Stable Isotope Analysis Research Articles

Combining biotracer and stomach contents analysis to understand trophic dynamics in the northern Gulf of Mexico

The northern Gulf of Mexico (nGOM) is a taxonomically rich ecosystem. Previous work based on a meta-analysis of stomach contents data has shown the trophic connectivity of predators and prey to be substantial. However, the trophic dynamics of many economically and ecologically important species are still not well understood. Sportfish, such as Spotted Seatrout (Cynoscion nebulosus), Red Drum (Sciaenops ocellatus), and Southern Flounder (Paralichthys lethostigma), support recreational fisheries throughout the region. Gulf Menhaden (Brevoortia patronus), a high biomass forage fish with the region's largest commercial fishery, is considered an important forage species. In this study, we use information from meta-analysis of stomach contents and stable isotope analysis to investigate the importance of prey taxa, including Gulf Menhaden, for nGOM nearshore predators. Stomach contents and stable isotope analyses are generally evaluated independently, with stomach contents used to directly identify trophic interactions, while stable isotopes of carbon (δ13C) and nitrogen (δ15N) provide insight into a consumer's long-term feeding habits. We used a multispecies trophic model, EcoDiet, developed by Hernvann et al. (2022), that integrates information of both stomach contents and stable isotopes into a single framework to estimate trophic link probabilities and diet proportions. Data in the model include n = 41 predators, n = 173 prey, and n = 497 unique predator and prey interactions. The results indicate that nGOM nearshore predators are generalists using the diverse prey base, and in concordance with previous findings, there is no single Menhaden-dependent predator. Our findings better quantify the trophic interactions of the highly diverse nGOM region and have important implications regarding future ecosystem modeling and management considerations for the Gulf Menhaden stock.

Spatio‐temporal differences in the diet and trophic ecology of Kelp Gulls (Larus dominicanus) in South Africa

Opportunistic species, including some gulls (Laridae), can benefit from urbanization and increased anthropogenic food resources. Knowledge of the level of exploitation of anthropogenic resources by gulls is crucial to understand how changes in food availability might affect their populations, which in turn may impact other species. The Kelp Gull Larus dominicanus is widely distributed in the southern hemisphere and consumes a wide variety of resources, from waste in landfills to seabirds. We assessed if proximity to landfills influenced the diet and trophic ecology of Kelp Gull incubating adults and chicks using a combination of conventional diet sampling (stomach contents, regurgitated pellets), and carbon and nitrogen stable isotope analysis of blood plasma in seven colonies over two consecutive years in South Africa. Kelp Gulls consumed a variety of resources, including marine (e.g. fish), coastal (e.g. molluscs), terrestrial natural (e.g. insects, mammals), or terrestrial anthropogenic (e.g. chicken, bread) items. Inter‐annual differences were most apparent among gulls breeding next to a seabird colony, whereas gulls breeding at other colonies were more consistent in diet and trophic ecology. At most colonies, chicks were fed more natural and higher trophic level food items than were consumed by incubating adults. Overall, distance to landfill did not strongly affect the diet and trophic ecology and our results suggest that the Kelp Gull's broad feeding ecology allows it to buffer changes in food availability by switching among food resources. However, such dietary changes triggered by human actions may impact other species, including threatened seabirds.

Diet of Andean leaf-eared mice (Phyllotis) living at extreme elevations on Atacama volcanoes: insights from metagenomics, DNA metabarcoding, and stable isotopes.

