Detrimental effects of carotenoid pigments: the dark side of bright coloration

Carotenoid pigments produce yellow, orange, and red integumentary color displays that can serve as reliable signals of health and condition. In many birds and fish, individuals gain competitive or mating advantages by ingesting and utilizing large quantities of carotenoid pigments. Carotenoid pigments serve as antioxidants, performing important functions as free-radical scavengers. The beneficial effects of carotenoid pigments are well documented, but rarely have researchers considered potential detrimental effects of high-level accumulation of carotenoids. We maintained American goldfinches (Carduelis tristis) on high- or low-carotenoid diets through molt and tested for damage to the liver and skeletal muscle. High intake of carotenoids had no measurable effect on liver enzymes but caused an increase in creatine kinase, an indicator of skeletal muscle breakdown, and a reduction in vertical flight performance, a measure of skeletal muscle integrity. The detrimental effects of high-level carotenoid accumulation were approximately equivalent to the negative effects of removing carotenoids from the diet. The adverse effects observed in this study have important implications for theories of the function and evolution of colorful plumage.     

Carotenoid coloration is related to fat digestion efficiency in a wild bird

Some of the most spectacular visual signals found in the animal kingdom are based on dietarily derived carotenoid pigments (which cannot be produced de novo), with a general assumption that carotenoids are limited resources for wild organisms, causing trade-offs in allocation of carotenoids to different physiological functions and ornamentation. This resource trade-off view has been recently questioned, since the efficiency of carotenoid processing may relax the trade-off between allocation toward condition or ornamentation. This hypothesis has so far received little exploratory support, since studies of digestive efficiency of wild animals are limited due to methodological difficulties. Recently, a method for quantifying the percentage of fat in fecal samples to measure digestive efficiency has been developed in birds. Here, we use this method to test if the intensity of the carotenoid-based coloration predicts digestive efficiency in a wild bird, the house finch (Haemorhous mexicanus). The redness of carotenoid feather coloration (hue) positively predicted digestion efficiency, with redder birds being more efficient at absorbing fats from seeds. We show for the first time in a wild species that digestive efficiency predicts ornamental coloration. Though not conclusive due to the correlative nature of our study, these results strongly suggest that fat extraction might be a crucial but overlooked process behind many ornamental traits.     

Polymorphic ROS scavenging revealed by CCCP in a lizard

Ingestion of antioxidants has been argued to scavenge circulating reactive molecules (e.g., free radicals), play a part in mate choice (by mediating access to this important resource), and perhaps increase life span. However, recent work has come to question these relationships. We have shown elsewhere in the polychromatic lizard, Ctenophorus pictus, that diet supplementation of carotenoids as antioxidants does not depress circulating natural reactive oxygen species (ROS) levels and leads to no corresponding improvement of color traits. However, a much stronger test would be to experimentally manipulate the ROS levels themselves and assess carotenoid-induced ROS depression. Here, we achieve this by using carbonyl cyanide 3-chlorophenylhydrazone, which elevates superoxide (SO) formation approximately threefold at 10 μM in this model system. We then look for depressing effects on ROS of the carotenoids in order to assess whether ‘super-production’ of SO makes carotenoid effects on elevated ROS levels detectable. The rationale for this treatment was that if not even such elevated levels of SO are reduced by carotenoid supplementation, the putative link carotenoids, ROS depression, and mate quality (in terms of antioxidant capacity) is highly questionable. We conclude that there is no significant effect of carotenoids on mean SO levels even at the induced ROS levels. However, our results showed a significant interaction effect between carotenoid treatment and male color, with red males having higher ROS levels than yellow males. We suggest that this may be because different pigments are differently involved in the generation of the integumental colors in the two morphs with concomitant effects on ROS depletion depending on carotenoid uptake or allocation to coloration and antioxidation.     

