Requirements for carotenoids in fish diets

Carotenoids are among the most widespread and important pigment classes in living organisms, and in marine animals, astaxanthin is the most commonly occurring red carotenoid. Carotenoids are vitamin A precursors and are fundamental in photosynthesis and light protection in plants. Increasing attention has been drawn to a possible light protection, cancer prevention and immune enhancement by carotenoids in mammals. Reported functions in fish range from a general enhancement of performance to specific functions in reproduction and metabolism. In this paper, we show that astaxanthin is essential for growth and survival of fish and crawfish, and discuss this fact in relation to the use of purified and semipurified diets in nutritional studies. The similarity in action of astaxanthin and canthaxanthin compared to α-tocopherol (vitamin E) and retinol (vitamin A) suggests that these two carotenoids should be listed among the fat soluble vitamins.     

Metabolism of carotenoids in sea-urchin Pseudocentrotus depressus

• 1.1. Feeding experiments with β,β-carotene, canthaxanthin and astaxanthin on the sea urchin Pseudocentrotus depressus were investigated.
• 2.2. In the case of β,β-carotene group, β-carotene was accumulated, β-isocryptoxanthin appeared and β-echinenone increased 6.8 times as much as the control group. On the other hand, in canthaxanthin and astaxanthin groups, canthaxanthin and astaxanthin increased significantly, respectively. The metabolic products of these carotenoids could not be found.
• 3.3. It was concluded that β,β-carotene was bioconverted to β-echinenone via β-isocryptoxanthin in P. depressus and could not be oxidatively metabolized beyond β-echinenone.

     

Pathway engineering strategies for production of beneficial carotenoids in microbial hosts

Carotenoids, such as lycopene, β-carotene, zeaxanthin, canthaxanthin and astaxanthin have many benefits for human health. In addition to the functional role of carotenoids as vitamin A precursors, adequate consumption of carotenoids prevents the development of a variety of serious diseases. Biosynthesis of carotenoids is a complex process and it starts with the common isoprene precursors. Condensation of these precursors and subsequent modifications, by introducing hydroxyl- and keto-groups, leads to the generation of diversified carotenoid structures. To improve carotenoid production, metabolic engineering has been explored in bacteria, yeast, and algae. The success of the pathway engineering effort depends on the host metabolism, specific enzymes used, the enzyme expression levels, and the strategies employed. Despite the difficulty of pathway engineering for carotenoid production, great progress has been made over the past decade. We review metabolic engineering approaches used in a variety of microbial hosts for carotenoid biosynthesis. These advances will greatly expedite our efforts to bring the health benefits of carotenoids and other nutritional compounds to our diet.     

Reduction of oxidized guanosine by dietary carotenoids: A pulse radiolysis study

Time-resolved pulse radiolysis investigations reported herein show that the carotenoids β-carotene, lycopene, zeaxanthin and astaxanthin (the last two are xanthophylls - oxygen containing carotenoids) are capable of both reducing oxidized guanosine as well as minimizing its formation. The reaction of the carotenoid with the oxidized guanosine produces the radical cation of the carotenoid. This behavior contrasts with the reactions between the amino acids and dietary carotenoids where the carotenoid radical cations oxidized the amino acids (tryptophan, cysteine and tyrosine) at physiological pH.     

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.     

Dietary carotenoids predict plumage coloration in wild house finches

Carotenoid pigments are a widespread source of ornamental coloration in vertebrates and expression of carotenoid-based colour displays has been shown to serve as an important criterion in female mate choice in birds and fishes. Unlike other integumentary pigments, carotenoids cannot be synthesized; they must be ingested. Carotenoid-based coloration is condition-dependent and has been shown to be affected by both parasites and nutritional condition. A controversial hypothesis is that the expression of carotenoid-based coloration in wild vertebrates is also affected by the amount and types of carotenoid pigments that are ingested. We tested this carotenoid-limitation hypothesis by sampling the gut contents of moulting house finches and comparing the concentration of carotenoid pigments in their gut contents with the colour of growing feathers. We found a positive association: males that ingested food with a higher concentration of carotenoid pigments grew brighter ornamental plumage. We also compared the concentration of carotenoids in the gut contents of males from two subspecies of house finches with small and large patches of carotenoid-based coloration. Consistent with the hypothesis that carotenoid access drives the evolution of carotenoid-based colour displays, males from the population with limited ornamentation had much lower concentrations of carotenoids in their gut contents than males from the population with extensive ornamentation. These observations support the idea that carotenoid intake plays a part in determining the plumage brightness of male house finches.     

