On the role of small estuaries in retaining buoyant particles.

Estuaries, as connectors between land and ocean, have complex interactions of river and tidal flows that affect the transport of buoyant materials like floating plastics, oil spills, organic matter, and larvae. This study investigates surface-trapped buoyant particle transport in estuaries by using idealized and realistic numerical simulations along with a theoretical model. While river discharge and estuarine exchange flow are usually expected to export buoyant particles to the ocean over subtidal timescales, this study reveals a ubiquitous physical transport mechanism that causes retention of buoyant particles in estuaries. Tidally varying surface convergence fronts affect the aggregation of buoyant particles, and the coupling between particle aggregation and oscillatory tidal currents leads to landward transport at subtidal timescales. Landward transport and retention of buoyant particles is greater in small estuaries, while large estuaries tend to export buoyant particles to the ocean. A dimensionless width parameter incorporating the tidal radian frequency and lateral velocity distinguishes small and large estuaries at a transitional value of around 1. Additionally, higher river flow tends to shift estuaries toward seaward transport and export of buoyant particles. These findings provide insights into understanding the distribution of buoyant materials in estuaries and predicting their fate in the land-sea exchange processes.

Improving Estimates of Dissolved Organic Carbon (DOC) Concentration from In Situ Fluorescence Measurements across Estuaries and Coastal Wetlands.

The use of optical proxies is essential to the sustained monitoring of dissolved organic carbon (DOC) in estuaries and coastal wetlands, where dynamics occur on subhour time scales. In situ dissolved organic matter (DOM) fluorescence, or FDOM, is now routinely measured along with ancillary water-quality indicators by commercial sondes. However, its reliability as an optical proxy of DOC concentration is often limited by uncertainties caused by in situ interferences and by variability in DOM composition and water matrix (ionic strength, pH) that are typical at the land-ocean interface. Although corrections for in situ interferences already exist, validated strategies to account for changes in the DOM composition and water matrix in these systems are still lacking. The transferability of methods across systems is also poorly known. Here, we used a comprehensive data set of laboratory-based excitation-emission matrix fluorescence and DOC concentration matched to in situ sonde measurements to develop and compare approaches that leverage ancillary water-quality indicators to improve estimates of DOC concentration from FDOM. Our analyses demonstrated the validity of in situ interference correction schemes, the importance of ancillary water-quality indicators to account for DOM composition and water matrix change, and the good transferability of the proposed methods.

Investigation of alkyl, aryl, and chlorinated OPFRs in sediments from estuarine systems: Seasonal variation, spatial distribution and ecological risks assessment.

Estuaries in South Africa are very important for biodiversity conservation and serve as focal points for leisure and tourism activities. The organophosphate flame retardants (OPFRs) levels in these aquatic systems haven't been documented in any studies as of yet. Due to the negative effects of persistent organic pollutants in South African estuaries, we examined the occurrence of eight OPFRs in sediments of two estuaries by studying their spatiotemporal distribution, season variation, and ecological risks. The Sundays Estuary (SDE), a semi-urbanized agricultural surrounding system, recorded an ∑8OPFR concentration in sediments that ranged from 0.71 to 22.5 ng/g dw, whereas Swartkops Estuary, a largely urbanized system, recorded a concentration that ranged from 0.61 to 119 ng/g dw. Alkyl-OPFRs were the prevalent homologue in both estuaries compared to the chlorinated and aryl groups. While TBP, TCPP, and TCrP were the most abundant compounds among the homologue groups. There was no distinct seasonal trend of ∑8OPFR concentration in either estuary, with summer and autumn seasons recording the highest concentrations in SDE and SWE, respectively. Ecological risks in the majority of the study sites for the detected compounds were at low (RQ < 0.1) and medium levels (0.1 ≤ RQ < 1) for certain species of fish, Daphnia magna and algae. However, the cumulative RQs for all the compounds had ∑RQs ≥1 for most sites in both estuaries, indicating that these organisms, if present in both estuaries, may be exposed to potential ecological concerns due to accumulated OPFR chemicals. The scope of future studies should be broadened to include research areas that are not only focus on the bioaccumulation patterns of these compounds but also find sustainable ways to reduce them from these estuarine environments.

