Mercury concentrations and differences in isotopic niches of fish from upstream and downstream of an Amazon reservoir dam.

Reservoir construction promotes many environmental impacts, including the enhancement of mercury concentrations in fish. The processes that can influence mercury concentrations in fish in Amazonian reservoirs are still little explored in depth, especially when we consider the possible particularities of the ecosystems in question. This study aims to investigate how mercury concentrations in fish could be influenced by the Tucuruí dam, considering possible changes in their feeding and trophic position according to the dam position (up or downstream). Fish were sampled upstream and downstream of the Tucuruí reservoir, and total mercury (THg) and stable isotopes of carbon and nitrogen (δ15N and δ13C) were measured in muscles. We observed three different Hg bioaccumulation patterns influenced by the dam. These differences occurred due to species trophic niche changes corroborated by the isotope analysis. Higher THg concentrations downstream compared to those upstream ones were only observed for Geophagus proximus. On the contrary, Plagioscion squamosissimus, from downstream, presented lower concentrations than upstream ones. The isotopic niche of these two species presented different changes according to the sampled site. THg biomagnification was higher upstream compared to downstream, considering that the regression slope was approximately two times higher upstream versus downstream. THg concentrations in fish were explained by the differences in their feeding habits according to their location in relation to the dam. The difference in THg biomagnification was able to reflect differences in structure of the food web chain in ecosystems under the dam's influence.

Comparative genomics and stable isotope analysis reveal the saprotrophic-pathogenic lifestyle of a neotropical fungus.

In terrestrial forested ecosystems, fungi may interact with trees in at least three distinct ways: (i) associated with roots as symbionts; (ii) as pathogens in roots, trunks, leaves, flowers, and fruits; or (iii) decomposing dead tree tissues on soil or even on dead tissues in living trees. Distinguishing the latter two nutrition modes is rather difficult in Hymenochaetaceae (Basidiomycota) species. Herein, we have used an integrative approach of comparative genomics, stable isotopes, host tree association, and bioclimatic data to investigate the lifestyle ecology of the scarcely known neotropical genus Phellinotus, focusing on the unique species Phellinotus piptadeniae. This species is strongly associated with living Piptadenia gonoacantha (Fabaceae) trees in the Atlantic Forest domain on a relatively high precipitation gradient. Phylogenomics resolved P. piptadeniae in a clade that also includes both plant pathogens and typical wood saprotrophs. Furthermore, both genome-predicted Carbohydrate-Active Enzymes (CAZy) and stable isotopes (δ13C and δ15N) revealed a rather flexible lifestyle for the species. Altogether, our findings suggest that P. piptadeniae has been undergoing a pathotrophic specialization in a particular tree species while maintaining all the metabolic repertoire of a wood saprothroph. IMPORTANCE: This is the first genomic description for Phellinotus piptadeniae. This basidiomycete is found across a broad range of climates and ecosystems in South America, including regions threatened by extensive agriculture. This fungus is also relevant considering its pathotrophic-saprotrophic association with Piptadenia goanocantha, which we began to understand with these new results that locate this species among biotrophic and necrotrophic fungi.

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.

Developing δ15N and δ13C isoscapes using whole blood from Magellanic penguins, Spheniscus magellanicus.

RATIONALE: Understanding the migration of marine animals is hindered by the limitations of traditional tracking methods. It is therefore crucial to develop alternative methods. Stable isotope-based tracking has proven useful for this task, although it requires detailed isoscapes in the focal area. Here, we present predator-based isoscapes of the coastal zone of the Patagonian Shelf Large Marine Ecosystem (PSLME), which offers a novel tool for geolocation. METHODS: Whole-blood samples from breeding Magellanic penguins nesting at 11 colonies were used to create δ15N and δ13C isoscapes. Isotopic values were assigned to random positions inside their corresponding foraging area. Spatial analysis and data interpolation resulted in δ15N and δ13C isoscapes for the coastal zone of the PSLME, which were validated through cross-validation. RESULTS: The isoscapes mean standard error ranged from 0.05 to 0.41 for δ15N and from 0.07 to 0.3 for δ13C, similar to the error range of the mass spectrometer used for measuring isotope ratios. Predictive surfaces reflected the latitudinal trends, with δ13C and δ15N values increasing northwards. δ13C values showed a strong latitudinal gradient, while δ15N values had two distinct domains, with higher values in the north. The error surface indicated the highest certainty within 130 km from the shore and within the reported Magellanic penguin foraging areas. CONCLUSIONS: Both isoscapes revealed strong spatial variation. The δ13C isoscape showed a latitudinal gradient, consistent with patterns in other oceans. The δ15N isoscape clearly separated northern and southern colonies, likely influenced by nitrogen sources. The error obtained fell within the measurement error ranges, adding credibility to the models.

