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Crop rotational diversity is an important part of sustainable agricultural and soil management to improve crop yield and soil fertility including enhancing soil organic matter (SOM) stabilization. Because of the physical protection via interactions with soil minerals, SOM in mineral-associated fractions is believed to be longer-lived and more stable relative to SOM in particulate (light) fractions. However, it is still unclear how crop rotational diversity alters soil carbon distribution, composition and stabilization in soil physical fractions. To address this, we studied a 37 years’ agricultural site with different crop rotational diversity (from continuous corn or alfalfa up to four species (corn, soybean, winter wheat, and red clover)). Soil carbon analysis, targeted compound analysis and nuclear magnetic resonance spectroscopy methods were used to obtain the distribution and degradation of SOM components in light and mineral-associated (F53–2000 µm, F2–53 µm, and F) fractions. Higher soil organic carbon (SOC) concentrations were observed in F with relatively high diversified crop rotations (three and four types of crops) compared to monoculture or two crops in the rotations, which suggests that carbon storage is enhanced in mineral-stabilized pools. Higher concentrations of long-chain aliphatic compounds as well as increased accumulation and preservation of lignin-derived compounds in fine aggregates (

Maternal exposure to polystyrene nanoplastics impact postnatal brain development in mouse offspring. This work motivates further studies to determine the levels of nanoplastics in the environment.

Environmental metabolomics provides insight into how anthropogenic activities have an impact on the health of an organism at the molecular level. Within this field, in vivo NMR stands out as a powerful tool for monitoring real-time changes in an organism’s metabolome. Typically, these studies use 2D 13C-1H experiments on 13C-enriched organisms. Daphnia are the most studied species, given their widespread use in toxicity testing. However, with COVID-19 and other geopolitical factors, the cost of isotope enrichment increased ~6–7 fold over the last two years, making 13C-enriched cultures difficult to maintain. Thus, it is essential to revisit proton-only in vivo NMR and ask, “Can any metabolic information be obtained from Daphnia using proton-only experiments?”. Two samples are considered here: living and whole reswollen organisms. A range of filters are tested, including relaxation, lipid suppression, multiple-quantum, J-coupling suppression, 2D 1H-1H experiments, selective experiments, and those exploiting intermolecular single-quantum coherence. While most filters improve the ex vivo spectra, only the most complex filters succeed in vivo. If non-enriched organisms must be used, then, DREAMTIME is recommended for targeted monitoring, while IP-iSQC was the only experiment that allowed non-targeted metabolite identification in vivo. This paper is critically important as it documents not just the experiments that succeed in vivo but also those that fail and demonstrates first-hand the difficulties associated with proton-only in vivo NMR.

Per- and polyfluoroalkyl substances (PFASs) have been widely used since the 1940s in industry and everyday household products. They also persist in the environment and bioaccumulate in humans and wildlife. Despite these concerns, the identities of most PFASs in environmental and biological samples are unknown. Herein, we describe a novel cyclic ion mobility mass spectrometer (cIMS), hyphenated with gas chromatography (GC) atmospheric pressure chemical ionization, that can reveal the presence of unknown PFASs on the basis of the ratio of their mass and collision cross section (CCS). Prediction of the CCS of ca. 20,000 chemicals used in industry and commerce indicates that most compounds characterized by CCS values that are less than the sum of 100 Å

Synergism between different phases gives rise to chemical, biological or environmental reactivity, thus it is increasingly important to study samples intact. Here, SASSY (SimultAneous Solid and Solution spectroscopY) is introduced to simultaneously observe (and differentiate) all phases in multiphase samples using standard, solid-state NMR equipment. When monitoring processes, the traditional approach of studying solids and liquids sequentially, can lead to information in the non-observed phase being missed. SASSY solves this by observing the full range of materials, from crystalline solids, through gels, to pure liquids, at full sensitivity in every scan. Results are identical to running separate 13C CP-MAS solid-state and 13C solution-state experiments back-to-back but requires only a fraction of the spectrometer time. After its introduction, SASSY is applied to process monitoring and finally to detect all phases in a living freshwater shrimp. SASSY is simple to implement and thus should find application across all areas of research.

