High-precision 14C measurements of parenchyma-rich Hymenolobium petraeum tree species confirm bomb-peak atmospheric levels and reveal local fossil-fuel CO2 emissions in the Central Amazon.

Atmospheric radiocarbon (14C) recorded in tree rings has been widely used for atmospheric 14C calibration purposes and climate studies. But atmospheric 14C records have been limited along tropical latitudes. Here we report a sequence from 1938 to 2007 of precisely measured 14C dates in tree rings of the parenchyma-rich Hymenolobium petraeum tree species (Porto Trombetas, 1°S, 56°W) from the Central Brazilian Amazon. H. petraeum has discernible growth ring boundaries that allow dating techniques to be employed to produce calendrical dates. Bomb-peak tree-ring 14C reconstruction coincides with the broader changes associated with reported values of the Southern Hemisphere atmospheric 14C curve (SH zone 3; values within the ±2σ interval), suggesting that inter-hemispheric air-mass transport of excess-14C injected into the stratosphere during intensive atmospheric nuclear tests is relatively uniform across distinct longitudinal regions. From the early 1980s onwards, H. petraeum had lower 14C values than other pantropical 14C records. Through 14C-based estimation, we found a strong influence of fossil-fuel CO2 contributions from Porto Trombetas mining operations and shipping traffic on inland waterways. An increase of at least 6.3 ± 0.8 ppm of fossil-fuel CO2 has been detected by 14C. Our findings invite further 14C analyses using tree rings of tropical tree species as a potential tracer for a wide range of environmental sources of atmospheric 14C-variability.

Smoke radiocarbon measurements from Indonesian fires provide evidence for burning of millennia-aged peat.

In response to a strong El Niño, fires in Indonesia during September and October 2015 released a large amount of carbon dioxide and created a massive regional smoke cloud that severely degraded air quality in many urban centers across Southeast Asia. Although several lines of evidence indicate that peat burning was a dominant contributor to emissions in the region, El Niño-induced drought is also known to increase deforestation fires and agricultural waste burning in plantations. As a result, uncertainties remain with respect to partitioning emissions among different ecosystem and fire types. Here we measured the radiocarbon content (14C) of carbonaceous aerosol samples collected in Singapore from September 2014 through October 2015, with the aim of identifying the age and origin of fire-emitted fine particulate matter (particulate matter with an aerodynamic diameter less than or equal to 2.5 µm). The Δ14C of fire-emitted aerosol was -76 ± 51‰, corresponding to a carbon pool of combusted organic matter with a mean turnover time of 800 ± 420 y. Our observations indicated that smoke plumes reaching Singapore originated primarily from peat burning (∼85%), and not from deforestation fires or waste burning. Atmospheric transport modeling confirmed that fires in Sumatra and Borneo were dominant contributors to elevated PM2.5 in Singapore during the fire season. The mean age of the carbonaceous aerosol, which predates the Industrial Revolution, highlights the importance of improving peatland fire management during future El Niño events for meeting climate mitigation and air quality commitments.

Does Cedrela always form annual rings? Testing ring periodicity across South America using radiocarbon dating.

KEY MESSAGE: Radiocarbon dating shows that Cedrela trees from Bolivia, Ecuador and Venezuela form one ring per year but Cedrela trees from Suriname form two rings per year. ABSTRACT: Tropical tree rings have the potential to yield valuable ecological and climate information, on the condition that rings are annual and accurately dated. It is important to understand the factors controlling ring formation, since regional variation in these factors could cause trees in different regions to form tree rings at different times. Here, we use 'bomb-peak' radiocarbon (14C) dating to test the periodicity of ring formation in Cedrela trees from four sites across tropical South America. We show that trees from Bolivia, Ecuador and Venezuela have reliably annual tree rings, while trees from Suriname regularly form two rings per year. This proves that while tree rings of a particular species may be demonstrably annual at one site, this does not imply that rings are formed annually in other locations. We explore possible drivers of variation in ring periodicity and find that Cedrela growth rhythms are most likely caused by precipitation seasonality, with a possible degree of genetic control. Therefore, tree-ring studies undertaken at new locations in the tropics require independent validation of the annual nature of tree rings, irrespective of how the studied species behaves in other locations.

