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CO2 storage in deep saline aquifers is an effective strategy for reducing greenhouse gas emission. However, salt precipitation triggered by evaporation of water into injected dry CO2 causes injectivity reduction. Predicting the distribution of precipitated salts and their impact on near-well permeability remains challenging. Therefore, a detailed investigation of the interactions between salt precipitation and porous domain is essential for of revealing the mechanisms of pore blockage due to salt crystallization. Through series of microfluidic experiments, direct observations, coupled with detailed imaging processing, form the basis for explaining these phenomena and provide a relationship between water and salt saturations, highlighting the critical roles played by local capillary-driven flow and water film along grains in influencing water relocation. The results reveal two distinct types of salt crystallization: occurring inside the brine with smooth edges and at the CO2-brine interface with rough edges. Furthermore, the impact of local heterogeneity and surface wettability on salt precipitation patterns is discussed. The transition region between the porous domains and inlet/outlet channels exhibits brine backflow and a larger amount of salt accumulation. This paper presents a comprehensive analysis of the dynamic process of salt dry-out occurring during CO2 injection at the pore scale.
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The class Limnochordia harbors a single cultivated member, the mesophilic Limnochorda pilosa, which was isolated from a meromictic lake. Despite numerous molecular signatures reported in various ecosystems, the ecophysiological versatility of this deeply branched lineage of Firmicutes (Bacillota) remains poorly understood. The objective of this study was to use targeted cultivation, based on metagenome-assembled genomes from a deep terrestrial aquifer in Western Siberia, to isolate two new thermophilic members of the class. These isolates, described as Geochorda subterranea gen. nov. sp. nov. and Carboxydochorda subterranea gen. nov. sp. nov. within the Geochordaceae fam. nov., were capable of both anaerobic and aerobic respiration using fumarate and O2, respectively, with simple sugars as electron donors. The cultivated Geochordaceae have demonstrated fermentative growth and degradation of various polymers, including starch, maltose, maltodextrin, xylan, and chitin. The carboxydotrophic C. subterranea sp. nov. exhibited autotrophic growth via the Calvin-Benson-Bassham cycle, using CO, H2, and formate as electron donors and O2 as an electron acceptor, adding metabolic flexibility to the bacterium in the nutrient-depleted "deep biosphere" and supporting the possibility of aerobic metabolism in the deep subsurface. The broad physiological potential deciphered from physiological experiments and comparative genomic data explains the widespread distribution of uncultivated members of the class Limnochordia in various ecosystems, where they can oxidize complex organic substrates through both aerobic and anaerobic respiration, as well as pursue a chemolithotrophic lifestyle through the oxidation of H2 or CO.
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Recent advances in subsurface microbiology have demonstrated the habitability of multi-million-year-old igneous rocks, despite the scarce energy supply from rock-water interactions. Given the minimal evolution coupled with exceedingly slow metabolic rates in subsurface ecosystems, spatiotemporally stable igneous rocks can sustain microbes over geological time scales. This study investigated a 2-billion-year-old mafic rock in the Bushveld Igneous Complex, South Africa, where ultradeep drilling is being executed by the International Continental Scientific Drilling Program (ICDP). New procedures were successfully developed to simultaneously detect indigenous and contaminant microbial cells in a drill core sample. Precision rock sectioning coupled with infrared, fluorescence, and electron microscopy imaging of the rock section with submicron resolution revealed microbial colonization in veins filled with clay minerals. The entry and exit of microbial cells in the veins are severely limited by tight packing with clay minerals, the formation of which supplies energy sources for long-term habitability. Further microbiological characterization of drilled rock cores from the Bushveld Igneous Complex will expand the understanding of microbial evolution in deep igneous rocks over 2 billion years.
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Bactérias , África do Sul , Bactérias/isolamento & purificação , Bactérias/classificação , Sedimentos Geológicos/microbiologia , Minerais/análise , Minerais/metabolismo , Argila/química , Microbiologia do SoloRESUMO
Mechanoluminescence (ML) and digital image correlation (DIC) have emerged as promising optical methods to visualize and measure deformation fields. In this study, a dual-modal sensing skin, called the ML-DIC skin is introduced, that is capable of emitting ML and facilitating DIC measurements under various lighting conditions, including daylight, night or darkness, and UV irradiation. Four ML-DIC skins are fabricated with or without carbon nanotubes (CNTs) using a composite powder consisting of SrAl2O4: Eu,Dy (SAO), and acrylic resin, with CNT milling times of 48, 72, and 96 h for three of four skins, respectively. DIC measurements are performed under multiple lighting conditions for measuring photoluminescence, persistence luminescence, and reflection. Uniaxial tension tests demonstrate the superior performance of ML-DIC skins with CNTs compared with pristine SAO skins, with the skin subjected to 48 h of CNT dispersion exhibiting optimal performance. Further investigations focus on ML emission and DIC measurements near the crack-tip vicinity of static and propagating cracks as well as on surfaces above subsurface cracks. The integration of ML and DIC techniques offers a versatile approach for comprehensive deformation analysis applicable to diverse environments, with implications for materials science, engineering, and structural health monitoring.