外源一氧化氮对亚心形扁藻的生长效应及光谱研究

对海洋绿藻亚心形扁藻(Platymonas subcordiformis)分别进行了不同一氧化氮(NO)浓度和光照强度的培养实验,并进行了叶绿素a和类胡萝卜素含量测定,及室温吸收光谱和荧光光谱测定.结果表明,不同NO浓度、不同加入方式对亚心形扁藻生长有明显不同的促进或抑制作用,NO对亚心形扁藻生长的影响存在阈值.不同浓度NO在不同光照下对微藻生长的作用是一致的,外源NO可以弥补弱光照对藻生长的限制.NO对亚心形扁藻光合色素含量的影响和对藻密度的影响是一致的.NO对藻细胞中叶绿素复合蛋白组成没有显著影响,但对色素含量及其相对组成会产生一系列影响.NO能使藻细胞激发能的传递效率提高,从而提高光合作用速率,表现为细胞生长加快,藻细胞生物量增加.     

Maternally derived carotenoid pigments affect offspring survival, sex ratio, and sexual attractiveness in a colorful songbird

In egg-laying animals, mothers can influence the development of their offspring via the suite of biochemicals they incorporate into the nourishing yolk (e.g. lipids, hormones). However, the long-lasting fitness consequences of this early nutritional environment have often proved elusive. Here, we show that the colorful carotenoid pigments that female zebra finches (Taeniopygia guttata) deposit into egg yolks influence embryonic and nestling survival, the sex ratio of fledged offspring, and the eventual ornamental coloration displayed by their offspring as adults. Mothers experimentally supplemented with dietary carotenoids prior to egg-laying incorporated more carotenoids into eggs, which, due to the antioxidant activity of carotenoids, rendered their embryos less susceptible to free-radical attack during development. These eggs were subsequently more likely to hatch, fledge offspring, produce more sons than daughters, and produce sons who exhibited more brightly colored carotenoid-based beak pigmentation. Provisioned mothers also acquired more colorful beaks, which directly predicted levels of carotenoids found in eggs, thus indicating that these pigments may function not only as physiological ‘damage-protectants’ in adults and offspring but also as morphological signals of maternal reproductive capabilities.     

Plumage coloration and nutritional condition in the great tit<Emphasis Type="Italic"> Parus major</Emphasis>: the roles of carotenoids and melanins differ

The size and coloration of some body characters seem to influence mate choice in many species. Most animal colours are either structural or based on melanin or carotenoid pigments. It has recently been suggested that carotenoid-based or structural coloration may be a condition-dependent trait, whereas melanin-based coloration is not; a difference that may be highly relevant when studying the evolution of multiple mating preferences. We tested this hypothesis in the great tit (Parus major). The size of the melanin breast band was not correlated to nutritional condition as estimated by the rate of tail growth (ptilochronology), controlling for locality, age, sex, year and season effects. However, the correlation was significant for the hue of yellow breast (carotenoid-based coloration), and the slopes of the regressions of the two pigments to growth bars differed significantly. These results suggest that the expression of the two traits may be regulated by different mechanisms.     

Astaxanthin and papilioerythrinone in the skin of birds: a chromatic convergence of two metabolic routes with different precursors?

Carotenoids are organic pigments involved in several important physiological functions and may serve as indicators of individual quality in animals. These pigments are only obtained by animals from the diet, but they can be later transformed into other carotenoids by specific enzymatic reactions. The diet of farm-reared and probably wild red-legged partridges (Alectoris rufa) is mainly based on cereals that contain high levels of lutein and zeaxanthin. These two carotenoids are also predominant in internal tissues and blood of red-legged partridges. However, in their integuments, astaxanthin and papilioerythrinone (the last one identified in this work) are mainly present in their free form and esterified with fatty acids. According to available literature about carotenoid metabolism in animals, we propose that astaxanthin (λ _max?=?478 nm) and papilioerythrinone (λ _max?=?452–478 nm) are the result of a chromatic convergence of the transformation of dietary zeaxanthin and lutein, respectively. Moreover, the results obtained in this work provide the first identification by liquid chromatography coupled to accurate mass quadrupole time-of-flight mass spectrometer system of papilioerythrinone (m/z 581.3989 [M?+?H]⁺) in the skin (i.e., not feathers) of a vertebrate. Astaxanthin and papilioerythrinone are very close in terms of chemical structure and coloration, and the combination of these two keto-carotenoids is responsible for the red color of the ornaments in red-legged partridges.     

