Nitrogen Isotope Values Research Articles

A Strong Link Between Oceanographic Conditions and Zooplankton δ13C and δ15N Values in the San Jorge Gulf, Argentina

Maps of (baseline) δ13C and δ15N values of primary producers or consumers near the base of food webs provide crucial information for interpreting patterns in the isotopic composition of consumers that occupy higher trophic levels. In marine systems, understanding how oceanographic variables influence these values enables the creation of dynamic isoscapes across time and space, providing insights into how ecosystems function. The San Jorge Gulf (SJG) in the southwest Atlantic Ocean (45° S–47° S) is an area of particular importance, as it is located on one of the most productive continental shelves in the world, supporting large fisheries and marine mammal and seabird populations. We reconstructed spatial variation in zooplankton δ13C and δ15N values across SJG and investigated their relationship with physical and chemical oceanographic conditions. During cruises in the austral spring of 2016 and 2017, we collected medium-sized copepods whose isotopic composition integrate short-term (days to weeks) variation in oceanographic conditions recorded by phytoplankton at the base of the food web. We also collected data on water column depth, surface and bottom temperatures, water column stability, and macronutrient (nitrate, phosphate, and silicic acid) concentrations. The results revealed significant variation in both δ13C and δ15N values of up to 7-8‰ over a relatively small spatial scale (200–300 km). Copepod δ13C values were lower at the center of the SJG, showing an inverse correlation with water column stability, surface nitrate concentration, and water column depth. δ15N values showed a strong and negative relationship with surface nitrate concentration and water column stability, increasing from south to north in the SJG. δ15N values also showed a positive relationship with surface silicic acid concentration. These spatial patterns in nutrient dynamics and copepod carbon and nitrogen isotope values are interpreted in the context of the dominant northward current and temporal development of the frontal systems in the SJG.

Just Hitching a Ride: Stable Isotopes Reveal Non-Feeding Behaviour of Anisakis simplex Within Its Host Fish.

Anisakis simplex larvae, commonly found in marine fish, cause anisakiasis in humans, resulting in gastric to gastro-allergic symptoms. Despite known health risks, the impact of Anisakidae larvae on fish hosts is less understood. This study aimed to investigate this interaction by assessing the feeding strategy of A. simplex. Anisakis larvae were isolated from North Sea Merluccius merluccius tissues (stomach, body cavity, liver and muscle) and were analysed for carbon (δ13C) and nitrogen (δ15N) isotope values. Significant differences in δ13C values were found among host tissues, with the liver differing from muscle and stomach tissues. In contrast, no differences were noted for the associated parasites. Additionally, δ15N values indicated that the host occupied a significantly higher relative trophic position than its parasite. This suggests a lack of direct nutrient transfer from host to parasite, as the parasite would typically exhibit higher stable isotope values than the tissue they feed on. Therefore, A. simplex's stable isotope values might reflect those of its previous host (crustacean and/or small fish), providing insights into diet and movement of the paratenic M. merluccius host. Further research is needed to confirm these findings across different fish species and to explore A. simplex as a proxy for trophic ecology.

Isotopic analysis of modern sorghum and finger millet from different altitudes in Ethiopia: implications for ancient farming practices

C4 crops such as sorghum (Sorghum bicolor) and finger millet (Eleusine coracana) have played a significant role in the economic livelihood in arid and semi-arid zones of tropical and sub-tropical Africa since prehistoric times. However, to date, our knowledge of their past management practices is limited. Stable isotope analysis of archaeobotanical remains has been recognized as a valuable tool for reconstructing past agricultural practices, e.g. water management, and fertilization. Nonetheless, our limited understanding of the isotopic variability of C4 plants calls for further research on modern plant before application to archaeobotanical remains. In this paper, we aim to enhance our understanding of modern C4 botanical remains' isotopic variability by analyzing sorghum and finger millet plants. These crops were cultivated according to traditional local practices and collected from ten villages located in the Konso Zone (South Ethiopia) and Tigray Regional State (North Ethiopia), where they are among the daily ingredients for food, and traditional alcoholic and non-alcoholic drinks. We analyzed carbon and nitrogen stable isotopes of seeds and biosilica content in chaff, as it has been suggested that a relationship can exist between silicon and C:N. Carbon isotope values show significant variability, positively correlated with altitude. By demonstrating the sensitivity of C4 grain carbon stable isotope to altitude variations, which are likely connected to water availability, this study offers invaluable insights for the accurate assessment of isotopic values derived from ancient C4 crops. The absence of significant correlations with δ15N suggests that nitrogen isotope values may be less effective for understanding environmental variations in this kind of context. This highlights the limitations of nitrogen isotope data for interpreting ancient agricultural practices and underscores the importance of relying more on carbon isotopes for insights related to environmental conditions and altitude. Furthermore, we confirm that the amount of assimilated carbon may depend also on the biosilica content, which is in turn modulated by environmental parameters such as water availability or soil silicon levels.