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Heat emitted by buildings and other infrastructure accumulates in the subsurface. This additional heat can cause a pronounced shift in thermal boundary conditions of the important groundwater ecosystem. Shallow groundwater systems in Central Europe are often inhabited by communities of fauna adapted to cold and stable conditions as well as microorganisms, whose activity is dependent on ambient temperatures. At a local groundwater temperature hotspot of up to 23⯰C, caused by a water park, we assessed the environmental impact of this thermal alteration on the shallow groundwater system. The results show that the overall groundwater quality at the site is influenced by anthropogenic land use, compared to wells in a nearby water protection zone. However, neither hydrochemical nor ecological characteristics of groundwater from wells in the vicinity of the water park indicate any significant dependence on temperature. Hence, we conclude that in this eutrophic and anoxic aquifer moderate heat stress does not lead to significant alterations in terms of hydrochemistry as well as microbiological properties. Due to the overall low oxygen concentrations (<1â¯mg/l), stygofauna is present only occasionally and cannot be used as bioindicators. These results have to be verified for other aquifer types and would benefit from a more in-depth analysis of microbial community composition.
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This research assesses the stability of sedimentary rock slopes in Teloi, Sik, Kedah, by focusing on the mechanical properties of the rock layers and their susceptibility to weathering. Key tests include the slake durability index (SDI), elastic modulus of knocking ball (Ekb), and electrical resistivity tomography (ERT). The incorporation of electrical resistivity tomography (ERT) data through the virtual reality platform facilitates the visualization of subsurface conditions. The variability of strength characteristic of interbedded sedimentary rocks leads to the differential weathering of rock layers, which causes deterioration on the slope structure. The testing revealed significant variability in rock strength, with sandstone displaying higher durability (Id > 17.1%) and elasticity (Ekb: 0.97 to 29.31 GPa) compared to shale and siltstone, which exhibited lower durability and elasticity (Id < 2.2%, Ekb: 0.2 to 2.2 GPa). Utilizing the Wenner array setup, three distinct electrical resistivity lines were established to evaluate subsurface anomalies. The ERT profiles revealed variations in electrical resistivity among different rock types, identifying areas of weaker material, which are siltstone and shale, while high resistivity areas indicate sandstone. Kinematic analysis through the stereonet process revealed direct toppling as the primary failure mechanism, driven by the critical orientations of joint sets J1, J2, and J3. This aligns with on-site observations of hanging sandstone blocks prone to toppling failure. The findings of this research show that the slake durability index (SDI) and the elastic modulus of the knocking ball (Ekb) enhance the assessment of mechanical properties and weathering resistance of interbedded sedimentary rocks. The virtual reality platform was particularly helpful in analyzing and visualizing the sub-surface conditions and enhancing the evaluation of complex geological data. As a conclusion, this integrated method was helpful in the comprehensive geotechnical evaluation of the slopes, enabling the selection of effective stabilization measures by assessing the differential weathering of interbedded sedimentary rock and identifying potential failure zones.
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The micro-phytoplankton (>20 µm) adaptations and resilience were assessed using morphological traits (shape, surface-to-volume ratio; S:V, and greatest-axial-linear-dimension; GALD) from sea-surface and different SCML-depths (shallow:20-50 m, intermediate:50-100 m, and deep:100-140 m) across different bioregions of Indian Ocean. The dominant simple elongated phytoplankton-geometric-shapes (PGSs) and morphological traits showed distinct north-south distribution and varied with light and nutrient availability. Further, SCML and corresponding sea-surface PGS will be similar or dissimilar if the former is located within or deeper than mixed-layer depth. Also, simple and complex PGS contribution gradually decreases and increases with increasing depth. Additionally, shallow SCML-PGS showed low-S:V and high-GALD while vice-versa for intermediate/deep SCML-PGS due to phenotypic plasticity behavior. Overall, only simple-PGS (cylinder, elliptic-prism, and prism-on-parallelogram) showed strong adaptive behavior through phenotypic plasticity and were highlighted as potential ecological tracers to address ecological impact of oceanographic processes (including coastal eutrophication, and aerosol deposition) linked to nutrient and light availability in predicted ocean change scenarios.
