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.

Carbon and nitrogen isotopic similarity between the endangered Darwin's fox (Lycalopex fulvipes) and sympatric free-ranging dogs in Chiloé Island, Chile.

Darwin's fox is an opportunistic omnivorous predator native to Chile classified as endangered by the IUCN Red List. Habitat use by Darwin's foxes can be negatively affected by the presence of free-ranging dogs that range freely across native and non-native habitats and can be a source of fox mortality. The objective of this study was to analyze the isotopic similarity of Darwin's fox and sympatric free-ranging dogs in Chiloé Island to determine the impact of anthropogenic environmental alterations on wild predators. We use hair samples to characterise and compare their δ13C and δ15N values and to evaluate isotopic similarity and isotope niches overlap. A generalised linear model was used to associate the isotope value with landscape variables (forest cover and vegetation type) and distance to the nearest house. We found no significant differences in δ13C or δ15N values between foxes and dogs, and a marginally significant isotope niche overlap (59.4 %). None of the selected variables at landscape and site scale were related to isotope values. Although our study is not a probe of direct contact between foxes and free-ranging dogs, the high isotopic similarity highlights the risk of pathogen spillover from free-ranging dogs to Darwin's foxes.

The Physiological Effect of a Holoparasite Over a Cactus Along an Environmental Gradient.

Echinopsis chiloensis is an endemic cactus from Chile, distributed in a temperature and rainfall gradient between 30° and 35° South latitude, with mean temperatures increasing and precipitation decreasing toward the north. It is the main host of the mistletoe Tristerix aphyllus, a holoparasite completely dependent on the cactus for water, carbon, and minerals. In this study, we investigated the consequences of parasitism over the fitness and physiology of this cactus throughout its distribution range and how it is affected by the environment. We measured five functional traits in eight populations latitudinally distributed, the first three only for the host: reproductive fitness, stomatal traits (density and size), and photosynthesis (during winter and summer); and the last two for the host and parasite: stable isotopes (∂13C and ∂15N), and nutrients (carbon and nitrogen content). The results showed a negative effect of parasitism over fitness of infected cacti. However, the higher nitrogen concentrations in cactus tissues toward the south improved overall fitness. Regarding photosynthesis, we only observed a negative effect of parasitism during the dry season (summer), which is also negatively affected by the increase in summer temperatures and decrease in winter rainfall toward the north. There were no differences in nutrient concentration or in the isotopic signature of healthy and infected cacti. Conversely, we observed a higher carbon and lower nitrogen concentration in mistletoes than in cacti regardless of latitude. The loss of temperature seasonality toward the north increases the C:N ratio, and the values between the parasite and its host diverge. ∂15N was similar between parasites and hosts while ∂13C of the parasite was enriched when compared to its host. Overall, the infection by T. aphyllus affects Echinopsis chiloensis fitness but showed no strong effects over the cactus physiology, except for the summer photosynthesis. Therefore, our data revealed that E. chiloensis response to T. aphyllus infection is sensitive to environmental changes in a way that could be strongly impacted by the desertification projected for this area due to climate change.

Comparative isotope ecology of western Amazonian rainforest mammals.

Closed-canopy rainforests are important for climate (influencing atmospheric circulation, albedo, carbon storage, etc.) and ecology (harboring the highest biodiversity of continental regions). Of all rainforests, Amazonia is the world's most diverse, including the highest mammalian species richness. However, little is known about niche structure, ecological roles, and food resource partitioning of Amazonian mammalian communities over time. Through analyses of δ13Cbioapatite, δ13Chair, and δ15Nhair, we isotopically characterized aspects of feeding ecology in a modern western Amazonian mammalian community in Peru, serving as a baseline for understanding the evolution of Neotropical rainforest ecosystems. By comparing these results with data from equatorial Africa, we evaluated the potential influences of distinct phylogenetic and biogeographic histories on the isotopic niches occupied by mammals in analogous tropical ecosystems. Our results indicate that, despite their geographical and taxonomic differences, median δ13Cdiet values from closed-canopy rainforests in Amazonia (-27.4‰) and equatorial Africa (-26.9‰) are not significantly different, and that the median δ13Cdiet expected for mammalian herbivores in any closed-canopy rainforest is -27.2‰. Amazonian mammals seem to exploit a narrower spectrum of dietary resources than equatorial African mammals, however, as depicted by the absence of highly negative δ13Cdiet values previously proposed as indicative of rainforests (<-31‰). Finally, results of keratin and bioapatite δ13C indicate that the predictive power of trophic relationships, and traditional dietary ecological classifications in bioapatite-protein isotopic offset expectations, must be reconsidered.