Per- and polyfluoroalkyl substances (PFAS) are a class of persistent organic pollutants used in industrial applications due to their physicochemical properties that results in their ubiquitous presence across environmental matrices. To date, legacy PFAS have been well-studied, however, the concentration of alternative PFAS may exceed the concentration of legacy pollutants and more information is needed regarding the sub-lethal toxicity at the molecular-level of aquatic model organisms, such as Daphnia magna. Perfluorobutanoic acid (PFBA), perfluorohexanoic acid (PFHxA), perfluorohexanesulfonic acid (PFHxS), and perfluorononanoic acid (PFNA), are four widely detected PFAS alternatives of varying chain length and polar functionality that are quantified in aquatic environments. This study examines the metabolic perturbations of PFAS with varying chemistries to D. magna using targeted mass spectrometry -based metabolomics. Daphnia were acutely exposed to sub-lethal concentrations of PFBA, PFHxA, PFHxS, and PFNA before the polar metabolite profile was extracted from single organisms. Multivariate analysis demonstrated significant separation between the sub-lethal concentrations of PFHxA, PFHxS, and PFNA relative to the controls; in sum, longer chain lengths demonstrated greater overall perturbations to the extracted metabolic profiles. Univariate statistics revealed significant perturbations in the concentrations of several amino acids, nucleotides/nucleosides, and neurotransmitters with exposure to PFAS. These metabolic perturbations are consistent with disruptions in energy metabolism (pantothenate and Coenzyme A metabolism, histidine metabolism) and protein synthesis (aminoacyl-tRNA biosynthesis and amino acid metabolism) which were identified through biochemical pathway analysis. These results provide evidence that while PFAS chemistry (chain length and polar functional group) invokes unique metabolic responses there is also an underlying toxic mode of action that is common with select PFAS exposure. Overall, this work highlights the capabilities of environmental metabolomics to elucidate the molecular-level perturbations of pollutants within the same chemical class to model aquatic organisms which can be used to prioritize risk assessment of substituted PFAS alternatives. This article is protected by copyright. All rights reserved. Environ Toxicol Chem 2022;00:0-0. © 2022 SETAC.

Forest soils are an important carbon (C) sink and critical component of the global C cycle. Warmer temperatures and increased atmospheric nitrogen (N) deposition are altering the biogeochemistry in forest soils and disrupting the intricate balance between C storage and C respired across the globe. The molecular biogeochemistry of soil organic matter (SOM) with warming, N-addition, and simultaneous warming and N-addition was analyzed in soil samples from the Soil Warming × Nitrogen Addition Study at the Harvard Forest Long-term Ecological Research Site using advanced techniques. The results unequivocally demonstrate that warming and N-addition alter the molecular composition of SOM as individual stressors uniquely and in combination. Warming alone and in combination with N-addition accelerated SOM decomposition while N-addition alone slowed SOM degradation. The two-factor N-addition and warming plots contain SOM more like the warming only plots but exhibited unique changes over time (from 4 to 10 years) that could not be predicted by studying N-addition or warming alone. The specific SOM components and the overall SOM decomposition suggests that N-addition and warming impacts are not additive. N-addition may hinder warming impacts antagonistically over time but not to the extent where advanced SOM decomposition from warming is supplanted. As such, the results from warming alone and N-addition alone are not necessarily additive compared to the observed SOM molecular compositional changes when these treatments are applied simultaneously. Marked evolution in the molecular biogeochemistry of SOM demonstrates the sensitivity of SOM trajectories to multiple interactive global environmental changes and the continued need to study long-term impacts more holistically.

Organic pollutants can be identified by comparing their electron ionization (EI) mass spectra with those in libraries or obtained from authentic standards. Nevertheless, libraries are incomplete; standards may be unavailable or too costly, or their synthesis may be too time-consuming. This study evaluates the performance of quantum chemical electron ionization mass spectrometry (QCEIMS) vis-a-vis competitive fragmentation modeling (CFM) for suspect screening and unknown identification. EI mass spectra of 35 compounds, including halogenated organics, organophosphorus flame retardants (OPFRs), and disinfection byproducts were computed. Computational results were compared with EI mass spectra compiled in the NIST Library as well as collision-induced dissociation (CID) mass spectra obtained from radical cations M•+ generated by charge-exchange atmospheric pressure chemical ionization (APCI). The results indicate that QCEIMS performs equivalently or better than CFM. Average match factors between computed and experimental (NIST) EI mass spectra were 656 vs 503 for the halogenated organics, and on average, QCEIMS predicted 55% of the products generated by CID vs 17% predicted by CFM. QCEIMS predicted 37% of the OPFR CID products whereas CFM predicted 29%. QCEIMS performed comparably to a commercial combinatorial fragmentation method for suspect screening of a dust sample, identifying 19/20 targets. Examples of unknown pollutants, whose reference spectra were unavailable at the time of discovery, are also presented. The computational results suggest that QCEIMS can help guide the analyst in obtaining authentic standards and raise the possibility that, with advances in computing, an unknown may eventually be confirmed in hours as opposed to the days or months required to obtain authentic standards.