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Fitoplâncton , Oceano Índico , Monitoramento Ambiental , EutrofizaçãoRESUMO
A thiosulfate-oxidizing bacterium, Thiomicrospira sp. strain V2501, was isolated from groundwater collected in a terrestrial deep subsurface environment. This strain was capable of chemolithoautotrophic growth on CO2 and thiosulfate. Here, we report the 2,240,851 bp complete genome sequence of strain V2501.
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This study numerically investigated the dynamic process of residual saltwater contamination in subterranean reservoirs (aquifer upstream of subsurface physical barrier) for the first time. The results indicated that the groundwater level elevation resulting from subsurface physical barriers induced a small hydraulic gradient in the subterranean reservoir, with residual saltwater convection driven by the density contrast between saltwater and freshwater. This led to persistent inland intrusion of residual saltwater, causing a gradual expansion of the contamination area (i.e., concentration higher than 0.25 g/L) and a decreased area of high-salinity zone. The area of low concentration residual saltwater (0.25 g/L) initially increased and then decreased over time. For large-scale aquifers, the proportion of the maximum contaminated area to the total aquifer area increased from 0.05 to 0.17. Subsequently, the low-concentration saltwater was considered as a relative high-concentration for ambient freshwater, which released salts into the surroundings and the contamination area reduced to 0.0003 at 160000 d. In small-scale aquifers, residual saltwater transport was affected by inland boundary, and the low-concentration saltwater reached inland boundary at approximately 2000 d, which limited its horizontal expansion and resulted in the polluted area reaching an inflection point early relative to large-scale aquifers. The salt transfer from high-concentration residual saltwater to low-concentration saltwater was closely related to aquifer parameters of hydraulic conductivity, dispersivity, and molecular diffusion coefficient. These findings suggest that residual saltwater contamination should not be overlooked when assessing the efficiency of subterranean reservoirs.
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The electrical resistivity of subsurface rocks is one of the important sensitive parameters characterizing the internal physics of the Earth. Currently, research on subsurface electrical structures using geomagnetic sounding methods primarily focuses on two approaches: the first is based on observations from a few geomagnetic stations, which have low spatial resolution and cannot effectively describe the distribution of anomalies; the second is based on mobile geomagnetic observations, which have low temporal resolution and cannot promptly reflect anomalies. To address these issues, this study deployed a dense geomagnetic array for long-term observation in the southern segment of the Xiaojiang Fault Zone in the Yuxi area of southern Yunnan. This setup aims to promptly capture seismic magnetic anomalies, providing more data support and fundamental information for short-term earthquake prediction. Based on the long-term observation data from the dense array, the study of the subsurface electrical structure is carried out. The results indicate that during the observation period, which was seismically quiet, the regional subsurface electrical structure remained stable. A large-scale subsurface low-resistivity body was observed in the region, and the electrical structures at the two ends of the southern segment of the Xiaojiang Fault Zone were found to be completely different.
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Nearly all molecular oxygen (O2) on Earth is produced via oxygenic photosynthesis by plants or photosynthetically active microorganisms. Light-independent O2 production, which occurs both abiotically, e.g., through water radiolysis, or biotically, e.g., through the dismutation of nitric oxide or chlorite, has been thought to be negligible to the Earth system. However, recent work indicates that O2 is produced and consumed in dark and apparently anoxic environments at a much larger scale than assumed. Studies have shown that isotopically light O2 can accumulate in old groundwaters; that strictly aerobic microorganisms are present in many apparently anoxic habitats; and that microbes and metabolisms that can produce O2 without light are widespread and abundant in diverse ecosystems. Analysis of published metagenomic data reveals that the enzyme putatively capable of nitric oxide dismutation (NOD) forms four major phylogenetic clusters and occurs in at least 16 bacterial phyla most notably the Bacteroidota. Similarly, a re-analysis of published isotopic signatures of dissolved O2 in groundwater suggests in-situ production in up to half of the studied ecosystems. Geochemical and microbiological data supports the conclusion that "dark oxygen" production is an important and widespread yet overlooked process in apparently anoxic environments with far-reaching implications for subsurface biogeochemistry and ecology.
