Validation model for Raman based skin carotenoid detection

Raman spectroscopy holds promise as a rapid objective non-invasive optical method for the detection of carotenoid compounds in human tissue in vivo. Carotenoids are of interest due to their functions as antioxidants and/or optical absorbers of phototoxic light at deep blue and near UV wavelengths. In the macular region of the human retina, carotenoids may prevent or delay the onset of age-related tissue degeneration. In human skin, they may help prevent premature skin aging, and are possibly involved in the prevention of certain skin cancers. Furthermore, since carotenoids exist in high concentrations in a wide variety of fruits and vegetables, and are routinely taken up by the human body through the diet, skin carotenoid levels may serve as an objective biomarker for fruit and vegetable intake. Before the Raman method can be accepted as a widespread optical alternative for carotenoid measurements, direct validation studies are needed to compare it with the gold standard of high performance liquid chromatography. This is because the tissue Raman response is in general accompanied by a host of other optical processes which have to be taken into account. In skin, the most prominent is strongly diffusive, non-Raman scattering, leading to relatively shallow light penetration of the blue/green excitation light required for resonant Raman detection of carotenoids. Also, sizable light attenuation exists due to the combined absorption from collagen, porphyrin, hemoglobin, and melanin chromophores, and additional fluorescence is generated by collagen and porphyrins. In this study, we investigate for the first time the direct correlation of in vivo skin tissue carotenoid Raman measurements with subsequent chromatography derived carotenoid concentrations. As tissue site we use heel skin, in which the stratum corneum layer thickness exceeds the light penetration depth, which is free of optically confounding chromophores, which can be easily optically accessed for in vivo RRS measurement, and which can be easily removed for subsequent biochemical measurements. Excellent correlation (coefficient R = 0.95) is obtained for this tissue site which could serve as a model site for scaled up future validation studies of large populations. The obtained results provide proof that resonance Raman spectroscopy is a valid non-invasive objective methodology for the quantitative assessment of carotenoid antioxidants in human skin in vivo.     

Diet quality and the attractiveness of male body odor

Human axillary sweat may provide information pertaining to genetic relatedness and health status. A significant contributor to good health, both in the short and longer term, is a diet rich in fruit and vegetables. In this study we tested whether dietary fruit and vegetable intake, assessed indirectly by skin spectrophotometry (assessing dietary carotenoid intakes) and subjectively by food frequency questionnaire, was associated with more pleasant smelling sweat. Male participants provided axillary sweat samples and dietary information. Female participants then evaluated these samples on several affective, qualitative and psychophysical dimensions. The skin spectrophotometry measure (CIELab b*), indicative of greater fruit and vegetable intake, was significantly associated with more pleasant smelling sweat (with more floral, fruity, sweet and medicinal qualities), independent of sweat intensity. Self-report dietary data revealed that fat, meat, egg and tofu intake was associated with more pleasant smelling sweat, and greater carbohydrate intake with stronger smelling less pleasant sweat. These data parallel facial judgments, in which yellower more carotenoid rich skin, is found to be more attractive.     

Cover Picture: J. Biophotonics 7/2012

The cover illustrates the measurement of human skin carotenoid levels based on pressure mediated reflection spectroscopy. Carotenoids have a widespread distribution in fruits and vegetables, are taken up through the diet, and play an important role in tissue health through their function as antioxidants. Their characteristic absorption in the blue/green spectral range can be quantified in human tissue such as the thumb and tracked upon dietary supplementation. (Picture: I. V. Ermakov and W. Gellermann, pp. 442–453, in this issue)     

Associations between eating frequency, adiposity, diet, and activity in 9-10 year old healthy-weight and centrally obese children