Authenticating the Geographical Origin of Jingbai Pear in Northern China by Multiple Stable Isotope and Elemental Analysis.

The Jingbai pear is one of the best pear species in China with high quality and nutrition values which are closely linked to its geographical origin. With the purpose of discriminating the PGI Mentougou Jingbai pear from three other producing regions, the stable isotope ratios and elemental profiles of the pears (n = 52) and the corresponding soils and groundwater were determined using isotope ratio mass spectrometry (IRMS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. The results revealed that δ15N, δ18OJ, and Li were significantly different (p < 0.05) in samples from different regions, which indicated their potential to be used in the geographical origin classification of the Jingbai pear. The nitrogen isotopic values of the pear pulp were positively correlated with the δ15N value and nitrogen content of the corresponding soils, whilst the B, Na, K, Cr, and Cd contents of the pear pulps were positively correlated with their corresponding soils. Orthogonal partial least squares discriminant analysis (OPLS-DA) was performed in combination with analysis of the stable isotopes and elemental profiles, making it possible to distinguish the cultivation regions from each other with a high prediction accuracy (a correct classification rate of 92.3%). The results of this study highlight the potential of stable isotope ratios and elemental profiles to trace the geographical origin of pears at a small spatial scale.

Utilization of stable isotopes in teeth to investigate pygmy sperm whales (Kogia breviceps) in the southeastern United States

AbstractThe pygmy sperm whale (Kogia breviceps) is currently the second‐most stranded cetacean in the southeastern United States (SEUS), but information concerning their population structure is severely limited. This study used stable isotope analysis to investigate the isotopic niches and population structure of K. breviceps among three regions in the SEUS determined by the distance of the continental shelf break and a fourth region in the Gulf of Mexico. Teeth from stranded individuals were subsampled via dental drill at four separate areas on each tooth, representing four different age classes: calf, juvenile, subadult, and adult. Stable isotope ratios of carbon and nitrogen were measured in the organic component of 42 teeth, and oxygen isotope ratios were measured in the inorganic (hydroxyapatite) component of 19 teeth. No differences were found in δ13C, δ15N, or δ18O values between regions. However, differences between age classes were significant for adults and subadults for δ13C and adults and juveniles for δ15N. Differences between males and females were also significant for δ15N. Results from this study provide the first carbon and nitrogen isotope values from different age classes of pygmy sperm whales as well as the first reported oxygen isotope values for this species.

Multiple controls on the preservation of organic matter in the lower Mississippian Luzhai Formation black shale in southern China

The early Carboniferous experienced profound climate cooling that drove the Earth's climate from a mid-Paleozoic greenhouse into the Late Paleozoic Ice Age. Several climate cooling events have been reported, including the Tournaisian and Visean (early to mid-Early Carboniferous). In this study, we perform multi-proxy analyses (organic carbon isotopes, nitrogen isotopes, major and trace elements and organic petrology/geochemistry) of samples from an early Carboniferous section in Nandan region (Guangxi, China). Our goal is to investigate the global carbon‑nitrogen cycle, and associated oceanic productivity and redox perturbations, during a key climatic transition interval, as well as the controls on organic matter enrichment in the sediments. The carbon and nitrogen isotope profiles of the Nandan section record major perturbations during the mid-Tournaisian and early Visean. The mid-Tournaisian carbon isotope excursion (TICE) is marked by a positive δ13Corg shift of 1.8 ‰ (from −27.7 ‰ to −25.9 ‰), correlating with a positive δ15N shift. The early Visean carbon isotope excursion (VICE) is characterized by a positive shift in δ13Corg (from −28.2 to −25.5 ‰, with an excursion magnitude of 2.7 ‰), but associated with a negative shift in δ15N (from +5 ‰ to +4 ‰). The positive δ13Corg excursions during these events most likely reflect enhanced organic matter burial with expansion of anoxic seafloor in the global ocean. The drop in nitrogen isotope values in the early Visean is interpreted to be linked with less denitrification under more oxic conditions. The decrease of organic matter contents up section is consistent with the shift to more oxic conditions and increased sedimentary dilution caused by sea-level fall, which is ultimately controlled by orogenic events and climate cooling.

