Measurement Uncertainty and Risk of False Compliance Assessment Applied to Carbon Isotopic Analyses in Natural Gas Exploratory Evaluation.

The concept of uncertainty in an isotopic analysis is not uniform in the scientific community worldwide and can compromise the risk of false compliance assessment applied to carbon isotopic analyses in natural gas exploratory evaluation. In this work, we demonstrated a way to calculate one of the main sources of this uncertainty, which is underestimated in most studies focusing on gas analysis: the δ13C calculation itself is primarily based on the raw analytical data. The carbon isotopic composition of methane, ethane, propane, and CO2 was measured. After a detailed mathematical treatment, the corresponding expanded uncertainties for each analyte were calculated. Next, for the systematic isotopic characterization of the two gas standards, we calculated the standard uncertainty, intermediary precision, combined standard uncertainty, and finally, the expanded uncertainty for methane, ethane, propane, and CO2. We have found an expanded uncertainty value of 1.8‰ for all compounds, except for propane, where a value of 1.6‰ was obtained. The expanded uncertainty values calculated with the approach shown in this study reveal that the error arising from the application of delta calculation algorithms cannot be neglected, and the obtained values are higher than 0.5‰, usually considered as the accepted uncertainty associated with the GC-IRMS analyses. Finally, based on the use of uncertainty information to evaluate the risk of false compliance, the lower and upper acceptance limits for the carbon isotopic analysis of methane in natural gas are calculated, considering the exploratory limits between -55‰ and -50‰: (i) for the underestimated current uncertainty of 0.5‰, the lower and upper acceptance limits, respectively, are -54.6‰ and -50.4‰; and (ii) for the proposed realistic uncertainty of 1.8‰, the lower and upper acceptance limits would be more restrictive; i.e., -53.5‰ and -51.5‰, respectively.

Accumulation of Per- and Polyfluoroalkyl Substances (PFAS) in Coastal Sharks from Contrasting Marine Environments: The New York Bight and The Bahamas.

Per- and polyfluoroalkyl substances (PFAS) enter the marine food web, accumulate in organisms, and potentially have adverse effects on predators and consumers of seafood. However, evaluations of PFAS in meso-to-apex predators, like sharks, are scarce. This study investigated PFAS occurrence in five shark species from two marine ecosystems with contrasting relative human population densities, the New York Bight (NYB) and the coastal waters of The Bahamas archipelago. The total detected PFAS (∑PFAS) concentrations in muscle tissue ranged from 1.10 to 58.5 ng g-1 wet weight, and perfluorocarboxylic acids (PFCAs) were dominant. Fewer PFAS were detected in Caribbean reef sharks (Carcharhinus perezi) from The Bahamas, and concentrations of those detected were, on average, ∼79% lower than in the NYB sharks. In the NYB, ∑PFAS concentrations followed: common thresher (Alopias vulpinus) > shortfin mako (Isurus oxyrinchus) > sandbar (Carcharhinus plumbeus) > smooth dogfish (Mustelus canis). PFAS precursors/intermediates, such as 2H,2H,3H,3H-perfluorodecanoic acid and perfluorooctanesulfonamide, were only detected in the NYB sharks, suggesting higher ambient concentrations and diversity of PFAS sources in this region. Ultralong-chain PFAS (C ≥ 10) were positively correlated with nitrogen isotope values (δ15N) and total mercury in some species. Our results provide some of the first baseline information on PFAS concentrations in shark species from the northwest Atlantic Ocean, and correlations between PFAS, stable isotopes, and mercury further contextualize the drivers of PFAS occurrence.

Using Stable Isotope Techniques to Analyze the Trophic Relationship between Argentine Hake (Merluccius hubbsi) and Anisakidae.

