Preparation of the red yeast,Xanthophyllomyces dendrorhous,as feed additive with increased availability of astaxanthin

Xanthophyllomyces dendrorhous (Phaffia rhodozyma) is used as a colorant for aquaculture, egg yolks, and crustaceans but its carotenoids can only be absorbed by animals when its cell wall is degraded. Conditions that degraded the cell wall of X. dendrorhous were developed. To measure the degrees of cell wall degradation, the carotenoid extractability (extracted carotenoid by acetone/total carotenoid) unit was used. Treatment with HCl (0.2 M, 9 h, 90 °C) followed by neutralization to pH 3 by NaOH and spray-drying increased carotenoid extractability to 100% with minimal decomposition.     

Biorefinery approach and environment-friendly extraction for sustainable production of astaxanthin from marine wastes

Microorganisms (microalgae and fungi) are currently the main sources of astaxanthin; however, this carotenoid also accumulates in crustaceans, salmonids, and birds. Seafood (derived from marine animals) processing wastes are significant sources of astaxanthin and can be employed as feed and for nutraceutical applications, where shrimp wastes are the most exploited seafood industry waste employed for astaxanthin extraction. This review discusses different sources, efficient environment-friendly extraction methods employed for astaxanthin extraction, biorefinery approaches for efficient extraction and future aspects of the application of these waste sources for commercial preparation of astaxanthin complexes. It also includes a brief overview of the advantages, disadvantages, and challenges for obtaining astaxanthin from various sources and various case scenarios integrating different biorefinery approaches.     

Unique carotenoid lactoside, P457, in Symbiodinium sp. of dinoflagellate

The dinoflagellates are a large group of unicellular alge in marine and fresh water. Some are an endosymbiont of marine animals. Photosynthetic dinoflagellates have peridinin, a light-harvesting carotenoid. In addition, a unique carotenoid, P457, was found from Amphinidium. The presence of P457 in Symbiodinium derived from marine animals has not been reported. We reconfirmed the molecular structure of P457, a neoxanthin-like carotenoid with an aldehyde group and a lactoside, from Symbiodinium sp. NBRC 104787 isolated from a sea anemone. In addition, we investigated the distribution of P457 and peridinin in various Symbiodinium and scleractinian coral species, and possible biosynthetic pathways of these carotenoids are proposed.     

Biomineralizations: insights and prospects from crustaceans

For growing, crustaceans have to molt cyclically because of the presence of a rigid exoskeleton. Most of the crustaceans harden their cuticle not only by sclerotization, like all the arthropods, but also by calcification. All the physiology of crustaceans, including the calcification process, is then linked to molting cycles. This means for these animals to find regularly a source of calcium ions quickly available just after ecdysis. The sources of calcium used are diverse, ranging from the environment where the animals live to endogenous calcium deposits cyclically elaborated by some of them. As a result, crustaceans are submitted to an important and energetically demanding calcium turnover throughout their life. The mineralization process occurs by precipitation of calcium carbonate within an organic matrix network of chitin-proteins fibers. Both crystalline and stabilized amorphous polymorphs of calcium carbonate are found in crustacean biominerals. Furthermore, Crustacea is the only phylum of animals able to elaborate and resorb periodically calcified structures. Notably for these two previous reasons, crustaceans are more and more extensively studied and considered as models of choice in the biomineralization research area.     

Relationships between dietary carotenoids, body tissue carotenoids, parasite burden, and health state in wild mallard (Anas platyrhynchos) ducklings

