Different responses of oxygen and hydrogen isotopes in leaf and tree-ring organic matter to lethal soil drought.

The oxygen and hydrogen isotopic composition (δ18O, δ2H) of plant tissues are key tools for the reconstruction of hydrological and plant physiological processes and may therefore be used to disentangle the reasons for tree mortality. However, how both elements respond to soil drought conditions before death has rarely been investigated. To test this, we performed a greenhouse study and determined predisposing fertilization and lethal soil drought effects on δ18O and δ2H values of organic matter in leaves and tree rings of living and dead saplings of five European tree species. For mechanistic insights, we additionally measured isotopic (i.e. δ18O and δ2H values of leaf and twig water), physiological (i.e. leaf water potential and gas-exchange) and metabolic traits (i.e. leaf and stem non-structural carbohydrate concentration, carbon-to-nitrogen ratios). Across all species, lethal soil drought generally caused a homogenous 2H-enrichment in leaf and tree-ring organic matter, but a low and heterogenous δ18O response in the same tissues. Unlike δ18O values, δ2H values of tree-ring organic matter were correlated with those of leaf and twig water and with plant physiological traits across treatments and species. The 2H-enrichment in plant organic matter also went along with a decrease in stem starch concentrations under soil drought compared with well-watered conditions. In contrast, the predisposing fertilization had generally no significant effect on any tested isotopic, physiological and metabolic traits. We propose that the 2H-enrichment in the dead trees is related to (i) the plant water isotopic composition, (ii) metabolic processes shaping leaf non-structural carbohydrates, (iii) the use of carbon reserves for growth and (iv) species-specific physiological adjustments. The homogenous stress imprint on δ2H but not on δ18O suggests that the former could be used as a proxy to reconstruct soil droughts and underlying processes of tree mortality.

The stable isotope hydrology of Sable Island, Nova Scotia, Canada with implications for evaluating the water budget of wild horses.

We investigated the stable isotope hydrology of Sable Island, Nova Scotia, Canada over a five year period from September, 2017 to August, 2022. The δ2H and δ18O values of integrated monthly precipitation were weakly seasonal and ranged from -66 to -15 ‰ and from -9.7 to -1.9 ‰, respectively. Fitting these monthly precipitation data resulted in a local meteoric water line (LMWL) defined by: δ2H = 7.22 ± 0.21 · δ18O + 7.50 ± 1.22 ‰. Amount-weighted annual precipitation had δ2H and δ18O values of -36 ± 11 ‰ and -6.1 ± 1.4 ‰, respectively. Deep groundwater had more negative δ2H and δ18O values than mean annual precipitation, suggesting recharge occurs mainly in the winter, while shallow groundwater had δ2H and δ18O values more consistent with mean annual precipitation or mixing of freshwater with local seawater. Surface waters had more positive values and showed evidence of isolation from the groundwater system. The stable isotopic compositions of plant (leaf) water, on the other hand, indicate plants use groundwater as their source. Fog had δ2H and δ18O values that were significantly more positive than those of local precipitation, yet had similar 17O-excess values. δ2H values of horsehair from 4 individuals lacked seasonality, but had variations typical to those of precipitation on the island. Differences in mean δ2H values of horsehair were statistically significant and suggest variations in water use may exist between spatially disparate horse communities. Our results establish an important initial framework for ongoing isotope studies of feral horses and other wildlife on Sable Island.

Seasonality of hydrogeochemical evolutions and isotopic variabilities (δ18O, δ2H and d-excess) in the surface water as well as groundwater from tropical central-south Mexico.

Due to adverse impact of the global warming on hydrological resources, we intended to document the hydrogeochemical evolutions of surface and groundwater at tropical central-south Mexico in terms of seasonality of rock-water interaction, precipitation/evaporation variation and moisture source by evaluating the major ion chemistry in Piper and Gibbs plots, Durov diagram and through estimation of the chloro-alkaline indices as well as assessing the stable isotope compositions (δ18O and δ2H) in samples from different seasons of a year. Surface water of the Lake Coatetelco shifted from mostly Ca-Mg-HCO3 facies in wet summer-autumn to Na-HCO3-Cl facies in the dry spring due to elevated Na, Cl and HCO3. Greater evaporation in spring led to a maximum δ18O enrichment of ca.7‰ compared to the other seasons, and much depleted deuterium excess (-40.92‰ to -39.20‰). Interaction of the lake water with subsurface carbonate lithologies, and comparable isotopic compositions reflected the enhanced interaction between the surface water body and aquifers in the wet autumn. Effect of seasonality, however, was unclear on the groundwater facies, and its heterogenous composition (Ca-Mg-HCO3, Na-HCO3-Cl and Na-HCO3) reflected the interactions with different lithologies. Fractionations in isotope compositions of the groundwater were caused from recharge at different elevations, seasonality of moisture sources and moisture recycling. The water-mineral saturation index was an efficient proxy of seasonality as the lake water and groundwater (avg SIcalcite > 0.5) of the dry autumn were saturated with calcite. This vital information about carbonate precipitation, pCO2 and chemical facies would be useful for the better interpretation of paleoclimate archives in this region.