The Argentine hake (Merluccius hubbsi) is a vital fishery species in the Southwest Atlantic, recognized for its substantial economic importance. Previous studies have identified Anisakidae larvae as common parasites of M. hubbsi. However, the nutritional relationships between these parasites and their host remain poorly understood. This study employs stable isotope techniques to investigate the specific nutritional relationships between Anisakidae larvae and different tissues of M. hubbsi. The findings reveal notable differences in δ13C and δ15N compositions between the parasites and their host. The lower δ13C values in parasites compared to host tissues indicate the utilization of different carbon sources. The δ15N values of the parasites partially overlap with those of the host's stomach, indicating that the parasites primarily derive nutrients from the host's stomach. Nutritional niche indicators show that parasites have a broad carbon range (CR) and nitrogen range (NR), suggesting a high diversity in nutritional sources. The trophic discrimination factor (ΔTDF), which represents the difference in stable isotope values between host tissues and parasites, was analyzed for both δ13C and δ15N. The ΔTDFδ13C between the host liver and the parasites showed the greatest variation, indicating a strong dependence of the parasites on the liver's carbon sources. In contrast, variations in ΔTDFδ15N between host tissues and parasites were minimal. Analyzing ΔTDF across different stages of gonadal maturity in the host fish indicates that, as the gonads of the host fish mature, ΔTDFδ13C between host tissues and parasites significantly decreases (p < 0.01). The Kruskal-Wallis test showed significant differences in ΔTDFδ13C values among different parasite infection levels in muscle, liver, and stomach tissues, while no significant differences were found for ΔTDFδ15N values. These findings offer valuable insights into the nutritional relationships between parasites and hosts, aiding in a better understanding of the growth conditions and habitats of M. hubbsi.

Chemical and isotopic tracers combined with mixing models for tracking nitrate contamination in the Pampa de Pocho aquifer, Argentina.

In recent years, it has become evident that human activities have significantly disrupted the nitrogen cycle surpassing acceptable environmental thresholds. In this study, chemical and isotopic tracers were combined with a mathematical mass balance model (EMMA), PHREEQC inverse mixing model, and statistical analyses to evaluate groundwater quality, across an area experiencing substantial human activities, with a specific focus on tracing the origin of nitrate (NO3-) with potential water mixing processes. This multi-technique approach was applied to an unconfined aquifer underlying an agricultural area setting in an inter-mountain depression (i.e., the "Pampa de Pocho Plain" in Argentina). Here, the primary identified geochemical processes occurring in the investigated groundwater system include the dissolution of carbonate salts, cation exchange, and hydrolysis of alumino-silicates along with incorporating ions from precipitation. It was observed that the chemistry of groundwater, predominantly of sodium bicarbonate with sulfate water types, is controlled by the area's geology, recharge from precipitation, and stream water infiltration originating from the surrounding hills. Chemical results reveal that 60% of groundwater samples have NO3- concentrations exceeding the regional natural background level, confirming the impact of human activities on groundwater quality. The dual plot of δ15NNO3 versus δ18ONO3 values indicates that groundwater is affected by NO3- sources overlapping manure/sewage with organic-rich soil. The mathematical EMMA model and PHREEQC inverse modeling, suggest organic-rich soil as an important source of nitrogen in the aquifer. Here, 64 % of samples exhibit a main mixture of organic-rich soil with manure, whereas 36 % of samples are affected mainly by a mixture of manure and fertilizer. This study demonstrates the utility of combining isotope tracers with mathematical modeling and statistical analyses for a better understanding of groundwater quality deterioration in situations where isotopic signatures of contamination sources overlap.

Trace metals pollution and trophic position of three sea urchin species in the Gulf of California.

Sea urchin species are ecologically important in the Gulf of California and are becoming popular as a local fishery due to their commercial value. The most abundant species are Echinometra vanbrunti, Eucidaris thouarsii, and Tripneustes depressus. The objective of this study was to evaluate cadmium, lead, copper, zinc, and iron concentrations, as well as stable isotope values in these sea urchin species in the Santa Rosalia mining area (STR), in three sites: Punta Gorda, Punta el Aterrizaje, and Punta Salina. The highest Fe concentration (100.2 mg kg-1) was found in E. vanbrunti, while the highest concentrations of Pb (15.1 mg kg-1), Cu (14.5 mg kg-1), and Zn (347.7 mg kg-1) were recorded in E. thouarsii, and the highest Cd concentration (10.8 mg kg-1) was found in T. depressus. The main health risk of trace metal pollution in STR may be caused by Cd and Pb. δ15N and δ13C values were higher in E. thouarsii and T. depressus, respectively; E. thouarsii has the highest trophic position. Specimen size was not related to metal concentrations, but a positive relationship was observed between specimen size and isotopic values in T. depressus. The three species showed different bioaccumulation patterns for the metals analyzed. Additionally, collection sites and seasons play an important role in the variability of metal concentration.