The rising prevalence of childhood obesity is a key public health issue worldwide. Increased eating frequency (EF) is one aspect of diet that has been beneficially associated with obesity, although the mechanisms are unclear. The aims of the current study were to determine whether increased EF was associated with improved adiposity in children, and if this was due to differences in dietary and activity behaviors. Cross-sectional data from 1,700 children aged 9-10 year were analyzed to examine the associations between EF, as estimated from diet diaries, measures of adiposity, and activity measured by accelerometer. Analyses were stratified by obesity status using waist-to-height ratio to define obesity as it has been shown to be a good predictor of adverse health outcomes. Mean EF was 4.3 occasions/day and after adjustment for underreporting, energy intake (EI), and activity significant relative mean differences of -2.4% for body weight (P = 0.001), -1.0% for BMI (P = 0.020), -33% for BMI z-score (P = 0.014), and -0.6% for waist circumference (P = 0.031) per increase in eating occasion were found in healthy-weight but not centrally obese children. Differences between the extreme quartiles of EF were observed for total fat intake at breakfast (-18%, P < 0.001), fruit and vegetables from snacks (201% healthy-weight and 209% centrally obese children, P < 0.01), and for healthy-weight children, vigorous activity (4%, P = 0.003). Increased EF was favorably associated with adiposity, diet quality, and activity behaviors in healthy-weight but not centrally obese children. Future obesity interventions should consider the mediating role of diet quality and activity in the relationship between EF and adiposity in children.     

Urinary Chromium Excretion in Response to an Insulin Challenge Is Not a Biomarker for Chromium Status

Over 50 years ago, chromium (Cr) was proposed to be an essential trace element; however, recent studies indicate that this status should be removed as the effects of Cr supplementation appear to be pharmacological rather than nutritional. The pharmacological basis for Cr's effects can explain the inability of investigators to discover a biomarker for Cr status. One potential biomarker has not been examined to date. Cr is known to be mobilized in the body in response to insulin (or insulin release in response to a glucose challenge), resulting in an increase in urinary Cr excretion. The magnitude of increase in urinary Cr loss as a function of dietary Cr intake was tested as a potential biomarker for Cr. Zucker lean rats housed in carefully controlled metal-free conditions were provided a series of purified diets containing variable Cr contents (from 16 μg/kg diet to 2,000 μg/kg) for 23 weeks. The 16 μg/kg diet contained less Cr than any diet examined to date. Urine samples were collected before and after insulin and glucose challenges (0, 2, 6, and 12 h postinjection). Urinary Cr levels were analyzed by the standard method of addition using graphite furnace atomic absorption. The rate of urinary Cr loss after a glucose or insulin challenge was found to not be dependent on the Cr content of the rats' diets. Blood iron levels of the rats were also measured to determine if the addition of Cr to the diet altered iron status. The Cr content of the diet was found to have no affect on blood iron levels. Overall, the study demonstrated that insulin-stimulated urinary Cr excretion cannot be used as a biomarker for Cr status.     

Photoperiod Regulates Lean Mass Accretion,but Not Adiposity,in Growing F344 Rats Fed a High Fat Diet

In this study the effects of photoperiod and diet, and their interaction, were examined for their effects on growth and body composition in juvenile F344 rats over a 4-week period. On long (16L:8D), relative to short (8L:16D), photoperiod food intake and growth rate were increased, but percentage adiposity remained constant (ca 3-4%). On a high fat diet (HFD), containing 22.8% fat (45% energy as fat), food intake was reduced, but energy intake increased on both photoperiods. This led to a small increase in adiposity (up to 10%) without overt change in body weight. These changes were also reflected in plasma leptin and lipid levels. Importantly while both lean and adipose tissue were strongly regulated by photoperiod on a chow diet, this regulation was lost for adipose, but not lean tissue, on HFD. This implies that a primary effect of photoperiod is the regulation of growth and lean mass accretion. Consistent with this both hypothalamic GHRH gene expression and serum IGF-1 levels were photoperiod dependent. As for other animals and humans, there was evidence of central hyposomatotropism in response to obesity, as GHRH gene expression was suppressed by the HFD. Gene expression of hypothalamic AgRP and CRH, but not NPY nor POMC, accorded with the energy balance status on long and short photoperiod. However, there was a general dissociation between plasma leptin levels and expression of these hypothalamic energy balance genes. Similarly there was no interaction between the HFD and photoperiod at the level of the genes involved in thyroid hormone metabolism (Dio2, Dio3, TSHβ or NMU), which are important mediators of the photoperiodic response. These data suggest that photoperiod and HFD influence body weight and body composition through independent mechanisms but in each case the role of the hypothalamic energy balance genes is not predictable based on their known function.     