On the flanks of >6000 m Andean volcanoes that tower over the Atacama Desert, leaf-eared mice (Phyllotis vaccarum) live at extreme elevations that surpass known vegetation limits. What the mice eat in these barren, hyperarid environments has been the subject of much speculation. According to the arthropod fallout hypothesis, sustenance is provided by windblown insects that accumulate in snowdrifts ('aolian deposits'). It is also possible that mice feed on saxicolous lichen or forms of cryptic vegetation that have yet to be discovered at such high elevations. We tested hypotheses about the diet of mice living at extreme elevations on Atacama volcanoes by combining metagenomic and DNA metabarcoding analyses of gut contents with stable-isotope analyses of mouse tissues. Genomic analyses of contents of the gastrointestinal tract of a live-captured mouse from the 6739 m summit of Volcán Llullaillaco revealed evidence for an opportunistic but purely herbivorous diet, including lichens. Although we found no evidence of animal DNA in gut contents of the summit mouse, stable isotope data indicate that mice native to elevations at or near vegetation limits (~5100 m) include a larger fraction of animal prey in their diet than mice from lower elevations. Some plant species detected in the gut contents of the summit mouse are known to exist at lower elevations at the base of the volcano and in the surrounding Altiplano, suggesting that such plants may occur at higher elevations beneath the snowpack or in other cryptic microhabitats.

At the base of deep-sea food webs: Assemblage and trophic structure of suprabenthos and zooplankton in submarine canyons

Submarine canyons act as hotspots of biodiversity, hosting vulnerable marine ecosystems, and playing a fundamental role in bridging coastal zones with deeper areas. Here, we investigated the suprabenthic and Deep Scattering Layer (DSL) zooplankton fauna, that play a key role in deep-sea food webs, as main resources for both mobile and sessile megafauna, in two submarine canyons (Squillace and Amendolara) of the Ionian Sea (Central Mediterranean Sea). Our results highlighted different taxonomic and functional diversity between the two adjacent canyons: (i) biomass and abundance of suprabenthos followed an opposite trend in the two canyons, increasing both with depth in Amendolara (higher abundance and biomass in the lower part of the canyon), and decreasing with depth in Squillace (greater in the head of the canyon); (ii) DSL zooplankton abundance and biomass followed a spatial distribution, decreasing with increasing distance from the coast for both canyons (i.e. lower offshore than at the head of the canyon). Food-web structure investigated by means of stable isotope analysis of δ13C and δ15N showed a more diverse trophic niche for suprabenthos than for zooplankton. Furthermore, possible feeding modes of species with unknown feeding behaviour have been proposed. The results of the current article highlight the different ecological processes occurring within each canyon. Understanding the spatial variations of communities inhabiting submarine canyons, especially those at the base of deep-sea food webs which can act as driver of megafaunal communities (both sessile and mobile-commercial species), is essential to focalise future conservation efforts.

Stable isotope analysis reveals diet niche partitioning between native species and the invasive black bullhead (Ameiurus melas Rafinesque, 1820)

Stable isotope analysis reveals diet niche partitioning between native species and the invasive black bullhead (Ameiurus melas Rafinesque, 1820)

Quantifying remediation of chlorinated volatile compounds by sulfidated nano zerovalent iron treatment using numerical modeling and CSIA

Sulfidated nanoscale zerovalent iron (S-nZVI) has demonstrated promising reactivity and longevity for remediating chlorinated volatile compounds (cVOC) contaminants in laboratory tests. However, its effectiveness in field applications remains inadequately evaluated. This study provides the first quantitative evaluation of the long-term effectiveness of carboxymethyl cellulose-stabilized S-nZVI (CMC-S-nZVI) at a cVOC-contaminated field site. A reactive transport model-based numerical approach delineates the change in cVOC concentrations and carbon isotope values (i.e., δ13C from compound-specific stable isotope analysis (CSIA)) caused by dissolution of dense non-aqueous phase liquid, sorption, and pathway-specific degradation and production, respectively. This delineation reveals quantitative insights into remediation effectiveness typically difficult to obtain, including extent of degradation, contributions of different degradation pathways, and degradation rate coefficients. Significantly, even a year after CMC-S-nZVI application, degradation remains an important process effectively removing various cVOC contaminants (i.e., chlorinated ethenes, 1,2-dichloroethanes, and chlorinated methanes) at an extent varying from 5 %-62 %. Although the impacts of CMC-S-nZVI abundance on degradation vary for different cVOC and for different sampling locations at the site, for the primary site contaminants of tetrachloroethene and trichloroethene, their predominance of dichloroelimination pathway (≥ 88 %), high degradation rate coefficient (0.4–1.7 d-1), and occurrence at locations with relatively high CMC-S-nZVI abundance strongly indicate the effectiveness of abiotic remediation. These quantitative assessments support that CMC-S-nZVI supports sustainable ZVI-based remediation. Further, the novel numerical approach presented in this study provides a powerful tool for quantitative cVOC remediation assessments at complex field sites where multiple processes co-occur to control both concentration and CSIA data.