While rodents and other mammals have traditionally served as models for studying carotenoid physiology, many wild animals from a variety of other taxa utilize carotenoids for self-maintenance and reproduction and accumulate far greater concentrations than those found in mammals. Though we have basic understandings of the control and value of carotenoids in some wild animal systems, many gaps remain. For example, parasites and pathogens impose severe survival constraints on free-ranging organisms, but little is known of how carotenoids work in concert with the immune system to combat natural infectious challenges. Furthermore, due to the high mortality rate from which many young animals suffer, health and carotenoid status during the early stages of development may be critical to survival. The relative importance of dietary versus physiological mechanisms for carotenoid uptake has also been understudied in the wild. To begin to shed light on these issues, we studied relationships between dietary and tissue carotenoids, hematological immune parameters, and endoparasitism of wild mallard (Anas platyrhynchos) ducklings at a variety of ages. Lutein, zeaxanthin, β-cryptoxanthin, β-carotene, and canthaxanthin were the most common carotenoids in liver, plasma, and gut contents. We found that, early in development (when food intake is limited), carotenoids were comparatively concentrated in internal tissue (e.g., liver), presumably a carry-over from maternal contributions in yolk, but as ducklings approached independence (and increasingly fed on their own) concentrations were greatest in gut contents. Canthaxanthin concentrations were lower in the plasma and liver of older individuals compared to younger ducklings, even though gut canthaxanthin concentration did not change with age. Additionally, β-carotene was nearly absent from circulation, despite moderate levels within the gut, suggesting a high rate of conversion to retinol. Using principal components analysis, we revealed a correlation between an increased ability to assimilate dietary carotenoids and lower levels of chronic stress (as assessed by lower heterophil-to-lymphocyte ratios) and a correlation between a reduced carotenoid status and increased investment in the immune system (as assessed by higher total leukocyte count). We also found that individuals without parasites had an overall reduced carotenoid status. Thus, we demonstrate age-specific differences in carotenoid allocation in growing animals from a precocial bird species and provide correlational evidence that parasitism and health in wild animals are related to carotenoid status and assimilation ability.     

Storage of Carotenoids in Crustaceans as an Adaptation to Modulate Immunopathology and Optimize Immunological and Life‐History Strategies

Why do some invertebrates store so much carotenoids in their tissues? Storage of carotenoids may not simply be passive and dependent on their environmental availability, as storage variation exists at various taxonomic scales, including among individuals within species. While the strong antioxidant and sometimes immune‐stimulating properties of carotenoids may be beneficial enough to cause the evolution of features improving their assimilation and storage, they may also have fitness downsides explaining why massive carotenoid storage is not universal. Here, the functional and ecological implications of carotenoid storage for the evolution of invertebrate innate immune defenses are examined, especially in crustaceans, which massively store carotenoids for unclear reasons. Three testable hypotheses about the role of carotenoid storage in immunological (resistance and tolerance) and life‐history strategies (with a focus on aging) are proposed, which may ultimately explain the storage of large amounts of these pigments in a context of host–pathogen interactions.     

Carotenoprotein complexes in entomostracan crustaceans (Streptocephalus dichotomus and Moina micrura)

The carotenoprotein complexes of a freshwater fairy shrimp (Streptocephalus dichotomus) and a daphnid (Moina micrura) were characterised and compared. Based on thin layer chromatography and mass spectral analysis, a variety of cartenoprotein complexes such as astaxanthin, canthaxanthin, antheraxanthin, lutein, beta-cryptoxanthin and violaxanthin were found. Both crustaceans had astaxanthin and canthaxanthin as predominant prosthetic groups. Amino acid analysis of the complexes further revealed high levels of asparagine, glutamine and glycine in both species. Our study highlights the presence of naturally available carotenoid species in both crustaceans and their possible inter-conversion in anostracans.     

Surrogate biochemistry: use of <Emphasis Type="Italic">Escherichia coli</Emphasis> to identify plant cDNAs that impact metabolic engineering of carotenoid accumulation

Carotenoids synthesized in plants but not animals are essential for human nutrition. Therefore, ongoing efforts to metabolically engineer plants for improved carotenoid content benefit from the identification of genes that affect carotenoid accumulation, possibly highlighting potential challenges when pyramiding traits represented by multiple biosynthetic pathways. We employed a heterologous bacterial system to screen for maize cDNAs encoding products that alter carotenoid accumulation either positively or negatively. Genes encoding carotenoid biosynthetic enzymes from the bacterium Erwinia uredovora were introduced into Escherichia coli cells that were subsequently transfected with a maize endosperm cDNA expression library; and these doubly transformed cells were then screened for altered carotenoid accumulation. DNA sequencing and characterization of one cDNA class conferring increased carotenoid content led to the identification of maize cDNAs encoding isopentenyl diphosphate isomerase. A cDNA that caused a reduced carotenoid content in E. coli was also identified. Based on DNA sequence analysis, DNA hybridization, and further functional testing, this latter cDNA was found to encode the small subunit of ADP-glucose pyrophosphorylase, a rate-controlling enzyme in starch biosynthesis that has been of interest for enhancing plant starch content.     