Early Colonial Diet in El Japón, Xochimilco, Mexico: Examining dietary continuity through stable isotope analysis of bone collagen and bioapatite.

OBJECTIVES: Early colonial documents from central Mesoamerica detail raising and planting of European livestock and crops alongside native ones. The extent to which Indigenous people, especially of the rural commoner class, consumed newly introduced foods is less known. This gap in knowledge is addressed through stable isotope analysis and comparison to published archaeological botanical, human, and faunal data. MATERIALS AND METHODS: Stable isotope analysis of bone collagen and bioapatite is applied to 74 skeletal samples of Indigenous human remains representing Colonial period individuals from El Japón-a farming hamlet in the Xochimilco area-to provide insight into long-term individual dietary practices in the context of a rapidly transforming Mesoamerican world. RESULTS: Carbon isotope ratios in collagen (δ13Ccollagen) average -8.10/00 VPDB (SD 0.55), while δ15N averages 8.90/00 AIR (SD 0.50). δ13Cbioapatite averages -2.90/00 VPDB (SD 0.60). Modest increase in carbon isotopic diversity is observed among more recent males from El Japón when compared to earlier males and females. DISCUSSION: Based on the isotopic results, it is estimated that the individuals of El Japón consumed maize or other C4 plants as a central source of carbohydrates. Dietary protein was largely supplied through domestic maize-fed fauna but potentially supplemented by wild terrestrial and aquatic fauna and fowl. Similarity in skeletal isotopic composition between precontact Mesoamericans from other sites and El Japón individuals of both earlier and later stratigraphy is interpreted as continuity in local diets and foodways despite potentially available European alternatives. Colonial taxation demands on preexisting agricultural regimes may have incentivized maize production, thus indirectly contributing to the maize-centered aspect of local foodways.

OBJETIVOS: Los textos de la época colonial temprana del centro de México documentan la producción de cultivos y ganado europeo a la par de los productos agropecuarios nativos. La magnitud a la cuál las comunidades indígenas consumieron estos productos se conoce con menos certeza en especial dentro de los asentamientos rurales. En este trabajo, se analiza la variabilidad de datos de isótopos estables en el sitio El Japón, Xochimilco y los resultados se comparan con respecto al sexo biológico y la cronología; así como también con datos publicados de muestras humanas y faunísticas. MATERIALES Y MÉTODOS: Se aplican los estudios de isotopos estables en colágeno y bioapatita a 74 muestras esqueléticas de El Japón de la época colonial temprana, una aldea agrícola del área de Xochimilco, con tal de abordar las practicas dietéticas en el contexto de un mundo Mesoamericano en transformación tras el contacto europeo. RESULTADOS: Los isótopos estables de carbono en colágeno (δ13Ccollagen) producen un promedio de −8.10/00 VPDB (DE 0.55), mientras tanto los isótopos estables de nitrógeno en el mismo tejido producen un promedio de 8.90/00 AIR (DE 0.50). Los isótopos estables de carbono en la bioapatita (δ13Cbioapatite) producen un promedio de −2.90/00 VPDB (DE 0.60). Se observa un incremento mínimo en la diversidad isotópica entre los individuos de sexo masculino en comparación a los individuos de sexo femenino de la etapa temprana y tardía del sitio. DISCUSIÓN: Con base en los resultados isotópicos, y con base en comparación a muestras humanas de contextos arqueológicos europeos y norteamericanos se estima que los individuos de El Japón consumieron maíz u otros cultivos tipo C4 como fuentes principales de carbohidratos. Las fuentes de proteína dietética posiblemente fueron fauna alimentada con maíz, pero también se pudieron haber suplementado con alimentos silvestres incluyendo aves silvestres, y fauna terrestre o acuática. La similitud en variación isotópica entre sitios mesoamericanos que preceden el contacto europeo y El Japón de ambas etapas (temprana y tardía) se interpretan como persistencia en fuentes de alimentación y tradiciones culinarias a pesar de las posibles alternativas europeas. Las demandas tributarias coloniales sobre la producción agrícola chinampera pudiesen haber contribuido indirectamente a la continuidad del maíz como fuente alimenticia principal.