Reconstructing dietary practices at Tell Kamid el-Loz (Lebanon) during the Bronze and Iron Age III / Persian to Hellenistic periods using plant micro-remains from dental calculus and stable isotope analysis of bone collagen

Tell Kamid el-Loz (Lebanon) was an important Bronze Age urban center that dominated one of the central crossroads of the Ancient Near East, connecting Egypt and the Levant with northern Mesopotamia, Anatolia, and Syria, as well as the interior with the Mediterranean coast. However, by the early Iron Age, the site had shrunk to a small rural settlement. Later, in the Iron Age III / Persian-Hellenistic, only enigmatic pits and a large cemetery remained. In this paper, we analyzed plant micro-remains from the dental calculus of 15 individuals (3 from the Middle Bronze Age II and 12 from the Iron Age III / Persian-Hellenistic) and δ13C and δ15N stable isotope data from tbulk bone collagen of 74 individuals (10 from the Middle Bronze Age II and 64 from the Iron Age III / Persian-Hellenistic) and 13 Late Bronze Age animal bones (7 Ovis/Capra and 6 Bos). Our results indicate general stability of human diet throughout the Middle Bronze Age II and the Iron III / Persian-Hellenistic periods, with a reliance on C3 plant crops and terrestrial animals also consuming C3 plants. In the later period, the plant micro-remains indicate the consumption of C4 plants and sedges, and the stable isotope analysis indicates differences in diet between males and females.

Variability in resource use of sympatric cowfish Aracana aurita and A. ornata in southeastern Australian waters

Sympatric species that share morphological similarities tend to diversify their prey resources to coexist. The striped cowfish (Aracana aurita) and ornate cowfish (A. ornata) live together on the continental shelf from western to southern Australia including Tasmanian waters. The present study analyzed stomach contents and stable isotope ratios (δ13C and δ15N) to investigate the use of food resource between the two species collected in Tasmanian waters, Australia. In total, 139 A. aurita and 46 A. ornata were collected from 2014 to 2018. The small size class of A. aurita (<140 mm SL) mainly consumed polychaetes and amphipods, while its large size class (≥140 mm SL) preyed more on bivalves and crustacean decapods. The main prey items of the small size class of A. ornata (<140 mm SL) were amphipods and crabs. No differences in diet were observed between the sexes of either species. Stable isotope analyses showed significant differences in δ15N and δ13C between the two species, but not between the two size classes of A. aurita (ANOVA, P < 0.05). Integrating analyses of stomach contents and stable isotopes indicated that the two species preferred different food resources based on different trophic sources. Consequently, although the habitats of the two species overlap, both species can co-exist through dietary segregation.

Tracing the geographic origin of Atlantic cod products using stable isotope analysis.

