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Canthaxanthin fixation in rainbow trout
Oncorhynchus mykiss : effect of dietary
phospholipid content

 ° Laboratoire d’écophysiologie des invertébrés, Université de Montpellier II, CP092, Place Eugène Bataillon, F-34095 Montpellier
   e-mail : castille@univ-montp2.fr
°° Laboratoire de Nutrition des Poissons, Unité mixte INRA-INFREMER, Station d’Hydrobiologie, F-64310 Saint-Pée-sur-Nivelle
   e-mail : choubert@st-pee.inra.fr

SUMMARY                                                                                     RÉSUMÉ

   Colour and canthaxanthin concentration in the muscle of rainbow trout                    Fixation de la canthaxanthine dans la truite arc-en-ciel Oncorhynchus
were studied in a feeding trial with three different phospholipid levels.                   mykiss : effet de la concentration en phospholipides de l’aliment. Par A.
Rainbow trout with an initial mean body weight of 190 g were fed, ad libi-                  ABAT, R. CASTILLO and G. CHOUBERT.
tum, 2 meals/day in triplicate, the experimental diets supplemented at a rate
of 80 mg canthaxanthin/kg diet combined with three different phospholipid                      La couleur et la concentration en canthaxanthine du muscle de truite
levels (0, 4, and 8 %) were furnished during 6 weeks. At each sample time                   arc-en-ciel ont été étudiées en relation avec trois différents taux de phos-
(2, 4, 6 weeks) canthaxanthin concentration in the muscle of trout showed                   pholipide dans l’aliment. Des truites arc-en-ciel, d’un poids moyen initial de
no significant differences (P ≥ 0.05) between the different dietary groups.                 190 g ont reçu, ad libitum, 2 repas/jour en triplicat, des aliments expéri-
Phospholipids added to the diet were not found to enhance the deposition of                 mentaux supplémentés en canthaxanthine (au taux de 80 mg canthaxanthi-
canthaxanthin in the muscle of the trout. Colour measurements gave results                  ne/kg d’aliment) combinés à trois différents taux de phospholipides (0, 4,
somewhat different except for the a* colour parameter. Higher values                        and 8 %) pendant 6 semaines. A chaque prélèvement (2, 4, 6 semaines) les
(P ≤ 0.05) for lightness, chroma and hue angle were always obtained for                     concentrations en canthaxanthine dans le muscle de truite n’étaient pas
trout fed the high dietary phospholipid content (8 %). On the other hand                    significativement (P ≥ 0.05) différentes entre les groupes. Les phospholi-
colour of the muscle of trout fed the low dietary phospholipid level (4 %)                  pides ajoutés à l’aliment n’ont pas eu d’effet sur le dépôt en canthaxanthine
showed no significant differences (P ≥ 0.05) with control fish for lightness                dans le muscle de truite. En ce qui concerne les mesures de couleur, les
and chroma, but a significant difference was noted for the hue angle. It has                résultats obtenus diffèrent sensiblement, excepté pour le paramètre a*. Des
been concluded that the addition of phospholipids to the diet of trout may                  valeurs plus élevées (P ≤ 0.05) pour la luminosité, la saturation et la teinte
have a positive effect on colour parameters without any effect on can-                      ont toujours été obtenues pour les truites recevant l’aliment riche en phos-
thaxanthin concentration.                                                                   pholipides (8 %). Par contre, les paramètres de la couleur du muscle de trui-
                                                                                            te recevant l’aliment contenant 4 % de phospholipides n’étaient pas diffé-
                                                                                            rents (P ≥ 0.05) de ceux des animaux recevant l’aliment témoin (sans phos-
                                                                                            pholipides ajoutés) pour la luminosité et la saturation. Toutefois une diffé-
                                                                                            rence a été notée pour la teinte. Il semble que l’addition de phospholipides
                                                                                            à un aliment supplémenté en canthaxanthine pour truite arc-en-ciel a un
                                                                                            effet positif sur les paramètres de la couleur du muscle, sans entrainer
                                                                                            d’augmentation notable de la concentration en canthaxanthine musculaire.

KEY-WORDS : phospholipids - canthaxanthin - flesh                                           MOTS-CLÉS : phospholipides - canthaxanthine - couleur
colour - trout.                                                                             musculaire - truite.

Introduction                                                                                hydrophilic (water affinity) phosphate chain and the hydro-
                                                                                            phobic fatty acid chains. They are therefore surface active
  Canthaxanthin, a carotenoid pigment, is commonly used in                                  and play an important role as emulsifying agent, thus promo-
intensive rearing fish farms to enhance the natural colour of                               ting the absorption of lipids and lipid soluble compounds
the flesh of rainbow trout. This lipid soluble compound is not                              (O’DOHERTY et al., 1973).
synthesized by the fish and must be supplied with the diet.                                   The purpose of this study was to investigate whether die-
The digestibility of canthaxanthin is low (20 - 40 %,                                       tary phospholipids may enhance the absorption of canthaxan-
CHOUBERT and LUQUET, 1979).                                                                 thin and therefore may increase flesh pigmentation in the
  Phospholipds combine within the same molecule both the                                    rainbow trout.