Trophic plasticity of a tropical seabird revealed through DNA metabarcoding and stable isotope analyses.

DNA metabarcoding and stable isotope analysis have significantly advanced our understanding of marine trophic ecology, aiding systematic research on foraging habits and species conservation. In this study, we employed these methods to analyse faecal and blood samples, respectively, to compare the trophic ecology of two Red-billed Tropicbird (Phaethonaethereus; Linnaeus, 1758) colonies on Mexican islands in the Pacific. Trophic patterns among different breeding stages were also examined at both colonies. Dietary analysis reveals a preference for epipelagic fish, cephalopods, and small crustaceans, with variations between colonies and breeding stages. Isotopic values (δ15N and δ13C) align with DNA metabarcoding results, with wider niches during incubation stages. Differences in diet are linked to environmental conditions and trophic plasticity among breeding stages, influenced by changing physiological requirements and prey availability. Variations in dietary profiles reflect contrasting environmental conditions affecting local prey availability.

Body size predicts ontogenetic nitrogen stable-isotope (δ15N) variation, but has little relationship with trophic level in ectotherm vertebrate predators.

Large predators have disproportionate effects on their underlying food webs. Thus, appropriately assigning trophic positions has important conservation implications both for the predators themselves and for their prey. Large-bodied predators are often referred to as apex predators, implying that they are many trophic levels above primary producers. However, theoretical considerations predict both higher and lower trophic position with increasing body size. Nitrogen stable isotope values (δ15N) are increasingly replacing stomach contents or behavioral observations to assess trophic position and it is often assumed that ontogenetic dietary shifts result in higher trophic positions. Intraspecific studies based on δ15N values found a positive relationship between size and inferred trophic position. Here, we use datasets of predatory vertebrate ectotherms (crocodilians, turtles, lizards and fishes) to show that, although there are positive intraspecific relationships between size and δ15N values, relationships between stomach-content-based trophic level (TPdiet) and size are undetectable or negative. As there is usually no single value for 15N trophic discrimination factor (TDF) applicable to a predator species or its prey, estimates of trophic position based on δ15N in ectotherm vertebrates with large size ranges, may be inaccurate and biased. We urge a reconsideration of the sole use of δ15N values to assess trophic position and encourage the combined use of isotopes and stomach contents to assess diet and trophic level.

Comparison of the precursor, amino acid oxidation, and end-product methods for the evaluation of protein turnover in senior dogs.

Stable isotope methods have been used to study protein metabolism in humans; however, there application in dogs has not been frequently explored. The present study compared the methods of precursor (13C-Leucine), end-products (15N-Glycine), and amino acid oxidation (13C-Phenylalanine) to determine the whole-body protein turnover rate in senior dogs. Six dogs (12.7 ± 2.6 years age, 13.6 ± 0.6 kg bodyweight) received a dry food diet for maintenance and were subjected to all the above-mentioned methods in succession. To establish 13C and 15N kinetics, according to different methodologies blood plasma, urine, and expired air were collected using a specifically designed mask. The volume of CO2 was determined using respirometry. The study included four methods viz. 13C-Leucine, 13C-Phenylalanine evaluated with expired air, 13C-Phenylalanine evaluated with urine, and 15N-Glycine, with six dogs (repetitions) per method. Data was subjected to variance analysis and means were compared using the Tukey test (P<0.05). In addition, the agreement between the methods was evaluated using Pearson correlation and Bland-Altman statistics. Protein synthesis (3.39 ± 0.33 g.kg-0,75. d-1), breakdown (3.26 ± 0.18 g.kg-0.75.d-1), and flux estimations were similar among the four methods of study (P>0.05). However, only 13C-Leucine and 13C-Phenylalanine (expired air) presented an elevated Pearson correlation and concordance. This suggested that caution should be applied while comparing the results with the other methodologies.

Seeing further into the early steps of the endangered atlantic goliath grouper (Epinephelus itajara): Eye lenses high resolution isotopic profiles reveal ontogenetic trophic and habitat shifts.