Effect of Dietary Carotenoid Supplementation on Food Intake and Immune Function in a Songbird with no Carotenoid Coloration

Studies of ornamental carotenoid coloration suggest that animals may have evolved specialized mechanisms for maximizing color expression and advertising their potential worth as a mate. For example, when given a choice of foods, many carotenoid‐pigmented fishes and birds select the more colorful, presumably carotenoid‐rich foods, and then accumulate these pigments at high levels in both the integument and systemically, in order to boost their immune system and hence directly advertise their health state with their colors. The majority of animals, however, do not exhibit sexually selected carotenoid coloration, which raises the question of whether they still optimize pigment intake and allocation in ways that boost endogenous accumulation and health. We tested the effect of carotenoid supplementation on food intake, carotenoid accumulation in blood, and both innate and adaptive immunity in male society finches (Lonchura domestica) – a non‐carotenoid‐colored estrildid finch relative of the zebra finch (Taeniopygia guttata; a species in which males do display sexually attractive and health‐revealing carotenoid color). Males fed a carotenoid‐rich diet for 2 wk did not consume more food than control males. Still, consumption of the carotenoid‐rich diet for 2 wk significantly elevated circulating levels of carotenoids in blood in male society finches, yielding the potential for immune enhancement. In fact, carotenoid‐enriched finches performed significantly better than control birds in our assay of constitutive innate immunity (bacterial‐killing activity of whole blood), although not in our test of inducible adaptive immunity (response to a mitogenic challenge with phytohemagglutinin). These results suggest that affinities for carotenoid‐rich foods may be particular to species with sexually selected carotenoid pigmentation, but that, as in humans and other mammals (e.g. mice, rats) without carotenoid color, the immune‐boosting action of carotenoids is conserved regardless of the strength of sexual selection on pigment use.     

Fruit Juice Supplementation Alters Human Skin Antioxidant Levels <Emphasis Type="Italic">In Vivo</Emphasis>: Case Study of Korean Adults by Resonance Raman Spectroscopy

Carotenoids, found in many fruits and vegetables, are antioxidants that protect human skin from UV radiation. In humans, fruit and vegetable intake increases carotenoid contents in skin, which are conventionally assessed by invasive blood tests. In this study, 47 healthy Korean subjects (volunteers) consumed fruit juice containing tomato, apple, strawberry, or grape three times per week for 6 weeks. Skin antioxidant levels were measured by non-invasive resonance Raman spectroscopy. The correlation between skin carotenoid (SC) score with demographic data (age, height, weight) and juice supplementation and changes in SC scores among groups were analyzed. Variations in skin antioxidant levels increased with juice supplementation (p < 0.05). Fruit juice intake was significantly correlated with SC score, indicating increased skin antioxidant levels. Grape and tomato increased skin antioxidant levels and showed higher antioxidant activity than other fruits. Fruit juices containing high levels of carotenoids and antioxidants may provide modest benefits to human health.     

Beyond thriftiness: independent and interactive effects of genetic and dietary factors on variations in fat deposition and distribution across populations

The thrifty genotype hypothesis initiated speculation that feast and famine cycling throughout history may have led to group-specific alterations of the human genome, thereby augmenting the capacity for excessive fat mass accrual when immersed in the modern-day obesogenic environment. Contemporary work, however, suggests alternative mechanisms influencing fuel utilization and subsequent tissue partitioning to be more relevant in the etiology of population-based variation in adipose storage. The objective of this study was to evaluate the independent and interactive contribution of ancestral admixture as a proxy for population-based genetic variation and diet on adipose tissue deposition and distribution in peripubertal children and to identify differences in racial/ethnic and sex groups. Two-hundred seventy-eight children (53% male) aged 7-12 years, categorized by parental self-report as African- (n = 91), European- (n = 110), or Hispanic American (n = 77), participated. Ancestral genetic admixture was estimated using 140 ancestry informative markers. Body composition was evaluated by dual-energy X-ray absorptiometry; energy expenditure by indirect calorimetry and accelerometry; and diet by 24-h-recall. Admixture independently contributed to all adiposity parameters; i.e., estimates of European and Amerindian ancestries were positively associated with all adiposity parameters, whereas African genetic admixture was inversely associated with adiposity. In boys, energy intake was associated with adiposity, irrespective of macronutrient profile, whereas in girls, the relationship was mediated by carbohydrate. We also observed moderating effects of energy balance/fuel utilization of the interaction between ancestral genetic admixture and diet. Interactive effects of genetic and non-genetic factors alter metabolic pathways and underlie some of the present population-based differences in fat storage.     