Improved radiocarbon analyses of modern human hair to determine the year-of-death by cross-flow nanofiltered amino acids: common contaminants, implications for isotopic analysis, and recommendations.

RATIONALE: In forensic investigation, radiocarbon ((14)C) measurements of human tissues (i.e., nails and hair) can help determine the year-of-death. However, the frequent use of cosmetics can bias hair (14)C results as well as stable isotope values. Evidence shows that hair exogenous impurities percolate beyond the cuticle layer, and therefore conventional pretreatments are ineffective in removing them. METHODS: We conducted isotopic analysis ((14)C, δ(13)C, δ(15)N and C/N) of conventionally treated and cross-flow nanofiltered amino acid (CFNAA)-treated samples (scalp- and body-hair) from a single female subject using fingernails as a reference. The subject studied frequently applies a permanent dark-brown dye kit to her scalp-hair and uses other care products for daily cleansing. We also performed pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) analyses of CFNAA-treated scalp-hair to identify contaminant remnants that could possibly interfere with isotopic analyses. RESULTS: The conventionally treated scalp- and body-hair showed (14)C offsets of ~21‰ and ~9‰, respectively. These offsets confirm the contamination by petrochemicals in modern human hair. A single CFNAA extraction reduced those offsets by ~34%. No significant improvement was observed when sequential extractions were performed, as it appears that the procedure introduced some foreign contaminants. A chromatogram of the CFNAA scalp-hair pyrolysis products showed the presence of petroleum and plant/animal compound residues, which can bias isotopic analyses. CONCLUSIONS: We have demonstrated that CFNAA extractions can partially remove cosmetic contaminants embedded in human hair. We conclude that fingernails are still the best source of keratin protein for year-of-death determinations and isotopic analysis, with body-hair and/or scalp-hair coupled with CFNAA extraction a close second.

Blank corrections for ramped pyrolysis radiocarbon dating of sedimentary and soil organic carbon.

Ramped pyrolysis (RP) targets distinct components of soil and sedimentary organic carbon based on their thermochemical stabilities and allows the determination of the full spectrum of radiocarbon ((14)C) ages present in a soil or sediment sample. Extending the method into realms where more precise ages are needed or where smaller samples need to be measured involves better understanding of the blank contamination associated with the method. Here, we use a compiled data set of RP measurements of samples of known age to evaluate the mass of the carbon blank and its associated (14)C signature, and to assess the performance of the RP system. We estimate blank contamination during RP using two methods, the modern-dead and the isotope dilution method. Our results indicate that during one complete RP run samples are contaminated by 8.8 ± 4.4 µg (time-dependent) of modern carbon (MC, fM ∼ 1) and 4.1 ± 5.5 µg (time-independent) of dead carbon (DC, fM ∼ 0). We find that the modern-dead method provides more accurate estimates of uncertainties in blank contamination; therefore, the isotope dilution method should be used with caution when the variability of the blank is high. Additionally, we show that RP can routinely produce accurate (14)C dates with precisions ∼100 (14)C years for materials deposited in the last 10,000 years and ∼300 (14)C years for carbon with (14)C ages of up to 20,000 years.

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

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

A novel post-1950 CE atmospheric 14C record for the tropics using absolutely dated tree rings in the equatorial Amazon.

In this study, we present a comprehensive atmospheric radiocarbon (14C) record spanning from 1940 to 2016, derived from 77 single tree rings of Cedrela odorata located in the Eastern Amazon Basin (EAB). This record, comprising 175 high-precision 14C measurements obtained through accelerator mass spectrometry (AMS), offers a detailed chronology of post-1950 CE (Common Era) 14C fluctuations in the Tropical Low-Pressure Belt (TLPB). To ensure accuracy and reliability, we included 14C-AMS results from intra-annual successive cuts of the tree rings associated to the calendar years 1962 and 1963 and conducted interlaboratory comparisons. In addition, 14C concentrations in 1962 and 1963 single-year cuts also allowed to verify tissue growth seasonality. The strategic location of the tree, just above the Amazon River and estuary areas, prevented the influence of local fossil-CO2 emissions from mining and trade activities in the Central Amazon Basin on the 14C record. Our findings reveal a notable increase in 14C from land-respired CO2 starting in the 1970s, a decade earlier than previously predicted, followed by a slight decrease after 2000, signaling a transition towards the fossil fuel era. This shift is likely attributed to changes in reservoir sources or global atmospheric dynamics. The EAB 14C record, when compared with a shorter record from Muna Island, Indonesia, highlights regional differences and contributes to a more nuanced understanding of global 14C variations at low latitudes. This study not only fills critical spatial gaps in existing 14C compilations but also aids in refining the demarcation of 14C variations over South America. The extended tree-ring 14C record from the EAB is pivotal for reevaluating global patterns, particularly in the context of the current global carbon budget, and underscores the importance of tropical regions in understanding carbon-climate feedbacks.