Topographic influence on ecohydrology in volcanic watersheds of the western Pacific monsoon area: evidence from water stable isotope composition of meteoric water, thermal water, and plants.

In Taiwanese volcanic watersheds, we investigated stable water isotopes in meteoric water, plants, and thermal water. Meteoric water exhibited a seasonal cycle, with heavier isotopes in winter and lighter ones in summer, especially in the southern region. The northern monsoon signal lagged the south by two weeks. In the Tatun mountains, young water fractions indicated prevalent old water sources. In the northern watershed, streamwater mainly came from the winter monsoon, while the southern one was influenced by alternating monsoons. Both indices indicated that winter plants depended on summer rainfall. Streamwater and plants had distinct sources in winter, supporting ecohydrological separation. Thermal spring water's d-excess helped identify water-rock interactions, with low d value signaling such interactions. The topographic wetness index showed a higher summer monsoon contribution to southern streamwater but a lower one to plants. The mean linear channel direction significantly affected the monsoon contribution fraction, with northeast-oriented channels vulnerable to northeastward winter monsoons. Finally, we developed a model illustrating hydrological processes on short and long timescales. Our findings enhance our understanding of hydrological disturbances' impact on water resources and ecosystems.

Cholesterol Stiffening of Lipid Membranes.

Biomembrane order, dynamics, and other essential physicochemical parameters are controlled by cholesterol, a major component of mammalian cell membranes. Although cholesterol is well known to exhibit a condensing effect on fluid lipid membranes, the extent of stiffening that occurs with different degrees of lipid acyl chain unsaturation remains an enigma. In this review, we show that cholesterol locally increases the bending rigidity of both unsaturated and saturated lipid membranes, suggesting there may be a length-scale dependence of the bending modulus. We review our published data that address the origin of the mechanical effects of cholesterol on unsaturated and polyunsaturated lipid membranes and their role in biomembrane functions. Through a combination of solid-state deuterium NMR spectroscopy and neutron spin-echo spectroscopy, we show that changes in molecular packing cause the universal effects of cholesterol on the membrane bending rigidity. Our findings have broad implications for the role of cholesterol in lipid-protein interactions as well as raft-like mixtures, drug delivery applications, and the effects of antimicrobial peptides on lipid membranes.

TRITIUM CONCENTRATION IN MONTHLY PRECIPITATION NEAR THE FUSION TEST FACILITY IN JAPAN BEFORE AND AFTER THE DEUTERIUM PLASMA EXPERIMENT.

In Japan, the deuterium plasma experiment using the Large Helical Device was started at the National Institute for Fusion Science (NIFS) in March 2017 to investigate high-temperature plasma physics and hydrogen isotope effects in research leading towards the realisation of fusion energy. The deuterium plasma experiment produces small amount of tritium by fusion reactions. To understand any impacts by the experiment to the surrounding environment, monthly precipitation samples have been collected at the NIFS site since November 2013 to assess the relationship between isotope composition and chemical species in precipitation including tritium. By comparing data before and after the deuterium plasma experiment start, it was found that tritium released from the main stack of the fusion test facility had no impact on the environment surrounding NIFS.

LEVELS OF ATMOSPHERIC TRITIUM IN THE SITE OF FUSION TEST FACILITY.

In the deuterium plasma experiment using Large Helical Device at the National Institute for Fusion Science (NIFS), a small amount of tritium is produced by the D-D fusion reaction. Then, a part of produced tritium is discharged into the environment via a stack. Thus, the atmospheric tritium in the site of NIFS has been monitored before starting the deuterium plasma experiment. The atmospheric tritium concentrations at NIFS were indicated to be background levels in Japan. To investigate the impact of tritium discharged from the stack, the correlation between the atmospheric tritium concentration and the tritium concentration observed in the stack was evaluated, and no significant correlation was found. In addition, the atmospheric tritium concentration at NIFS ranged within the background levels in Japan. Therefore, the impact of discharged tritium from the stack would be negligible in the environment at NIFS.