Increasing demand for fish and seafood means that the traceability of marine products is becoming ever more important for consumers, producers and regulators. Highly complex and globalised supply networks create challenges for verifying a stated catch region. Atlantic cod is one of the most commercially important species in the northeast Atlantic. Several regional fisheries supply cod into the trade network, of which some are at greater risk of overexploitation than others. Tools allowing retrospective testing of spatial origin would significantly assist sustainable harvesting of fish, reducing incentives for illegal fishing and fraud. Here, we investigate whether stable isotope ratios of carbon, nitrogen and sulphur can be used to retrospectively identify the catch region of Atlantic cod (Gadus morhua). We measured the isotopic composition of muscle tissue from 377 cod from 10 catch regions across the northeast Atlantic and then applied three different assignment methods to classify cod by region of most likely origin. The assignment method developed was subsequently tested using independently sourced, known-origin samples. Individual cod could be traced back to their true origin with an average assignment accuracy of 70-79% and over 90% accuracy for certain regions. Assignment success rates comparable to those using genetic techniques were achieved when assigning among restricted and pre-selected regions. However, assignment accuracy to the fishery region estimated from independent samples across the whole geographic range of cod averaged ~25% overall, highlighting the need for careful application of isotope-based approaches. Stable isotope techniques can provide effective tools to test for origin in Atlantic cod, but not all catch regions are isotopically distinct. Stable isotopes could be combined with genetic techniques to result in higher assignment accuracy than could be achieved using either method independently. Assignment potential can be estimated from reference datasets, but estimates of realistic assignment accuracy require independently collected data.

Sample preservation methods for nitrous oxide concentration and isotope ratio measurements in aquatic environments

AbstractThe nitrogen (N) and oxygen (O) stable isotope analysis of dissolved nitrous oxide (N2O) can provide important constraints on the sources and cycling of N2O in aquatic environments. The isotopic composition of aqueous N2O, both in field (natural abundance) or experimental (15N‐labeling) samples, however, may be altered by abiotic reactions involving nitrite () or hydroxylamine (NH2OH) and microbial activity during sample storage, if samples are not adequately preserved. Here we tested five different preservatives, mercuric chloride (HgCl2), copper sulfate (CuSO4), zinc chloride (ZnCl2), hydrochloric acid (HCl) mixed with sulfamic acid (SFA), and sodium hydroxide (NaOH), for fixing natural water samples from an estuary and a lake with different concentrations over a range of different storage times for N2O analyses. ZnCl2 and CuSO4 decreased the pH, and led to abiotic N2O production from , shifting the N2O isotopic composition significantly. Removal of with a mixture of SFA and HCl did not always prevent the alteration of the original N and O isotope composition of N2O, confirming the requirement for complete removal, and underscoring the biasing effects of at very low pH, even at trace levels. At low background, HgCl2 preservation led to robust and accurate results. However, in light of its toxicity and bioaccumulation potential in food webs, this fixative should be avoided. The addition of NaOH reduced abiotic side reactions involving , and yielded the most reliable and reproducible results for N2O concentration and isotope ratio measurements across tested settings and storage times.

A late Pleistocene nest cave of Gymnogyps californianus (California Condor) in Texas: New radiocarbon and stable isotope analyses

ABSTRACT Fossil remains of Gymnogyps californianus (California Condor) from Mule Ears Peak Cave, Big Bend National Park, Texas, recovered in the 1930s were reexamined to determine a precise age for nesting condors in this region. Bones of at least 6 prefledged chicks account for most (15, or 65%) of the 23 skeletal elements from this cave and a new osteology collection of known-age condor chicks at the U.S. National Museum, Division of Birds, now allows accurate estimate of the age of death of these fossil condor chicks based on their bone development and porosity. Current and previous radiocarbon dates on juvenile and adult bones, respectively, indicate the presence of condors at this site beginning at ~15,000 calendar years before present (cal yrs BP), with definite nesting occurring at ~13,000 cal yrs BP. Stable isotope analysis (δ13C and δ15N) of bone collagen on 2 bones of adult condors reflects a diet similar to other fossil condors previously analyzed from the inland western U.S. The δ13C values in the Mule Ears Peak condors indicate a diet of megafauna that subsisted on C4 plants in a desert grassland ecosystem. These results support the hypothesis that condors were extirpated from the inland west with the loss of megafauna at the end of the Pleistocene. Further, Big Bend National Park with its vast open space and cliffs and canyons for nesting condors should be considered as an additional release site for captive-reared condors as part of the current Condor Recovery Program.