Demystifying the tropics: FTIR characterization of pantropical woods and their α-cellulose extracts for past atmospheric 14C reconstructions.

To ensure unbiased tree-ring radiocarbon (14C) results, traditional pretreatments carefully isolate wood cellulose from extractives using organic solvents, among other chemicals. The addition of solvents is laborious, time-consuming, and can increase the risk of carbon contamination. Tropical woods show a high diversity in wood-anatomical and extractive composition, but the necessity of organic-solvent extraction for the 14C dating of these diverse woods remains untested. We applied a chemical treatment that excludes the solvent step on the wood of 8 tropical tree species sampled in South-America and Africa, with different wood-anatomical and extractive properties. We analyzed the success of the extractive removal along with several steps of the α-cellulose extraction procedure using Fourier Transform Infrared (FTIR) spectroscopy and further confirmed the quality of 14C measurements after extraction. The α-cellulose extracts obtained here showed FTIR-spectra free of signals from various extractives and the 14C results on these samples showed reliable results. The chemical method evaluated reduces the technical complexity required to prepare α-cellulose samples for 14C dating, and therefore can bolster global atmospheric 14C applications, especially in the tropics.

Longevity Estimates of Canary Palms and Dragon Trees via Radiocarbon Dating: Initial Results.

Correctly estimating the maximum lifespan of plant species is a necessary component of demographic and life-history studies, which, in turn, are needed for understanding climatic impacts. Arboreal monocotyledons, which can grow to >30 m in height and >5 m in trunk perimeter, are difficult to age because they do not undergo seasonal dormancy; hence, their longevity has been estimated using various size-related methods. In this study, we tested radiocarbon (14C) dating with Accelerator Mass Spectrometry (AMS) as an additional tool for determining the age of two iconic monocotyledons: the Canary Island palm and the dragon tree. A total of 25 samples were collected from the basal stem of four palms and five dragon trees on Gran Canaria and Tenerife and then processed using the most advanced 14C-AMS analysis available. Calibration curves provided by the "IntCal group" were used to determine the oldest possible age of each sample, and 16 of them were found to be "modern", i.e., formed after the 1950s. Nine samples that were either collected from exterior, but lignified, palm tissues or from interior, and lignified, dragon tree tissues suggested ages > 300 years. Given the constant improvement of 14C-AMS tools, they can contribute to the further refinement of existing scientific knowledge on Macaronesian charismatic megaflora.

Radiocarbon and wood anatomy as complementary tools for generating tree-ring records in Bolivia.