The Mycobacterium tuberculosis ORF Rv0654 encodes a carotenoid oxygenase mediating central and excentric cleavage of conventional and aromatic carotenoids

Mycobacterium tuberculosis, the causative agent of tuberculosis, is assumed to lack carotenoids, which are widespread pigments fulfilling important functions as radical scavengers and as a source of apocarotenoids. In mammals, the synthesis of apocarotenoids, including retinoic acid, is initiated by the β-carotene cleavage oxygenases I and II catalyzing either a central or an excentric cleavage of β-carotene, respectively. The M. tuberculosis ORF Rv0654 codes for a putative carotenoid oxygenase conserved in other mycobacteria. In the present study, we investigated the corresponding enzyme, here named M. tuberculosis carotenoid cleavage oxygenase (MtCCO). Using heterologously expressed and purified protein, we show that MtCCO converts several carotenoids and apocarotenoids in vitro. Moreover, the identification of the products suggests that, in contrast to other carotenoid oxygenases, MtCCO cleaves the central C15-C15' and an excentric double bond at the C13-C14 position, leading to retinal (C(20)), β-apo-14'-carotenal (C(22)) and β-apo-13-carotenone (C(18)) from β-carotene, as well as the corresponding hydroxylated products from zeaxanthin and lutein. Moreover, the enzyme cleaves also 3,3'-dihydroxy-isorenieratene representing aromatic carotenoids synthesized by other mycobacteria. Quantification of the products from different substrates indicates that the preference for each of the cleavage positions is determined by the hydroxylation and the nature of the ionone ring. The data obtained in the present study reveal MtCCO to be a novel carotenoid oxygenase and indicate that M. tuberculosis may utilize carotenoids from host cells and interfere with their retinoid metabolism.     

Absorption and metabolism of dietary carotenoids

A number of carotenoids with diverse structures are present in foods and have beneficial effects on human health due to their common antioxidant activity and their respective biological activities. The major carotenoids found in human tissues, however, are limited to several including such as β-carotene, lycopene, and lutein. We have little knowledge of whether carotenoids are selectively absorbed in intestine and metabolized discriminately in the body. Moreover, the metabolic transformation of carotenoids in mammals other than vitamin A formation has not been fully elucidated. Here, the intestinal absorption and oxidative metabolism of dietary carotenoids are reviewed with a focus on dietary xanthophylls.     

Selective absorption of carotenoids in the common green iguana (Iguana iguana)

In the animal kingdom, species-specific differences with regard to the absorption of intact carotenoids are observed. The causes of these differences are not entirely understood. To investigate the absorption of selected carotenoids, 20 juvenile green iguanas (Iguana iguana) were fed a carotenoid deficient basal diet for 56 days. Thereafter, the iguanas were assigned to receive a basal diet supplemented with different carotenoids (80 mg/kg diet) such as beta-carotene, canthaxanthin and apo-8'-carotenoic acid ethyl ester for 28 days. Changes in plasma carotinoid concentrations associated with the individual diets were used as indicators of carotenoid absorption. In both the experimental and control groups, only the oxygenated carotenoids (xanthophylls), lutein, zeaxanthin and canthaxanthin, were found in the plasma. Canthaxanthin and apo-8'-carotenoic acid ethyl ester were readily absorbed and recovered from the plasma. However, the supplementation of beta-carotene caused no increase in plasma beta-carotene concentration. Additionally, beta-carotene, canthaxanthin or apo-8'-carotenoic acid ethyl ester did not affect the concentrations of retinol and alpha-tocopherol in plasma. In conclusion, the study demonstrates that iguanas appear to be selective accumulators of polar xanthophylls. The iguana might, therefore, be a valuable model to investigate the selectiveness of carotenoid absorption as well as the function of xanthophylls in animals.     