Conserving soil carbon (C) and harnessing the potential for soil C sequestration requires an improved understanding of the processes through which organic material accumulates in soil. Currently, competing hypotheses exist regarding the dominant mechanisms that control soil C accumulation and transfers to mineral-associated pools. Long-standing hypotheses rely upon an assumed strong relationship between the quantity of organic inputs and soil C accumulation, while more recent hypotheses have shifted the focus towards the more complex controls of root activity, microbial processing and priming, and organo-mineral complexation. The Detrital Input and Removal Treatment (DIRT) experiment can test these competing hypotheses through field manipulations of detrital inputs. After 20 years of detrital manipulations in the wet, temperate forest of the H.J. Andrews Experimental Station, we found that with the termination of live root activity, the significant influx of dead root material and absence of soil priming by roots led to decreases in particulate organic matter (POM), but increases in stable mineral associated organic matter (MAOM). This suggests that soil mineral particles in undisturbed soils are not saturated with C in the presence of live roots and that pools of MAOM are sensitive to the balance between microbial-induced stabilization and microbial-induced priming and destabilization. Twenty years of aboveground litter removal did not change bulk soil C stocks or pools. Soil C stabilization did not increase in response to increases in high quality litter inputs, in contrast to recent theory, but in accordance with other empirical results. In contrast, increases in low quality wood litter led to a large increase in bulk soil C, with gains over 20 years confined to increases in POM. These findings offer insight into the pathways controlling soil C contents and provide potential explanations for the often-limited potential to increase mineral associated soil C in many vegetated soils and observed buffered responses of soil C stocks to disturbances such as drought, fire, and timber harvest.

Endocrine disruption is intimately linked to controlling the population of pollutant-exposed organisms through reproduction and development dysregulation. This study investigated how endocrine disruption in a predator organism could affect prey species biology through infochemical communication. Daphnia magna and Chlorella vulgaris were chosen as model prey and predator planktons, respectively, and fenoxycarb was used for disrupting the endocrine system of D. magna. Hormones as well as endo- and exometabolomes were extracted from daphnids and algal cells and their culture media and analyzed using liquid chromatography with tandem mass spectrometry. Biomolecular perturbations of D. magna under impaired offspring production and hormone dysregulation were observed. Differential biomolecular responses of the prey C. vulgaris, indicating changes in methylation and infochemical communication, were subsequently observed under the exposure to predator culture media, containing infochemicals released from the reproducibly normal and abnormal D. magna, as results of fenoxycarb exposure. The observed cross-species transfer of the endocrine disruption consequences, initiated from D. magna, and mediated through infochemical communication, demonstrates a novel discovery and emphasizes the broader ecological risk of endocrine disruptors beyond reproduction disruption in target organisms.

Comprehensive multiphase NMR combines the ability to study and differentiate all phases (solids, gels, and liquids) using a single NMR probe. The general goal of CMP-NMR is to study intact environmental and biological samples to better understand conformation, organization, association, and transfer between and across phases/interfaces that may be lost with conventional sample preparation such as drying or solubilization. To date, all CMP-NMR studies have used 4 mm probes and rotors. Here, a larger 7 mm probehead is introduced which provides ∼3 times the volume and ∼2.4 times the signal over a 4 mm version. This offers two main advantages: (1) the additional biomass reduces experiment time, making 13C detection at natural abundance more feasible; (2) it allows the analysis of larger samples that cannot fit within a 4 mm rotor. Chicken heart tissue and Hyalella azteca (freshwater shrimp) are used to demonstrate that phase-based spectral editing works with 7 mm rotors and that the additional biomass from the larger volumes allows detection with 13C at natural abundance. Additionally, a whole pomegranate seed berry (aril) and an intact softgel capsule of hydroxyzine hydrochloride are used to demonstrate the analysis of samples too large to fit inside a conventional 4 mm CMP probe. The 7 mm version introduced here extends the range of applications and sample types that can be studied and is recommended when 4 mm CMP probes cannot provide adequate signal-to-noise (S/N), or intact samples are simply too big for 4 mm rotors.