Taxonomy of oxalotrophic <Emphasis Type="Italic">Methylobacterium</Emphasis> strains

Most of the oxalotrophic bacteria are facultative methylotrophs and play important ecological roles in soil fertility and cycling of elements. This study gives a detailed picture of the taxonomy and diversity of these bacteria and provides new information about the taxonomical variability within the genus Methylobacterium. Twelve mesophilic, pink-pigmented, and facultatively methylotrophic oxalate-oxidizing strains were included in this work that had been previously isolated from the soil and some plant tissues by the potassium oxalate enrichment method. The isolates were characterized using biochemical tests, cellular lipid profiles, spectral characteristics of carotenoid pigments, G+C content of the DNA, and 16S rDNA sequencing. The taxonomic similarities among the strains were analyzed using the simple matching (S _SM) and Jaccard (S _J) coefficients, and the UPGMA clustering algorithm. The phylogenetic position of the strains was inferred by the neighbor-joining method on the basis of the 16S rDNA sequences. All isolates were Gram-negative, facultatively methylotrophic, oxidase and catalase positive, and required no growth factors. Based on the results of numerical taxonomy, the strains formed four closely related clusters sharing ≥85% similarity. Analysis of the 16S rDNA sequences demonstrated that oxalotrophic, pink-pigmented, and facultatively methylotrophic strains could be identified as members of the genus Methylobacterium. Except for M. variabile and M. aquaticum, all of the Methylobacterium type strains tested had the ability of oxalate utilization. Our results indicate that the capability of oxalate utilization seems to be an uncommon trait and could be used as a valuable taxonomic criterion for differentiation of Methylobacterium species.     

Frequent summer nuptial flights of ants provide a primary food source for bats

In many ant species, nuptial flight tends to be short in time and assumed to be synchronous across a large area. Here, we report that, in the upper Jordan Valley, northern Israel, massive nuptial flights of Carpenter ants (Camponotus sp.) occur frequently throughout the summer, and their alates form up to 90% of the diet of the greater mouse-tailed bat (Rhinopoma microphyllum) during this period. This fat and protein-rich diet enables female bats to lactate during summer, and the large amount of fat that both sexes accumulate may serve as an energy source for their following winter hibernation and posthibernation mating in early spring (March–April). We suggest that the annual movement of these bats to the Mediterranean region of Israel may have evolved in order to enable them to exploit the extremely nutritious forms of ant alates when the bats’ energetic demands are highest.     

Integument coloration signals reproductive success,heterozygosity, and antioxidant levels in chick-rearing black-legged kittiwakes

Carotenoid pigments are important for immunity and as antioxidants, and carotenoid-based colors are believed to provide honest signals of individual quality. Other colorless but more efficient antioxidants such as vitamins A and E may protect carotenoids from bleaching. Carotenoid-based colors have thus recently been suggested to reflect the concentration of such colorless antioxidants, but this has rarely been tested. Furthermore, although evidence is accruing for multiple genetic criteria for mate choice, carotenoid-based colors have rarely been shown to reflect both phenotypic and genetic quality. In this study, we investigated whether gape, tongue, eye-ring, and bill coloration of chick-rearing black-legged kittiwakes Rissa tridactyla reflected circulating levels of carotenoids and vitamins A and E. We further investigated whether integument coloration reflected phenotypic (body condition and fledging success) and genetic quality (heterozygosity). We found that the coloration of fleshy integuments was correlated with carotenoid and vitamin A levels and fledging success but only in males. Furthermore, the coloration of tongue and eye-ring was correlated with heterozygosity in both males and females. Integument colors might therefore be reliable signals of individual quality used by birds to adjust their parental care during the chick-rearing period.     