Tree of life metabarcoding can serve as a biotic benchmark for shifting baselines in urbanized estuaries.

Urbanization of estuaries drastically changed existing shorelines and bathymetric contours, in turn modifying habitat for marine foundational species that host critical biodiversity. And yet we lack approaches to characterize a significant fraction of the biota that inhabit these ecosystems on time scales that align with rates of urbanization. Environmental DNA (or eDNA) metabarcoding that combines multiple assays targeting a broad range of taxonomic groups can provide a solution, but we need to determine whether the biological communities it detects ally with different habitats in these changing aquatic environments. In this study, we tested whether tree of life metabarcoding (ToL-metabarcoding) data extracted from filtered seawater samples correlated with four known geomorphic habitat zones across a heavily urbanized estuary (Sydney Harbour, Australia). Using this method, we substantially expanded our knowledge on the composition and spatial distribution of marine biodiversity across the tree of life in Sydney Harbour, particularly for organisms where existing records are sparse. Excluding terrestrial DNA inputs, we identified significant effects of both distance from the mouth of Sydney Harbour and geomorphic zone on biological community structure in the ToL-metabarcoding dataset (entire community), as well as in each of the taxonomic subgroups that we considered (fish, macroinvertebrates, algae and aquatic plants, bacteria). This effect appeared to be driven by taxa as a collective versus a few individual taxa, with each taxon explaining no more than 0.62% of the variation between geomorphic zones. Similarly, taxonomic richness was significantly higher within geomorphic zones with large sample sizes, but also decreased by 1% with each additional kilometer from the estuary mouth, a result consistent with a reduction in tidal inputs and available habitat in upper catchments. Based on these results, we suggest that ToL-metabarcoding can be used to benchmark biological monitoring in other urbanized estuaries globally, and in Sydney Harbour at future time points based on detection of bioindicators across the tree of life. We also suggest that robust biotic snapshots can be archived following extensive curation of taxonomic assignments that incorporates ecological affinities, supported by records from relevant and regional biodiversity repositories.

Examining the Effects of Land Use, Salinity Gradient, and Water Pollution on Greenhouse Gas Emissions in Urbanized Estuaries.

Estuaries are significant contributors to greenhouse gases (GHGs) in waterways. However, the effects of human activities and ecological variables on GHG emissions in estuaries remain poorly understood. This study examines the patterns and causes of GHG emissions in the Scheldt Estuary, focusing on the roles of salinity, water contamination, and land use. The findings indicate that salinity negatively impacts the release of carbon dioxide (CO2) and nitrous oxide (N2O), likely due to reduced salt levels and cleaner water upstream. Water contamination's influence on GHG emissions was more pronounced in cleaner, upriver sites compared to saltier downstream locations. Specifically, CO2 emissions quadrupled, and N2O emissions tripled as water conditions worsened from healthy (near the mouth, bordered by agricultural land) to polluted (farther downstream, bordered by urban areas). Methane (CH4) emissions were significantly higher in aquatic locations than in salty sites. The reduced impact of contamination from downstream to the river mouth may be due to increasing population density. Urban sites emitted about twice as much CO2 and N2O as those in natural and industrial areas. Machine learning analysis also showed that fertilizers and organic enrichment, along with salinity, significantly increased GHG emissions. These results highlight the importance of understanding the interplay of salinity, water contamination, and land use in influencing GHG emissions in coastal ecosystems.

Simulating implications of fish behavioral response for managing hypoxia in estuaries with spatial dissolved oxygen variability.