Estuarine mangroves are often considered nurseries for the Atlantic Goliath grouper juveniles. Yet, the contributions of different estuarine primary producers and habitats as sources of organic matter during early ontogenetic development remain unclear. Given the species' critically endangered status and protection in Brazil, obtaining biological samples from recently settled recruits in estuaries is challenging. In this study, we leveraged a local partnership with fishers and used stable isotope (C and N) profiles from the eye lenses of stranded individuals or incidentally caught by fishery to reconstruct the trophic and habitat changes of small juveniles. The eye lens grows by the apposition of protein-rich layers. Once these layers are formed, they become inert, allowing to make inferences on the trophic ecology and habitat use along the development of the individual until its capture. We used correlations between fish size and the entire eye lens size, along with estuarine baselines, to reconstruct the fish size and trophic positions for each of the lens layers obtained. We then used dominant primary producers and basal sources from mangrove sheltered, exposed estuarine and marine habitats to construct an ontogenetic model of trophic and habitat support changes since maternal origins. Our model revealed marine support before the juveniles reached 25 mm (standard length), followed by a rapid increase in reliance on mangrove sheltered sources, coinciding with the expected size at settlement. After reaching 60 mm, individuals began to show variability. Some remained primarily supported by the mangrove sheltered area, while others shifted to rely more on the exposed estuarine area around 150 mm. Our findings indicate that while mangroves are critical for settlement, as Goliath grouper juveniles grow, they can utilize organic matter produced throughout the estuary. This underscores the need for conservation strategies that focus on seascape connectivity, as protecting just one discrete habitat may not be sufficient to preserve this endangered species and safeguard its ecosystem functions.

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.

Enameloid-bound δ15 N reveals large trophic separation among Late Cretaceous sharks in the northern Gulf of Mexico.

The nitrogen isotopic composition (15 N/14 N ratio, or δ15 N) of enameloid-bound organic matter (δ15 NEB ) in shark teeth was recently developed to investigate the biogeochemistry and trophic structures (i.e., food webs) of the ancient ocean. Using δ15 NEB , we present the first nitrogen isotopic evidence for trophic differences between shark taxa from a single fossil locality. We analyze the teeth of four taxa (Meristodonoides, Ptychodus, Scapanorhynchus, and Squalicorax) from the Late Cretaceous (83-84 Ma) Trussells Creek site in Alabama, USA, and compare the N isotopic findings with predictions from tooth morphology, the traditional method for inferring shark paleo-diets. Our δ15 NEB data indicate two distinct trophic groups, with averages separated by 6.1 ± 2.1‰. The lower group consists of Meristodonoides and Ptychodus, and the higher group consists of Scapanorhynchus and Squalicorax (i.e., lamniforms). This δ15 NEB difference indicates a 1.5 ± 0.5 trophic-level separation between the two groups, a finding that is in line with paleontological predictions of a higher trophic level for these lamniforms over Meristodonoides and Ptychodus. However, the δ15 NEB of Meristodonoides is lower than suggested by tooth morphology, although consistent with mechanical tests suggesting that higher trophic-level bony fishes were not a major component of their diet. Further, δ15 NEB indicates that the two sampled lamniform taxa fed at similar trophic levels despite their different inferred tooth functions. These two findings suggest that tooth morphology alone may not always be a sufficient indicator of dietary niche. The large trophic separation revealed by the δ15 NEB offset leaves open the possibility that higher trophic-level lamniforms, such as those measured here, preyed upon smaller, lower trophic-level sharks like Meristodonoides.

Differentiating wild from captive animals: an isotopic approach.