Pattern of water use by Tamarix ramosissima seedlings in floodplains under varied groundwater depths in the hinterland of the Taklimakan Desert

The water use of Tamarix ramosissima Ledeb. seedlings after flooding were analyzed both to explore the maintenance mechanism and pattern of natural regeneration of riparian forest, which will provide a scientific basis for restoration of desert riparian forest and ecosystem stability in the lower reaches of inland rivers of arid regions. This study area was located in the hinterland of the Taklimakan Desert in Xinjiang, China. Tamarix ramosissima seedlings growing on different groundwater depths at the river floodplain were used as the study system. The rooting depths of T. ramosissima seedlings with different basal stem diameters were ascertained by the root excavation method. The water source for the T. ramosissima seedlings was clarified using hydrogen and oxygen stable isotope methods, and the water use efficiency of T. ramosissima seedlings was investigated by stable carbon isotope (δ13C) analysis. As the basal stem diameter classes of the T. ramosissima seedlings increased, their root depths increased. As the groundwater depth increased, the seedlings changed from primarily utilizing deep soil water to utilizing shallow soil water. In the three sample sites, the average depth of water uptake of the seedlings with basal stem diameters of 0–5 mm was 110.5, 44.1 and 39.1 cm, respectively, and that of seedlings with basal stem diameters of 5–11 mm was 83.8, 73.6 and 37.7 cm, respectively. As groundwater depth increased, the average water uptake depth of the seedlings gradually became shallower. There was no significant difference in the δ13C values of leaves under different groundwater depths, indicating that the seedlings were not subjected to water stress. Thus, surface water played a greater role than groundwater in T. ramosissima seedling water utilization. Therefore, when analyzing ecological water conveyance patterns, attention should be paid to T. ramosissima located in areas with deep groundwater. Shallow-rooted seedlings with small basal stem diameters face an increased risk of wilting if they do not receive timely recharges of surface water. Data AvailabilityAll data generated or analyzed during this study are included in this published article.

Source differentiation of BTEX compounds in groundwater contaminated due to refinery activities

This study aims to identify sources of groundwater contamination in a refinery area using integrated compound-specific stable isotope analysis (CSIA), oil fingerprinting techniques, hydrogeological data, and distillation analysis. The investigations focused on determination of the origin of benzene, toluene, ethylbenzene, and xylenes (BTEX), and aliphatic hydrocarbons as well. Groundwater and floating oil samples were collected from extraction wells for analysis. Results indicate presence of active leaks in both the northern and southern zones. In the northern zone, toluene was found to primarily originate from oil products like aviation turbine kerosene (ATK or aviation fuel), kerosene, regular gasoline, and diesel fuel. Additionally, stable isotope ratios of carbon and hydrogen for ethylbenzene, o-xylene (ortho xylene) and p-xylene (para xylene) in zone A suggested the pollution originated from gasoline within the northern zone. The origin of super gasoline (with higher octane) identified in southern zone using δ13C and δ2H values of toluene in the floating oil and groundwater samples. Further, biodegradation of toluene likely occurred in southern zone according to δ13C and δ2H. The findings underscore the critical importance of integrating CSIA and fingerprinting techniques to effectively address the challenges of source identification and relying solely on each method independently is insufficient. Accordingly, comparing the GC-MS results of floating oil samples with ATK and jet fuel (JP4) standards can be effectively utilized for source differentiation. However, this method showed no practical application to distinguish different types of diesel or gasoline. The accuracy and reliability of source identification of BTEX compounds may significantly improve when hydrogeological data incorporates with stable isotopes analysis. Additionally, the results of this study will elevate the procedures for fuel-related contaminants source identification of the polluted groundwater that is crucial to develop effective remediation strategies.