Research progress on isotope tracing on the sources and transformations of reactive nitrogen in the earth-atmosphere system.

Since the Industrial Revolution, human activities have led to a rapid and sustained increase in reactive nitrogen production, resulting in nitrogen enrichment at the Earth's surface and triggering many ecological and environmental issues. Stable isotopes are effective tools for tracing the sources and mechanisms of environmental processes. The nitrogen isotope values in surface environments integrate the isotope signatures of different nitrogen sources and the isotope fractionation effects of transformation processes. The composition of nitrogen isotopes can thus be utilized to trace the sources and cycling of nitrogen at the surface, aiding the development of strategies to reduce reactive nitrogen emissions, and assess the ecological effects of nitrogen enrichment. We reviewed the research progress on nitrogen isotope in the sources of reactive nitrogen in atmospheric systems, plant nitrogen utilization, and tracking of nitrogen processes in forest ecosystems. We further discussed how to gain a more systematic and accurate understanding of nitrogen cycle within and between the various spheres of the surface environment.

Isotope values reveal “canopy effect” in deer territoriality and maize consumption for dogs from archaeological sites in Kentucky dating to the Middle Woodland through Late Fort Ancient time periods

This research analyzes stable carbon, nitrogen, and oxygen isotope values in tooth collagen and enamel to investigate foraging and territorial behavior in white-tailed deer (Odocoileus virginianus Zimmermann) and maize (Zea mays Linneaus) consumption in dogs (Canis familiaris Linneaus). The study analyzed 22 deer teeth from 13 archaeological sites in Kentucky covering a span of approximately 1,500 years of human occupation. The article presents evidence of foraging and territorial behavior in white-tailed deer, identified through the “canopy effect” observed in deer stable carbon isotope values in remains spanning from the Middle Woodland (200 BCE – CE 500) to Late Fort Ancient (CE 1400 – 1680) time periods in Kentucky. Additionally, teeth samples from nine dog and one wolf (Canis lupus Linneaus) were analyzed and indicate significant consumption of maize in seven of the dog samples. These specimens came from seven archaeological sites in Kentucky dating from the Early through Late Fort Ancient (CE 1400 – 1680) time periods. The study also contributes to the growing database of isotope studies in the Eastern Woodlands by presenting carbon and nitrogen isotope values on deer bone collagen from nine deer bone samples from three of the same archaeological sites dating to the Middle Woodland to Early Late Woodland.

Modern history of hypoxia in Narragansett Bay: The geochemical record

Increased inputs of nitrogen from agricultural runoff, urbanization and suburbanization have resulted in degradation of water quality, including increased frequency and severity of hypoxia, in estuarine ecosystems. Much work has been conducted in recent years to characterize the spatial and temporal extent of hypoxia in coastal systems, but the historical record of hypoxia in such systems is much less well known. The current work examines the history of hypoxia in upper Narragansett Bay, an urbanized estuary in the northeastern U.S., through vertical profiles of geochemical markers in sediment cores. Concentrations of authigenic molybdenum indicate more frequent/ longer periods of hypoxia that are related to changes in population and anthropogenic inputs to the Bay from the surrounding watersheds. Cores from the urbanized upper bay, greatly affected by wastewater treatment facilities (WWTFs), indicate greater duration of hypoxia in the 20th century, with periods of hypoxia decreasing through mid-century and recurring thereafter. Trends of hypoxia are closely related to improvements and failures of WWTFs in surrounding communities. In Greenwich Bay, with a suburban watershed and only one WWTF, hypoxia increased substantially in parallel with growth of population in the surrounding watershed. Carbon and nitrogen concentrations and isotope values reflect increased nitrogen enrichment and productivity in the Bay in the 2nd half of the 20th century. These results can help inform study of the environmental responses to societal activities that may affect water quality.