Background: Wildlife farming can be an important but complex tool for conservation. To achieve conservation benefits, wildlife farming should meet a variety of criteria, including traceability conditions to identify the animals' origin. The traditional techniques for discriminating between wild and captive animals may be insufficient to prevent doubts or misdeclaration, especially when labels are not expected or mandatory. There is a pressing need to develop more accurate techniques to discriminate between wild and captive animals and their products. Stable isotope analysis has been used to identify animal provenance, and some studies have successfully demonstrated its potential to differentiate wild from captive animals. In this literature review, we examined an extensive collection of publications to develop an overall picture of the application of stable isotopes to distinguish between wild and captive animals focusing on evaluating the patterns and potential of this tool. Survey methodology: We searched peer-reviewed publications in the Web of Science database and the references list from the main studies on the subject. We selected and analyzed 47 studies that used δ13C, δ15N, δ2H, δ18O, and δ34S in tissues from fish, amphibians, reptiles, birds, and mammals. We built a database from the isotope ratios and metadata extracted from the publications. Results: Studies have been using stable isotopes in wild and captive animals worldwide, with a particular concentration in Europe, covering all main vertebrate groups. A total of 80.8% of the studies combined stable isotopes of carbon and nitrogen, and 88.2% used at least one of those elements. Fish is the most studied group, while amphibians are the least. Muscle and inert organic structures were the most analyzed tissues (46.81% and 42.55%). δ13C and δ15N standard deviation and range were significantly higher in the wild than in captive animals, suggesting a more variable diet in the first group. δ13C tended to be higher in wild fishes and in captive mammals, birds, reptiles, and amphibians. δ15N was higher in the wild terrestrial animals when controlling for diet. Only 5.7% of the studies failed to differentiate wild and captive animals using stable isotopes. Conclusions: This review reveals that SIA can help distinguish between wild and captive in different vertebrate groups, rearing conditions, and methodological designs. Some aspects should be carefully considered to use the methodology properly, such as the wild and captivity conditions, the tissue analyzed, and how homogeneous the samples are. Despite the increased use of SIA to distinguish wild from captive animals, some gaps remain since some taxonomic groups (e.g., amphibians), countries (e.g., Africa), and isotopes (e.g., δ2H, δ18O, and δ34S) have been little studied.

Feeding ecology based on stable carbon and nitrogen isotopes: A comparative study on different Guiana dolphin tissues.

Different tissues are used for stable isotope analysis in cetacean investigations. However, variation in the isotopic composition of tissues with different turnover rates has been reported for cetaceans. To better understand stable carbon and nitrogen isotopes (δ13C and δ15N) in skin compared to other tissues, this study assessed the isotopic variation among the liver, muscle, and skin of Guiana dolphins (Sotalia guianensis), as well as the influence of sex on these variations. No differences were found in δ13C among male tissues, but females showed lower values in the liver compared to muscle and skin. Differences in δ15N were observed among all tissues, with different variation patterns for males and females. Four females were distinguished from males and other females by their 13C depletion in all tissues and δ15N variation pattern. We conclude that skin and muscle may be equivalent in δ13C values for Guiana dolphins. The multiple-tissue analysis brings new insights into their feeding ecology and provides background for stable isotope analysis using non-destructive sampling techniques in small cetaceans.

Sources of intraspecific variation in the isotopic niche of a semi-aquatic predator in a human-modified landscape.

Intraspecific variation modulates patterns of resource use by species, potentially affecting the structure and stability of food webs. In human-modified landscapes, habitat disturbance modifies trophic interactions and intraspecific niche variation, impacting population persistence. Here, we investigated the relationship of sex, ontogeny, and habitat factors with the trophic niche of Caiman crocodilus in an agricultural landscape. We evaluated temporal variation in the trophic niche parameters using carbon and nitrogen stable isotope analysis from different body tissues. We found that caimans exploit the same carbon and nitrogen pools through time, with low isotopic variability between seasons, partly due to the slow isotope turnover rates of tissues in crocodilians. Conversely, the trophic niche of caimans varied across habitats, but with no evidence of a difference between natural and anthropogenic habitats. It apparently results from the influence of habitat suitability, connectivity, and caiman movements during the foraging. Our findings highlight the broader niches of juvenile caimans relative to adults, possibly in response of territorialism and opportunistic foraging strategy. Although using similar resources, females had a larger niche than males, probably associated with foraging strategies during nesting. Considering the sex and body size categories, caimans occupied distinct isotopic regions in some habitats, indicating apparent niche segregation. Ontogenetic trophic shifts in the isotopes (δ13C and δ15N) depended on sex, leading to resource partitioning that can potentially reduce intraspecific competition. Decision-makers and stakeholders should consider the trophic dynamics of sex and body size groups for the sustainable management and conservation of caiman populations, which implies in the maintenance of wetland habitats and landscape heterogeneity in the Formoso River floodplain.

Colonial waterbirds provide persistent subsidies to swamp forests along an estuarine island food chain.