Comparative Toxicity of Gonyaulax monilata and Gymnodinium breve to Annelids, Crustaceans, Molluscs and a Fish

SYNOPSIS. Toxicities of laboratory cultures of the dinoflagellates Gonyaulax monilata and Gymnodinium breve were compared using fish, annelids, crustaceans and molluscs. Toxicity was evaluated as mortality over a 48-hr test period in 100%, 75% 50%, 25% and 10% concentrations of dinoflagellate cultures. Responses of the assay animals indicated that the dinoflagellates do not produce the same toxins. Effects of the toxins seemed uniform within each major group of animals tested. Fish were most sensitive to both G. breve and Gonyaulax monilata; crustaceans were resistant to both; annelids and molluscs were more sensitive to G. monilata than to G. breve.     

Carotenoid scarcity, synthetic pteridine pigments and the evolution of sexual coloration in guppies (Poecilia reticulata).

Carotenoid-based sexual coloration is the classic example of an honest signal of mate quality. Animals cannot synthesize carotenoid pigments and ultimately depend on dietary sources. Thus, in carotenoid-poor environments, carotenoid coloration may be a direct indicator of foraging ability and an indirect indicator of health and vigour. Carotenoid coloration may also be affected, more directly, by parasites in some species. Carotenoids are not, however, the only conspicuous pigments available to animals. Pteridine pigments, with similar spectral properties, are displayed in the exoskeletons and wings of insects, the irides of birds and the skins of fishes, lizards and amphibians. Unlike carotenoids, pteridines are synthesized de novo by animals. We report that the orange spots that male guppies (Poecilia reticulata) display to females contain red pteridine pigments (drosopterins) in addition to carotenoids. We also examined the relationship between drosopterin production by males and carotenoid availability in the field. The results contrasted sharply with the hypothesis that males use drosopterins to compensate for carotenoid scarcity: males used more, not less, drosopterins in streams with higher carotenoid availability. The positive association between drosopterin use and carotenoid availability could reflect the costs of drosopterin synthesis or it could be a consequence of females preferring a particular pigment ratio or hue. Male guppies appear to use drosopterin pigments in a manner that dilutes, but does not eliminate, the indicator value of carotenoid coloration.     

Salinity-induced changes in the fine structure of the gills of the semiterrestrial estuarian crab, Uca uruguayensis (Nobili, 1901) (Decapoda, Ocypodidae)

The posterior gills of Uca uruguayensis are mostly lined with a thick tissue which presents the characteristics of a typical salt-transporting epithelium. Electron microscope analysis of gill tissue from crabs acclimated to both low (2.5 per thousand) and high (44 per thousand) salinity showed significant development of the basolateral membrane interdigitations with numerous mitochondria and conspicuous apical membrane infoldings. In high-salinity acclimated crabs, the basolateral interdigitations extended to the apical membrane. Under these conditions, apical infoldings were expanded laterally (forming wide subcuticular spaces), while the apical infoldings of low-salinity adapted animals appeared as regular leaflets. Septate desmosomes were also much more developed in low-salinity exposed animals than in those kept under high-salinity conditions. These morphological observations were analyzed for correlation with the currently-accepted ion hyporegulation model for crustaceans, which is mainly based on transcellular sodium flow. In this study, we propose an ion hyporegulation model involving apical paracellular sodium flux.     