Melanin- and carotenoid-dependent signals of great tits (<Emphasis Type="Italic">Parus major</Emphasis>) relate differently to metal pollution

Due to their high phenotypic plasticity, the expression of secondary sexual characteristics is particularly sensitive to stress. Here, we investigated the expression of two conspicuous visual signals in great tits (Parus major) in a metal pollution gradient. In three study sites with marked differences in metal contamination (mainly lead, cadmium, copper and zinc), we compared melanin and carotenoid colouration of great tits. While carotenoid colouration (yellow breast) was negatively related to metal pollution, the size of a melanin trait (breast stripe) was larger in the most polluted sites. Environmental pollutants not only affect the expression of conspicuous signals but may even enhance, directly or indirectly, a signal of male quality such as breast stripe. Our results also support the multiple messages hypothesis predicting that different signals highlight different aspects of geno- and phenotypic condition of the bearer.     

Biological treatment of chicken feather waste for improved biogas production

A two-stage system was developed which combines the biological degradation of keratin-rich waste with the production of biogas. Chicken feather waste was treated biologically with a recombinant Bacillus megaterium strain showing keratinase activity prior to biogas production. Chopped, autoclaved chicken feathers (4%, W/V) were completely degraded, resulting in a yellowish fermentation broth with a level of 0.51 mg/mL soluble proteins after 8 days of cultivation of the recombinant strain. During the subsequent anaerobic batch digestion experiments, methane production of 0.35 Nm³/kg dry feathers (i.e., 0.4 Nm³/kg volatile solids of feathers), corresponding to 80% of the theoretical value on proteins, was achieved from the feather hydrolyzates, independently of the pre-hydrolysis time period of 1, 2 or 8 days. Cultivation with a native keratinase producing strain, Bacillus licheniformis resulted in only 0.25 mg/mL soluble proteins in the feather hydrolyzate, which then was digested achieving a maximum accumulated methane production of 0.31 Nm³/kg dry feathers. Feather hydrolyzates treated with the wild type B. megaterium produced 0.21 Nm³ CH₄/kg dry feathers as maximum yield.     

氧化石墨烯对淡水微藻生长及生物活性物质的影响

为掌握氧化石墨烯（GO）的水环境风险,以斜生栅藻（Scenedesmus obliquus）和湖泊微拟球藻（Nannochloropsis limnetica）为研究对象,探究了GO对淡水微藻生长及其生物活性物质（碳水化合物、总蛋白质、总脂）的影响.结果表明,GO对2种微藻具有中等毒性,72h EC₅₀值分别为25.63和48.44mg/L.透射电镜（TEM）观察发现,GO纳米片层既能附着于藻细胞表面也能进入藻细胞内部,造成藻细胞超微结构明显变化,包括：质壁分离;叶绿体收缩;淀粉粒数量减少甚至消失.较低浓度（10mg/L）GO会促进微藻中光合色素（叶绿素a、叶绿素b、类胡萝卜素）合成;而较高浓度（100mg/L）GO暴露下,2种微藻的类胡萝卜素和斜生栅藻叶绿素a的含量显著降低.2种浓度的GO总体上刺激了藻细胞内生物活性物质的合成,这是污染胁迫下的一种主动防御机制;而较高浓度GO造成碳水化合物含量显著降低,可能原因是细胞中储能物质由淀粉向中性脂转化.     

