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Background Molecular stable isotope ratios are a novel type of dietary biomarker with high sensitivity and specificity for certain foods. Among these, fatty acid carbon isotope ratios (CIRs) have strong potential but have not been investigated as dietary biomarkers. Objectives We evaluated whether fatty acid CIRs and mass proportions were associated with meat, fish, and sugar-sweetened beverage (SSB) intake. Methods Thirty-two men [aged 46.2 ± 10.5 y; BMI (kg/m2): 27.2 ± 4.0] underwent a 12-wk inpatient dietary intervention at the National Institute of Diabetes and Digestive and Kidney Diseases in Phoenix, Arizona. Men were randomly assigned to 1 of 8 dietary treatments varying the presence/absence of dietary meat, fish, and SSBs in all combinations. Fatty acid CIRs and mass proportions were measured in fasting blood samples and adipose tissue biopsies that were collected pre- and postintervention. Dietary effects were analyzed using multivariable regression and receiver operating characteristic AUCs were calculated using logistic regression. Results CIRs of the several abundant SFAs, MUFAs and arachidonic acid (20:4n-6) in plasma were strongly associated with meat, as were a subset of these fatty acids in RBCs. Effect sizes in plasma ranged from 1.01‰ to 1.93‰ and were similar but attenuated in RBCs. Mass proportions of those fatty acids were not associated with diet. CIRs of plasma dihomo-γ-linolenic acid (20:3n–6) and adipose palmitic acid (16:0) were weakly associated with SSBs. Mass proportions of plasma odd-chain fatty acids were associated with meat, and mass proportions of plasma EPA and DHA (20:5n–3 and 22:6n–3) were associated with fish. Conclusions CIRs of plasma and RBC fatty acids show promise as sensitive and specific measures of dietary meat. These provide different information from that provided by fatty acid mass proportions, and are informative where mass proportion is not. This trial is registered at www.clinicaltrials.gov as NCT01237093. [ABSTRACT FROM AUTHOR]

Background: Objective dietary biomarkers are urgently needed for a wider range of foods and nutrients. The breath carbon isotope ratio (CIR; measured as δ13C values) has potential as a noninvasive measure of short-term added sugar (AS) intake but has not been evaluated in a controlled-feeding study.Objective: The aim was to evaluate the effect of short-term AS intake on breath CIR in a dose-response, randomized, crossover feeding study.Methods: Six men and 6 women, aged 25 to 60 y, were randomly assigned to a balanced sequence of 5 dietary treatments. Three treatments delivered low (0 g/d), medium (75 g/d), or high (150 g/d) amounts of AS over the course of a single day's breakfast and lunch and 2 switched high and low intake amounts between breakfast and lunch. Experimental meals delivered 60% of daily energy and added-sugar targets. There was a washout period of 1-2 wk between treatments. Breath was collected at 2-h intervals from 08:00 (fasting) to 16:00 h. Breath CIR was measured using cavity ring-down spectroscopy, and the effects of dietary treatments and baseline were evaluated using multivariate linear regression.Results: Breath CIR showed a significant response to increasing AS intake at all sampling time points (all P < 0.0001), with a dose-response of 0.030 (95% CI: 0.024, 0.037) ‰/g. Fasting breath CIR (baseline) influenced postfeeding breath CIR at all sampling time points (P < 0.0001); however, effect sizes were largest in the morning. For afternoon-collected samples (14:00 and 16:00), the effect of recent AS intake (lunch) was 4-fold greater than the effect of previous added-sugar intake (breakfast).Conclusions: These findings support the potential of the breath CIR as a biomarker of short-term AS intake in healthy US adults. More work is needed to evaluate other potential dietary effects and whether multiple breath collections could capture daily AS intake. [ABSTRACT FROM AUTHOR]