A long-term study of stable isotope ratios of fingernail keratin and amino acids in a mother-infant dyad.

To evaluate the potential of compound-specific isotope analysis of amino acids (CSIA-AA) for investigating infant feeding practices, we conducted a long-term study that compared infant and maternal amino acid (AA) nitrogen isotope ratios. Fingernail samples were collected from a single mother-infant dyad over 19 months postpartum. Carbon and nitrogen stable isotope ratios were measured in the bulk keratin of the fingernail samples. Selected samples were then hydrolyzed and derivatized for compound-specific nitrogen isotope analysis of keratin AAs. As in previous studies, infant bulk keratin nitrogen isotope values increased during exclusive breastfeeding and fell with the introduction of complementary foods and eventual cessation of breastfeeding. Infant trophic AAs had elevated nitrogen isotope values relative to the mother, while the source AAs were similar between the mother and infant. Proline and threonine appeared to track the presence of human milk in the infant's diet as the isotopic composition of these AAs remained offset from maternal isotope values until the cessation of breastfeeding. Although CSIA-AA is costly and labor intensive, it appears to hold potential for estimating the duration of breastfeeding, even after the introduction of complementary foods. Through the analysis of a full suite of AAs, it may also yield insights into infant physiology and AA synthesis.

Stable isotope spatial patterns for the Southwest Atlantic Ocean towards polar waters

Mapping stable isotope gradients (isoscapes) has become a powerful tool to understand and forecast the status and variability of marine ecosystems at different levels of ecological organization. To differentiate five marine areas from the Southwest Atlantic Ocean towards oceanic and polar waters, a key foraging area for many marine consumers, we built isoscapes at different spatial scales using carbon (δ13C) and nitrogen (δ15N) isotope values of phytoplankton, zooplankton and particulate organic matter in sediment. We analyzed the isotopic variability between marine areas in relation with oceanographic parameters (e.g. temperature, salinity) and geographical sampling site data (e.g. latitude, longitude). We collected samples during 6 oceanographic surveys conducted in spring and autumn between 2014 and 2019 at the Beagle Channel, the Atlantic coast of Tierra del Fuego and Burdwood Bank. We included also published isotopic data of zooplankton from two other oceanic areas (the Polar Frontal Zone and Polar Antarctic Peninsula waters) to construct large-scale isoscapes. We found that the marine areas analyzed have substantially different δ13C and δ15N baselines; some differences exist between spring and autumn but the general pattern of isotopic variability remains similar. Combining different biological components and spatial scale analysis, isotopic variability was found to be related to variables such as seawater temperature, depth, chlorophyll and nutrients. The generated data will enhance the efficacy of isoscapes in long-term monitoring initiatives that documents alterations in attributes and features across marine expanses. This is particularly pertinent to areas under legal protection, such as the oceanic Marine Protected Areas (MPAs) established in Argentine waters.

Health inequality in medieval Cambridge, 1200-1500 CE.

Health inequality is not only a major problem today; it left its mark upon past societies too. For much of the past, health inequality has been poorly studied, mostly because bioarchaeologists have concentrated upon single sites rather than a broader social landscape. This article compares 476 adults in multiple locations of medieval Cambridge (UK). Samples include ordinary townspeople (All Saints), people living in a charitable institution (the Hospital of St. John), and members of a religious order (the Augustinian Friary). These groups shared many conditions of life, such as a similar range of diseases, risk of injury, and vertebral disk degeneration. However, people living on charity had more indicators of poor childhood health and diet, lower adult stature, and a younger age at death, reflecting the health effects of poverty. In contrast, the Augustinian friars were members of a prosperous, well-endowed religious house. Compared with other groups, they were taller (perhaps a result of a richer diet during their adolescent growth period); their adult carbon and nitrogen isotope values are higher, suggesting a diet higher in terrestrial and/or marine animal protein; and they had the highest prevalence of foot problems related to fashionable late medieval footwear. As this illustrates, health inequality will take particular forms depending upon the specificities of a social landscape; except in unusual circumstances where a site and its skeletal samples represent a real cross-section of society, inequality is best investigated by comparison across sites.