Extraction and analysis of carotenoids from the New Zealand sea urchin Evechinus chloroticus gonads

Sea urchin gonad (roe) is a highly valued food in Japan and North America. Gonad price is strongly influenced by quality, with appearance, especially colour being a major determinant. Previous attempts to extract a carotenoid profile from the New Zealand sea urchin species Evechinus chloroticus have been challenging due to the large amount of lipid present in the gonad. A carotenoid extraction and high performance liquid chromatography (HPLC) analysis method was developed to reduce lipid contamination by incorporating a saponification and lipid cold precipitation in the extraction procedure. This method enabled greater carotenoid purity and enhanced analysis by HPLC. Echinenone was found to be the main carotenoid present in all E. chloroticus gonads. Dark coloured gonads contained higher levels of fucoxanthin/fucoxanthinol, β-carotene and xanthophylls such as astaxanthin and canthaxanthin. This information on the modification and deposition of carotenoids will help in the development of diets to enhance gonad colour.     

Strong and weak plasma response to dietary carotenoids identified by cluster analysis and linked to beta-carotene 15,15'-monooxygenase 1 single nucleotide polymorphisms

The mechanisms as well the genetics underlying the bioavailability and metabolism of carotenoids in humans remain unclear. To begin to address these questions, we used cluster analysis to examine individual temporal responses of plasma carotenoids from a controlled-diet study of subjects who consumed carotenoid-rich beverages. Treatments, given daily for 3 weeks, were watermelon juice at two levels (20-mg lycopene, 2.5-mg β-carotene, n=23 and 40-mg lycopene, 5-mg β-carotene, n=12) and tomato juice (18-mg lycopene, 0.6-mg β-carotene, n=10). Cluster analysis revealed distinct groups of subjects differing in the temporal response of plasma carotenoids and provided the basis for classifying subjects as strong responders or weak responders for β-carotene, lycopene, phytoene and phytofluene. Individuals who were strong or weak responders for one carotenoid were not necessarily strong or weak responders for another carotenoid. Furthermore, individual responsiveness was associated with genetic variants of the carotenoid metabolizing enzyme β-carotene 15,15'-monooxygenase 1. These results support the concept that individuals absorb or metabolize carotenoids differently across time and suggest that bioavailability of carotenoids may involve specific genetic variants of β-carotene 15,15'-monooxygenase 1.     

The effect of an acute phase response on tissue carotenoid levels of growing chickens (Gallus gallus domesticus)

Plasma, liver and skin carotenoids decrease following infectious disease challenges. Since these challenges often involve substantial host pathology and chronic immune responses, the mechanism underlying altered carotenoid deposition is unclear. Therefore, changes in tissue carotenoid levels were examined during an acute phase response induced by lipopolysaccharide (LPS) or interleukin-1 (IL-1). In two experiments, chicks were hatched from carotenoid-deplete eggs (n=28, n=64, respectively) and fed 0, 8 or 38 mg carotenoids (lutein+canthaxanthin)/kg diet. For chicks fed 38 mg carotenoids, but not those fed 0 or 8 mg, LPS generally reduced plasma lutein, canthaxanthin and total carotenoids (P<0.05), and liver lutein, zeaxanthin, canthaxanthin and total carotenoids (P<0.05). Additionally, LPS reduced thymic total carotenoids (P=0.05) and increased thymocyte lutein (P=0.07), zeaxanthin (P=0.07) and total carotenoids (P=0.07). Finally, LPS increased bursal canthaxanthin (P<0.01), but had no effect on shank carotenoids (P>0.5). In chicks hatched from carotenoid-replete eggs (n=36) and fed dietary lutein (38 mg/kg diet), LPS reduced plasma and liver zeaxanthin and liver total carotenoids (P<0.05); IL-1 reduced plasma and liver lutein, zeaxanthin and total carotenoids (P<0.05). Therefore, an acute phase response plays a role in reduced tissue carotenoids during infectious disease.     

Northern berries as a source of carotenoids

Carotenoids are bioactive substances in human diet. The aim of the study was to determine β-carotene and xanthophylls in four berries species. An HPLC gradient elution system were used for separation and quantification of the carotenoids. The highest total carotenoid content among the berries studied was found in cloudberry (2840 μg/100g dw), followed by blueberry (2140 μg/100 g). All berries had lutein but it was a predominant carotenoid in blueberry. The highest β-carotene levels were found in cloudberry (83% of total Car content). Cranberry and cowberry were the poorest sources of carotenoids. Our data will be included in the regional database of resources with the increased nutritional value.     