The science of tropical dendrochronology is now emerging in regions where tree-ring dating had previously not been considered possible. Here, we combine wood anatomical microsectioning techniques and radiocarbon analysis to produce the first tree-ring chronology with verified annual periodicity for a new dendrochronological species, Neltuma alba (commonly known as "algarrobo blanco") in the tropical Andes of Bolivia. First, we generated a preliminary chronology composed of six trees using traditional dendrochronological methods (i.e., cross-dating). We then measured the 14C content on nine selected tree rings from two samples and compared them with the Southern Hemisphere (SH) atmospheric 14C curves, covering the period of the bomb 14C peak. We find consistent offsets of 5 and 12 years, respectively, in the calendar dates initially assigned, indicating that several tree rings were missing in the sequence. In order to identify the tree-ring boundaries of the unidentified rings we investigated further by analyzing stem wood microsections to examine anatomical characteristics. These anatomical microsections revealed the presence of very narrow terminal parenchyma defining several tree-ring boundaries within the sapwood, which was not visible in sanded samples under a stereomicroscope. Such newly identified tree rings were consistent with the offsets shown by the radiocarbon analysis and allowed us to correct the calendar dates of the initial chronology. Additional radiocarbon measurements over a new batch of rings of the corrected dated samples resulted in a perfect match between the dendrochronological calendar years and the 14C dating, which is based on good agreement between the tree-ring 14C content and the SH 14C curves. Correlations with prior season precipitation and temperature reveal a strong legacy effect of climate conditions prior to the current Neltuma alba growing season. Overall, our study highlights much potential to complement traditional dendrochronology in tree species with challenging tree-ring boundaries with wood anatomical methods and 14C analyses. Taken together, these approaches confirm that Neltuma alba can be accurately dated and thereby used in climatic and ecological studies in tropical and subtropical South America.

Radiocarbon bomb-peak signal in tree-rings from the tropical Andes register low latitude atmospheric dynamics in the Southern Hemisphere.

South American tropical climate is strongly related to the tropical low-pressure belt associated with the South American monsoon system. Despite its central societal role as a modulating agent of rainfall in tropical South America, its long-term dynamical variability is still poorly understood. Here we combine a new (and world's highest) tree-ring 14C record from the Altiplano plateau in the central Andes with other 14C records from the Southern Hemisphere during the second half of the 20th century in order to elucidate the latitudinal gradients associated with the dissemination of the bomb 14C signal. Our tree-ring 14C record faithfully captured the bomb signal of the 1960's with an excellent match to atmospheric 14C measured in New Zealand but with significant differences with a recent record from Southeast Brazil located at almost equal latitude. These results imply that the spreading of the bomb signal throughout the Southern Hemisphere was a complex process that depended on atmospheric dynamics and surface topography generating reversals on the expected north-south gradient in certain years. We applied air-parcel modeling based on climate data to disentangle their different geographical provenances and their preformed (reservoir affected) radiocarbon content. We found that air parcel trajectories arriving at the Altiplano during the bomb period were sourced i) from the boundary layer in contact with the Pacific Ocean (41%), ii) from the upper troposphere (air above the boundary layer, with no contact with oceanic or continental carbon reservoirs) (38%) and iii) from the Amazon basin (21%). Based on these results we estimated the ∆14C endmember values for the different carbon reservoirs affecting our record which suggest that the Amazon basin biospheric 14C isoflux could have been reversed from negative to positive as early as the beginning of the 1970's. This would imply a much faster carbon turnover rate in the Amazon than previously modelled.

HPLC purification of higher plant-dervied lignin phenols for compound specific radiocarbon analysis.

The ability to measure the radiocarbon content of compounds isolated from complex mixtures has begun to revolutionize our understanding of carbon transformations on earth. Because samples are often small, each new compound isolation method must be tested for background carbon contamination (C(ex)). Here, we present a new method for compound-specific radiocarbon analysis (CSRA) of higher plant-derived lignin phenols. To test for C(ex), we compared the Δ(14)C values of unprocessed lignin phenol containing standard materials (woods, leaves, natural vanillin, and synthetic vanillin) with those of lignin phenols liberated by CuO oxidation and purified by two-dimensional high-pressure liquid chromatography (HPLC) coupled to mass spectrometry (MS) and UV detection. We assessed C(ex) associated with (1) microwave assisted CuO oxidation of bulk samples to lignin phenol monomers, (2) HPLC purification, and (3) accelerator mass spectrometry (AMS) sample preparation. The Δ(14)C of purified compounds (corrected for C(ex)) agreed, within error, with those of bulk materials for samples that were >10 µg C. This method will allow routine analysis of the Δ(14)C of lignin phenols isolated from terrestrial, aquatic, and marine settings, revealing the time scale for the processing of one of the single largest components of active organic carbon reservoirs on earth.

Source signatures from combined isotopic analyses of PM2.5 carbonaceous and nitrogen aerosols at the peri-urban Taehwa Research Forest, South Korea in summer and fall.