Seasonal oxygen isotope variations in freshwater bivalve shells as recorders of Amazonian rivers hydrogeochemistry.

Freshwater bivalve shell oxygen isotope values (δ18OS) may act as a recorder of river δ18O variations that can then be interpreted in terms of hydrology (e.g. precipitation-evaporation balance, precipitation and river discharge patterns). We investigated the potential of this proxy measured across the hinge of South American unionid shells: Anodontites elongatus collected in Peru and A. trapesialis in Brazil. The isotopic signatures were reproducible between individuals of the same species. A. trapesialis clearly showed a strong δ18OS cyclicity in accordance with its growth patterns while A. elongatus presented less clear δ18OS with lower amplitude. We confirm that the deposition of successive growth lines and increments is annual, with growth line corresponding to the wet season. Also, we suggest that low amplitude of δ18OS in the A. elongatus shells indicates a habitat close to the river while large amplitude of δ18OS cycles observed in A. trapesialis shells would reflect a floodplain lake habitat, seasonally disconnected from the river and thus subjected to higher seasonal fluctuations in water δ18O. Considering these promising first results, future studies could be directed towards the use of fossil shells to reconstruct the past and present hydrological and geochemical conditions of the Amazon.

Does faecal matter reflect location? An initial assessment of isotopic variability between consumed prey remains and faecal matter for wild jaguars.

Faecal isotopic analysis may complement other non-invasive wildlife survey tools for monitoring landscape use by carnivores, such as motion-detecting cameras and non-invasive genetic sampling. We analysed carbon, nitrogen, and strontium isotopes in faecal matter produced by jaguars (Panthera onca) as well as bones from consumed prey at the Mountain Pine Ridge Forest Reserve (MPR) in Belize, Central America. The MPR is ideally suited for a spatial isotope study as vegetation and geology both vary considerably. The isotopic composition of faecal matter should reflect the habitat and geology where consumed prey lived. We used bone from consumed prey recovered from jaguar scats as a proxy for diet. Faecal matter and bone showed comparable spatial isotopic trends, suggesting that the isotopic composition of jaguar faeces can be used to detect foraging in different habitats (pine forest versus broadleaf forest) or on different geologies (Mesozoic carbonates; Palaeozoic granite, contact metamorphics, and metasediments). This result is reassuring as bones are not always present in carnivore scats. Studying landscape use by cryptic and wide-ranging carnivore species like jaguars remains challenging. Isotopic analysis of faecal matter complements the existing array of non-invasive spatial monitoring tools.

Stable isotope analysis of white-tailed deer teeth as a paleoenvironmental proxy at the Maya site of La Joyanca, northwestern Petén, Guatemala.

Carbon and oxygen isotopes ratios from herbivore teeth have previously been used as paleo-environmental proxies in temperate zones. However, their utility in tropical zones remains uncertain. In this study, sequential sub-samples from white-tailed deer (Odocoileus virginianus) teeth (second and third molars) from the Maya archaeological site of La Joyanca, located in northwestern Petén, Guatemala, show that δ18O of enamel carbonate corresponds broadly to modern observed precipitation δ18O over the 10-month period of tooth formation, capturing rainfall seasonality. The analyses also detect significant diachronic differences in the δ18O between the periods 1100-1000 BP (850-950 A.D.) and 1000-900 BP (950-1050 A.D.) at La Joyanca. The δ13C in both periods are indicative of a C3-plant based diet, which suggests cultivation of maize did not differentially affect deer diet during this period.

Does trawl fisheries affect the diet of fishes? A stable isotope analysis approach.

Trawling is the main fishing practice worldwide and its ecosystem effects have been raising concern over the past decades. Long-term impacts can be monitored through changes in the trophic structure, and several studies evaluated trophic level (TL) shifts in fish populations between trawled and untrawled environments. However, published results are contrasting. We performed a metanalysis integrating all available studies that evaluated TL shifts in fishes between trawled and untrawled environments and conducted a local study comparing several features of the trophic ecology in two species of fishes. According to the metanalysis, TL does not change significantly with trawling. In contrast, the local study showed higher TLs and broader isotopic niches in the trawled environment. Diet reconstruction indicated a potential consumption of hake, the main discard component, at the trawled environment. All the studies used in the metanalysis were conducted in the Northern Hemisphere, whereas the local study represents the first data available from the Southern Hemisphere. As industrial commercial fisheries in Argentina are relatively recent, it is possible that our data are capturing the initial stage of ecological changes induced by trawling, compared with the historical fisheries located at the Northern Hemisphere.

The relative isotopic abundance (δ13C, δ15N) during composting of agricultural wastes in relation to compost quality and feedstock.