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Apple tree productivity is influenced by the quantity of water and nutrients that are supplied during planting. To enhance resource utilization efficiency and optimize yields, a suitable strategy for supplying water and nitrogen must be established. A field experiment was conducted using a randomized block group design on five-year-old apple trees in Ningxia, with two irrigation lower limit levels (55%FC (W1) and 75%FC (W2)) and four N application levels (0 (N1), 120 (N2), 240 (N3), and 360 (N4) kg·ha-1). Our findings showed that leaf N content increased with a higher irrigation lower limit, but the difference was not statistically significant. However, the leaf N content significantly increased with increasing N application. The growth pattern of new shoots followed logistic curve characteristics, with the maximum new shoot growth rate and time of new shoot growth being delayed under high water and high nitrogen treatments. Apple yield and yield components (weight per fruit and number of fruits per plant) were enhanced under N application compared to no N application. The maximum apple yields were 19,405.3 kg·ha-1 (2022) and 29,607 kg·ha-1 (2023) at the N3 level. A parabolic relationship was observed between apple yield and N application level, with the optimal range of N application being 230-260 kg⸱ha-1. Apple quality indicators were not significantly affected by the irrigation lower limit but were significantly influenced by N application levels. The lower limit of irrigation did not have a significant impact on the quality indicators of the apples. Water and N utilization efficiencies improved with the W2 treatment at the same N application level. A negative relationship was observed between the amount of nitrogen applied and the biased productivity of nitrogen fertilizer. The utilization of nitrogen fertilizer was 127.6 kg·kg-1 (2022) and 200.3 kg·kg-1 (2023) in the W2N2 treatment. The apple yield was sustained, the quality of the fruit improved, and a substantial increase in water productivity was achieved with the W2N3 treatment. The findings of this study can be used as a reference for accurate field irrigation.
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The terrestrial subsurface harbors unique microbial communities that play important biogeochemical roles and allow for studying a yet unknown fraction of the Earth's biodiversity. The Saint-Leonard cave in Montreal City (Canada) is of glaciotectonic origin. Its speleogenesis traces back to the withdrawal of the Laurentide Ice Sheet 13,000 years ago, during which the moving glacier dislocated the sedimentary rock layers. Our study is the first to investigate the microbial communities of the Saint-Leonard cave. By using amplicon sequencing, we analyzed the taxonomic diversity and composition of bacterial, archaeal and eukaryote communities living in the groundwater (0.1 µm- and 0.2 µm-filtered water), in the sediments and in surface soils. We identified a microbial biodiversity typical of cave ecosystems. Communities were mainly shaped by habitat type and harbored taxa associated with a wide variety of lifestyles and metabolic capacities. Although we found evidence of a geochemical connection between the above soils and the cave's galleries, our results suggest that the community assembly dynamics are driven by habitat selection rather than dispersal. Furthermore, we found that the cave's groundwater, in addition to being generally richer in microbial taxa than sediments, contained a considerable diversity of ultra-small bacteria and archaea.
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With climate warming and accelerated urbanisation, severe urban flooding has become a common problem worldwide. Frequent extreme rainfall events and the siltation of drainage pipes further increase the burden on urban drainage networks. However, existing studies have not fully considered the effects of rainfall and pipeline siltation on the response characteristics of flooding when constructing numerical models of urban flooding simulations. To solve this problem, a surface-subsurface coupling model was constructed by combining the Saint-Venant equation, Manning equation, a one-dimensional pipeline model (SWMM), and a two-dimensional surface overflow model (LISFLOOD-FP). Then, the SWMM model considering pipeline siltation and the two-dimensional surface overflow model (LISFLOOD-FP) were coupled with the flow exchange governing equation, and the urban flooding response characteristics considering the coupling effect of "rainfall and drainage pipeline siltation" were analysed. To enhance the solvability of waterlogging prediction, an intelligent prediction model of urban flooding based on Bayes-CNN-BLSTM was established by combining a convolutional neural network (CNN), bidirectional long short-term memory neural network (BLSTM), Bayesian optimisation (Bayes), and an interpretable loss function error correction method. The actual rainfall events and flooding processes recorded by the monitoring equipment at Huizhou University were used to calibrate and verify the model. The results show that in the Rainfall 1 and Rainfall 2 scenarios, the overload rates of the pipelines in the current siltation scenario were 60.06 % and 68.37 %, respectively, and the proportions of overflow nodes were 24.87 % and 25.89 %, respectively. When the drainage network was initially put into operation, the overload rates of the pipeline were 36.67 % and 41.16 %, and the overflow nodes accounted for 3.05 % and 4.06 %, respectively. The inundated area and volume of urban flooding increased when the combined siltation coefficient (CSC) was 0.2; therefore, two desilting schemes were determined. Under Rainfall 1, Rainfall 2, and the four rainfall recurrence periods, the Bayes-CNN-BLSTM model had clear advantages in terms of accuracy, reliability, and robustness.