Detrimental metabolic effects of combining long-term cigarette smoke exposure and high-fat diet in mice

Obesity and cigarette smoking are both important risk factors for insulin resistance, cardiovascular disease, and cancer. Smoking reduces appetite, which makes many people reluctant to quit. Few studies have documented the metabolic impact of combined smoke exposure (se) and high-fat-diet (HFD). Neuropeptide Y (NPY) is a powerful hypothalamic feeding stimulator that promotes obesity. We investigated how chronic se affects caloric intake, adiposity, plasma hormones, inflammatory mediators, and hypothalamic NPY peptide in animals fed a palatable HFD. Balb/c mice (5 wk old, male) were exposed to smoke (2 cigarettes, twice/day, 6 days/wk, for 7 wk) with or without HFD. Sham-exposed mice were handled similarly without se. Plasma leptin, hypothalamic NPY, and adipose triglyceride lipase (ATGL) mRNA were measured. HFD induced a 2.3-fold increase in caloric intake, increased adiposity, and glucose in both sham and se cohorts. Smoke exposure decreased caloric intake by 23%, with reduced body weight in both dietary groups. Fat mass and glucose were reduced only by se in the chow-fed animals. ATGL mRNA was reduced by HFD in se animals. Total hypothalamic NPY was reduced by HFD, but only in sham-exposed animals; se increased arcuate NPY. We conclude that although se ameliorated hyperphagia and reversed the weight gain associated with HFD, it failed to reverse fat accumulation and hyperglycemia. The reduced ATGL mRNA expression induced by combined HFD and se may contribute to fat retention. Our data support a powerful health message that smoking in the presence of an unhealthy Western diet increases metabolic disorders and fat accumulation.     

Selective Leptin Insensitivity and Alterations in Female-Reproductive Patterns Linked to Hyperleptinemia during Infancy

The dramatic increase in the prevalence of childhood obesity worldwide makes the investigation of its early developmental stages and effective prevention strategies an urgent issue. CCK₁ deficient OLETF rats are a model of obesity previously used to study the early phases of this disorder. Here, we exposed wild type (LETO) females to an early obesogenic environment and genetically obese OLETF females to a lean postnatal environment, to assess long term alterations in leptin sensitivity, predisposition to diet induced obesity and adult female health. We found that genetically lean females reared by obese mothers presented early postnatal hyperleptemia, selectively reduced response to leptin and sensitivity to diet induced obesity when exposed to a high palatable diet as adults. The estrous cycle structure and intake profile were permanently disrupted, despite presenting normal adiposity/body weight/food intake. Genetically obese females reared by lean dams showed normalized early levels of leptin and reduced body weight, food intake and body fat at adulthood; normalized estrous cycle structure and food intake across the cycle, improved hormonal profile and peripheral leptin sensitivity and a remarkable progress in self-control when exposed to a high fat/palatable diet. Altogether, it appears that the early postnatal environment plays a critical role in determining later life coping with metabolic challenges and has an additive effect on the genetic predisposition that makes OLETF females morbidly obese as adults. This work also links, for the first time, alterations in the leptin system during early development to later life abnormalities related to female reproduction and health.     

Skin carotenoid concentration correlates with male hunting skill and territory quality in the kestrel Falco tinnunculus

In the context of sexual selection carotenoid based signals are candidates for indicator traits: they have to be taken up in the diet by animals, they can indicate the ability of the bearer to accumulate a limited resource, and they help in maintaining the health status. We investigated the yellow-orange colouration of the tarsi of the kestrel Falco tinnunculus in relation to sex, diet and to different aspects of male reproductive behaviour. The colouration of the tarsi (estimated as hue) was more intense in males than in females. Among males, the tarsi hue was associated with the intake of invertebrates; this was true also if the population diet was based mainly on voles. Carotenoid based colouration was positively associated with the number of vertebrate preys delivered to the nest per time unit and with territory quality (calculated on the basis of home-range size, habitat extension and prey availability). These results are consistent with predictions derived from good-parent models of sexual selection, suggesting that in the common kestrel carotenoid based colouration is important as an indicator of male quality.     

Evaluation of a Diet Quality Index Based on the Probability of Adequate Nutrient Intake (PANDiet) Using National French and US Dietary Surveys

Background

Existing diet quality indices often show theoretical and methodological limitations, especially with regard to validation.

Objective

To develop a diet quality index based on the probability of adequate nutrient intake (PANDiet) and evaluate its validity using data from French and US populations.