Trophic effects of jellyfish blooms on fish populations in ecosystems of the coastal waters of China

Jellyfish play an important role in the material cycling and energy flow of food webs, and massive aggregations may have deleterious consequences for local fisheries; yet a theoretical framework of the trophic effects of jellyfish blooms on coastal fisheries is unclear. To address this knowledge gap, we assessed the trophic interactions between cooccurring bloom jellyfish and dominant fish groups (omnivorous fish and piscivorous fish) in the coastal waters of China (CWC) via stable isotope analysis; we subsequently discussed how jellyfish blooms may affect energy flow through coastal ecosystems. Our results indicate a considerable degree of trophic overlap (mean ratio > 65 %) between jellyfish and small omnivorous fish (< 10 cm), highlighting a similarity in feeding habits, while the overlap ratio decreased to <55 % of the large omnivorous fish group (> 10 cm). Relatively higher trophic levels and smaller overlaps of large omnivorous fish were found in the ecosystem with high jellyfish biomass, which suggested that they may reinforce the ontogenetic trophic shift pattern to alleviate the potential for resource competition with jellyfish under conditions of jellyfish explosion. The smallest trophic overlap (< 20 %) highlighted the strong trophic differentiation between jellyfish and piscivorous fish. Additionally, our study suggested that a massive aggregation of jellyfish can negatively influence zooplankton but may not transfer energy further up efficiently, implying a weak trophic coupling between jellyfish and upper-trophic levels in CWC ecosystems. Thus, we speculate that jellyfish play an important role in shaping pathways involved in the energy transfer of food webs and that large blooms may negatively affect fisheries through bottom-up control affecting prey availability. In general, these results hold strong potential to further improve the understanding of the trophic interactions between jellyfish and fish populations. Furthermore, this study provides valuable data for predicting the consequences of jellyfish blooms on ecosystems, and is crucial for ecosystem-based management of coastal fisheries.

A taste of youth: Seasonal changes in the diet of immature white sharks in eastern Australia

White sharks (Carcharodon carcharias) play a crucial ecological role, shaping ecosystems through direct predation and risk effects. On the east coast of Australia, immature white sharks are broadly distributed, inhabiting a wide range of habitats and ecosystems from temperate Tasmania to tropical North Queensland. Using stable isotopes and fatty acids of muscle and plasma, we examined the diet and habitat use of 136 immature white sharks (152–388 cm total length) captured on SMART drumlines on the East Australian coast. This facilitated the temporal assessment of white shark trophic ecology from a few weeks to approximately a year. Biochemistry of muscle samples showed that white sharks predominantly feed on low trophic level prey from coastal environments. A seasonal shift in diet was evident, with the increasing proportions of essential fatty acids in muscle tissues during spring and summer suggesting a greater consumption of high-nutrition preys during those months compared to autumn and winter. By combining stable isotope and fatty acid analysis, we gain a comprehensive understanding of immature white shark diet in eastern Australia. Our results confirm that white sharks are generalist predators that exhibit seasonal shifts in their diet. Their high use of coastal habitats reinforces the importance of these areas for foraging, which is crucial for growth and development during this critical life stage.

Applications of zinc stable isotope analysis in environmental and biological systems: a review

Zinc (Zn) is an essential trace element that is involved in both biotic and abiotic processes in Earth's surface environments. Over the last 20 years, advances in mass spectrometry instrumentation have enabled unprecedented high-precision Zn isotope abundance ratio determinations in natural and anthropogenic samples. Supported by a solid theoretical background inherited from the traditional non-metal stable isotopes (C, N, H, O, S), the understanding of the controlling factors of Zn isotope fractionation has rapidly evolved. In this article, we review the main applications and findings of Zn isotopes in a variety of scientific domains, including metal contamination, magmatic differentiation, plant uptake, weathering, global climate change, dietary and trophic chains, and biomedicine. The analytical aspects of Zn isotope determination in solid and water samples and the principles and mechanisms of Zn isotope fractionation are also reviewed. This work aims to provide a general yet in-depth panorama of Zn isotope chemistry and to demonstrate the versatility and potential of this isotope system for studying the biological, geological and chemical cycling of Zn.

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

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.