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Despite the remoteness of the Antarctic continent and Southern Ocean, microplastic (MPs) contamination has been evidenced in recent years. However, the deep-sea compartments of the Southern Ocean are yet to be investigated. In the present study, we conducted a baseline MP assessment of the deep-sea waters and bottom sediments of the Bransfield Strait, Antarctica. A low abundance of suspected MPs was found. The average MP abundances in bottom sediments and water samples were 0.09 MP/g (range of 0-0.2 MP/g) and 7.00 MP/L (range of 0-16 MP/L), respectively. The majority of the particles were fibers identified as cellulose, although polyethylene terephthalate (PET) and polyacrylonitrile (PAN) was also detected. These results suggest low MP contamination levels in the Southern Ocean's deepest environmental compartments. However, future studies must aim to investigate the smallest MP fractions and, if possible, nanoplastic (<1 µm) contamination in these remote compartments.
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Monitoramento Ambiental , Sedimentos Geológicos , Microplásticos , Água do Mar , Poluentes Químicos da Água , Sedimentos Geológicos/química , Regiões Antárticas , Microplásticos/análise , Poluentes Químicos da Água/análise , Água do Mar/química , Polietilenotereftalatos/química , Resinas Acrílicas/químicaRESUMO
Subsurface chlorophyll maxima (SCM) significantly contributes to oceanic primary productivity, emphasizing the need to study its dynamics and governing mechanisms. We used datasets from various platforms to investigate relationships between the SCM characteristics (SCM depth (ZSCM), SCM magnitude (Chlmax), SCM thickness (TSCM)) and environmental variables modulated by various physical processes in the Northern Indian Ocean (NIO). In the Arabian Sea (western NIO), seasonal processes like convective mixing and upwelling, primarily regulated the SCM characteristics. In the Bay of Bengal (eastern NIO), SCM characteristics were jointly influenced by fresh water influx, barrier layer formation, presence of eddies, and the propagation of Kelvin and Rossby waves. Any changes in these oceanic processes, potentially driven by climate change, could therefore impact oceanic primary production. Additionally, a positive association obtained between Chlmax and downward CO2 flux, while a shallower ZSCM, associated with higher concentrations of DMS, indicated SCM's role in regulating atmospheric gases.
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Clorofila , Monitoramento Ambiental , Água do Mar , Clorofila/análise , Oceano Índico , Água do Mar/química , Mudança Climática , Estações do AnoRESUMO
The understanding of mixing-controlled reactive dynamics in heterogeneous porous media remains limited, presenting significant challenges for modeling subsurface contaminant transport processes and for designing cost-effective environmental remedial efforts. The complexity of accurately observing, measuring, and modeling mixing-limited reactive transport has led to inadequate exploration of these critical processes. This study investigates the mixing and reaction kinetics affected by stagnant zones, which are commonly found in alluvial aquifers-aquitards and fracture-matrix systems. By conducting experiments involving conservative and bimolecular reactive transport through porous media within translucent chambers filled with two sizes of glass beads and under varying flow rates, we explored the effects of grain size and hydrodynamic conditions. Using a high-resolution camera, we monitored the concentration changes of conservative and reactive tracers, with subsequent interpretation through three-dimensional numerical simulations. The outcomes revealed the emergence of distinct mixing interfaces within both mobile and stagnant zones, culminating in a bi-peaked plume formation. Notably, the mixing and reaction times in media containing stagnant zones were found to be approximately 10 times longer than in homogeneous media. These findings, through experimental and modeling efforts, advance our understanding of mixing-limited reactive transport phenomena within heterogeneous media, underscoring the significant role of stagnant zones-a topic previously underexplored.