Revue Méd. Vét., 2002, 153, 10, 665-668
666                                                                                                         ABAT (A.) AND COLLABORATORS

Materials and methods                                                         The measurement of colour attributes was carried out by
                                                                              applying the chromameter onto the muscle. After flashing,
  Fish. Rainbow trout Oncorhynchus mykiss with a mean                         L*, a*, and b* reflected light values were recorded. From a*
body weight of 190 g were settled in 9 tanks (1m in diame-                    and b* values the hue (H°ab = tan-1b*/a*) and the chroma
ter), set in parallel (30 fish/tank). The tanks were gravity fed              (C*ab = (a*2+b*2)1/2) were calculated according to
with spring water (Temperature = 17 ± 1°C, O2 = 8-9 mg/l,                     WYSZECKI and STILES (1967).
Ph = 7.4, Cl- = 22.5 mg/l, Ca2+ = 75 mg/l) at a rate of
                                                                                Calculation. Data were processed through ANOVA (inde-
5 volume changes per hour.
                                                                              pendant group analysis) followed by the Tukey multiple
  Diet. The composition of the experimental diets is given in                 comparison test (SAS, 1985) ; the significance level was
Table I. Phospholipids (soja lecithin, DAFA LPR, Nikerson
                                                                              P ≤ 0.05.
s.a., Marne-la-Vallée, France) were added to the diet at a rate
of 0, 4, and 8 %. The diets were supplemented with 80 mg of
canthaxanthin (Carophyll red, F. Hoffmann-La Roche,                          Results
Basel, Switzerland)/kg of diet. Phospholipids were mixed
with the other ingredients before pelleting through a 4.5 mm                     Data concerning colour measurements are given in Table II
dye without steam (Simon Heesen pelleting machine, Boxtel,                    No phospholipid effect (P ≥ 0.05) was reported for the a*
The Netherlands). Fish were hand fed ad libitum 2 meals per                   colour parameter while higher data (P ≤ 0.05) for luminosity,
day during 6 weeks.                                                           chroma and hue were always obtained for trout fed the
  Analyses. At each sampling time (2, 4, 6 weeks) 15 fish per                 highest dietary phospholipid content (8 %). On the other
diet were randomly sampled. The latero-dorsal muscle was                      hand, the colour of the muscle of trout fed the lower dietary
removed. Colour measurement was then processed and the                        phospholipid level (4 %), compared with control, showed no
sample was frozen (-20°C) until canthaxanthin analyses.                       significant difference for luminosity and chroma, but a signi-
                                                                              ficant difference was noted for the hue.
  The colour of the flesh was processed with a chromameter
(Minolta Co. Ltd, Osaka, Japan) at three different points of                    Canthaxanthin concentration in the muscle of trout fed dif-
the muscle, above the lateral line : 1) behind the head, 2)                   ferent amount of phospholipids are reported for each sam-
below the dorsal fin and 3) near the tail. The measures have                  pling time (Figure 1). Canthaxanthin concentration in the
been accomplished at the beginning and at the end of the                      muscle of trout showed no significant difference (P ≥ 0.05)
experiment, according to CHOUBERT et al. (1997). The                          between the 3 dietary groups. Therefore there was a tendency
apparatus was calibrated prior the measurements with a white                  for high dietary phospholipid level (8 %) to enhance can-
plate standard reference (Minolta Co. Ltd, Osaka, Japan).                     thaxanthin concentration in the flesh during the first 2 weeks.

                    (1) DAFA LPR, Nickerson s.a., Marne-la-Vallée, France
                    (2) Labbé et al., 1993.
                    (3) Carophyll(r) red, F. Hoffmann - La Roche, Basel, Switzerland
                                                TABLE I. — Composition of the experimental diets.
                                         TABLEAU I. — Composition des régimes alimentaires expérimentaux.

                                                                                                              Revue Méd. Vét., 2002, 153, 10, 665-668
CANTHAXANTHIN FIXATION IN RAINBOW TROUT ONCORHYNCHUS MYKISS                                                                                  667

                   * Amount of phospholipids added to the diet: 0 %, 4 %, 8 %.
                   Results are Mean ± SE, n=15. Means of an attribute within a column with no common superscripts differ significantly
                   TABLE II. — Colour measurements of muscle of fish fed canthaxanthin and different amounts of phospholipids during
                     6 weeks.
                   TABLEAU II. — Paramètres de la couleur du muscle de truite recevant dans son alimentation de la canthaxanthine et dif-
                     férents taux de phospholipides pendant 6 semaines.