Effects of pre-treatment, historical age, and sample characteristics on the stable isotope analyses of killer whale (Orcinus orca) bone.

Stable isotope analysis of bone provides insight into animal foraging and allows for ecological reconstructions over time, however pre-treatment is required to isolate collagen. Pre-treatments typically consist of demineralization to remove inorganic components and/or lipid extraction to remove fats, but these protocols can differentially affect stable carbon (δ13C) and nitrogen (δ15N) isotope values depending on the chemicals, tissues, and/or species involved. Species-specific methodologies create a standard for comparability across studies and enhance understanding of collagen isolation from modern cetacean bone. Elemental analyzers coupled to isotope ratio mass spectrometers were used to measure the δ13C and δ15N values of powdered killer whale (Orcinus orca) bone that was intact (control) or subjected to one of three experimental conditions: demineralized, lipid-extracted, and both demineralized and lipid-extracted. Additionally, C:N ratios were evaluated as a proxy for collagen purity. Lastly, correlations were examined between control C:N ratios vs. historical age and control C:N ratios vs. sample characteristics. No significant differences in the δ15N values were observed for any of the experimental protocols. However, the δ13C values were significantly increased by all three experimental protocols: demineralization, lipid extraction, and both treatments combined. The most influential protocol was both demineralization and lipid extraction. Measures of the C:N ratios were also significantly lowered by demineralization and both treatments combined, indicating the material was closer to pure collagen after the treatments. Collagen purity as indicated via C:N ratio was not correlated with historical age nor sample characteristics. If only the δ15N values from killer whale bone are of interest for analysis, no pre-treatment seems necessary. If the δ13C values are of interest, samples should be both demineralized and lipid-extracted. As historical age and specimen characteristics are not correlated with sample contamination, all samples can be treated equally.

Further investigation into the impact of manuring on stable carbon (δ13C) and nitrogen isotope (δ15N) values in pulses: a four-year experiment examining Celtic bean (Vicia faba)

Plant stable carbon (δ13C) and nitrogen (δ15N) isotope values can be used to directly investigate crop husbandry practices such as manuring; a key variable in understanding the scale and intensity of past farming practices. We present new results from a four-year experiment examining the impact of manuring on the δ13C and δ15N values of a heritage cultivar of the broad or faba bean (Vicia faba), the ‘Celtic black broad bean’. This paper builds upon our previous pilot study (Treasure ER, Church MJ, Gröcke DR (2016) The influence of manuring on stable isotopes (δ13C and δ15N) in Celtic bean (Vicia faba L.): archaeobotanical and palaeodietary implications. Archaeol Anthropol 8:555–562. https://doi.org/10.1007/s12520-015-0243-6). Our results indicate that manuring increased δ15N values in beans, with a mean value of 0.5 ± 0.4‰ in control samples compared to a mean value of 2.1 ± 1.3‰ in the most intensively manured plots. It was not always possible to distinguish between an unmanured and manured crop on the basis of δ15N values, although manured crops consistently exhibited larger variation in δ15N. Bean δ13C values show no clear relationship with manuring, although large variability (> 3‰) was found within crops cultivated under the same conditions. Manuring also increased plant biomass production, seed-size and the bean per plant ratio. Overall, we suggest that high δ15N values (> 1.5‰) in archaeobotanical remains of pulses can be attributed to small-scale, intensive cultivation that is typical of gardens where manuring rates are very intensive.