Birds are excellent vectors of allochthonous matter and energy due to their high mobility, with more intense flow when waterbirds congregate in breeding colonies, feeding in surrounding aquatic and terrestrial areas, and promoting nutritional pulses to nutrient-poor environments. In southern Brazil, a swamp forest on an estuarine island is used by waterbirds for breeding, providing an opportunity to investigate the potential effects of transport of matter between nutrient-rich environments. Soil, plants, invertebrates, and blood from terrestrial birds were collected and stable isotopes compared to similar organisms in a control site without heronries. Values of δ15N and δ13C from waterbirds were higher in the colony in comparison to the control site (spatial effect). The enrichment of 15N and 13C provided during the active colony period persisted after the breeding period, especially for δ15N, which was higher in all compartments (temporal effect). Moreover, the enrichment of 15N occurred along the entire trophic chain (vertical effect) in the colony environment, including different guilds of invertebrates and land birds. The enrichment in 13C seems to lose strength and was mostly explained by factors such as trophic guild rather than site, especially in birds. Bayesian mixture models with terrestrial vs. estuarine endpoints demonstrated that all organisms from both colony and control environments had assimilated estuarine matter. Finally, detritivorous invertebrates showed greater assimilation when compared to other guilds. This study demonstrates that adjacent nutrient-rich environments, such as palustrine forests and estuaries, are nutritionally enriched in several dimensions from nearby autochthonous subsidies that are maintained throughout the year.

Movement and population dispersal of dolphinfish (Coryphaena hippurus) across the Eastern Tropical Pacific inferred from carbon and nitrogen stable isotope analyses.

The dolphinfish (Coryphaena hippurus) is a globally distributed marine predator that supports one of the most important coastal fisheries along the Eastern Tropical Pacific (ETP), but its spatial movements in this area are poorly understood. Stable isotope values (δ13 C and δ15 N) of white muscle from dolphinfish (n = 220) captured at different locations across the ETP (i.e., Mexico, Costa Rica, Ecuador, Peru and oceanic areas) were normalized to copepod baseline stable isotope values to estimate dolphinfish trophic position, movements and population dispersal. Movement or residence patterns were inferred from the difference in δ15 N values (Δ15 Ndolphinfish-copepod ‰) between copepods and dolphinfish muscle. Baseline corrected isotope values (δ13 Cdolphinfish-copepod and δ15 Ndolphinfish-copepod ) of dolphinfish muscle were used to estimate isotopic niche metrics and infer population dispersal across isoscapes. Values of δ13 C and δ15 N differed between juvenile and adult dolphinfish and across the ETP. Trophic position estimates ranged from 3.1 to 6.0 with a mean of 4.6. Adults and juveniles had similar trophic position estimates, whereas isotopic niche areas (SEA ‰2 ) of adults were greater relative to juveniles in every location. Adult dolphinfish showed "moderate movement by some individuals" in all locations based on Δ15 Ndolphinfish-copepod values, except for Costa Rica where adults were classified with "high degree of movement by some individuals" whereas juveniles showed "limited movement" in all areas except Mexico. Population dispersal based on Δ15 Ndolphinfish-copepod values showed "moderate" and "high" dispersal for adults and "no dispersal" for most juveniles, except for Mexico. This study provides insight into potential dolphinfish spatial mobility across an area of interest for multiple nations, which can help to improve stock assessments and management of the species.

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.

Nitrate isotopes (δ15N, δ18O) in precipitation: best practices from an international coordinated research project.

Stable isotope ratios of nitrogen and oxygen (15N/14N and 18O/16O) of nitrate (NO3-) are excellent tracers for developing systematic understanding of sources, conversions, and deposition of reactive atmospheric nitrogen (Nr) in the environment. Despite recent analytical advances, standardized sampling of NO3-) isotopes in precipitation is still lacking. To advance atmospheric studies on Nr species, we propose best-practice guidelines for accurate and precise sampling and analysis of NO3- isotopes in precipitation based on the experience obtained from an international research project coordinated by the International Atomic Energy Agency (IAEA). The precipitation sampling and preservation strategies yielded a good agreement between the NO3- concentrations measured at the laboratories of 16 countries and at the IAEA. Compared to conventional methods (e.g., bacterial denitrification), we confirmed the accurate performance of the lower cost Ti(III) reduction method for isotope analyses (15N and 18O) of NO3- in precipitation samples. These isotopic data depict different origins and oxidation pathways of inorganic nitrogen. This work emphasized the capability of NO3- isotopes to assess the origin and atmospheric oxidation of Nr and outlined a pathway to improve laboratory capability and expertise at a global scale. The incorporation of other isotopes like 17O in Nr is recommended in future studies.