Rationale: Dried blood spots (DBSs) are gaining popularity for biomarker analyses in ecological research due to their advantages for use in field‐based research and in remote settings; however, many DBS biomarkers remain unvalidated. We validated the application of compound‐specific stable nitrogen isotope analyses of amino acids (CSIA‐AAs) to field‐prepared DBSs for determining trophic positions of wild‐caught Arctic char (Salvelinus alpinus). Methods: Whole blood and muscle from Arctic char were collected, and DBSs were created in the field. We measured the stable nitrogen isotope ratios (expressed as δ15N values) of the amino acids glutamic acid (Glu) and phenylalanine (Phe) isolated from Arctic char samples using CSIA‐AAs. We then compared amino acid δ15N values from DBSs and the other sample types (whole blood and muscle) from the same specimens. We calculated and compared trophic position estimates generated from whole blood, DBSs, and muscle. Results: The δ15N values of Glu and Phe, as well as trophic position estimates from DBSs, were highly correlated with δ15N values and estimates from both whole blood and muscle. The DBS amino acid δ15N values and trophic position estimates agreed well with those from whole blood. Although mean differences between amino acid δ15N values from DBSs and muscle were noted, the offsets were small and resulted in a 0.2 mean difference between trophic position estimates for DBSs and muscle. Conclusions: We demonstrate that the application of CSIA‐AAs to field‐prepared DBSs of Arctic char generates similar trophic position estimates to those from whole blood and muscle. We suggest that DBSs could be developed as a minimally invasive sampling technique to study feeding ecology of wild fish and perhaps other organisms of interest. [ABSTRACT FROM AUTHOR]

Dried blood spots (DBS), created by applying and drying a whole blood sample onto filter paper, provide a simple and minimally invasive procedure for collecting, transporting, and storing blood. Because DBS are ideal for use in field and resource‐limited settings, we aimed to develop a simple and accurate DBS‐based approach for assessing mercury (Hg) exposure and dietary carbon sources for landlocked Arctic char, a sentinel fish species in the Arctic. We collected liquid whole blood (from the caudal vein), muscle, liver, and brains of Arctic char (n = 36) from 8 lakes spanning a Hg gradient in the Canadian High Arctic. We measured total Hg concentrations ([THg]) of field‐prepared DBS and Arctic char tissues. Across a considerable range, [THg] of DBS (0.04–3.38 μg/g wet wt) were highly correlated with [THg] of all tissues (r2 range = 0.928–0.996). We also analyzed the compound‐specific carbon isotope ratios (expressed as δ13C values) of essential amino acids (EAAs) isolated from DBS, liquid whole blood, and muscle. The δ13C values of 5 EAAs (δ13CEAAs; isoleucine [Ile], leucine [Leu], phenylalanine [Phe], valine [Val], and threonine [Thr]) from DBS were highly correlated with δ13CEAAs of liquid whole blood (r2 range = 0.693–0.895) and muscle (r2 range = 0.642–0.881). The patterns of δ13CEAAs of landlocked Arctic char were remarkably consistent across sample types and indicate that EAAs are most likely of algal origin. Because a small volume of blood (~50 µL) dried on filter paper can be used to determine Hg exposure levels of various tissues and to fingerprint carbon sources, DBS sampling may decrease the burdens of research and may be developed as a nonlethal sampling technique. Environ Toxicol Chem 2020;39:893–903. © 2020 SETAC [ABSTRACT FROM AUTHOR]