Mercury concentrations in Seaside Sparrows and Marsh Rice Rats differ across the Mississippi River Estuary

Mercury (Hg) concentrations and their associated toxicological effects in terrestrial ecosystems of the Gulf of Mexico are largely unknown. Compounding this uncertainty, a large input of organic matter from the 2010 Deepwater Horizon oil spill may have altered Hg cycling and bioaccumulation dynamics. To test this idea, we quantified blood concentrations of total mercury (THg) in Seaside Sparrows (Ammospiza maritima) and Marsh Rice Rats (Oryzomys palustris) in marshes west and east of the Mississippi River in 2015 and 2016. We also tested for a difference in THg concentrations between oiled and non-oiled sites. To address the potential confounding effect of diet variation on Hg transfer, we used stable nitrogen (δ15N) and carbon (δ13C) isotope values as proxies of trophic position and the source of primary production, respectively. Our results revealed that five to six years after the spill, THg concentrations were not higher in sites oiled by the spill compared to non-oiled sites. In both species, THg was higher at sites east of the Mississippi River compared to control and oiled sites, located west. In Seaside Sparrows but not in Marsh Rice Rats, THg increased with δ15N values, suggesting Hg trophic biomagnification. Overall, even in sites with the most elevated THg, concentrations were generally low. In Seaside Sparrows, THg concentrations were also lower than previously reported in this and other closely related passerines, with only 7% of tested birds exceeding the lowest observed effect concentration associated with toxic effects across bird species (0.2 µg/g ww). The factors associated with geographic heterogeneity in Hg exposure remain uncertain. Clarification could inform risk assessment and future restoration and management actions in a region facing vast anthropogenic changes.

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.

Kinetic nitrogen isotope effects of 18 amino acids degradation during burning processes

Understanding the nitrogen isotopic variations of individual amino acids (AAs) is essential for utilizing the nitrogen isotope values of individual amino acids (δ15N-AA) as source indicators to identify proteinaceous matter originating from biomass combustion processes. However, the nitrogen isotope effects (ε) associated with the degradation of individual amino acids during combustion processes have not been previously explored. In this study, we measured the nitrogen isotope values of residual free amino acids -following a series of controlled combustion experiments at temperatures of 160–240 °C and durations of 2 min to 8 h, as described in Part 1. δ15N values of proline, aspartate, alanine, valine, glycine, leucine, and isoleucine are more positive than their initial δ15N values after prolonged combustion. Variations in δ15N values of the most AAs conform to the Rayleigh fractionation during combustion and their nitrogen isotope effects (ε) are greatly impacted by their respective combustion degradation pathways. This is the first time the ε values associated with the degradation pathways of AAs during combustion have been characterized. Only the ε values associated with Pathway 1 (dehydration to form dipeptide) and 2 (simultaneous deamination and decarboxylation) are found to be significant and temperature-dependent, ranging from + 2.9 to 6.4‰ and + 0.9‰ to + 3.8‰, respectively. Conversely, ε values associated with other pathways are minor. This improves the current understanding on the degradation mechanisms of protein nitrogen during biomass burning.

C, N, Hg isotopes and elemental chemostratigraphy across the Ordovician–Silurian transition in the Argentine Precordillera: Implications for the link between volcanism and extinctions

Global geological processes that occurred during the Ordovician–Silurian transition (OST) were investigated exploring sections from the Argentine Precordillera (Cerro La Chilca, Baños de Talacasto and Villicum) which were deposited close to polar zones of Gondwana during the oldest Phanerozoic glaciation. C-N-Hg isotopes and redox-sensitive trace metal chemostratigraphy allowed to assess the relative importance of volcanism, glaciation, post-glacial sedimentation and extinction events during the OST. At Cerro La Chilca and Baños de Talacasto sections, Katian and early Hirnantian strata are missing. Hirnantian and Rhuddanian strata occur in the all three sections. At Cerro La Chilca, coeval positive δ13Corg shift, Hg spike and Δ199Hg ∼ 0 ‰ were recorded in the Sandbian Los Azules Formation, suggesting volcanic Hg loading. Coeval Hg/TOC spikes at the Baños de Talacasto and Villicum sections and Δ199Hg ∼ 0 ‰ in the late Hirnantian (M. persculptus Zone) recorded the second pulse of the Late Ordovician mass extinction (LOME 2), for which volcanism was postulated as the cause of warming and anoxia. Another Hg/TOC spike in the early Rhuddanian at Baños de Talacasto and Villicum is coeval with slightly positive Δ199Hg values, an enrichment related, perhaps, to an enhanced continental runoff that followed the Late Ordovician glacial cycle (LOGC-3). At the Cerro La Chilca, positive δ15N values in the Sandbian suggest less intense water-column denitrification while negative nitrogen isotope values in the LOME 2 period indicate denitrifying conditions. At the Baños de Talacasto and Villicum sections, sea-level fluctuations and nitrogen upwelling helped shaping the δ15N pathway within the LOME 2 and early Rhuddanian intervals, with negative δ15N values attesting that conditions became less reducing closer to the OST. The odd-MIF Hg isotope pattern within the LOME 2 time interval was probably induced by coeval volcanism, glaciation and sea-level fall, and in the early Rhuddanian by deglaciation and sea-level rise.