Patagonian partnerships: the extinct Dusicyon avus and its interaction with prehistoric human communities.

The southern Mendoza province, located in the northern region of Patagonia, was inhabited by hunter-gatherer groups until historic times. Previous archaeological studies have reported canid remains among faunal assemblages, which were assumed to be part of the human diet. However, the taxonomic identification and significance of these canids within human groups have raised questions. In this study, we used ancient DNA analysis, morphological examination and stable isotope analysis (δ13Ccol and δ15N) to re-evaluate the taxonomic assignment of a canid discovered at the Late Holocene burial site of Cañada Seca. Previous morphological identifications suggested that it belonged to the genus Lycalopex, but our results conclusively demonstrate that the individual belongs to the extinct fox species Dusicyon avus. This finding expands Dusicyon avus' known geographical distribution to Patagonia's northern extremity. Furthermore, statistical predictions based on genetic divergence undermine the hypothesis that hybridization between Canis and Dusicyon, facilitated by the introduction of domestic dogs, played a role in the extinction of Dusicyon species. On the other hand, our findings indicate that a Dusicyon avus individual shared a similar diet and was probably buried alongside humans, suggesting a close relationship between the two species during their lives and deaths.

Methods for Intracellular Peptidomic Analysis.

Peptides have broad biological significance among different species. Intracellular peptides are considered a particular class of bioactive peptides, whose generation is initiated by proteasomal degradation of cytosolic, nuclear, or mitochondrial proteins. To extract and purify intracellular peptides, which may apply for biological peptides in general, it is important to consider the initial source: tissue, cell, or fluid. First, it is important to proceed fast with inactivation of proteases and/or peptidases commonly present in the biological source of peptides, which might rapidly degrade peptides during the initial process of extraction. The incubation of biological tissues, cells, and fluids at 80 °C for up to 20 min have been sufficient to fully inactivate proteases or peptidases activities. It is particularly important not to acidify the samples at high temperature, because it can lead to nonspecific hydrolysis reactions; particularly, the Asp-Pro peptide bond can be cleaved at acidic environments and elevated temperatures. Unfortunately, not every sample can have proteinases and peptidases denatured by heating the biological source of intracellular peptides. Plasma, for example, when heated at temperatures higher than 55 °C can clot and trap peptides within the fibrin net. Therefore, alternative conditions for inactivating proteinases and peptidases must apply for plasma samples. In this chapter, the most successful methods used in our laboratory to extract intracellular peptides are described.

A green-footprint approach for parallel multiclass analysis of contaminants in roasted coffee via LC-HRMS.

The development and validation of a simple, comprehensive, and environment-friendly procedure to determine pesticide residues, naturally occurring and processing contaminants in roasted coffee is presented. A solid-liquid extraction of pesticides and mycotoxins with ethyl acetate and the concurrent partition of acrylamide to an aqueous phase follows a parallel analytical strategy that requires a single analytical portion to determine contaminants that are typically analyzed by dedicated single residue methods. The partition rules the lipids out of the aqueous extract before an "in-tube" dispersive solid phase microextraction (dSPME) for acrylamide retention. This is followed by the elution with buffer prior to injection. This extract is independently introduced into the system front end followed by the injection of the compounds from the organic phase, yet all spotted in the same run. A novel liquid chromatography high-resolution mass spectrometry (LC-HRMS) method setup enables the quantification of 186 compounds at 10 µg/kg, 226 at 5 µg/kg, and the acrylamide at 200 µg/kg for a total of 414 molecules, with acceptable recoveries (70-120%) and precision (RSD < 20%) making this strategy significantly faster and cost-effective than the dedicated single residue methods. Even though the presence of chlorpyrifos, acrylamide, and ochratoxin A was confirmed on samples of different origins, the findings were below the limit of quantification. During the storage of raw coffee, no proof of masking of OTA was found; however, condensation with glucose was evidenced during thermal processing experiments with sucrose by using stable isotope labeling (SIL). No detected conjugates were found in roasted nor in commercial sugar-added torrefacto samples, an industrial processing usually carried out above the decomposition temperature of the disaccharide.