城市大气环境下绿化植物叶片比叶重和光合色素含量

研究了大叶女贞(Ligustrum lucidum Ait.)和小叶女贞(Ligustrum quihoui Carr.)在不同大气环境条件下叶片中Pb、Cd、叶绿素a (Chl a)、叶绿素b(Chl b)、类胡萝卜素(Car)的含量及比叶重(LMA),分析了因素之间的相互关系.不同环境条件下叶片中Pb、Cd、Chl a、Chl b、Chl (a+b)、Car的含量及LMA具有显著差异.随污染程度的加剧,叶片中Pb、Cd的含量、LMA和Chl a/Chl b升高,而Chl a、Chl b和Chl (a+b)含量则呈现相反的变化趋势.大叶女贞叶片中Car含量变化不明显,而小叶女贞叶片中Car含量在污染严重的采样点明显较其他采样点高.2种植物叶片中Pb、Cd的含量对叶片LMA和光合色素含量均具有明显影响,Chl a、Chl b及Chl (a+b)含量与叶片中Pb、Cd的含量呈显著负相关(P<0.05).叶片中Cd含量的增大导致Car含量增大,而Pb的作用与物种有关.叶片中Pb含量的增大导致2物种LMA增大,而Cd含量导致大叶女贞LMA增大而小叶女贞LMA变化不明显.污染胁迫下植物叶片会增大LMA、Chl a/Chl b,减小Chl a、Chl b和Chl (a+b)含量,据此可对大气污染进行生物监测.     

Assortative mating by carotenoid-based plumage colour: a quality indicator in American goldfinches,<Emphasis Type="Italic"> Carduelis tristis</Emphasis>

We investigated the mating patterns in 22 breeding pairs of wild American goldfinches (Carduelis tristis) with respect to their body size, condition, and carotenoid-based, yellow plumage colour. Using reflectance spectrometry, we objectively quantified plumage colours across the bird-visible wavelengths, revealing a unexpected UV peak in the reflectance spectrum from yellow feathers. We summarized our colour measurements using a principal components analysis to create a single variable, carotenoid PC1, that represents the intensity of this carotenoid-based yellow colour, a measure of phenotypic quality in this species. We found no evidence of assortative mating with respect to measures of body size or condition but there was positive assortative mating by carotenoid PC1, such that the yellow plumage colours of males and females were significantly correlated within pairs. We argue that the yellow carotenoid coloration of goldfinches may be important in mutual mate choice and, thus, that sexual selection in this species may act upon female ornamentation, as well as the more obvious plumage signals of males. Because assortative mating results in an increase in genetic variance, we suggest that this might be a mechanism that maintains variance in ornamental traits in spite of the variance-eroding effects of sexual selection.     

Carotenoid maintenance handicap and the physiology of carotenoid-based signalisation of health

Despite a reasonable scientific interest in sexual selection, the general principles of health signalisation via ornamental traits remain still unresolved in many aspects. This is also true for the mechanism preserving honesty of carotenoid-based signals. Although it is widely accepted that this type of ornamentation reflects an allocation trade-off between the physiological utilisation of carotenoids (mainly in antioxidative processes) and their deposition in ornaments, some recent evidence suggests more complex interactions. Here, we further develop the models currently proposed to explain the honesty of carotenoid-based signalisation of heath status by adding the handicap principle concept regulated by testosterone. We propose that under certain circumstances carotenoids may be dangerous for the organism because they easily transform into toxic cleavage products. When reserves of other protective antioxidants are insufficient, physiological trade-offs may exist between maintenance of carotenoids for ornament expression and their removal from the body. Furthermore, we suggest that testosterone which enhances ornamentation by increasing carotenoid bioavailability may also promote oxidative stress and hence lower antioxidant reserves. The presence of high levels of carotenoids required for high-quality ornament expression may therefore represent a handicap and only individuals in prime health could afford to produce elaborate colourful ornaments. Although further testing is needed, this ‘carotenoid maintenance handicap’ hypothesis may offer a new insight into the physiological aspects of the relationship between carotenoid function, immunity and ornamentation.     