Lipid extraction alters amino acid composition and bulk, but not amino acid, carbon and nitrogen isotope values.

Concerns exist over observed shifts in value and variance of nitrogen isotopes following physicochemical extraction of lipids from organic matter. The mechanisms behind these apparent changes in bulk tissue δ15N values are not fully understood yet have major implications for analytical costs and integrity of data interpretations. Changes in proximate analysis, amino acid composition, C:N ratios, bulk tissue and amino acid δ13C and δ15N values, and resulting isotope-based food web metrics were compared between lipid-intact and lipid-extracted muscle tissue of fishes spanning <1% to >20% muscle fat content to identify mechanisms of nitrogen isotope fractionation associated with physicochemical lipid extraction. Bulk δ13C and δ15N values increased and %N, C:N ratios and crude protein content decreased following lipid extraction. Resulting bulk isotope niche spacing and overlap varied significantly between lipid-intact and lipid-extracted tissues. While amino acid composition significantly changed during lipid extraction, particularly for lipid-associated amino acids (e.g., Glu, Lys, Ser), individual amino acid δ13C and δ15N values, and their associated compound-specific isotope analysis of amino acids (CSIA-AA)-based food web metrics, did not. Physicochemical lipid extraction caused significant tissue composition changes (e.g., leaching of amino acids and 15N-deplete nitrogenous waste) that affected δ13C and δ15N values and tissue %C and %N beyond simply removing lipids. However, lipid extraction did not alter individual amino acid δ13C or δ15N values or their associated CSIA-AA-based food web metrics.

Land cover and space use influence coyote carnivory: evidence from stable-isotope analysis

For many species, the relationship between space use and diet composition is complex, with individuals adopting varying space use strategies such as territoriality to facilitate resource acquisition. Coyotes (Canis latrans) exhibit two disparate types of space use; defending mutually exclusive territories (residents) or moving nomadically across landscapes (transients). Resident coyotes have increased access to familiar food resources, thus improved foraging opportunities to compensate for the energetic costs of defending territories. Conversely, transients do not defend territories and are able to redirect energetic costs of territorial defense towards extensive movements in search of mates and breeding opportunities. These differences in space use attributed to different behavioral strategies likely influence foraging and ultimately diet composition, but these relationships have not been well studied. We investigated diet composition of resident and transient coyotes in the southeastern United States by pairing individual space use patterns with analysis of stable carbon (δ13C) and nitrogen (δ15N) isotope values to assess diet. During 2016–2017, we monitored 41 coyotes (26 residents, 15 transients) with GPS radio-collars along the Savannah River area in the southeastern United States. We observed a canopy effect on δ13C values and little anthropogenic food in coyote diets, suggesting 13C enrichment is likely more influenced by reduced canopy cover than consumption of human foods. We also observed other land cover effects, such as agricultural cover and road density, on δ15N values as well as reduced space used by coyotes, suggesting that cover types and localized, resident-like space use can influence the degree of carnivory in coyotes. Finally, diets and niche space did not differ between resident and transient coyotes despite differences observed in the proportional contribution of potential food sources to their diets. Although our stable isotope mixing models detected differences between the diets of resident and transient coyotes, both relied mostly on mammalian prey (52.8%, SD = 15.9 for residents, 42.0%, SD = 15.6 for transients). Resident coyotes consumed more game birds (21.3%, SD = 11.6 vs 13.7%, SD = 8.8) and less fruit (10.5%, SD = 6.9 vs 21.3%, SD = 10.7) and insects (7.2%, SD = 4.7 vs 14.3%, SD = 8.5) than did transients. Our findings indicate that coyote populations fall on a feeding continuum of omnivory to carnivory in which variability in feeding strategies is influenced by land cover characteristics and space use behaviors.