DEUTERIUM TRANSFER ANALYSIS INCLUDING FOOD CHAIN FROM SEAWATER INTO ABALONE.

Tritium is released into the ocean from nuclear facilities located at coastal areas. In addition, tritiated water is decided to be released into the ocean from the Fukushima Dai-ichi Nuclear Power Plant. Although released tritium concentration would be strictly controlled, impact of tritium on the marine products is major concern for the public. In this study, deuterium transfers from seawater into seaweed (ulva) and abalone were measured. In addition, organically bound deuterium (OBD) transfer from ulva into abalone was measured. OBD concentrations in ulva were saturated in 2 weeks and those in abalone were saturated in 6 months. Ulva and abalone were exposed to seawater containing 0.2% (mol-D/mol-H) deuterium. Maximum OBD concentrations in ulva were ~0.1% (mol-D/mol-H) and those in abalone muscle were ~0.035% (mol-D/mol-H). Numerical deuterium transfer model was constructed. Obtained numerical model well represented the OBD-enriched ulva feeding experiment.

A novel deuterium-based model for measurement of exogenous surfactant using deuterium-depleted water.

Stable isotope tracers, like 13 C, can be used for the measurement of the partition between the endogenous and exogenous pulmonary disaturated-phosphatidylcholine (DSPC). Deuterium labeling methods are still not fully explored. Our aim was to investigate the feasibility of using deuterium-depleted water (DDW) and deuterium-enriched water (DEW) to measure endogenous and exogenous pulmonary DSPC in a rabbit model of surfactant depletion. Data obtained from the 13 C dilution method were used as a reference. We studied 9 adult rabbits: 4 drank DDW and 5 DEW for 5 days. Lung surfactant depletion was induced at Day 5 by repeated saline bronchoalveolar lavages (BAL), which were stored as a pool (BAL pool). After endogenous surfactant depletion, rabbits received exogenous surfactant followed by a second BAL depletion procedure (End-Experiment Pool). DSPC quantity, and palmitic acid (PA)-DSPC 2 H/1 H (δ2 H) and 13 C/12 C ratios (δ13 C) of exogenous surfactant batches and of BAL pools were measured by High-Resolution Mass Spectrometry. The amount of exogenous surfactant recovered from the lungs ranged from 45% to 81% and, it was highly correlated with those obtained with the use of the 13 C (r = 0.9844, p < 0.0001). We demonstrated that commercially available purified DDW and even low doses of DEW can be used to modify the deuterium background of endogenous surfactants with the purpose of measuring the contribution of exogenous surfactants to the endogenous alveolar surfactant pool.

Comparative cryogenic extraction rehydration experiments reveal isotope fractionation during root water uptake in Gramineae.

Determining whether isotope fractionation occurs during root water uptake is a prerequisite for using stem or xylem water isotopes to trace water sources. However, it is unclear whether isotope fractionation occurs during root water uptake in gramineous crops. We conducted prevalidation experiments to estimate the isotope measurement bias associated with cryogenic vacuum distillation (CVD). Next, we assessed isotope fractionation during root water uptake in two common agronomic crops, wheat (Triticum aestivum L.) and maize (Zea mays L.), under flooding after postdrought stress conditions. Cryogenic vacuum distillation caused significant depletion of 2 H but negligible effects on 18 O for both soil and stem water. Surprisingly CVD caused depletion of 2 H and enrichment of 18 O in root water. Stem and root water δ18 O were more than soil water δ18 O, even considering the uncertainty of CVD. Soil water 18 O was depleted compared with irrigation water 18 O in the pots with plants but enriched relative to irrigation water 18 O in the pots without plants. These results indicate that isotope fractionation occurred during wheat and maize root water uptake after full irrigation and led to a heavy isotope enrichment in stem water. Therefore, the xylem/stem water isotope approach widely used to trace water sources should be carefully evaluated.

First use of triply labelled water analysis for energy expenditure measurements in mice.