Ecological, morphological and phylogenetic correlates of interspecific variation in plasma carotenoid concentration in birds

Carotenoids are important as pigments for bright coloration of animals, and as physiologically active compounds with a wide array of health-related benefits. However, the causes of variation in carotenoid acquisition and physiology among species are poorly known. We measured the concentration of carotenoids in the blood of 80 wild bird species differing in diet, body size and the extent of carotenoid-based traits. Preliminary analyses showed that diet significantly explains interspecific variability in plasma carotenoids. However, dietary influences were apparently overridden by phylogenetic relationships among species, which explained most (65%) of this variability. This phylogenetic effect could be due partly to its covariation with diet, but may also be caused by interspecific differences in carotenoid absorption from food to the blood stream, mediated, for example by endothelial carriers or gut parasites. Carotenoid concentrations also decreased with body size (which may be explained by the allometric relationship between ingestion rate and body mass), and correlated positively with the extent of carotenoid-dependent coloration of plumage and bare parts. Therefore, the acquisition of carotenoids from the diet and their use for both health and display functions seem to be constrained by ecological and physiological aspects linked to the phylogeny and size of the species.     

The carotenoids of wild and blue disease affected farmed tiger shrimp (Penaeus monodon, Fabricus)

1. The main carotenoids in wild Penaeus monodon exoskeleton were astaxanthin di- and mono-esters, astaxanthin, and beta-carotene. 2. Wild P. monodon exoskeleton contained on average 26.3 ppm total carotenoid; normally pigmented farmed shrimp had a similar concentration (25.3 ppm). 3. Exoskeletons of farmed "blue" P. monodon (i.e. blue-coloured, as opposed to the normally red-blue/black banded shrimp) contained significantly less total carotenoid (4.3-7 ppm). The only major carotenoid being astaxanthin. 4. Commercially available diets contained only trace quantities of canthaxanthin. 5. Nutritional deficiency with respect to carotenoids is suggested as the cause of blue disease in farmed P. monodon.     

Synthesis of highly 13C enriched carotenoids: access to carotenoids enriched with 13C at any position and combination of positions

Carotenoids and their metabolites are essential factors for the maintenance of important life processes such as photosynthesis. Animals cannot synthesize carotenoids de novo, they must obtain them via their food. In order to make intensive animal husbandry possible and maintain human and animal health synthetic nature identical carotenoids are presently commercially available at the multi-tonnes scale per year. Synthetically accessible (13)C enriched carotenoids are essential to apply isotope sensitive techniques to obtain information at the atomic level without perturbation about the role of carotenoids in photosynthesis, nutrition, vision, animal development, etc. Simple highly (13)C enriched C(1), C(2) and C(3) building blocks are commercially available via 99% (13)CO. The synthetic routes for the preparation of the (13)C enriched building blocks starting from the commercially available systems are discussed first. Then, how these building blocks are used for the synthesis of the various (13)C enriched carotenoids and apocarotenoids are reviewed next. The synthetic Schemes that resulted in (13)C enriched β-carotene, spheroidene, β-cryptoxanthin, canthaxanthin, astaxanthin, (3R,3'R)-zeaxanthin and (3R,3'R,6'R)-lutein are described. The Schemes that are reviewed can also be used to synthetically access any carotenoid and apocarotenoid in any (13)C isotopically enriched form up to the unitarily enriched form.     

One-electron reduction potentials of dietary carotenoid radical cations in aqueous micellar environments

The one-electron reduction potentials of the radical cations of five dietary carotenoids (β-carotene, canthaxanthin, zeaxanthin, astaxanthin and lycopene) in aqueous micellar environments have been obtained from a pulse radiolysis study of electron transfer between the carotenoids and tryptophan radical cations as a function of pH, and lie in the range of 980–1060 mV. These values are consistent with our observation that the carotenoid radical cations oxidise tyrosine and cysteine. The decays of the carotenoid radical cations in the absence of added reactants suggest a distribution of exponential lifetimes. The radicals persist for up to about 1 s, depending on the medium.     