                   FIGURE 1. — Canthaxanthin concentration in the muscle of trout fed different amount of phospholipids.
                   FIGURE 1. — Concentration en canthaxanthine dans le muscle de truite ayant ingéré différents taux de phospholipides.

Discussion                                                                        hand, in poultry, the use of phospholipids in the diet of broi-
                                                                                  ler gave conflicting results: the addition of 2 % lecithin to
   It is well known that canthaxanthin is less absorbed than                      broiler ration did not significantly influence their pigmenta-
astaxanthin by rainbow trout. It is for this reason that can-                     tion (RATCLIFF et al., 1959), while soybean lecithin added
thaxanthin was choosen to study the effect of phospholipids,                      to broiler diets depressed the deposition of carotenoid pig-
if any, on a compound which is not well absorbed by the fish.                     ments in the flesh (WILLIAMS, 1962). More recently it was
Lecithin increased the digestibility and absorption of fat in                     claimed that inclusion of lysolecithin in the diet of laying
man (O’DOHERTY et al., 1973).                                                     hens led to the increasing of pigment conversion from 16.1 %
   In our experiment, phospholipids added to the diet were                        to 26.8 % (CALTRON et al., 1998).
not found to enhance the deposition of canthaxanthin in the
muscle of rainbow trout. Moreover phospholipids did not                              It has been concluded from our experiment, that the addi-
have any effect on the a* colour parameter while the higher                       tion of phospholipids to the diet of the trout may have a posi-
amount of phospholipids (8 %) in the diet led to higher colour                    tive effect on colour parameters except a*, however, there is
parameter.                                                                        no response concerning canthaxanthin concentration in the
   Literature does not report comparative studies on the effect                   muscle of the fish.
of phospholipids on flesh pigmentation in fish. On the other

Revue Méd. Vét., 2002, 153, 10, 665-668
668                                                                                                           ABAT (A.) AND COLLABORATORS

Acknowledgements                                                               CHOUBERT G. and LUQUET P. : Influence de l’agglomération et du stoc-
                                                                                     kage des aliments composés sur leur teneur en canthaxanthine :
                                                                                     Conséquence sur la digestibilité et la fixation de ce pigment chez la
   The authors wish to thank Y. Hontang, F. Terrier and F. Sandres for the           truite arc-en-ciel. Ann. Zootech., 1979, 28 : 145-157.
maintenance of the experimental animals. The authors are grateful to           LABBÉ C., LOIR M., KAUSHIK S. and MAISSE G : The influence of both
Nikerson s.a. (Marne-la-Vallée, France) for providing us with the soja leci-         rearing temperature and dietary lipid origin on fatty acid composition
                                                                                     of spermatozoan polar lipids in rainbow trout Oncorhynchus mykiss.
thin (DAFA LPR), Produits Roche France (Neuilly-sur-Seine, France ), for
                                                                                     Effect on sperm cryopreservation tolerance. In: S Kaushik and P.
the canthaxanthin beadlets, and to F. HOFFMAN-LA ROCHE (Basel,                       Luquet (Eds). Fish Nutrition in Practice, Biarritz (France), June 24-
Switzerland) for carotenoid standards.                                               27, 1991. Ed. INRA, Paris 1993 (Les Colloques, N° 61), 49-59.
                                                                               O’DOHERTY P.J., KAKIS G. and KUKSIS A. : Role of luminal lecithin in
                                                                                     intestinal fat absorption. Lipids, 1973, 8 : 241-328.
                                                                               OSADCA M., ARAUJO M. and DE RITTER E. : Determination of can-
References                                                                           thaxanthin in concentrates and feeds. J. Assoc. Offic. Anal. Chem.,
                                                                                     1972, 55 : 110-113.
                                                                               RATCLIFF R.G., DAY E.J. and HILL J.E. : Broiler pigmentation as influen-
CALTRON D., SANDERS D. and BRINKHAUS F. : Lisolecitina e pigmen-                     ced by dietary modifications. Poutry Sci., 1959, 38 : 1039-1048.
    tazione del tuorlo d’uovo. Riv. Avicolt., 1998, 67 : 36-37.                SAS - SAS/DSTAT : Guide for personal computers. Ver.6., SAS Institute
CHOUBERT G., BLANC J-M. and VALLÉE F. : Colour measurement,                          Inc., Cary, 1985, USA.
    using the CIE LCH colour space of muscle of rainbow trout,                 WILLIAMS W.P. : Feedstuffs, 1962, 34 : 34-35, 52, 54.
    Oncorhynchus mykiss (Walbaum) fed astaxanthin: effect of family,           WYSZECKI G. and STILES W.S. : Color Science. John Wiley and Sons,
    ploidy, sex, and location of reading. Aquac Res., 1997, 28 : 15-22.              New York, 1967, USA.

                                                                                                                 Revue Méd. Vét., 2002, 153, 10, 665-668

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