Background Naturally occurring carbon and nitrogen stable isotope ratios [13C/12C (CIR) and 15N/14N (NIR)] are promising dietary biomarkers. As these candidate biomarkers have long tissue residence times, long-term feeding studies are needed for their evaluation. Objective Our aim was to evaluate plasma, RBCs, and hair CIR and NIR as biomarkers of fish, meat, and sugar-sweetened beverage (SSB) intake in a 12-wk dietary intervention. Methods Thirty-two men (aged 46.2 ± 10.5 y; BMI: 27.2 ± 4.0 kg/m2) underwent a 12-wk inpatient dietary intervention at the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) in Phoenix, Arizona. The effects of fish, meat, and SSB intake on CIR and NIR were evaluated using a balanced factorial design, with each intake factor at 2 levels (present/absent) in a common, background diet (50% carbohydrate, 30% fat, 20% protein). Fasting blood samples were taken biweekly from baseline, and hair samples were collected at baseline and postintervention. Data were analyzed using multivariable regression. Results The postintervention CIR of plasma was elevated when diets included meat (β = 0.89, 95% CI: 0.73,1.05) and SSBs (β = 0.48, 95% CI: 0.32, 0.64). The postintervention NIR of plasma was elevated when diets included fish (β = 0.85, 95% CI: 0.64, 1.05) and meat (β = 0.61, 95% CI: 0.42, 0.8). Results were similar for RBCs and hair. Postintervention RBC CIR and NIR had strong associations with baseline, suggesting that turnover to the intervention diets was incomplete after 12 wk. Estimates of isotopic turnover rate further confirmed incomplete turnover of RBCs. Conclusions CIR was associated with meat and SSBs, and more strongly with meat. NIR was associated with fish and meat, and more strongly with fish. Overall, CIR and NIR discriminated between dietary fish and meat, and to a lesser extent SSBs, indicating their potential utility as biomarkers of intake in US diets. Approaches to make these biomarkers more specific are needed. This trial was registered at clinicaltrials.gov as NCT01237093. [ABSTRACT FROM AUTHOR]

Benthic invertebrates are a crucial trophic link in Arctic marine food webs. However, estimates of the contribution of different primary production sources sustaining these organisms are not well characterised. We measured the stable carbon isotope values (δ13C) of essential amino acids (EAAs) in muscle tissue from two common bivalve genera (Macoma spp. and Astarte spp.) collected in Hanna Shoal in the northeastern Chukchi Sea. Mixing models comparing the δ13CEAA fingerprints of the bivalves to a suite of primary production endmembers revealed relatively high contributions of EAAs from phytoplankton and bacteria in both species. We also examined whether δ13CEAA fingerprints could be produced from the EAAs preserved in bivalve shells, which could allow primary production sources to be estimated from ancient bivalve shells. The δ13CEAA fingerprints from a suite of paired modern bivalve shells and muscle from Macoma calcarea from across the Chukchi Sea revealed a correspondence between the estimates of the dominant primary production source of EAAs derived from analyses of these two tissue types. Our findings indicate that δ13CEAA fingerprinting of marine bivalves can be used to examine dominant organic matter sources in the Arctic marine benthos in recent years as well as in deeper time. [ABSTRACT FROM AUTHOR]

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The carbon isotope ratio (δ13C) is elevated in corn- and cane sugar-based foods and has recently shown associations with sweetener intake in multiple U.S. populations. However, a high carbon isotope ratio is not specific to corn- and sugar cane-based sweeteners, as other foods, including meats and fish, also have elevated δ13C. This study examines whether the inclusion of a second marker, the nitrogen isotope ratio (δ15N), can control for confounding dietary effects on δ13C and improve the validity of isotopic markers of sweetener intake. The study participants are from the Yup'ik population of southwest Alaska and consume large and variable amounts of fish and marine mammals known to have elevated carbon and nitrogen isotope ratios. Sixty-eight participants completed 4 weekly 24-h recalls followed by a blood draw. RBC δ13C and δ15N were used to predict sweetener intake, including total sugars, added sugars, and sugar-sweetened beverages. A model including both δ13C and δ15N explained more than 3 times as much of the variation in sweetener intake than did a model using only δ13C. Because carbon and nitrogen isotope ratios are simultaneously determined in a single, high-throughput analysis, this dual isotope marker provides a simple method to improve the validity of stable isotope markers of sweetener intake with no additional cost. We anticipate that this multi-isotope approach will have utility in any population where a stable isotope biomarker is elevated in several food groups and there are appropriate "covariate" isotopes to control for intake of foods not of research interest. [ABSTRACT FROM AUTHOR]

The article discusses the results of a study that investigates the potential relationship between diet and beak deformities in Black-capped Chickadees or Poecile atricapillus and other resident avian species in Alaska. Birds with beak deformities have lower nitrogen values and more variable carbon values than those with normal beaks. There was no detection of a difference in natural dietary items across the study sites.