Hypoxia, or low dissolved oxygen (DO), is a widespread water quality problem affecting estuaries and coastal waters around the world. Water quality criteria for DO have been established for every estuary in the US and are an important part of the regulatory response to nutrient pollution and associated anthropogenic eutrophication. Experimental studies examining effects of low DO exposure have been to quantify outcomes based on hypoxia effects observed in individuals, such as increased mortality or growth impairment. Although laboratory exposure tests provide useful benchmarks for policy development, most of those considered in policy development did not consider behavioral responses to low DO. However, experimental research has shown that behavioral responses occur, and that behavior modifies exposure to low DO conditions. Here we begin development of a spatially explicit individual based model (SEIBM) intended to project behavioral outcomes of exposure to spatially variable hypoxia in estuaries. Our goal is to consider the responsiveness of an SEIBM to both different behavioral hypotheses, as well as realistic spatial patterns in hypoxia. A sensitivity analysis was used to explore responsiveness based on two movement strategies: avoidance and behavioral switching. We tested the sensitivity of a suite of movement parameters to changes in spatial patterns representative of an index estuary. The sensitivity analysis demonstrated that model responses to changes in movement strategies include biologically meaningful changes in site occupancy and movement distance centered on individual behavior near a normoxic-hypoxic boundary. Further, the model demonstrated important sensitivity to realistic changes in movement parameters, including the size and shape of the individual neighborhood describing knowledge useful for movement decisions. These results support the utility of the developed SEIBM for exploring behavioral responses of fish to hypoxia in estuaries. The sensitivity analysis also demonstrates parameter values that must be set based on empirical data and are sensitive to data quality. These results will be used to further develop the model and to plan field and laboratory studies to support model parametrization. The end goal is a model framework that can inform policy decisions regarding hypoxia resulting from anthropogenic nutrient loading in estuaries.

Scaling of fast-start performance and its thermal dependence in mummichog Fundulus heteroclitus.

Fast-start predator-escape performance and its sensitivity to temperature (24, 30, and 36°C) were evaluated in mummichog Fundulus heteroclitus across a range of body sizes spanning YOY to adult (35-68 mm standard length). Mummichogs exhibit isometry of body dimensions and areas of the dorsal and anal fins but negative allometry of the caudal fin area. These scaling relationships are consistent with observed decreases in fast-start angular velocities with increasing body size. Linear velocity, on the contrary, does not vary with size, and both large and small mummichogs are capable of traversing similar distances in a given amount of time. In addition, temperature influences fast-start performance in similar ways over the size range, though the magnitude of the effect varies with size for some performance measures. In general, fast-start performance increases with test temperature, but mummichogs acclimated to warmer temperatures exhibit lower performance at each test temperature. Altogether, our results suggest that mummichogs across the adult size range may suffer decreases in their predator-escape performance as increasing sea temperatures combine with short-term temperature fluctuations in the estuaries these fish occupy.

What makes a habitat a home? Habitat associations of juvenile European sea bass, Dicentrarchus labrax, in estuarine nurseries.

Selection of nursery habitats by marine fish, such as European sea bass (Dicentrarchus labrax), is poorly understood. Identifying and protecting the full range of juvenile nursery habitats is vital to supporting resilient fish populations and economically important fisheries. We examined how the condition, stomach fullness, and diet of juvenile European sea bass, along with their abundance, differ at high or low tide between the following estuarine habitats: saltmarsh, oyster reefs, shingle, sand, and mud edge habitats. Using a combination of fyke and seine netting we found no difference in sea bass abundance or condition across high-tide habitats, suggesting that rather than differentially selecting between them, juvenile sea bass use all available shallow habitats at high tide. Stomach fullness was significantly higher on saltmarsh and sand compared to mud, and thus these habitats may support better foraging. Dietary DNA metabarcoding revealed that sand and saltmarsh diets mostly comprised Hediste polychaetes, whereas zooplanktonic taxa dominated diets over mud. At low tide, sea bass abundance was highest in shingle and oyster reefs, where stomach fullness and condition were lowest. This may indicate a potential trade-off between using habitats for foraging and refuge. Although sea bass abundance alone does not capture productivity, the high abundance across all estuarine habitats at high tide suggests that it is important to consider the protection of a mosaic of interconnected habitats to support nursery functions rather than focus on individual habitat types.

Ecological assessment of microplastic contamination in surface water and commercially important edible fishes off Kadalundi estuary, Southwest coast of India.