Seasonality of hydrogeochemical evolutions and isotopic variabilities (δ18O, δ2H and d-excess) in the surface water as well as groundwater from tropical central-south Mexico.

Due to adverse impact of the global warming on hydrological resources, we intended to document the hydrogeochemical evolutions of surface and groundwater at tropical central-south Mexico in terms of seasonality of rock-water interaction, precipitation/evaporation variation and moisture source by evaluating the major ion chemistry in Piper and Gibbs plots, Durov diagram and through estimation of the chloro-alkaline indices as well as assessing the stable isotope compositions (δ18O and δ2H) in samples from different seasons of a year. Surface water of the Lake Coatetelco shifted from mostly Ca-Mg-HCO3 facies in wet summer-autumn to Na-HCO3-Cl facies in the dry spring due to elevated Na, Cl and HCO3. Greater evaporation in spring led to a maximum δ18O enrichment of ca.7‰ compared to the other seasons, and much depleted deuterium excess (-40.92‰ to -39.20‰). Interaction of the lake water with subsurface carbonate lithologies, and comparable isotopic compositions reflected the enhanced interaction between the surface water body and aquifers in the wet autumn. Effect of seasonality, however, was unclear on the groundwater facies, and its heterogenous composition (Ca-Mg-HCO3, Na-HCO3-Cl and Na-HCO3) reflected the interactions with different lithologies. Fractionations in isotope compositions of the groundwater were caused from recharge at different elevations, seasonality of moisture sources and moisture recycling. The water-mineral saturation index was an efficient proxy of seasonality as the lake water and groundwater (avg SIcalcite > 0.5) of the dry autumn were saturated with calcite. This vital information about carbonate precipitation, pCO2 and chemical facies would be useful for the better interpretation of paleoclimate archives in this region.

Feeding behavior of yellowfin tuna around two insular regions of the western Atlantic Ocean.

The yellowfin tuna is a very abundant tropical tuna species in the western equatorial Atlantic Ocean and an important fishery resource for the Brazilian tuna fleet. In this study we performed stable isotope analysis to better understand the spatial trophodynamics and dietary changes in yellowfin tuna around two insular marine protected areas in Brazil. A total of 65 yellowfin tuna specimens measuring between 47 and 138 cm LT (total length) were sampled around the archipelagos of Fernando de Noronha (FNA; n = 34) and Saint Peter and Saint Paul (SPSPA; n = 31) between July 2018 and September 2019. Bayesian mixing models and generalized additive models were used to investigate the contributions of four different prey items (zooplankton, cephalopods, fish larvae, and flying fish) to yellowfin tuna diet in each area and their potential changes in relation to predator growth. The four prey items were found to have different overall contributions between the two studied areas, with zooplankton being the most important prey in FNA, whereas flying fish was the most relevant prey to the species' diet in SPSPA. Significant changes in the species diet by size were also found, with fish smaller than 90 cm (TL) having a more generalist diet and larger animals relying more on consuming larger and more nutritious prey (i.e., flying fish). Our results suggest that these two marine protected areas play an important role in ocean dynamics, providing important and different foraging grounds for the development of this predator species.

Isotopic niche overlap among foraging marine turtle species in the Gulf of Mexico.