Forest ecosystems are important global soil carbon (C) reservoirs, but their capacity to sequester C is susceptible to climate change factors that alter the quantity and quality of C inputs. To better understand forest soil C responses to altered C inputs, we integrated three molecular composition published data sets of soil organic matter (SOM) and soil microbial communities for mineral soils after 20 years of detrital input and removal treatments in two deciduous forests: Bousson Forest (BF), Harvard Forest (HF), and a coniferous forest: H.J. Andrews Forest (HJA). Soil C turnover times were estimated from radiocarbon measurements and compared with the molecular-level data (based on nuclear magnetic resonance and specific analysis of plant- and microbial-derived compounds) to better understand how ecosystem properties control soil C biogeochemistry and dynamics. Doubled aboveground litter additions did not increase soil C for any of the forests studied likely due to long-term soil priming. The degree of SOM decomposition was higher for bacteria-dominated sites with higher nitrogen (N) availability while lower for the N-poor coniferous forest. Litter exclusions significantly decreased soil C, increased SOM decomposition state, and led to the adaptation of the microbial communities to changes in available substrates. Finally, although aboveground litter determined soil C dynamics and its molecular composition in the coniferous forest (HJA), belowground litter appeared to be more influential in broadleaf deciduous forests (BH and HF). This synthesis demonstrates that inherent ecosystem properties regulate how soil C dynamics change with litter manipulations at the molecular-level. Across the forests studied, 20 years of litter additions did not enhance soil C content, whereas litter reductions negatively impacted soil C concentrations. These results indicate that soil C biogeochemistry at these temperate forests is highly sensitive to changes in litter deposition, which are a product of environmental change drivers.

Sulphate and dissolved organic matter (DOM) in freshwater systems may regulate the formation of methylmercury (MeHg), a potent neurotoxin that biomagnifies in aquatic ecosystems. While many boreal lakes continue to recover from decades of elevated atmospheric sulphate deposition, little research has examined whether historically high sulphate concentrations can result in persistently elevated MeHg production and accumulation in aquatic systems. This study used sediment from a historically sulphate-impacted lake and an adjacent reference lake in northwestern Ontario, Canada to investigate the legacy effects of sulphate pollution, as well as the effects of newly added sulphate, natural organic matter (NOM) of varying sulphur content and a sulphate reducing bacteria (SRB) inhibitor on enhancing or inhibiting the Hg methylation and demethylation activity (

Per- and polyfluoroalkyl substances (PFAS) are a class of pollutants of concern due to their ubiquitous presence, persistence, and toxicity in aquatic environments. Legacy PFAS pollutants such as perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) have been more widely studied in aquatic environments. However, replacement PFAS, such as ammonium perfluoro (2-methyl-3-oxahexanoate; GenX) are increasingly being detected with little known information surrounding their toxicity. Here, Daphnia magna, a model organism for freshwater ecotoxicology was used to compare the acute sub-lethal toxicity of PFOS, PFOA, GenX, and PFAS mixtures. Using liquid chromatography with tandem mass spectrometry (LC-MS/MS), the targeted polar metabolic profile extracted from single Daphnia was quantified to investigate perturbations in the exposure groups versus the unexposed organisms. Multivariate statistical analyses demonstrated significant non-monotonic separation in PFOA, GenX, and PFAS mixture exposures. Sub-lethal exposure to concentrations of PFOS did not lead to significant separation in multivariate analyses. Univariate statistics and pathway analyses were used to elucidate the mode of action of PFAS exposure. Exposure to all individual PFAS led to significant perturbations in many amino acids including cysteine, histidine, tryptophan, glycine, and serine. These perturbations are consistent with biochemical pathway disruptions in the pantothenate and Coenzyme A (CoA) biosynthesis, thiamine metabolism, histidine metabolism, and aminoacyl-tRNA biosynthesis pathways. Overall, the collected metabolomic data is consistent with disruptions in energy metabolism and protein synthesis as the primary mode of action of sub-lethal PFAS exposure. Secondary modes of action among individual pollutant exposures demonstrated that the structural properties (carboxylic acid vs. sulfonic acid group) may play a role in the metabolic perturbations observed. Sub-lethal exposure to PFAS mixtures highlighted a mixed response when compared to the individual pollutants (PFOS, PFOA, and GenX). Overall, this study emphasizes the niche capability of environmental metabolomics to differentiate secondary modes of action from metabolic perturbations in both single pollutant and pollutant mixtures within the same chemical class.