This study focuses on the Kadalundi estuary, Kerala's first community reserve, investigating the prevalence and impacts of microplastics on both the estuarine environment and selected fish species. This study presents the initial evidence indicating the consumption of microplastic particles by 12 commercially important edible fish species inhabiting the Kadalundi estuary. Analysis revealed significant accumulations of microplastic fibers within the surface water. In examining 12 fish species from demersal and pelagic habitats, microplastics were found in both the gastrointestinal tracts and gills. In the digestive tracts, microplastic fragments constituted the highest proportion (46%), while in the gills, microplastic fibers were dominant (52.4%). This study observed a prevalence of blue microplastics over other colors in both water and fish samples. Notably, demersal species showed a higher incidence of ingested microplastics. Polymer analysis identified Polypropylene (PP), Nylon, Low-Density Polyethylene (LDPE), Polyethylene (PE), Polypropylene isotactic (iPP), PE 1 Octene copolymer, and Rayon in water samples, while fish samples predominantly contained LDPE, PP, PE, and Nylon. Risk assessment utilizing the Polymer Hazard Index (PHI) categorized certain polymers as posing minor to moderate risks. Pollution Load Index (PLI) computations indicated moderate to high levels of microplastic contamination across various sampling sites in the estuary. Principal Component Analysis (PCA) revealed a lack of correlation between fish size and microplastic ingestion, underscoring environmental factors' influence on microplastic intake. The study emphasizes the implications of microplastic pollution on the fragile ecosystem of the Kadalundi estuary, posing potential risks to biodiversity and human health.

Diversity and salinity adaptations of ammonia oxidizing archaea in three estuaries of China.

Ammonia-oxidizing archaea (AOA) are ubiquitously found in diverse habitats and play pivotal roles in the nitrogen and carbon cycle, especially in estuarine and coastal environments. Despite the fact that the diversity and distribution of AOA are thought to be tightly linked to habitats, little is known about the relationship that underpins their genomic traits, adaptive potentials, and ecological niches. Here, we have characterized and compared the AOA community in three estuaries of China using metagenomics. AOA were the dominant ammonia oxidizers in the three estuaries. Through phylogenetic analyses, five major AOA groups were identified, including the Nitrosomarinus-like, Nitrosopumilus-like, Aestuariumsis-like, Nitrosarchaeum-like, and Nitrosopelagicus-like groups. Statistical analyses showed that the aquatic and sedimentary AOA communities were mainly influenced by spatial factors (latitude and water depth) and environmental factors (salinity, pH, and dissolved oxygen) in estuaries, respectively. Compared to AOA dwelling in terrestrial and marine habitats, estuarine AOA encoded more genes involved in glucose and amino acid metabolism, transport systems, osmotic control, and cell motility. The low proteome isoelectric points (pI), high content of acidic amino acids, and the presence of potassium ion and mechanosensitive channels suggest a "salt-in" strategy for estuarine AOA to counteract high osmolarity in their surroundings. Our findings have indicated potential adaptation strategies and highlighted their importance in the estuarine nitrogen and carbon cycles. KEY POINTS: • Spatial and environmental factors influence water and sediment AOA respectively. • Estuarine AOA share low proteome isoelectric value and high acid amino acids content. • AOA adaptation to estuaries is likely resulted from their unique genomic features.

Development of a depuration protocol for commercially important edible bivalve molluscs of India: Ensuring microbiological safety.

We investigated the effect of depuration of three naturally contaminated commercially important tropical edible bivalve molluscs by varying temperature, salinity and body-size of animals harvested from Ashtamudi and Vembanad estuaries, India using a static depuration system to ensure microbiological food safety. Before depuration, the levels of faecal indicators and pathogens were above the acceptable limits for live consumption. The depuration water temperature had a significant effect on bacterial elimination. Log reduction of faecal coliforms (FC) and E. coli varied between room-temperature (RTDS) and low-temperature depuration system (LTDS) and it was in the range of 1.39-2.44 and 1.88-2.82 log MPN, respectively under RTDS and LTDS. The elimination of bacterial pathogens such as Vibrio and Salmonella spp. was rapid in RTDS compared to LTDS. The highest elimination of FC and E. coli (2.39 and 2.92 log) was at 35 psµ depuration and the lowest (0.87 and 1.65 log) at 15 psµ depuration. The reduction of FC and E. coli was higher in the medium-sized animals compared to the small animals. Based on the results it is recommended that depuration using the static system under room-temperature (30 ± 1 °C); a salinity range of (25-35 psµ) and using medium-size bivalves (clam>30 mm; mussel >45 mm, and oyster >65 mm length) as optimum conditions for producing microbiologically safe bivalves for live consumption within 36 h.