Sympatric species may overlap in their use of habitat and dietary resources, which can increase competition. Comparing the ecological niches and quantifying the degree of niche overlap among these species can provide insights into the extent of resource overlap. This information can be used to guide multispecies management approaches tailored to protect priority habitats that offer the most resources for multiple species. Stable isotope analysis is a valuable tool used to investigate spatial and trophic niches, though few studies have employed this method for comparisons among sympatric marine turtle species. For this study, stable carbon, nitrogen, and sulfur isotope values from epidermis tissue were used to quantify isotopic overlap and compare isotopic niche size in loggerhead (Caretta caretta), green (Chelonia mydas), and Kemp's ridley (Lepidochelys kempii) turtles sampled from a shared foraging area located offshore of Crystal River, Florida, USA. Overall, the results revealed high degrees of isotopic overlap (>68%) among species, particularly between loggerhead and Kemp's ridley turtles (85 to 91%), which indicates there may be interspecific competition for resources. Samples from green turtles had the widest range of isotopic values, indicating they exhibit higher variability in diet and habitat type. Samples from loggerhead turtles had the most enriched mean δ34S, suggesting they may forage in slightly different micro-environments compared with the other species. Finally, samples from Kemp's ridley turtles exhibited the smallest niche size, which is indicative of a narrower use of resources. This is one of the first studies to investigate resource use in a multispecies foraging aggregation of marine turtles using three isotopic tracers. These findings provide a foundation for future research into the foraging ecology of sympatric marine turtle species and can be used to inform effective multispecies management efforts.

Oxygen isotope variation in drinking water in Oaxaca, Mexico and its implications for forensic provenancing efforts.

Forensic anthropologists have used oxygen isotopic analyses (δ18O) in recent decades because of its ability to help estimate probable regions of origin of unidentified individuals based on isotopic composition of bodily tissues. Prior research has found that drinking water is geographically patterned and that tap water is often representative of drinking water in assessing these links between human tissues and geography. Researchers have begun applying the method in a variety of global contexts. This research critically assesses the assumption that tap water is isotopically representative of drinking water in Oaxaca, Mexico. Drinking water samples from Oaxaca were analyzed and compared with predicted δ18O values from a previously constructed tap water isoscape of Mexico. Hair samples from Oaxaca were also analyzed to compare against various water sources. Tap water δ18O values do not reliably reflect drinking water δ18O in sampled regions of Oaxaca, Mexico. Further, the models relating hair keratin to drinking water, built on this assumption, fail to hold predictive power. With an incomplete understanding of tap water's ability to represent drinking water isotopically, researchers need to study the role of other key factors in δ18O values. If the individual's being identified with δ18O are from non-Western communities, forensic practitioners run the risk of incorrectly predicting region of origin because of the method's assumptions. Recognizing the varying sociocultural realities of the communities forensic practitioners seek to aid is paramount as we grow and develop our methods moving forward.

Miocene surface uplift and orogenic evolution of the southern Andean Plateau (central Puna), northwestern Argentina.

We present stable hydrogen-isotope analyses of volcanic glass ([Formula: see text]Dg) and radiometric ages (U-Pb zircon, U-Th calcite, AMS14C) from deformed sedimentary deposits in the vicinity of the intermontane Pocitos Basin in the central Puna of the Andean Plateau at about 24.5°S. Our results demonstrate 2-km surface uplift since the middle to late Miocene and protracted shortening that persists until the present day, while other sectors of the Puna show evidence for tectonically neutral and/or extensional settings. These findings are at odds with previous studies suggesting near-modern elevations (4 km) of the Puna Plateau since the late Eocene and formation of the intermontane Miocene Arizaro-Pocitos Basin associated with gravitational foundering of a dense lithosphere. Geophysical and geochemical data support the removal of continental lithosphere beneath the Puna, but the timing and mechanisms by which this removal occurs have remained controversial. We hypothesize that intermontane basin formation in the central Puna is the result of crustal shortening since about 20 Ma, followed by rapid surface uplift, likely related to lithospheric delamination.

Biological invasions alter the structure of a tropical freshwater food web.

Biological invasions are expected to alter food web structure, but there are limited empirical data directly comparing invaded versus uninvaded food webs, particularly in species-rich, tropical systems. We characterize for the first time the food web of Lake Gatun-a diverse and highly invaded tropical freshwater lake within the Panama Canal. We used stable isotope analysis to reconstruct the trophic structure of the fish community of Lake Gatun and to compare it to that of a minimally invaded reference lake, Lake Bayano. We found significant differences between the trophic structures of these two Neotropical lakes, notably that Lake Gatun's fish community was characterized by a longer food chain, greater isotopic diversity, a broader range of trophic positions and body sizes, and shifts in the isotopic positions of several native taxa relative to Lake Bayano. The degree of isotopic overlap between native and non-native trophic guilds in Lake Gatun was variable, with herbivores exhibiting the lowest (20%-29%) overlap and carnivores the greatest (81%-100%). Overall, our results provide some of the first empirical evidence for the ways in which multiple introduced and native species may partition isotopic space in a species-rich tropical freshwater food web.