The transition of a society from traditional to market-based diets (termed the nutrition transition) has been associated with profound changes in culture and health. We are developing biomarkers to track the nutrition transition in the Yup'ik Eskimo population of Southwest Alaska based on naturally occurring variations in the relative abundances of carbon and nitrogen stable isotopes (δ15N and δ13C values). Here, we provide three pieces of evidence toward the validation of these biomarkers. First, we analyzed the δ15N and δ13C values of a comprehensive sample of Yup'ik foods. We found that δ15N values were elevated in fish and marine mammals and that δ13C values were elevated in market foods containing corn or sugar cane carbon. Second, we evaluated the associations between RBC δ15N and δ13C values and self-reported measures of traditional and market food intake (n = 230). RBC δ15N values were correlated with intake of fish and marine mammals (r = 0.52; P < 0.0001). RBC δ13C values were correlated with intake of market foods made from corn and sugar cane (r = 0.46; P < 0.0001) and total market food intake (r = 0.46; P < 0.0001). Finally, we assessed whether stable isotope ratios captured population-level patterns of traditional and market intake (n = 1003). Isotopic biomarkers of traditional and market intake were associated with age, community location, sex, and cultural identity. Self-report methods showed variations by age and cultural identity only. Thus, stable isotopes show potential as biomarkers for monitoring dietary change in indigenous circumpolar populations. [ABSTRACT FROM AUTHOR]

It is a nutritional challenge for nectar-feeding insects to meet the amino acid requirements of oviposition. Here we investigate whether egg amino acids derive from larval diet or are synthesized from nectar sugar in four species of butterfly: Colitis eurytheme, Speyeria mormonia, Euphydryas chalcedona, and Heliconius charitonia. These species exhibit a range of life history and differ in degree of shared phylogeny. We use 13C differences among plants to identify dietary sources of amino acid carbon, and we measure amino acid 13C using compound-specific stable isotope analysis. Egg essential amino acids derived solely from the larval diet, with no evidence for metabolic carbon remodeling. Carbon in nonessential amino acids from eggs derived primarily from nectar sugars, with consistent variation in amino acid turnover. There was no relationship between the nonessential amino acids of eggs and host plants, demonstrating extensive metabolic remodeling. Differences between species in carbon turnover were reflected at the molecular level, particularly by glutamate and aspartate. Essential amino acid 13C varied in a highly consistent pattern among larval host plants, reflecting a common isotopic "fingerprint" associated with plant biosynthesis. These data demonstrate conservative patterns of amino acid metabolism among Lepidoptera and the power of molecular stable isotope analyses for evaluating nutrient metabolism in situ. [ABSTRACT FROM AUTHOR]

Abstract: Soil animal detritivores play an important role in facilitating decomposition processes but little information is available on how the quality of dietary resources affects their stoichiometry of carbon (C) nitrogen (N) and phosphorus (P), and turnover of C and N. This study investigated how a fungal diet, Fusarium culmorum, with a low N content and imbalanced amino acid (AA) composition affected the physiology of three soil-dwelling collembolans (Folsomia candida, Protaphorura fimata and Proisotoma minuta) in comparison to a control diet, Saccharomyces cerevisiae, with a high N content and balanced AA composition. We compared the elemental composition of animals, their growth rates and tissue replacement of C and N. We also measured the individual AA δ13C to investigate the extent that Collembola may rely on endogenous sources to compensate for scarcity of essential AAs. The results showed that animal''s N content tracked closely the composition of their diets, decreasing from around 10 to 7% N from the high to low N diet. They also had a significant increase of C and a decrease of P. P. fimata was less affected than F. candida and P. minuta. The total incorporation of C and N in the animals due to growth and tissue replacement decreased from 11–17 to 6–12% DM d−1 on the high and low N diet respectively with P. fimata experiencing the smallest change. Essential AAs δ13C did not always match perfectly between Collembola species and their diets; particularly on the low N diet. Isotope patterns of AAs indicate that bacteria may have been the alternative source of essential AAs. While the results of this study cannot be extrapolated directly to the dynamics of Collembola populations in the field, they serve to demonstrate their flexibility in adapting physiologically to the temporal and spatial patchiness of the soil environment. [Copyright &y& Elsevier]