Isotopes are essential tools to apportion major sources of aerosols. We measured the radiocarbon, stable carbon, and stable nitrogen isotopic composition of PM2.5 at Taehwa Research Forest (TRF) near Seoul Metropolitan Area (SMA) during August-October 2014. PM2.5, TC, and TN concentrations were 19.4 ±â€¯10.1 µg m-3, 2.6 ±â€¯0.8 µg C m-3, and 1.4 ±â€¯1.4 µg N m-3, respectively. The δ13C of TC and the δ15N of TN were - 25.4 ±â€¯0.7‰ and 14.6 ±â€¯3.8‰, respectively. EC was dominated by fossil-fuel sources with Fff (EC) of 78 ±â€¯7%. In contrast, contemporary sources were dominant for TC with Fc (TC) of 76 ±â€¯7%, revealing the significant contribution of contemporary sources to OC during the growing season. The isotopic signature carries more detailed information on sources depending on air mass trajectories. The urban influence was dominant under stagnant condition, which was in reasonable agreement with the estimated δ15N of NH4+. The low δ15N (7.0 ±â€¯0.2‰) with high TN concentration was apparent in air masses from Shandong province, indicating fossil fuel combustion as major emission source. In contrast, the high δ15N (16.1 ±â€¯3.2‰) with enhanced TC/TN ratio reveals the impact of biomass burning in the air transported from the far eastern border region of China and Russia. Our findings highlight that the multi-isotopic composition is a useful tool to identify emission sources and to trace regional sources of carbonaceous and nitrogen aerosols.

Dynamic Nuclear Polarization NMR as a new tool to investigate the nature of organic compounds occluded in plant silica particles.

The determination of the chemical nature of the organic matter associated with phytoliths remains a challenge. This difficulty mainly stems from amounts of organic carbon (C) that are often well below the detection limit of traditional spectroscopic tools. Conventional solid-state 13C Nuclear Magnetic Resonance (NMR) is widely used to examine the nature and structure of organic molecules, but its inherent low sensitivity prohibits the observation of diluted samples. The recent advent of commercial microwave source in the terahertz range triggered a renewed interest in the Dynamic Nuclear Polarization (DNP) technique to improve the signal to noise ratio of solid-state NMR experiments. With this technique, the 13C spectrum of a phytolith sample containing 0.1% w/w C was obtained overnight with sufficient quality to permit a semi-quantitative analysis of the organic matter, showing the presence of peptides and carbohydrates as predominant compounds. Considering the natural abundance of the 13C isotope, this experiment demonstrates that DNP NMR is sufficiently sensitive to observe spin systems present in amounts as low as a few tens of ppm.

Nonannual tree rings in a climate-sensitive Prioria copaifera chronology in the Atrato River, Colombia.

In temperate climates, tree growth dormancy usually ensures the annual nature of tree rings, but in tropical environments, determination of annual periodicity can be more complex. The purposes of the work are as follows: (1) to generate a reliable tree-ring width chronology for Prioria copaifera Griseb. (Leguminoceae), a tropical tree species dwelling in the Atrato River floodplains, Colombia; (2) to assess the climate signal recorded by the tree-ring records; and (3) to validate the annual periodicity of the tree rings using independent methods. We used standard dendrochronological procedures to generate the P. copaifera tree-ring chronology. We used Pearson correlations to evaluate the relationship of the chronology with the meteorological records, climate regional indices, and gridded precipitation/sea surface temperature products. We also evaluated 24 high-precision 14C measurements spread over a range of preselected tree rings, with assigned calendar years by dendrochronological techniques, before and after the bomb spike in order to validate the annual nature of the tree rings. The tree-ring width chronology was statistically reliable, and it correlated significantly with local records of annual and October-December (OND) streamflow and precipitation across the upper river watershed (positive), and OND temperature (negative). It was also significantly related to the Oceanic Niño Index, Pacific Decadal Oscillation, and the Southern Oscillation Index, as well as sea surface temperatures over the Caribbean and the Pacific region. However, 14C high-precision measurements over the tree rings demonstrated offsets of up to 40 years that indicate that P. copaifera can produce more than one ring in certain years. Results derived from the strongest climate-growth relationship during the most recent years of the record suggest that the climatic signal reported may be due to the presence of annual rings in some of those trees in recent years. Our study alerts about the risk of applying dendrochronology in species with challenging anatomical features defining tree rings, commonly found in the tropics, without an independent validation of annual periodicity of tree rings. High-precision 14C measurements in multiple trees are a useful method to validate the identification of annual tree rings.