Ultrastructure of the peduncular aesthetascs of Speleonectes tanumekes (Crustacea, Remipedia) in comparison with crustaceans from stressful environments

Abstract. Osmotic stress associated with the freshwater environment and desiccation stress associated with the terrestrial environment may have a shortening effect on the length of the innervation of crustacean aesthetascs. Physical stress of the littoral environment may have a similar effect on the length of the cuticular portion of aesthetascs. The aesthetascs of crustaceans that inhabit these environments share a similar ultrastructural feature, which may help animals cope with these environmental stresses. This ultrastructural feature, the position of the basal bodies proximal to the lumen of the aesthetasc, is absent from the aesthetascs of crustaceans that occur in the typical marine environment. Interestingly, the ultrastructural feature associated with these stressful habitats is present in the peduncular aesthetascs of the remipede Speleonectes tanumekes , even though the environmental stresses that may invoke the reduction of aesthetascs are absent in the marine-cave environment where this animal occurs. The importance of the sensitivity of aesthetascs for survival in this lightless environment may result in a selective pressure that favors basal bodies to be positioned proximal to the lumen of the aesthetasc.     

Pinching forces in crayfish and fiddler crabs,and comparisons with the closing forces of other animals

The pinching forces of crustaceans are in many respects analogous to the biting forces of vertebrates. We examined the effects of body size and chelae size and shape, on the closing forces of the fiddler crab, Uca pugilator, and the crayfish, Procambarus clarkii. We hypothesized that the allometric relationships would be similar among species, and comparable to those reported for other decapod crustaceans. We further hypothesized that the scaling of the closing forces of crustaceans, with respect to body size and with the geometry of the pinching or biting structures, would be similar to that of vertebrates. We found that pinching forces increased with body mass, claw dimensions, and claw mass in U. pugilator, but only with claw height and claw mass in P. clarkii. Contraction time increased with body mass for both species combined, whereas contraction speed decreased. Pooled data for these and 17 other species of decapod crustacean revealed a positive correlation between the pinching force and body mass with a scaling exponent of 0.71. These data are remarkably comparable to the values on closing forces of vertebrate jaws, with the pooled data having a scaling exponent of 0.58, slightly below the value of 0.67 predicted for geometric similarity. Maximum closing forces vary tremendously among both crustaceans and animals in general, with body size and food habits being among the most important determining factors.     

Some studies on the biosynthesis of ubiquinone, isoprenoid alcohols, squalene and sterols by marine invertebrates

The ability of fourteen marine invertebrates to utilize [(14)C]mevalonate for the biosynthesis of isoprenoid compounds was investigated. Several of the animals, in particular crustaceans, bivalve molluscs, a coelenterate and a sponge, were unable to synthesize squalene and sterols, whereas gastropod molluscs, echinoderms, an annelid and a sponge could. Regardless of sterol-synthesizing ability the animals (with the exception of a sponge) always made dolichol and ubiquinone, and thus a specific block in squalene and sterol synthesis was indicated in some animals. Radioactivity accumulated in relatively large amounts in farnesol and geranylgeraniol in those animals incapable of making sterols.     

A carotenoid oxygenase is responsible for muscle coloration in scallop

As lipid microconstituents mainly of plant origin, carotenoids are essential nutrients for humans and animals, and carotenoid coloration represents an important meat quality parameter for many farmed animals. Currently, the mechanism of carotenoid bioavailability in animals is largely unknown mainly due to the limited approaches applied, the shortage of suitable model systems and the restricted taxonomic focus. The mollusk Yesso scallop (Patinopecten yessoensis) possessing orange adductor muscle with carotenoid deposition, provides a unique opportunity to research the mechanism underlying carotenoid utilization in animals. Herein, through family construction and analysis, we found that carotenoid coloration in scallop muscle is inherited as a recessive Mendelian trait. Using a combination of genomic approaches, we mapped this trait onto chromosome 8, where PyBCO-like 1 encoding carotenoid oxygenase was the only differentially expressed gene between the white and orange muscles (FDR = 2.75E-21), with 11.28-fold downregulation in the orange muscle. Further functional assays showed that PyBCO-like 1 is capable of degrading β-carotene, and inhibiting PyBCO-like 1 expression in the white muscle resulted in muscle coloration and carotenoid deposition. In the hepatopancreas, which is the organ for digestion and absorption, neither the scallop carotenoid concentration nor PyBCO-like 1 expression were significantly different between the two scallops. These results indicate that carotenoids could be taken up in both white- and orange-muscle scallops and then degraded by PyBCO-like 1 in the white muscle. Our data suggest that PyBCO-like 1 is the essential gene for carotenoid metabolism in scallop muscle, and its downregulation leads to carotenoid deposition and muscle coloration.     