Differences in the intensity of past forest fire events inferred from stable oxygen isotope analysis of charred bark

Understanding past fire regimes requires reliable proxy data that record fire conditions and preserve them over time. The objective of this study was to determine if the oxygen isotope composition of charred bark samples (pyrogenic organic matter) could be used as proxy data to differentiate wildfires based on burn intensity. We collected charred and uncharred bark samples from three fire sites in northern Ontario, Canada that represented a gradient of fire intensity as depicted by Fire Weather Index (FWI) data. We hypothesized that the mean Δ18Obark-char (the difference between δ18O of uncharred bark and a charred sample) would be greater for fires with higher intensities. Analysis of variance of Δ18Obark-char indicated a significant effect of fire event ( F = 73.6, p &lt; 0.001), which explained 57.0% of the variance. A prescribed surface fire treatment (mean FWI = 9.5) had significantly lower Δ18Obark-char than two natural crown fires (FWI = 21 and 27). These results demonstrate that Δ18Obark-char differentiated moderate from high intensity fires in a similar manner to the FWI data.

Microplastic pollution and nutrient enrichment shift the diet of freshwater macroinvertebrates

Microplastic pollution poses a global threat to freshwater ecosystems, with laboratory experiments indicating potential toxic impacts through chemical toxicity, physical abrasion, and false satiation. Bioplastics have emerged as a potential greener alternative to traditional oil-based plastics. Yet, their environmental effects remain unclear, particularly at scales relevant to the natural environment. Additionally, the interactive impacts of microplastics with other environmental stressors, such as nutrient enrichment, are poorly understood and rarely studied. Under natural conditions organisms might be able to mitigate the toxic effects of microplastics by shifting their diet, but this ability may be compromised by other stressors. This study combines an outdoor mesocosm experiment and stable isotope analysis to determine changes in the trophic niches of three freshwater invertebrate species exposed to conventional (HDPE) and bio-based biodegradable (PLA) microplastics at two concentrations, both independently and combined with nutrient enrichment. Exposure to microplastics altered the isotopic niches of two of the invertebrate species, with nutrient enrichment mediating this effect. Moreover, the effects of microplastics were consistent regardless of their type or concentration. Under enriched conditions, two of the species exposed to microplastics shifted to a specialised diet compared with controls, whereas little difference was observed between the isotopic niches of those exposed to microplastic and controls under ambient nutrient conditions. Additionally, PLA was estimated to support 24 % of the diet of one species, highlighting the potential assimilation of bioplastics by biota and possible implications. Overall, these findings suggest that the toxic effects of microplastics suggested from laboratory studies might not manifest under real-world conditions. However, this study does demonstrate that subtle sublethal effects occur even at environmentally realistic microplastic concentrations. The crucial role of nutrient enrichment in mediating microplastic effects underscores the importance of considering microplastic pollution in the context of other environmental stressors.

Using Stable Nitrogen Isotope Ratios from Human Skeletal Muscle Tissue for Postmortem Interval (PMI) Estimation (Part 1)

Human skeletal muscle tissue was analyzed for stable nitrogen isotope analysis (δ15N) to test the applicability of changes in values (Δ15N) through the putrefaction process for postmortem interval (PMI) estimation. Seasonal differences in Δ15N values of decom-posing skeletal muscle tissue were analyzed from 20 human donors at the University of Tennessee Anthropology Research Facility. This study revealed that change in the δ15N ratios of decomposing muscle tissue has potential as a novel method for PMI estimation. In winter months, when identifiable muscle tissue can be retained on a body for months, the data indicate that there is a systematic increase in Δ15N values with increasing temperatures when putrefaction progresses faster with values shifting as much as 3.0‰. This initial study indicates there is value in further method development of this quantitative approach to PMI estimation for recently deceased individuals, but further work is needed. While the analysis only requires a small amount of tissue, has a user- friendly collection and analysis protocol, and is less subjective than visual methods, there is still significant research to be done prior to widespread applications as a PMI estimation method. Understanding variability in δ15N across decomposing muscle tissue groups, individual variation (i.e., muscle mass, disease, body chem-istry), and an accurate approximation for a perimortem δ15N value necessary to calculate Δ15N all need to be further researched prior to generation of a reliable regression formula and error rates.