The doubly labelled water (DLW) method is widely used to determine energy expenditure. In this work, we demonstrate the addition of the third stable isotope, 17O, to turn it into triply labelled water (TLW), using the three isotopes measurement of optical spectrometry. We performed TLW (2H, 18O and17O) measurements for the analysis of the CO2 production (rCO2) of mice on different diets for the first time. Triply highly enriched water was injected into mice, and the isotope enrichments of the distilled blood samples of one initial and two finals were measured by an off-axis integrated cavity output spectroscopy instrument. We evaluated the impact of different calculation protocols and the values of evaporative water loss fraction. We found that the dilution space and turnover rates of 17O and 18O were equal for the same mice group, and that values of rCO2 calculated based on 18O-2H, or on 17O-2H agreed very well. This increases the reliability and redundancy of the measurements and it lowers the uncertainty in the calculated rCO2 to 3% when taking the average of two DLW methods. However, the TLW method overestimated the rCO2 compared to the indirect calorimetry measurements that we also performed, much more for the mice on a high-fat diet than for low-fat. We hypothesize an extra loss or exchange mechanism with a high fractionation for 2H to explain this difference.

Deuterium-depleted water: A new tracer to label pulmonary surfactant lipids in adult rabbits.

Stable isotope tracing can be safely used for metabolic studies in animals and humans. The endogenous biosynthesis of lipids (lipogenesis) is a key process throughout the entire life but especially during brain and lung growth. Adequate synthesis of pulmonary surfactant lipids is indispensable for life. With this study, we report the use of deuterium-depleted water (DDW), suitable for human consumption, as metabolic precursor for lipogenesis. We studied 13 adult rabbits for 5 days. Four rabbits drank tap water (TW) and served as controls; in four animals, DDW was substituted to drinking water, whereas five drank deuterium-enriched water (DEW). After 5 days, a blood sample and a bronchoalveolar lavage (BAL) sample were collected. The 2 H/1 H (δ2 H) of BAL palmitic acid (PA) desaturated phosphatidylcholine (DSPC), the major phospholipid of pulmonary surfactant, and of plasma water was determined by high-resolution mass spectrometry. We found that the δ2 H values of DDW, DEW and TW were -984 ± 2‰, +757 ± 2‰ and -58 ± 1‰, respectively. After 5 days, plasma water values were -467 ± 87‰, +377 ± 56‰ and -53 ± 6‰, and BAL DSPC-PA was -401 ± 27‰, -96 ± 38‰ and -249 ± 9‰ in the DDW, DEW and TW, respectively. With this preliminary study, we demonstrated the feasibility of using DDW to label pulmonary surfactant lipids. This novel approach can be used in animals and in humans, and we speculate that it could be associated with more favourable study compliance than DEW in human studies.

Formula Assignment Algorithm for Deuterium-Labeled Ultrahigh-Resolution Mass Spectrometry: Implications of the Formation Mechanism of Halogenated Disinfection Byproducts.

Ultrahigh-resolution mass spectrometry (UHR-MS) coupled with isotope labeling has attracted significant attention in elucidating the mechanisms of the transformation of dissolved organic matter (DOM). Herein, we developed a novel formula assignment algorithm based on deuterium (D)-labeled UHR-MS, namely, FTMSDeu, for the first time. This algorithm was employed to determine the precursor molecules of halogenated disinfection byproducts (Xn-DBPs) and to evaluate the relative contribution of electrophilic addition and substitution reactions in Xn-DBP formation according to the H/D exchange of DOM molecules. Further, tandem mass spectrometry with homologous-based network analysis was used to validate the formula assignment accuracy of FTMSDeu in the identification of iodinated disinfection byproducts. Electrophilic substitution accounted for 82-98, 71-89, and 43-45% of the formation for Cl-, Br-, and I-containing Xn-DBPs, respectively, indicating the dominant role of the electrophilic substitution in chlorinated disinfection byproducts with low Br and I concentrations. The absence of putative precursors in some Xn-DBPs also suggests that Xn-DBP formation includes secondary reactions (e.g., oxidation and hydrolysis) in addition to the electrophilic addition and/or substitution of halogens. These findings highlight the significance of isotopically labeled UHR-MS techniques in revealing the transformation of DOM in natural and engineered systems.

A high-temperature water vapor equilibration method to determine non-exchangeable hydrogen isotope ratios of sugar, starch and cellulose.