Changes of major carotenoids in gonads of sea urchins (Strongylocentrotus intermedius and S. nudus) at maturation

Changes in the carotenoid content in gonads of two sea urchins species were investigated during maturation. The content of echinenones and carotenes, the two major carotenoid fractions in gonads, is highest for Strongylocentrotus intermedius at the spawning gametogenic stage of gonad maturation for both sexes. For S. nudus, the content of these pigments is highest at stages of active gametogenesis and spawning for males and at the growth stage for females. A comparison of the carotenoid content dynamics during maturation of gonads for males, females and animals at the resting (sexual inactivity) stage was also carried out.     

Association Mapping of Total Carotenoids in Diverse Soybean Genotypes Based on Leaf Extracts and High-Throughput Canopy Spectral Reflectance Measurements

Carotenoids are organic pigments that are produced predominantly by photosynthetic organisms and provide antioxidant activity to a wide variety of plants, animals, bacteria, and fungi. The carotenoid biosynthetic pathway is highly conserved in plants and occurs mostly in chromoplasts and chloroplasts. Leaf carotenoids play important photoprotective roles and targeted selection for leaf carotenoids may offer avenues to improve abiotic stress tolerance. A collection of 332 soybean [Glycine max (L.) Merr.] genotypes was grown in two years and total leaf carotenoid content was determined using three different methods. The first method was based on extraction and spectrophotometric determination of carotenoid content (eCaro) in leaf tissue, whereas the other two methods were derived from high-throughput canopy spectral reflectance measurements using wavelet transformed reflectance spectra (tCaro) and a spectral reflectance index (iCaro). An association mapping approach was employed using 31,253 single nucleotide polymorphisms (SNPs) to identify SNPs associated with total carotenoid content using a mixed linear model based on data from two growing seasons. A total of 28 SNPs showed a significant association with total carotenoid content in at least one of the three approaches. These 28 SNPs likely tagged 14 putative loci for carotenoid content. Six putative loci were identified using eCaro, five loci with tCaro, and nine loci with iCaro. Three of these putative loci were detected by all three carotenoid determination methods. All but four putative loci were located near a known carotenoid-related gene. These results showed that carotenoid markers can be identified in soybean using extract-based as well as by high-throughput canopy spectral reflectance-based approaches, demonstrating the utility of field-based canopy spectral reflectance phenotypes for association mapping.     

Carotenoids, immunocompetence, and the information content of sexual colors: an experimental test

Many male birds use carotenoid pigments to acquire brilliant colors that advertise their health and condition to prospective mates. The direct means by which the most colorful males achieve superior health has been debated, however. One hypothesis, based on studies of carotenoids as antioxidants in humans and other animals, is that carotenoids directly boost the immune system of colorful birds. We studied the relationship between carotenoid pigments, immune function, and sexual coloration in zebra finches (Taeniopygia guttata), a species in which males incorporate carotenoid pigments into their beak to attract mates. We tested the hypotheses that increased dietary carotenoid intake enhances immunocompetence in male zebra finches and that levels of carotenoids circulating in blood, which also determine beak coloration, directly predict the immune response of individuals. We experimentally supplemented captive finches with two common dietary carotenoid pigments (lutein and zeaxanthin) and measured cell-mediated and humoral immunity a month later. Supplemented males showed elevated blood-carotenoid levels, brighter beak coloration, and increased cell-mediated and humoral immune responses than did controls. Cell-mediated responses were predicted directly by changes in beak color and plasma carotenoid concentration of individual birds. These experimental findings suggest that carotenoid-based color signals in birds may directly signal male health via the immunostimulatory action of ingested and circulated carotenoid pigments.