Considered as a promising source of sustainable energy, biofuel produced from algae holds many advantages. However, to truly understand the production process and assess the potential for further o...

The interactions of the different components of soils, such as living biomass and organic matter, with mineral surfaces are of considerable interest due to their environmental reactivity and potent...

Evaluate crop rotation diversity, perenniality, carbon (C) inputs, and C input quality as predictors of soil organic carbon (SOC) concentration and aggregate mean weight diameter (MWD). At a crop rotation trial in its 37th year in Ontario, Canada, species in rotations included corn (C, Zea mays L.), alfalfa (A, Medicago sativa L.), soybean (S, Glycine max (L.) Merr.), winter wheat (W, Triticum aestivum L.), and red clover (rc, Trifolium pratense L.). Rotations were: CC, CCAA, CCSS, CCSW, CCSWrc, AA. Soils (0–20 cm) were analyzed for aggregate MWD, aggregate C, and SOC concentrations. We estimated C inputs from historical yields and C input quality (C:N, lignin: N, or NMR-derived index) from structural plant tissues. C stabilization efficiency was estimated as the ratio of SOC stock per unit total or root C input. Crop rotation diversity failed to increase SOC concentrations or aggregate MWD. Perennialized rotations (CCSWrc, CCAA, AA) maintained the numerically highest SOC concentrations, and root C input increased SOC concentration. Out of 12 statistical tests relating C input quality to C stabilization efficiency, only 3 indicated a positive effect and 6 tests indicated a negative effect. Including a perennial forage such as alfalfa and limiting soybean frequency promotes aggregate MWD and SOC concentration more so than optimizing crop rotation diversity. Quality of structural plant inputs does not explain differences in C stabilization efficiency, possibly due to overriding influence of living root inputs.

The environment has many anthropogenic chemicals which, in sublethal concentrations, can alter the biochemical function of organisms. Environmental metabolomics uses analytical techniques such as nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) to uncover how the presence of environmental contaminants may alter the metabolic profile of organisms. Metabolomic experiments follow a typical workflow and can be applied to a gamut of organisms and contaminants or other environmental stressors. Following sublethal exposure, metabolic shifts and the contaminant toxic mode of action can be elucidated. Environmental metabolomics is a useful tool in ecotoxicology and may have a future role within environmental risk assessment strategies.

Daphnia (freshwater fleas) are among the most widely used organisms in regulatory aquatic toxicology/ecology, while their recent listing as an NIH model organism is stimulating research for understanding human diseases and processes. Daphnia are small enough to fit inside high field NMR spectrometers and can be kept alive indefinitely using flow systems that deliver food and oxygen. As such, in vivo NMR holds the potential to monitor when/if environmental stress is occurring, understand "why" chemicals are toxic (biochemical pathways impacted and toxic-mode-of-action), and differentiate between a temporary flux response (i.e. return to homeostasis) and a permanent change in biochemistry (likely a precursor to disease). At present however, such studies are limited as the in vivo NMR data of Daphnia are highly complex and the lack of spectral assignments makes extracting metabolic information difficult. In this study, Daphnia are 13C enriched to >97% 13C and numerous 1H and 13C 1D, 2D, and 3D NMR approaches are combined to provide, as complete as possible, an assignment of the Daphnia magna metabolome in vivo. Assignments are transferred (where possible) back to line narrowed susceptibility suppressed 1H 1D NMR spectra in order to permit the maximum amount of information to be gained in the future without the need for 13C enrichment. To our knowledge, this work represents the first time a comprehensive metabolic assignment of any small living organism has been performed using high field flow-based NMR.

Forest soil dissolved organic matter (DOM) is a major source of terrigenous dissolved organic carbon (DOC) that is an important component of biogeochemical cycles. While many studies have shown tha...