Material and Methods

The PANDiet is composed of adequacy probabilities for 24 nutrients grouped into two sub-scores. The relationship between the PANDiet score and energy intake were investigated. We evaluated the construct validity of the index by comparing scores for population sub-groups with ‘a priori’ differences in diet quality, according to smoking status, energy density, food intakes, plasma folate and carotenoid concentrations. French and US implementations of the PANDiet were developed and evaluated using national nutritional recommendations and dietary surveys.

Results

The PANDiet was not correlated with energy for the French implementation (r = −0.02, P>0.05) and correlated at a low level for the US implementation (r = −0.11, P<0.0001). In both implementations, a higher PANDiet score (i.e. a better diet quality) was associated with not smoking, having a lower-energy-dense diet, consuming higher amounts of fruits, vegetables, fish, milk and other dairy products and lower amounts of cheese, pizza, eggs, meat and processed meat, and having higher plasma folate and carotenoid concentrations after controlling for appropriate factors (all P<0.05, carotenoid data for US not available).

Conclusions

The PANDiet provides a single score that measures the adequacy of nutrient intake and reflects diet quality. This index is adaptable for use in different countries and relevant at the individual and population levels.     

The Self-Reported Clinical Practice Behaviors of Australian Optometrists as Related to Smoking,Diet and Nutritional Supplementation

Objective

The primary aim of this study was to examine the self-reported, routine clinical practice behaviors of Australian optometrists with respect to advice regarding smoking, diet and nutritional supplementation. The study also sought to assess the potential influence of practitioner age, gender, practice location (major city versus regional), therapeutic-endorsement status and personal nutritional supplementation habits upon management practices in these areas.

Methods

A survey was electronically distributed to Australian optometrists (n = 4,242). Respondents anonymously provided information about their personal demographics and lifestyle behaviors (i.e., age, gender, practice location, therapeutic-endorsement status, smoking status, nutritional supplement intake) and routine patient management practices with respect to advice across three domains: smoking, diet and nutritional supplementation. Multivariate logistic regression analyses were performed to assess for potential effects of the listed factors on practitioner behavior.

Results

A total of 283 completed surveys were received (completed survey response rate: 6.7%). Fewer than half of respondents indicated routinely asking their patients about smoking status. Younger practitioners were significantly (p < 0.05) less likely to enquire about patients’ smoking behaviors, but this did not extend to counseling for smoking cessation. Almost two-thirds of respondents indicated routinely counseling patients about diet. About half of practitioners specified routinely asking their patients about nutritional supplement intake; this form of questioning was significantly more likely if the respondent was female (p < 0.05). Practitioners who recommended nutritional supplements most commonly did so for age-related macular degeneration (91.2%) and dry eye disease (63.9%). The primary source of evidence used to guide practitioners’ nutrition-related patient management was reported to be peer-reviewed publications.

Conclusions

These findings demonstrate that there are no clear predictors of practitioner behavior across the three domains. Overall, this study suggests that there is scope for Australian optometrists to improve their routine engagement by questioning patients, as well as providing evidence-based clinical advice, about smoking status, diet and nutritional supplement behaviors, being key modifiable lifestyle risk factors with long-term implications for eye health.     

The effect of acarbose on the food intake, weight gain, and adiposity of LA/N-cp rats.

1. Groups of lean and obese LA/N-cp rats were administered the intestinal glucosidase inhibitor acarbose at 150 or 300 mg/kg diet from 7 until 17 weeks of age and the effects of the drug on food intake patterns and adiposity determined. 2. Dose related effects on body weights, adiposity and feed efficiency ratio were observed (control greater than 150 mg greater than 300 mg drug/kg diet) following treatment in both phenotypes, with the greatest differences observed in the obese phenotype. 3. Acarbose at both dosages was associated with phenotype-specific alterations in food intake amount and feeding pattern, resulting in an attenuation of age-associated increases in food intake. The feed efficiency ratio decreased in both phenotypes, and approached normally fed lean controls in obese rats administered the greater dosage. 4. These results indicate that patterns of food intake and weight gain differ markedly between lean and obese rats of this strain, and acarbose brings about a dose-related attenuation of developing food intake patterns in both phenotypes and which are associated with decreases in weight gain and adiposity. Thus, this drug may have therapeutic potential as an adjunct agent in the treatment of obesity and/or other disorders of carbohydrate intolerance.     