The analysis of the non-exchangeable hydrogen isotope ratio (δ2 Hne ) in carbohydrates is mostly limited to the structural component cellulose, while simple high-throughput methods for δ2 Hne values of non-structural carbohydrates (NSC) such as sugar and starch do not yet exist. Here, we tested if the hot vapor equilibration method originally developed for cellulose is applicable for NSC, verified by comparison with the traditional nitration method. We set up a detailed analytical protocol and applied the method to plant extracts of leaves from species with different photosynthetic pathways (i.e., C3 , C4 and CAM). δ2 Hne of commercial sugars and starch from different classes and sources, ranging from -157.8 to +6.4‰, were reproducibly analysed with precision between 0.2‰ and 7.7‰. Mean δ2 Hne values of sugar are lowest in C3 (-92.0‰), intermediate in C4 (-32.5‰) and highest in CAM plants (6.0‰), with NSC being 2 H-depleted compared to cellulose and sugar being generally more 2 H-enriched than starch. Our results suggest that our method can be used in future studies to disentangle 2 H-fractionation processes, for improving mechanistic δ2 Hne models for leaf and tree-ring cellulose and for further development of δ2 Hne in plant carbohydrates as a potential proxy for climate, hydrology, plant metabolism and physiology.

Continuous in situ measurements of water stable isotopes in soils, tree trunk and root xylem: Field approval.

RATIONALE: New methods to measure stable isotopes of soil and tree water directly in the field enable us to increase the temporal resolution of obtained data and advance our knowledge on the dynamics of soil and plant water fluxes. Only few field applications exist. However, these are needed to further improve novel methods and hence exploit their full potential. METHODS: We tested the borehole equilibration method in the field and collected in situ and destructive samples of stable isotopes of soil, trunk and root xylem water over a 2.5-month experiment in a tropical dry forest under natural abundance conditions and following labelled irrigation. Water from destructive samples was extracted using cryogenic vacuum extraction. Isotope ratios were determined with IRIS instruments using cavity ring-down spectroscopy both in the field and in the laboratory. RESULTS: In general, timelines of both methods agreed well for both soil and xylem samples. Irrigation labelled with heavy hydrogen isotopes clearly impacted the isotope composition of soil water and one of the two studied tree species. Inter-method deviations increased in consequence of labelling, which revealed their different capabilities to cover spatial and temporal heterogeneities. CONCLUSIONS: We applied the novel borehole equilibration method in a remote field location. Our experiment reinforced the potential of this in situ method for measuring xylem water isotopes in both tree trunks and roots and confirmed the reliability of gas permeable soil probes. However, in situ xylem measurements should be further developed to reduce the uncertainty within the range of natural abundance and hence enable their full potential.

Growth-phase dependence of bacterial membrane lipid profile and labeling for in-cell solid-state NMR applications.

Cell labeling is a preliminary step in multiple biophysical approaches, including the solid-state nuclear magnetic resonance (NMR) study of bacteria in vivo. Deuterium solid-state NMR has been used in the past years to probe bacterial membranes and their interactions with antimicrobial peptides, following a standard labeling protocol. Recent results from our laboratory on a slow-growing bacterium has shown the need to optimize this protocol, especially the bacterial growth time before harvest and the concentration of exogenous labeled fatty acids to be used for both Escherichia coli and Bacillus subtilis. It is also essential for the protocol to remain harmless to cells while providing optimal labeling. We have therefore developed a fast and facile approach to monitor the lipid composition of bacterial membranes under various growth conditions, combining solution 31P NMR and GCMS. Using this approach, the optimized labeling conditions of Escherichia coli and Bacillus subtilis with deuterated palmitic acid were determined. Our results show a modification of B. subtilis phospholipid profile as a function of the growth stage, as opposed to E. coli. Our protocol recommends low concentrations of exogenous palmitic acid in the growth medium, and bacteria harvest after the exponential phase.

Hyperspectral CARS microscopy and quantitative unsupervised analysis of deuterated and non-deuterated fatty acid storage in human cells.

Coherent anti-Stokes Raman scattering (CARS) implemented as a vibrational micro-spectroscopy modality eradicates the need for potentially perturbative fluorescent labeling while still providing high-resolution, chemically specific images of biological samples. Isotopic substitution of hydrogen atoms with deuterium introduces minimal change to molecular structures and can be coupled with CARS microscopy to increase chemical contrast. Here, we investigate HeLa cells incubated with non-deuterated or deuterium-labeled fatty acids, using an in-house-developed hyperspectral CARS microscope coupled with an unsupervised quantitative data analysis algorithm, to retrieve Raman susceptibility spectra and concentration maps of chemical components in physically meaningful units. We demonstrate that our unsupervised analysis retrieves the susceptibility spectra of the specific fatty acids, both deuterated and non-deuterated, in good agreement with reference Raman spectra measured in pure lipids. Our analysis, using the cell-silent spectral region, achieved excellent chemical specificity despite having no prior knowledge and considering the complex intracellular environment inside cells. The quantitative capabilities of the analysis allowed us to measure the concentration of deuterated and non-deuterated fatty acids stored within cytosolic lipid droplets over a 24 h period. Finally, we explored the potential use of deuterium-labeled lipid droplets for non-invasive cell tracking, demonstrating an effective application of the technique for distinguishing between cells in a mixed population over a 16 h period. These results further demonstrate the chemically specific capabilities of hyperspectral CARS microscopy to characterize and distinguish specific lipid types inside cells using an unbiased quantitative data analysis methodology.