An improved folate stable isotope dilution assay of unexploited food sources from Brazil.

Brazil has a diverse plant community, including underutilized non-conventional food crops (PANCs), which have the potential to be a rich source of food and contribute to food security. For assessing the folate content in a range of Brazilian PANCs, we extended the validation of an existing stable isotope dilution assay (SIDA) for the stably 13C-labelled 10-formyl-Pte[13C5]Glu (10-CHO-Pte[13C5]Glu). The SIDA method with an enzymatic treatment, purification step, and an LC-MS/MS measurement was validated regarding linearity, precision, LoD/LoQ, and recovery for 10-CHO-PteGlu. After successful validation, the study of some underutilized Brazilian non-conventional fruits and leaves from the São Paulo State University campus revealed them as an important source of folates. It provided the first insights into the folate content of unexploited food sources from Brazil. Pequi had the highest folate content among the fruits studied, with mean values of 333 µg/100 g based on fresh weight (FW). The analysis also shows that different cultivars of fruit or fruits from different growing locations have a high variability in folate content or other nutritional factors. In most fruits, the main vitamer was 5-CH3-H4folate, but jenipapo and taioba showed the highest content of 10-CHO-PteGlu with 28.22 µg/100 g (FW) in jenipapo peel and 75.64 µg/100 g (FW) in the taioba leaves. Thus, this study also provides results on the importance of the folate vitamer 10-CHO-PteGlu contributing to the total folate content.

Determination of Indispensable Amino Acid Digestibility of the Red Kidney Bean in Humans Using a Dual Stable Isotope Tracer Method.

BACKGROUND: Protein quality of the red kidney bean (RKB), a common source of dietary protein, has been assessed using the protein digestibility-corrected amino acid score (PDCAAS) determined in animal models using mainly oro-fecal digestibility. More recently, the FAO recommended to use digestible indispensable amino acid score (DIAAS) instead of PDCAAS but highlighted insufficient data on true ileal indispensable amino acid (IAA) digestibility of proteins because amino acids are absorbed in the ileum. OBJECTIVES: Using a recently developed dual stable isotope tracer method, we aimed to measure each IAA digestibility as representation of true ileal digestibility of the RKB, Phaseolus vulgaris, in humans consuming a typical Jamaican meal. METHODS: RKB-IAAs were intrinsically labeled by adding 2H2O to the plants. Uniformly labeled-[13C]-spirulina (standard protein) was added to a meal prepared with the labeled RKB and fed to 10 healthy adults (5 males, 5 females) in a nonrandomized trial as primed/intermittent doses to achieve a steady state IAA enrichment in plasma. Enrichment of 2H- and 13C-labeled IAA in plasma and the bean was measured by mass spectrometry. Each IAA digestibility (except tryptophan and histidine) was calculated as the ratio of plasma 2H-IAA to meal 2H-IAA divided by the ratio of plasma 13C-IAA to meal 13C-IAA adjusted for loss of 2H-atom during transamination and digestibility of spirulina. RESULTS: Adequate IAA labeling (>1000 ppm 2H excess) and plasma plateau isotopic enrichment were achieved. Mean RKB-IAA digestibility (%) was 79.4 ± 0.5, ranging from 69.0 ± 1.2 (threonine) to 85.7 ± 1.7 (lysine). CONCLUSION: The dual stable isotope tracer digestibility data are similar to published oro-fecal digestibility supporting substantial nitrogen exchange in the colon. The individual IAA digestibility is useful to derive DIAAS to replace PDCAAS. Using published RKB-IAA composition, extrapolated DIAAS was 0.63 based on the lowest score for methionine. CLINICAL TRIAL REGISTRATION: https://register. CLINICALTRIALS: gov; ID: NCT-04118517.

Intrinsic labelling of common beans with 2H2O to enable estimates of protein digestibility.