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Controlling atmospheric warming requires immediate reduction of carbon dioxide (CO2) emissions, as well as the active removal and sequestration of CO2 from current point sources. One promising proposed strategy to reduce atmospheric CO2 levels is geologic carbon sequestration (GCS), where CO2 is injected into the subsurface and reacts with the formation to precipitate carbonate minerals. Rapid mineralization has recently been reported for field tests in mafic and ultramafic rocks. However, unlike saline aquifers and depleted oil and gas reservoirs historically considered for GCS, these formations can have extremely low porosities and permeabilities, limiting storage volumes and reactive mineral surfaces to the preexisting fracture network. As a result, coupling between geochemical interactions and the fracture network evolution is a critical component of long-term, sustainable carbon storage. In this paper, we summarize recent advances in integrating experimental and modeling approaches to determine the first-order processes for carbon mineralization in a fractured mafic/ultramafic rock system. We observe the critical role of fracture aperture, flow, and surface characteristics in controlling the quantity, identity, and morphology of secondary precipitates and present where the influence of these factors can be reflected in newly developed thermo-hydro-mechanical-chemical models. Our findings provide a roadmap for future work on carbon mineralization, as we present the most important system components and key challenges that we are overcoming to enable GCS in mafic and ultramafic rocks.
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Groundwater aquifers are ecological hotspots with diverse microbes essential for biogeochemical cycles. Their ecophysiology has seldom been studied on a basin scale. In particular, our knowledge of chemosynthesis in the deep aquifers where temperatures reach 60 °C, is limited. Here, we investigated the diversity, activity, and metabolic potential of microbial communities from nine wells reaching ancient groundwater beneath Israel's Negev Desert, spanning two significant, deep (up to 1.5 km) aquifers, the Judea Group carbonate and Kurnub Group Nubian sandstone that contain fresh to brackish, hypoxic to anoxic water. We estimated chemosynthetic productivity rates ranging from 0.55 ± 0.06 to 0.82 ± 0.07 µg C L-1 d-1 (mean ± SD), suggesting that aquifer productivity may be underestimated. We showed that 60% of MAGs harbored genes for autotrophic pathways, mainly the Calvin-Benson-Bassham cycle and the Wood-Ljungdahl pathway, indicating a substantial chemosynthetic capacity within these microbial communities. We emphasize the potential metabolic versatility in the deep subsurface, enabling efficient carbon and energy use. This study set a precedent for global aquifer exploration, like the Nubian Sandstone Aquifer System in the Arabian and Western Deserts, and reconsiders their role as carbon sinks.
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Água Subterrânea , Água Subterrânea/microbiologia , Israel , Bactérias/metabolismo , Bactérias/genética , Bactérias/classificação , Adaptação Fisiológica , Microbiologia da Água , MicrobiotaRESUMO
BACKGROUND: Subsurface microorganisms contribute to important ecosystem services, yet little is known about how the composition of these communities is affected by small scale heterogeneity such as in preferential flow paths including biopores and fractures. This study aimed to provide a more complete characterization of microbial communities from preferential flow paths and matrix sediments of a clayey till to a depth of 400 cm by using 16S rRNA gene and fungal ITS2 amplicon sequencing of environmental DNA. Moreover, shotgun metagenomics was applied to samples from fractures located 150 cm below ground surface (bgs) to investigate the bacterial genomic adaptations resulting from fluctuating exposure to nutrients, oxygen and water. RESULTS: The microbial communities changed significantly with depth. In addition, the bacterial/archaeal communities in preferential flow paths were significantly different from those in the adjacent matrix sediments, which was not the case for fungal communities. Preferential flow paths contained higher abundances of 16S rRNA and ITS gene copies than the corresponding matrix sediments and more aerobic bacterial taxa than adjacent matrix sediments at 75 and 150 cm bgs. These findings were linked to higher organic carbon and the connectivity of the flow paths to the topsoil as demonstrated by previous dye tracer experiments. Moreover, bacteria, which were differentially more abundant in the fractures than in the matrix sediment at 150 cm bgs, had higher abundances of carbohydrate active enzymes, and a greater potential for mixotrophic growth. CONCLUSIONS: Our results demonstrate that the preferential flow paths in the subsurface are unique niches that are closely connected to water flow and the fluctuating ground water table. Although no difference in fungal communities were observed between these two niches, hydraulically active flow paths contained a significantly higher abundance in fungal, archaeal and bacterial taxa. Metagenomic analysis suggests that bacteria in tectonic fractures have the genetic potential to respond to fluctuating oxygen levels and can degrade organic carbon, which should result in their increased participation in subsurface carbon cycling. This increased microbial abundance and activity needs to be considered in future research and modelling efforts of the soil subsurface.