Quantifying euryhaline histories in red drum Sciaenops ocellatus: Scale isotope ratios.

Stable isotope analysis of fish muscle tissue has been used to quantify fish migratory behavior in many systems, and these chemical tracers are especially useful for euryhaline fishes that traverse significant salinities and gradients in baseline signatures. However, the removal of muscle tissues often requires lethal sampling. Fish scales may be removed non-lethally and offer potential alternatives to stable isotope ratios from muscle to indicate recent feeding histories. For this study, red drum Sciaenops ocellatus were collected in subtropical estuaries in the northwestern Gulf of Mexico and analyzed for stable isotope (𝛿13 C and 𝛿15 N) compositions of scale exteriors. Decalcification of scale samples was deemed unnecessary given the preservation of linearity and minimal offsets between paired decalcified and untreated scale samples. Stable isotope signatures of muscle tissue and scales were closely matched and indicated high degrees of residence within bays at fine spatial scales. This study also surveyed the existing body of literature comparing scale and muscle isotope values from the same individuals and found that the majority reports linear relationships with slopes close to unity, although species-specific offsets and intercepts vary. This work expands the body of literature indicating that scales are a viable non-lethal alternative for stable isotope assessments of dietary and habitat use histories for mobile fishes.

Methods for a composite ecological suitability measure to inform cumulative restoration assessments in Gulf of Mexico estuaries.

Ecosystem management requires a systematic, holistic approach that considers ecological and social outcomes. Effective restoration practices promote a balance of ecological and social goals by addressing ecological integrity, efficiently maximizing benefits while minimizing investment, and encompassing collaborative stakeholder engagement. Socio-ecological assessments can inform adaptive management and be utilized to prioritize restoration activities and monitor restoration effectiveness. In estuarine systems, socio-ecological assessments should evaluate the ability of habitats to support both ecologically and locally important species. The composite measure presented utilizes a combination of ecological and social measures to characterize ecological suitability for individual and multiple Gulf of Mexico estuarine species. The ecological suitability value (ES) for a given spatial unit is based on a suite of biophysical measures of the quality and extent of suitable habitat for each species, the species' trophic importance in a food web context, and the importance of each species in relation to stakeholder values and benefits. ES values for individual spatial units can be aggregated to estimate the distribution of ecological suitability at the estuarine scale. The ES values are calculated using examples for each step in the process. The information provided by ecological suitability characterizations can support restoration prioritization decisions for Gulf of Mexico estuaries and can provide a baseline measure to gauge restoration effectiveness over time to inform cumulative restoration assessments.

Biomarker responses in the Coilia dussumieri exposed to petroleum hydrocarbons contamination in urbanized estuaries along the west coast of India.

The estuaries of Maharashtra are under continuous pressure due to uncontrolled dumping of persistent toxic organic pollutants into these regions and their impact on the marine environment. In this study, total petroleum hydrocarbons (TPHs) in water, sediments, fish, and biomarker responses in the Coilia dussumieri exposed to TPHs were determined in seven urbanized tropical estuaries along the west coast of India during the winter and summer. The results of the cluster analysis highlighted that the concentration of TPHs in the water, sediment, and fish in the study area was spatially varied, with the highest concentration in the northern area of Maharashtra (NM) estuaries than in the southern region of Maharashtra (SM) estuaries during both seasons. The enrichment of TPHs in water and sediment in the middle part of most estuaries highlights the addition of anthropogenic organic matter. A higher concentration of TPHs in the muscle tissue of Coilia dussumieri observed in NM during the winter season, indicates the energetic intake and storage of large quantities of TPHs in their muscle tissue. The biochemical results showed decreased levels of total protein (PRT) under exposure to TPHs, under oxidative stress and reversing correlations was observed between catalase (CAT) and lipid peroxidation (LPO) activities with the respective TPHs. Similarly, reduced activity of the CAT antioxidant and increased glutathione-S-transferase (GST) under exposure to TPHs were more likely to occur under hydrocarbon stress. However, current results indicate that Coilia dussumieri actively produces oxidative stress and antioxidant reaction that can be used as biomarkers of pollution in the study area.