BACKGROUND: The use of plant protein intrinsically labelled with stable isotopes provides an innovative solution to assess the efficiency of protein intake by humans. Here, the incorporation of 2H has been applied to intrinsically labelled plant protein in the common bean. This study aimed to evaluate which is the best phenological phase of seed maturation to incorporate the heavy hydrogen isotope 2H into seed amino acids. Common beans (Phaseolus vulgaris L.) were grown in pots, then, after 50 days sowing, 2H2O dissolved in irrigation water was applied, then again at an interval of either 3, 6, 9, and 12 days. RESULTS: Applications of 2H2O at 6, 9, and 12 days after the first application, in the full-flowering stage, were the best treatments for enriching protein-bound amino acids in the bean seed with 2H. CONCLUSION: All treatments resulted in enrichment above 500 ppm, so the treatments (quantity and timing of 2H2O addition) were deemed successful for enriching bean seeds. This makes the intrinsically labelled seeds suitable for preparing test meals to assess the digestion and essential amino acid absorption of common bean amino acids in human subjects.

Plant invasion causes alterations in Darwin's finch feeding patterns in Galápagos cloud forests.

Invasive species pose a major threat to forest biodiversity, particularly on islands such as the Galapágos. Here, invasive plants are threatening the remnants of the unique cloud forest and its iconic Darwin's finches. We posit that food web disturbances caused by invasive Rubus niveus (blackberry), have contributed to the rapid decline of the insectivourous green warbler finch (Certhidae olivacea). We compared the birds' dietary changes in long-term management, short-term management and unmanaged areas. We measured C:N ratios, and δ15N­nitrogen and δ13C­carbon values in both consumer tissues (bird-blood) and food sources (arthropods), as indicators of resource use change, and collected mass abundance, and arthropod diversity data. We characterised the birds' diets using isotope mixing models. The results revealed that finches in (blackberry-invaded) unmanaged areas foraged more on abundant, yet lower quality, arthropods present in the invaded understory. This suggests that blackberry encroachment leads to a decrease in food source quality with physiological consequences for green warbler finch chicks. Results also implied that blackberry control has a short-term impact on food source quantity, which led to a decrease in chick recruitment that we observed in our previous studies; despite this, in the long-term, these managed systems show signs of recovery within three years of restoration.

Feeding Strategies and Trophic Niche Divergence of Three Groups of Dosidicus gigas off Peru: Based on Stable Carbon and Nitrogen Isotopes and Morphology of Feeding Apparatuses.

Dosidicus gigas (D. gigas) is a pelagic cephalopod of ecological and economic importance widely distributed in the eastern Pacific Ocean. Generally, small-, medium-, and large-sized groups of the squids have been respectively identified on the basis of the mantle length (ML) of adults. Intraspecific feeding variability maximizes the utilization of available food resources by D. gigas. However, the coexistence mechanism of three groups has not been fully understood yet. In our study, based on the analyses of beak morphology and stable carbon and nitrogen isotopes, the feeding strategies and coexistence patterns of large-, medium-, and small-sized groups of D. gigas were investigated. D. gigas had a wide range of 13C/12C (δ13C) and 15N/14N (δ15N) values in muscle tissue, variable feeding behaviors, and wide food sources. The δ13C or δ15N values showed no significant difference between the small- and medium-sized groups, which shared the same habitats and fed on preys with the similar trophic level. Compared to small- and medium-sized groups, the large-sized group had a smaller range of habitats and consumed more nearshore foods. Both isotopes and feeding apparatus morphology indicated a high degree of niche overlap between the small- and medium-sized groups, whereas the large-sized group differed significantly from other groups. In addition, the niche width of the female was larger than that of the male in all three groups. We inferred that the sex differences in body length and reproductive behavior led to the difference in niche width. The isotopic niche overlap between female and male samples was the most significant in large-sized group and the least significant in the small-sized group, indicating that different feeding strategies were adopted by the three groups. These findings proved that the three groups of D. gigas off Peruvian waters adopted a feeding strategy with inter- and intra-group regulation. This feeding strategy maximizes the use of food and habitat resources and ensures that different size groups can coexist in the same waters.