Time-resolved Rayleigh scattering measurements of methane clusters for laser-cluster fusion experiments.

We present a time-resolved analysis of Rayleigh scattering measurements to determine the average size of methane clusters and find the optimum timing for laser-cluster fusion experiments. We measure Rayleigh scattering and determine the average size of methane clusters varying the backing pressure (P0) from 11 bar to 69 bar. Regarding the onset of clustering, we estimate that the average size of methane clusters at the onset of clustering is Nc0≅20 at 11 bar. According to our measurements, the average cluster radius r follows the power law of r∝P01.86. Our ion time-of-flight measurements indicate that we have produced energetic deuterium ions with kT = 52±2 keV after laser-cluster interaction using CD4 gas at 50 bar. We find that this ion temperature agrees with the predicted temperature from CD4 clusters at 50 bar with r = 14 nm assuming the Coulomb explosion model.

Impact of Reduced Deuterium Intake on Thermoregulation.

The role of stable hydrogen isotopes in the thermoregulation and its regulation is poorly studied. We analyzed fluctuations in body temperature and changes in thermoregulation parameters in mice under conditions of reduced deuterium intake. The study was performed on male C57BL/6 mice that consumed water with a low (10 ppm) and normal (146 ppm) deuterium content. In 7 days, fluctuations of body temperature, locomotor activity, and oxygen uptake were assessed. Deuterium depletion in the body reduced the mean value of minute fluctuations of body temperature and the mean spectral density of minute fluctuations in body temperature in the 2-20-min periods. This attested to a stabilizing effect of deuterium depletion on the rhythms of body temperature fluctuations, without significant shifts in the thermogenesis parameters. Thus, drinking water with reduced deuterium content makes them less sensitive to external influences.

Variations in human body water isotope composition across the United States.

Accurate human provenancing using stable isotopes depends directly on solid understandings of the geographic and individual factors affecting isotope variability and incorporation into human tissues. Transfer of isotopic, and therefore spatial, information between environmental water and biological tissues is mediated by the isotopic composition of body water. Thus, there is a need to study body water isotope ratios at a population level and over a large isotopic and geographic range. We evaluated oxygen (δ18Obw) and hydrogen (δ2Hbw) isotope values of body water from 72 volunteers in 10 different cities across the US, and over a 5-10-day period. We analyzed covariates (e.g., water intake, physical activity, biometrics, gender) that might explain individual stable isotope ratio variations and tested a predictive model that incorporates the δ-values of drinking water, food, and O2 as well as individual variables to predict the δ-values of body water. The individual variability in body water isotope values overtime (mean 0.3‰ for δ18Obw and 2.3‰ for δ2Hbw) was lower than the intra-city variability (mean 0.9‰ for δ18Obw and 6.9‰ for δ2Hbw). Body water isotope values differed among cities (ANOVA: δ18ObwF = 97.2, p < 0.001; δ2HbwF = 176.2, p < 0.001). However, significant overlap among some cities with different drinking water was discovered. We detected significant covariation of measured drinking water and human body water isotope values (both isotope systems R2 ≥ 0.89, p < 0.001) and small but significant effects of the average daily exercise and amount of fluid intake. The differences between measured and model-predicted body water values (mean 0.12 ± 1.2‰ for Δδ18O and -1.2 ± 8.2‰ for Δδ2H) were statistically indistinguishable from zero (Δδ18O t = -0.751, p = 0.45; Δδ2H t = 1.133, p = 0.26). Here we show that community level variation exists in the δ18Obw and δ2Hbw values and the primary drivers are the regional differences in drinking water isotopes. Consistency of the body water isotope composition over the study period suggests that tissues would incorporate a stable isotope signal over time. The amount of drinking water and physical activity influence body water values, while the variation in the isotopic values of food may contribute to regional level variability, but that still remains to be assessed further. The human body water model provides accurate estimates for measured values, capturing and reproducing the main features of the body water isotope variation across space.