234U and 230Th determination by FIA-ICP-MS and application to uranium-series disequilibrium in marine samples.

A 234U and 230Th determination method based on an extraction chromatographic separation on a flow injection system coupled to a quadruple ICP-MS was developed. Two-milliliter UTEVA (Eichrom Co.) cartridges were applied as separation tool and 236U and 229Th as spikes. Loading and washing steps were carried out in 3 M HNO3 solution and 0.05 M ammonium oxalate applied to elute both uranium and thorium. The method was applied initially to the IAEA-327 soil reference sample and NIST SRM 4357 ocean sediment reference material, with the obtained 234U and 230Th concentrations in agreement with the reference levels. Samples from a deep-sea sediment core (2450 m water depth) were analyzed and based on 230Th/234U dating, a mean sedimentation rate of 3.3 cm ky(-1) was calculated. Samples from two sediment layers were also dated by 14C-AMS and the observed ages agree with the 230Th/234U results.

Plant growth conditions alter phytolith carbon.

Many plants, including grasses and some important human food sources, accumulate, and precipitate silica in their cells to form opaline phytoliths. These phytoliths contain small amounts of organic matter (OM) that are trapped during the process of silicification. Previous work has suggested that plant silica is associated with compounds such as proteins, lipids, lignin, and carbohydrate complexes. It is not known whether these compounds are cellular components passively encapsulated as the cell silicifies, polymers actively involved in the precipitation process or random compounds assimilated by the plant and discarded into a "glass wastebasket." Here, we used Raman spectroscopy to map the distribution of OM in phytoliths, and to analyze individual phytoliths isolated from Sorghum bicolor plants grown under different laboratory treatments. Using mapping, we showed that OM in phytoliths is distributed throughout the silica and is not related to dark spots visible in light microscopy, previously assumed to be the repository for phytolith OM. The Raman spectra exhibited common bands indicative of C-H stretching modes of general OM, and further more diagnostic bands consistent with carbohydrates, lignins, and other OM. These Raman spectra exhibited variability of spectral signatures and of relative intensities between sample treatments indicating that differing growth conditions altered the phytolith carbon. This may have strong implications for understanding the mechanism of phytolith formation, and for use of phytolith carbon isotope values in dating or paleoclimate reconstruction.

Quantifying archaeal community autotrophy in the mesopelagic ocean using natural radiocarbon.

An ammonia-oxidizing, carbon-fixing archaeon, Candidatus "Nitrosopumilus maritimus," recently was isolated from a salt-water aquarium, definitively confirming that chemoautotrophy exists among the marine archaea. However, in other incubation studies, pelagic archaea also were capable of using organic carbon. It has remained unknown what fraction of the total marine archaeal community is autotrophic in situ. If archaea live primarily as autotrophs in the natural environment, a large ammonia-oxidizing population would play a significant role in marine nitrification. Here we use the natural distribution of radiocarbon in archaeal membrane lipids to quantify the bulk carbon metabolism of archaea at two depths in the subtropical North Pacific gyre. Our compound-specific radiocarbon data show that the archaea in surface waters incorporate modern carbon into their membrane lipids, and archaea at 670 m incorporate carbon that is slightly more isotopically enriched than inorganic carbon at the same depth. An isotopic mass balance model shows that the dominant metabolism at depth indeed is autotrophy (83%), whereas heterotrophic consumption of modern organic carbon accounts for the remainder of archaeal biomass. These results reflect the in situ production of the total community that produces tetraether lipids and are not subject to biases associated with incubation and/or culture experiments. The data suggest either that the marine archaeal community includes both autotrophs and heterotrophs or is a single population with a uniformly mixotrophic metabolism. The metabolic and phylogenetic diversity of the marine archaea warrants further exploration; these organisms may play a major role in the marine cycles of nitrogen and carbon.