Variations in the relative isotopic abundance of C and N (δ13C and δ15N) were measured during the composting of different agricultural wastes using bench-scale bioreactors. Different mixtures of agricultural wastes (horse bedding manure + legume residues; dairy manure + jatropha mill cake; dairy manure + sugarcane residues; dairy manure alone) were used for aerobic-thermophilic composting. No significant differences were found between the δ13C values of the feedstock and the final compost, except for dairy manure + sugarcane residues (from initial ratio of -13.6 ± 0.2 ‰ to final ratio of -14.4 ± 0.2 ‰). δ15N values increased significantly in composts of horse bedding manure + legumes residues (from initial ratio of +5.9 ± 0.1 ‰ to final ratio of +8.2 ± 0.5 ‰) and dairy manure + jatropha mill cake (from initial ratio of +9.5 ± 0.2 ‰ to final ratio of +12.8 ± 0.7 ‰) and was related to the total N loss (mass balance). δ13C can be used to differentiate composts from different feedstock (e.g. C3 or C4 sources). The quantitative relationship between N loss and δ15N variation should be determined.

Oxygen isotope composition of North American bobcat (Lynx rufus) and puma (Puma concolor) bone phosphate: implications for provenance and climate reconstruction.

Feline carnivores are threatened by illegal wildlife trade. Tracing the provenance of unknown felid tissues via stable isotope analysis could provide important information in wildlife crime investigations. The oxygen isotope composition of mammalian skeletal phosphate (δ(18)Op) is widely applied to trace the origin of animal remains and to reconstruct migratory patterns in palaeontological, archaeological, ecological and wildlife forensic applications. Teeth and bones of terrestrial mammals form at constant body temperature in isotope equilibrium with body water, which is predominantly controlled by ingested meteoric water (δ(18)Ow) that varies systematically with latitude, altitude and climate. Here we analysed δ(18)Op of 106 North American puma and bobcat bones of known geographic origin to establish the first δ(18)Op-δ(18)Ow regression for feline carnivores: δ(18)Op = 0.40(±0.04) * δ(18)Ow + 20.10(±0.40) (R(2) = 0.46, n = 106). This was compared with those from their respective prey species (deer and rabbit), a canid carnivore (fox) and other placental mammals. Effects of species, sex and relative humidity on the feline δ(18)Op-δ(18)Ow correlation were analysed and additional intra-individual tissue comparisons (hair δ(18)Oh vs. bone δ(18)Op) were performed for some bobcat individuals. Bobcats and pumas exhibited only a moderate δ(18)Op-δ(18)Ow correlation, which differed from canid carnivores and other placental mammals. However, feline δ(18)Op values revealed a moderate relation with δ(18)Ow, which lacks for the δ(18)Oh of hair from the same bobcat individuals. This indicates a difference in oxygen isotope routing from body water to bioapatite and hair. Most herbivores and omnivores track δ(18)Ow in their bioapatite δ(18)Op values much better, whereas δ(18)Op and especially δ(18)Oh values of feline carnivores are less precise proxies for meteoric water δ(18)Ow values and thus for provenance determination in wildlife forensics and palaeoclimate reconstructions. Oxygen isotope fingerprinting of bobcat and puma is biased by factors related to their diet, behaviour and metabolism that need to be better understood.

Analysis of metabolic pools in broilers chicks.

This paper shows the possibility of obtaining new parameters for the mathematical modelling of data on stable isotopes in biological systems and its application in obtaining data on metabolic pools of blood plasma, blood serum, liver and muscle of broilers. This theory states that the modelling of turnover used for studies of isotopic incorporation when the metabolism has a single metabolic pool is feasible by the technique of setting an exponential. However, when the metabolism has more than one metabolic pool, it is necessary to apply the linearization technique, linear regression adjustment and evaluation of the assumptions of regression to obtain the kinetic parameters such as half-life (T1/2) and isotope exchange rate (k). The application of this technique on carbon-13 data from 100 one-day-old chicks, with the change of diet composed of grains of the photosynthetic cycle of plants from C4 to C3, in broilers has enabled the discovery that the liver, blood plasma and blood serum have a single metabolic pool; however, the pectoral muscle has two metabolic pools. For the liver, blood plasma and blood serum, the half-life values were found by the exponential fit being T1/2 = 1.4 days with the rate of exchange of k = 0.502, T1/2 = 2.4 days with k = 0.293 and T1/2 = 2.0 days with k = 0.348, respectively. For the pectoral muscle, after linearization, the half-life values were found for T1/2(1) = 1.7 and T1/2(2) = 3 days, with exchange rates of k1 = 0.405 and k2 = 0.235, representing approximately 66 and 34%, respectively.