Short-chain polychlorinated n-alkanes are ubiquitous industrial chemicals widely recognized as persistent organic pollutants. They represent only a small fraction of the 184,600 elemental compositions (C10-25) and the myriad isomers of all possible (mixed) halogenated n-alkanes (PXAs). This study prioritizes the PXAs on the basis of their potential to persist, bioaccumulate, and undergo long-range transport guided by quantitative structure-property relationships (QSPRs), density functional theory (DFT), chemical fate models, and partitioning space. The QSPR results narrow the list to 966 elemental compositions, of which 352 (23 Br, 83 Cl/F, 119 Br/Cl, and 127 Br/F) are likely constituents of substances used as lubricants, plasticizers, and flame retardants. Complementary DFT calculations suggest that an additional 1367 elemental compositions characterized by a greater number of carbon and fluorine atoms but fewer chlorine and bromine atoms may also pose a risk. The results of this study underline the urgent need to identify and monitor these suspected pollutants, most appropriately using mass spectrometry. We estimate that the resolving power required to distinguish ∼74% of the prioritized elemental compositions from the most likely interferents, i.e., chlorinated alkanes, is approximately 60,000 (full width at half-maximum). This indicates that accurate identification of the PXAs is achievable using most high-resolution mass spectrometers.

During pregnancy, appropriate placental metabolism is essential for fetuses to reach their growth potential. However, metabolic mechanisms during pregnancy remain poorly understood. Determination of the levels of placental metabolites in healthy pregnancy and how they change throughout gestation is critical for understanding placental function.To determine the effects of gestational age on placental metabolites using healthy pregnant mice.In the present study, we collected placental tissue samples from healthy pregnant mice at three timepoints in late gestation (n = 16 placentas per gestational age). Metabolite profiles were determined usingUsing HRMAS MRS, we identified 14 metabolites in murine placental tissue samples. The relative concentration of 12 of the 14 metabolites remains unchanged throughout late gestation. Lysine was found to decrease significantly (p = 0.04) and glucose showed an inverted U-shape relationship (p = 0.03) with gestational age.This study demonstrated the feasibility of HRMAS MRS to determine relative metabolite concentrations in murine placental tissue. These findings establish baseline levels of placental tissue metabolite profiles and will serve as reference ranges for future studies using mouse models of fetal distress.

Bisphenols are used in the production of polycarbonate plastics and epoxy resins. Bisphenol A (BPA) has been widely studied and is believed to act as an endocrine disruptor. Bisphenol F (BPF) and bisphenol S (BPS) have increasingly been employed as replacements for BPA, although previous studies suggested that they yield similar physiological responses to several organisms. Daphnia magna is a common model organism for ecotoxicology and was exposed to sub-lethal concentrations of BPA, BPF, and BPS to investigate disruption to metabolic profiles. Targeted metabolite analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to measure polar metabolites extracted from D. magna, which are linked to a range of biochemical pathways. Multivariate analyses and individual metabolite changes showed similar non-monotonic concentration responses for all three bisphenols (BPA, BPF, and BPS). Pathway analyses indicated the perturbation of similar and distinct pathways, mostly associated with protein synthesis, amino acid metabolism, and energy metabolism. Overall, we observed responses that can be linked to a chemical class (bisphenols) as well as distinct responses that can be related to each individual bisphenol type (A, F, and S). These findings further demonstrate the need for using metabolomic analyses in exposure assessment, especially for chemicals within the same class which may disrupt the biochemistry uniquely at the molecular-level.

Comprehensive multiphase (CMP) NMR, first described in 2012, combines all of the hardware components necessary to analyze all phases (solid, gel, and solution) in samples in their natural state. In combination with spectral editing experiments, it can fully differentiate phases and study the transfer of chemical species across and between phases, providing unprecedented molecular-level information in unaltered natural systems. However, many natural samples, such as swollen soils, plants, and small organisms, contain water, salts, and ionic compounds, making them electrically lossy and susceptible to RF heating, especially when using high-strength RF fields required to select the solid domains. While dedicated reduced-heating probes have been developed for solid-state NMR, to date, all CMP-NMR probes have been based on solenoid designs, which can lead to problematic sample heating. Here, a new prototype CMP probe was developed, incorporating a loop gap resonator (LGR) for decoupling. Temperature increases are monitored in salt solutions analogous to those in small aquatic organisms and then tested in vivo on Hyalella azteca (freshwater shrimp). In the standard CMP probe (solenoid), 80% of organisms died within 4 h under high-power decoupling, while in the LGR design, all organisms survived the entire test period of 12 h. The LGR design reduced heating by a factor of ∼3, which allowed 100 kHz decoupling to be applied to salty samples with generally ≤10 °C sample heating. In addition to expanding the potential for in vivo research, the ability to apply uncompromised high-power decoupling could be beneficial for multiphase samples containing true crystalline solids that require the strongest possible decoupling fields for optimal detection.