Effect of cadmium on photosynthetic pigments, lipid peroxidation, antioxidants, and artemisinin in hydroponically grown Artemisia annua

The effects of different cadmium (Cd) concentrations (0, 20, 60, and 100 μmol/L) on hydroponically grown Artemisia annua L. were investigated. Cd treatments applied for 0, 4, 12, 24, 72, 144, 216, and 336 hr were assessed by measuring the changes in photosynthetic pigments, electrolyte leakage, malondialdehyde (MDA) and antioxidants (ascorbic acid and glutathione), while the artemisinin content was tested after 0, 12, 144, 216, and 336 hr. A significant decrease was observed in photosynthetic pigment levels over time with increasing Cd concentration. Chlorophyll b levels were more affected by Cd than were chlorophyll a or carotenoid levels. The cell membrane was sensitive to Cd stress, as MDA content in all treatment groups showed insignificant differences from the control group, except at 12 hr treatment time. Ascorbic acid (AsA) content changed slightly over time, while glutathione (GSH) content took less time to reach a maximum as Cd concentration increased. Cd was found to promote synthesis and accumulation of artemisinin, especially at concentrations of 20 and 100 μmol/L. In conclusion, Cd stress can damage to photosynthetic pigments, and vigorously growing A. annua showed a strong tolerance for Cd stress. Appropriate amounts of added Cd aided synthesis and accumulation of artemisinin.     

Flower-visiting behavior of male bees is triggered by nectar-feeding insects

Bees are important pollinators for many flowering plants. Female bees are thought to be more effective pollinators than male bees because they carry much more pollen than males. Males of some solitary bee species are known to patrol near flowers that females visit. Because patrolling males visit flowers to mate or defend their territories, they may function as pollinators. However, the significance of patrolling males to pollination has not been studied. We studied males of a solitary bee, Heriades fulvohispidus (Megachilidae), patrolling near flowers and visiting flowers that attracted nectar-feeding insects, including conspecifics, on the Ogasawara (Bonin) Islands. To test the hypothesis that patrolling male bees may function as pollen vectors, we compared the frequency of visits by H. fulvohispidus to flowers of an endemic plant, Schima mertensiana (Theaceae); comparisons were made among flowers with a dead H. fulvohispidus, a dead beetle, a piece of plastic, and nothing (control flowers). Patrolling H. fulvohispidus more frequently visited flowers with a dead conspecific, a dead beetle, or a piece of plastic than the control flowers. Our experiment demonstrates that nectar-feeding insects (including conspecifics and other insects) enhance the flower-visiting frequency of patrolling H. fulvohispidus males on S. mertensiana flowers. Furthermore, we observed S. mertensiana pollen on patrolling males as well as females, suggesting that male bees may also function as pollen vectors.     

Bristle-like integumentary structures at the tail of the horned dinosaur Psittacosaurus

A specimen of the horned dinosaur Psittacosaurus from the early Cretaceous of China is described in which the integument is extraordinarily well-preserved. Most unusual is the presence of long bristle-like structures on the proximal part of tail. We interpret these structures as cylindrical and possibly tubular epidermal structures that were anchored deeply in the skin. They might have been used in display behavior and especially if one assumes that they were colored, they may have had a signal function. At present, there is no convincing evidence which shows these structures to be homologous to the structurally different integumentary filaments of theropod dinosaurs. Independent of their homology, however, the discovery of bristle-like structures in Psittacosaurus is of great evolutionary significance since it shows that the integumentary covering of at least some dinosaurs was much more complex than has ever been previously imagined.     

Immune challenges and visual signalling in tree frogs

In animals, mate-choice is often based on sexual signals that carry information and help the receiver make the best choice to improve the receiver’s fitness. Orange visual sexual signals have been hypothesised to carry immune information because they are often due to carotenoid pigments which are also involved in immunity response. Although many studies have focused on the direct relationships between coloration and immunocompetence, few studies have simultaneously studied immunocompetent response and coloration variation after an immune challenge. We tested this hypothesis on starved and ad libitum-fed males of the European tree frog Hyla arborea. Our results show that male coloration is not a reliable indicator of its immune response capacity in this species. However, after an immune challenge induced by a PHA (Phaseolus vulgaris phytohaemagglutinin) injection, starved males presented a significant coloration loss and this alteration was related to the immune response intensity. Taken together, these results suggest that the brighter (lighter) coloration may be used as a cue by female to exclude males with a recent immune challenge, due to diseases or parasites for example.