Carotenoids in fish. XXVI. Pungitius pungitius (L.) and Gasterosteus aculeatus L. (Gasterosteidae)

The author investigated the presence of various carotenoids in the different parts of the body of Pungitius pungitius (L.) and Gasterosteus aculeatus L. by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following carotenoids:

in Pungitius pungitius. α-carotene, β-carotene, β-cryptoxanthin, mutatochrome, zeaxanthin and astaxanthin;

in Gasterosteus aculeatus: β-carotene, β-cryptoxanthin, β-carotene epoxide, neothxanthin, canthaxanthin, mutatochrome, lutein, phoenicoxanthin, zeaxanthin, taraxanthin, tunaxanthin, astaxanthin, astaxanthin ester and α-doradexanthin. The total carotenoid content ranged from 2.229 to 138.504 µg/g wet weight.

     

Carotenoids and fatty liver disease: Current knowledge and research gaps

Carotenoids form an important part of the human diet, consumption of which has been associated with many health benefits. With the growing global burden of liver disease, increasing attention has been paid on the possible beneficial role that carotenoids may play in the liver. This review focuses on carotenoid actions in non-alcoholic fatty liver disease (NAFLD), and alcoholic liver disease (ALD). Indeed, many human studies have suggested an association between decreased circulating levels of carotenoids and increased incidence of NAFLD and ALD. The literature describing supplementation of individual carotenoids in rodent models of NAFLD and ALD is reviewed, with particular attention paid to β-carotene and lycopene, but also including β-cryptoxanthin, lutein, zeaxanthin, and astaxanthin. The effect of beta-carotene oxygenase 1 and 2 knock-out mice on hepatic lipid metabolism is also discussed. In general, there is evidence to suggest that carotenoids have beneficial effects in animal models of both NAFLD and ALD. Mechanistically, these benefits may occur via three possible modes of action: 1) improved hepatic antioxidative status broadly attributed to carotenoids in general, 2) the generation of vitamin A from β-carotene and β-cryptoxanthin, leading to improved hepatic retinoid signaling, and 3) the generation of apocarotenoid metabolites from β-carotene and lycopene, that may regulate hepatic signaling pathways. Gaps in our knowledge regarding carotenoid mechanisms of action in the liver are highlighted throughout, and the review ends by emphasizing the importance of dose effects, mode of delivery, and mechanism of action as important areas for further study. This article is part of a Special Issue entitled Carotenoids recent advances in cell and molecular biology edited by Johannes von Lintig and Loredana Quadro.     

A nondestructive method for extracting maternally derived egg yolk carotenoids

ABSTRACT.   Maternally deposited carotenoids are a prominent component of egg yolk and are vital for the development and growth of the embryo. In most studies of avian yolk carotenoids, eggs are destructively sampled and this may limit both the number of clutches studied and the research questions addressed. We describe an empirical field trial for a nondestructive biopsy method to extract small samples (0.05 ml) of egg yolk for high-performance liquid chromatography (HPLC) analysis of yolk carotenoid concentrations. We sampled 180 clutches ( N = 44 biopsies) of two species of introduced thrushes (genus Turdus ) from agricultural habitats in central North Island, New Zealand. Once the protocol was established, all biopsied eggs from clutches that were not depredated or deserted before candling were found to be developing normally after 3–5 d of incubation ( N = 28) and all hatched. Biopsy samples (>0.02 g) produced concentrations of yolk carotenoids (and variances) that were statistically indistinguishable from whole yolk destructive samples. In addition, our samples (>0.02 g) confirmed previously reported differences in yolk carotenoid concentrations between the two thrush species and revealed a significant decline in yolk carotenoid concentration with laying order. Further examination of how variability in yolk carotenoid concentration and identity influences offspring sex, success, and survival or, later in life, reproductive success and ability to efficiently incorporate dietary carotenoids into both integument and immune tissues will require larger sample sizes. Studies to date have been restricted by the number of destructive samples that investigators are willing (or permitted) to obtain from wild species. Thus, we hope that our nondestructive method of sampling yolk will promote further examination of the links between carotenoid uptake from the environment and maternal investment in the avian yolk.