Isotopes of nitrate and gadolinium fingerprints to assay human inputs in Guarani Aquifer System.

The use of environmental tracers brings comprehensive benefits to the management of water resources since it helps to prevent their pollution, minimize public health risks, and thus reduce the impact of urbanization. In Brazil, the Guarani Aquifer System (GAS) has strategic and environmental importance, making its preservation and sustainable exploitation mandatory. The present study aimed at evaluating sources of contamination in the GAS using the combination of geochemical data and two environmental tracers: nitrate isotopes (15NNO3 and 18ONO3) and one rare earth element (Gadolinium-Gd). For that, five wells-four exploiting the GAS and one the Bauru Aquifer System (BAS)-were selected to discuss the human inputs in groundwater used for public supply in an urban area. Traditional physicochemical analyses were conducted for six campaign samplings and nitrate monitoring for this period was evaluated on a time scale, also considering the accumulated rainfall. Besides that, the double isotopic method (DIM), e.g., δ18ONO3 e δ15NNO3, was applied to identify the fractionation and enable the distinction of the nitrate contamination source. In addition, the determination of anomalies of Gd, a wastewater-derived contaminant, was also performed to verify recent human inputs in groundwater. The results show that the local existence of nitrate in the GAS and BAS-even at low concentrations (values from 0.26 to 6.68 mg L-1)-originated from anthropogenic inputs (septic waste), as indicates the typical isotopic signals ratio in the isotopic approach. Associated with that, the evaluation of Gd permitted the separation of groundwater samples into older or more recent leakages. The use of environmental tracers to assess anthropogenic inputs in groundwater reiterates the importance of adopting more effective protection strategies for water resources management systems, in order to prevent contamination.

Comparison of markers to estimate microbial adherence and fibrolytic activity in forages after in vitro incubation.

The aims of this study were to evaluate the relationship between attached microbial markers and degradability of forage samples incubated in vitro and to compare these microbial markers. In Trial 1, the content of 15N, purines, and phosphorus (P) as well as xylanase activity in residue of different forage species were measured after 24 h of incubation in a conventional in vitro system at pH 6.8. Trial 2 used the same procedures as those of Trial 1 except that forage samples were incubated in media with different initial pH (5.5, 6.0, 6.5, or 7.0). There was no correlation (P > 0.10) between forage degradability and either microbial marker in Trial 1. Degradability of both, bermuda and ryegrass, and the content of all markers in the incubation residues was positively affected by increased pH (P < 0.05). The content of 15N in residues was linearly related to xylanase activity (P < 0.05) but not with P or purines content. In conclusion, the nutritional potential of different forage species may not to be compared, based on the content of microbial markers in the incubation residues. In other way, within a forage species, the in vitro degradation was directly associated to either marker. However, P presents analytical advantage over other markers.

What a few hairs can tell us about the resource use of giant armadillos.

Understanding of resource requirements of vulnerable species is key for conservation planning. Here, we used stable carbon (δ13 C) and nitrogen isotopes (δ15 N) of hair from giant armadillos (Priodontes maximus) to understand individual resource use. We collected hair from 19 giant armadillos analyzing 34 samples from the Brazilian Pantanal and compared the resource use between sexes and age classes. We also assessed the relationship of isotopic values with individuals' body mass and the habitat type used. We observed a wide variation in resource use by giant armadillos, showing that individuals consume prey that feeds on distinct resources and occupies different habitat types, indicating that there is individual variation in foraging behavior. Most giant armadillos presented a mixed diet (C3 /C4 resources, forests/open areas), but a quarter of individuals had strictly C4 diets (open areas). Males are more prone to forage on C4 resources, presenting an isotopic niche 2.6-times larger than that of females. Subadults presented diets more associated with habitats with high arboreal cover (C3 resources, forests), while adults foraged more in open areas (C4 resources). This result is mirrored by the positive relationship between δ13 C values and body mass, suggesting that larger giant armadillos tend to feed more in open areas. We observed that δ13 C values decreased as individuals increased the use of habitats with high arboreal cover. We stress the importance of conserving the natural mosaic in the Pantanal landscape for the species persistence, given that giant armadillos require all habitat types to fulfill their dietary, spatial, and developmental needs.