Effects of sedimentary heavy metals on meiobenthic community in tropical estuaries along eastern Arabian Sea.

The central west coast of India comprises the 720 km long coastline of Maharashtra state and houses widespread industrial zones along the eastern Arabian Sea. Sediments from seven industrial-dominated estuaries along the central west coast were studied for metal enrichment and benthic assemblages to determine sediment quality status and ecological effects in these areas. The suit of geochemical indices highlighted the contamination of sediment in the estuaries concerning heavy metals. Positive correlations of Hg with Co, Zn, Ni, Cr, and Pb indicated the source similarity and effect of anthropogenic activity. non-Metric Multidimensional Scaling (n-MDS) based on meiofaunal abundance showed a cleared separation of clusters through the gradient of heavy metal concentrations. The Canonical Correspondence Analysis (CCA) results with the Monte Carlo test signified those heavy metals influenced the meiobenthic community. Heavy metals (Cr, Ni, Zn, Cd, Pb, and Hg) were the main drivers shaping the meiofaunal community with a significant (p < 0.05) reduction in taxa richness, diversity, and evenness. Dominant meiofaunal assemblages evidence the tolerance of foraminiferans and nematodes. However, these taxa were affected by decreased abundance at impacted sites compared to other fauna. In conclusion, results demonstrated that impairment occurred in the meiofaunal community in most estuaries (except AB and KK).

Numerical modeling of wave attenuation: implications of representing vegetation found in coastal saltmarshes in the Chesapeake Bay.

Coastal communities are vulnerable to wave and storm surges during extreme events, highlighting the need to increase community resilience. The effectiveness of natural wetlands in attenuating waves is vital to designing strategies for protecting public safety. This study aimed to understand how vegetation attenuates waves and determine the best method for modeling vegetation's impact on wave dynamics. The researchers compared two different vegetation representations in numerical models, implicit and explicit, using SWAN and XBeach at varying spatial resolutions. The study focused on two marshes in the Chesapeake Bay, using field measurements to investigate the accuracy of each method in representing wave attenuation by vegetation and the implications of explicitly representing average characteristics of one vegetation species on a regional level. Results showed that explicit modeling using average vegetation characteristics provided more accurate results than the implicit model, which only showed wave attenuation due to topography. The finer scale resolution and site-specific vegetation characteristics further improved the accuracy of wave attenuation observed. Understanding the trade-offs between different vegetation representations in numerical models is essential to accurately represent wave attenuation and design effective protection strategies for coastal communities.

Risk assessments of microplastics accumulated in estuarine sediments at Cuddalore, Tamil Nadu, southeast coast of India.

In this study, the abundance of microplastics (MPs) in the Uppanar and Gadilam estuaries in Cuddalore, on the southeast coast of India, is reported. In the estuarine sediments, MP abundance ranged from 36.3 ± 3.39 to 51.6 ± 2.05 particles/Kg dw. Different types of MP shapes, such as fibers (41.7-47.9%), films (21.2-27.2%), and fragments (18.3-25.5%) were observed in the size range of 100-1000 µm. Diverse colours of MPs were observed, among which red (30.1-34.5%) was predominantly noticed in the estuarine sediments. Six polymers were identified by µ-FTIR, among which LDPE (39%) and PP (35%) were dominant. MPs pollution in these estuaries is composed of domestic, industrial, and fishing wastes. Risk assessments show that the area falls under hazard categories I to III, indicating low to high risk. This study improves knowledge on MPs contamination in Uppanar and Gadilam estuaries and provides impetus for further research to identify the actual sources and impacts of MPs on aquatic systems along the east coast of India.

Finds of Siboglinids (Annelida, Siboglinidae) in the Estuaries of the Largest Arctic Rivers Are Associated with Permafrost Gas Hydrates.

