OY2. According to ADC, carotenoid retention showed a marked lower value for the CX group. Muscle retentions were similar for astaxanthins from both sources.     

Pigmentation of salmonids — Genetical variation in carotenoid deposition in rainbow trout

Significant differences in the carotenoid, astaxanthin and cantaxanthin levels were observed between full-sib and half-sib of rainbow trout (Salmo gairdneri). The sex did not affect pigment deposition but a weak negative correlation between fish weight and level of carotenoids in the flesh was found.     

Chemical composition and physical properties of salted shrimp paste (Kapi) produced in Thailand

Chemical composition and physical properties of 11 salted shrimp pastes (Kapi) obtained from various places of Thailand were determined. Based on proximate composition, protein constituted the major component (29.44–53.27 %, dry wt. basis). All samples contained 22.77–35.47 % NaCl with A _w of 0.695–0.774. Various formal nitrogen contents (11.96–22.87 mg N/g sample) were in agreement with different degrees of hydrolysis (12.68–20.76 %), suggesting the varying cleavage of peptides among the samples. From electrophoretic study, salted shrimp paste contained a large amount of small molecular weight proteins and peptides. Different samples had different colors with \( \Delta E^{*} \) of 47.10–60.43 and \( \Delta C^{*} \) of 9.46–20.76. The samples had total carotenoid content of 0.54–1.97 mg/g sample. Free astaxanthin, astaxanthin diester and canthaxanthin were the major carotenoids in salted shrimp paste. Thus, salted shrimp paste is a good source of protein and serves as the nutritious condiment.     

Response of rainbow trout (Oncorhynchus mykiss) to varying dietary astaxanthin/canthaxanthin ratio: colour and carotenoid retention of the muscle

Rainbow trout with an average initial weight of 160 g were fed during 42 days diets containing varied keto‐carotenoids astaxanthin (Ax)/canthaxanthin (Cx) ratio, as follows: Ax 100% : Cx 0%; Ax 75% : Cx 25%; Ax 50% : Cx 50%; Ax 25% : Cx 75% and Ax 0% : Cx 100%. Muscle colour and carotenoid muscle retention were studied. Colour parameter values for mixed astaxanthin–canthaxanthin‐fed fish were intermediate between those obtained for Ax 0% : Cx 100% fed fish group and for Ax 100% : Cx 0% fed fish group. Concerning muscle carotenoid retention, it has been observed that as the level of canthaxanthin in diet increased, the muscle total carotenoid retention decreased. In the mean time, as the level of canthaxanthin in diet increased, the muscle astaxanthin retention decreased while that of canthaxanthin increased. The results reported here provide further evidence of non‐beneficial effects in terms of muscle colour and muscle carotenoid retention of the use of varying dietary astaxanthin/canthaxanthin ratio for feeding rainbow trout compared to values obtained for astaxanthin‐only feed.     

Conversion of β-carotene, canthaxanthin and astaxanthin to vitamin A in Atlantic halibut (Hippoglossus hippoglossus L.) juveniles

Larval Atlantic halibut fed Artemia has previously been shown to contain lower levels of Vitamin A compared to larvae fed zooplankton. The two types of live prey contain small or no amounts of vitamin A, but high levels of carotenoids that can be converted to vitamin A in other fish species. The purpose of this study was to investigate the ability of Atlantic halibut juveniles to convert β-carotene, astaxanthin and canthaxanthin to vitamin A. Three levels of each carotenoid and retinyl acetate were fed to Atlantic halibut juveniles for 60 days. A vitamin A and carotenoid deficient diet was fed in triplicate as control. A HPLC method modified from Nöll (1996) and validated for fish matrix was used to quantify both all-trans-retinol and 3,4-didehydro retinol. By comparing regression coefficients we observed that the increasing levels of carotenoids in the diets were reflected in increasing levels of vitamin A in both whole fish and liver samples. All carotenoids were converted to vitamin A, but to different degrees. Retinyl acetate and β-carotene resulted in whole fish vitamin A levels significantly higher than canthaxanthin and astaxanthin. 3,4-didehydro retinol was not detected when the overall level of all-trans-retinol was low. When 3,4-didehydro retinol appeared, it was always in lower levels than all-trans-retinol.