Galantamine alleviates inflammation and other obesity-associated complications in high-fat diet-fed mice

Obesity, a serious and growing health threat, is associated with low-grade inflammation that plays a role in mediating its adverse consequences. Previously, we have discovered a role for neural cholinergic signaling in controlling inflammation, and demonstrated that the cholinergic agent galantamine suppresses excessive proinflammatory cytokine release. The main objective of this study was to examine the efficacy of galantamine, a clinically-approved drug, in alleviating obesity-related inflammation and associated complications. After 8 wks on a high-fat diet, C57BL/6J mice were treated with either galantamine (4 mg/kg, intraperitoneally [i.p.]) or saline for 4 wks in parallel with mice on a low-fat diet and treated with saline. Galantamine treatment of obese mice significantly reduced body weight, food intake, abdominal adiposity, plasma cytokine and adipokine levels, and significantly improved blood glucose, insulin resistance and hepatic steatosis. In addition, galantamine alleviated impaired insulin sensitivity and glucose intolerance significantly. These results indicate a previously unrecognized potential of galantamine in alleviating obesity, inflammation and other obesity-related complications in mice. These findings are of interest for studying the efficacy of this clinically-approved drug in the context of human obesity and metabolic syndrome.     

Impact of fresh fruit smoothie consumption on apparent health of Asian faces

Skin carotenoid coloration has been proposed as a valid cue to health in humans, reflecting fruit and vegetable intake, and enhancing apparent health. Supplementation with a carotenoid-rich fruit and vegetable smoothie affects skin color, but it is not known if this skin color change enhances healthy appearance. In three experiments, we examine the effects of skin color change induced by supplementation with a carotenoid-rich fruit smoothie (25 mg carotenoids/d) on the apparent health of Malaysian Chinese faces. In experiment 1, observers were asked to identify the healthier looking of pairs of photographs of the same subject taken pre- and post-supplementation (or pre- and post-placebo), choosing the pre-supplementation (or pre-placebo) images. When confounding due to facial expression was eliminated in experiment 2, observers showed no preference for unmodified pre-supplementation photograph or the same image with skin color manipulated to simulate a level of smoothie-induced color change associated with 4 weeks of supplementation. In experiment 3, observers manipulated the skin color of face photographs along the smoothie-induced color change axis to optimize healthy appearance. Observers chose to induce a color change approximately equivalent to one third of the change induced by daily consumption of our carotenoid rich smoothie. This suggests that the skin color change induced by the supplementation enhanced apparent facial health, however the dose and duration of the supplementation overshot the optimal healthy-looking color of Malaysian Chinese skin. This suggests that there is an optimal carotenoid color for healthy appearance, and that this optimal level may be constrained by preferences for averageness, by the association between very yellow skin and ill health, or by negative health impacts of very high doses of carotenoids.     

Embryonic development within carotenoid-enriched eggs influences the post-hatch carotenoid status of the chicken

Carotenoids in the diet of the laying hen are incorporated into the egg yolk and subsequently into the liver and other tissues of the chicken embryo. Since these pigments are known to provide a range of health benefits to a variety of animals, it is of interest to know whether the effects of maternally derived carotenoids are strictly limited to the embryonic period or if they persist in the progeny after hatching. The aim of this study is to compare the effectiveness of pre-hatch (from the hen's diet) with that of post-hatch (from the progeny's diet) supplementation with carotenoids on the carotenoid status of the chick during the first 4 weeks of post-hatch life. Hens were fed a control diet or a diet supplemented with a carotenoid-rich extract of alfalfa. Eggs from the supplemented hens contained up to 22 times more carotenoids than the controls. The concentration of carotenoids in the livers of chicks hatching from the enriched eggs was initially 29 times greater than in the control chicks. Hepatic carotenoid concentrations in chicks from enriched eggs maintained post-hatch on the control diet were sustained at higher values compared with chicks from control eggs that were fed post-hatch on the carotenoid-supplemented diet, for at least the first 7 days. However, by 14 days, the latter group had overtaken the former in terms of liver carotenoid levels. Thus, under these conditions, maternal effects predominate for at least the first week after hatching, whereas from 2 weeks onwards, the progeny's diet becomes the main determinant of its carotenoid status. Since the antioxidant and immunostimulatory roles of carotenoids are likely to be especially important during the immediate post-hatch period, maternal dietary intake of carotenoids may have important ramifications for the viability of the offspring.