Gutless marine worms of the family Siboglinidae have been found in the estuaries of the largest Arctic rivers Yenisei, Lena, and Mackenzie. Siboglinid metabolism is provided by symbiotic chemoautotrophic bacteria. Strong salinity stratification is characteristic of the estuaries of the largest Arctic rivers and ensures a high salinity at depths of 25-36 m, where siboglinids were found. High methane concentrations, which are necessary for siboglinid metabolism, result from dissociation of permafrost gas hydrates under the influence of river runoff in the conditions of Arctic warming.

High spatial heterogeneity and low connectivity of bacterial communities along a Mediterranean subterranean estuary.

Subterranean estuaries are biogeochemically active coastal sites resulting from the underground mixing of fresh aquifer groundwater and seawater. In these systems, microbial activity can largely transform the chemical elements that may reach the sea through submarine groundwater discharge (SGD), but little is known about the microorganisms thriving in these land-sea transition zones. We present the first spatially-resolved characterization of the bacterial assemblages along a coastal aquifer in the NW Mediterranean, considering the entire subsurface salinity gradient. Combining bulk heterotrophic activity measurements, flow cytometry, microscopy and 16S rRNA gene sequencing we find large variations in prokaryotic abundances, cell size, activity and diversity at both the horizontal and vertical scales that reflect the pronounced physicochemical gradients. The parts of the transect most influenced by freshwater were characterized by smaller cells and lower prokaryotic abundances and heterotrophic production, but some activity hotspots were found at deep low-oxygen saline groundwater sites enriched in nitrite and ammonium. Diverse, heterogeneous and highly endemic communities dominated by Proteobacteria, Patescibacteria, Desulfobacterota and Bacteroidota were observed throughout the aquifer, pointing to clearly differentiated prokaryotic niches across these transition zones and little microbial connectivity between groundwater and Mediterranean seawater habitats. Finally, experimental manipulations unveiled large increases in community heterotrophic activity driven by fast growth of some rare and site-specific groundwater Proteobacteria. Our results indicate that prokaryotic communities within subterranean estuaries are highly heterogeneous in terms of biomass, activity and diversity, suggesting that their role in transforming nutrients will also vary spatially within these terrestrial-marine transition zones.

Global methane and nitrous oxide emissions from inland waters and estuaries.

Inland waters (rivers, reservoirs, lakes, ponds, streams) and estuaries are significant emitters of methane (CH4 ) and nitrous oxide (N2 O) to the atmosphere, while global estimates of these emissions have been hampered due to the lack of a worldwide comprehensive data set of CH4 and N2 O flux components. Here, we synthesize 2997 in-situ flux or concentration measurements of CH4 and N2 O from 277 peer-reviewed publications to estimate global CH4 and N2 O emissions from inland waters and estuaries. Inland waters including rivers, reservoirs, lakes, and streams together release 95.18 Tg CH4  year-1 (ebullition plus diffusion) and 1.48 Tg N2 O year-1 (diffusion) to the atmosphere, yielding an overall CO2 -equivalent emission total of 3.06 Pg CO2  year-1 . The estimate of CH4 and N2 O emissions represents roughly 60% of CO2 emissions (5.13 Pg CO2  year-1 ) from these four inland aquatic systems, among which lakes act as the largest emitter for both CH4 and N2 O. Ebullition showed as a dominant flux component of CH4 , contributing up to 62%-84% of total CH4 fluxes across all inland waters. Chamber-derived CH4 emission rates are significantly greater than those determined by diffusion model-based methods for commonly capturing of both diffusive and ebullitive fluxes. Water dissolved oxygen (DO) showed as a dominant factor among all variables to influence both CH4 (diffusive and ebullitive) and N2 O fluxes from inland waters. Our study reveals a major oversight in regional and global CH4 budgets from inland waters, caused by neglecting the dominant role of ebullition pathways in those emissions. The estimated indirect N2 O EF5 values suggest that a downward refinement is required in current IPCC default EF5 values for inland waters and estuaries. Our findings further indicate that a comprehensive understanding of the magnitude and patterns of CH4 and N2 O emissions from inland waters and estuaries is essential in defining the way of how these aquatic systems will shape our climate.