Abstract: Testes size often predicts the winner during episodes of sperm competition. However, little is known about the source of nutrients allocated to testes development, or testes plasticity under varying nutrient availability. Among many holometabolous insects, metabolic resources can derive from the larval or adult diet. Distinguishing the source of nutrients allocated to testes can shed light on life history factors (such as maternal influences) that shape the evolution of male reproductive strategies. Here we used an experimental approach to assess resource allocation to testes development in walnut flies (Rhagoletis juglandis) from differing nutritional backgrounds. We fed adult male walnut flies on sugar and yeast diets that contrasted with the larval diet in carbon and nitrogen stable isotope ratios. This design allowed us to assess the dietary source of testes carbon and nitrogen and its change over time. We found significant incorporation of adult dietary carbon into testes, implying that walnut flies are income breeders for carbon (relying more on adult resources). In contrast, we found little evidence that walnut flies incorporate adult dietary nitrogen into testes development. We discuss the implications of these allocation decisions for life history evolution in this species. [ABSTRACT FROM AUTHOR]

Stable carbon isotope ratios (δ 13 C) in breath show promise as an indicator of immediate metabolic fuel utilization in animals because tissue lipids have a lower δ 13 C value than carbohydrates and proteins. Metabolic fuel consumption is often estimated using the respiratory exchange ratio (RER), which has lipid and carbohydrate boundaries, but does not differentiate between protein and mixed fuel catabolism at intermediate values. Because lipids have relatively low δ 13 C values, measurements of stable carbon isotopes in breath may help distinguish between catabolism of protein and mixed fuel that includes lipid. We measured breath δ 13 C and RER concurrently in arctic ground squirrels ( Urocitellus parryii ) during steady-state torpor at ambient temperatures from − 2 to − 26 °C. As predicted, we found a correlation between RER and breath δ 13 C values; however, the range of RER in this study did not reach intermediate levels to allow further resolution of metabolic substrate use with the addition of breath δ 13 C measurements. These data suggest that breath δ 13 C values are 1.1‰ lower than lipid tissue during pure lipid metabolism. From RER, we determined that arctic ground squirrels rely on nonlipid fuel sources for a significant portion of energy during torpor (up to 37%). The shift toward nonlipid fuel sources may be influenced by adiposity of the animals in addition to thermal challenge. [ABSTRACT FROM AUTHOR]

Abstract: Metabolic resources in adults of holometabolous insects may derive either from larval or adult feeding. In Drosophila melanogaster, reproduction and lifespan are differently affected by larval vs. adult resource availability, and it is unknown how larval vs. adult acquired nutrients are differentially allocated to somatic and reproductive function. Here we describe the allocation of carbon derived from dietary sugar in aging female D. melanogaster. Larval and adult flies were fed diets contrasting in sucrose 13C/12C, from which we determined the extent to which carbon acquired at each stage contributed to adult somatic tissue and to egg manufacture. Dietary sugar is very important in egg provisioning; at every age, roughly one half of the carbon in eggs was derived from sugar, which turned over from predominantly larval to entirely adult dietary sources. Sucrose provided ∼40% of total somatic carbon, of which adult dietary sucrose came to supply ∼75%. Unlike in eggs, however, adult acquired sucrose did not entirely replace the somatic carbon from larvally acquired sucrose. Because carbon from larval sucrose appears to be fairly “replaceable”, larval sucrose cannot be a limiting substrate in resource allocation between reproduction and lifespan. [Copyright &y& Elsevier]