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Evaluation of submersible pressure transducers for streamflow monitoring in small streams

Compact streamgages requiring minimal infrastructure and equipped with submersible pressure transducers (PTs) are increasingly used to monitor small streams, yet disparate implementations obscure their accuracy under real-world conditions. This study isolated instrumentation-derived uncertainty in stage monitoring by co-locating various combinations of commercial vented and unvented PTs with seven U.S. Geological Survey (USGS) reference streamgages on small streams. Multi-year PT stage records from compact streamgages, collected and corrected following USGS protocols, were compared to concurrent reference observations. Vented PTs demonstrated an average measurement uncertainty of ±0.005 m and mean absolute percent error (MAPE) of ±0.2%. Unvented PTs exhibited higher uncertainty, averaging ±0.009 m and ±0.4% MAPE. Although both sensor types had stage errors up to ±0.3 m, 95% of vented PT and unvented PT observations were within 0.01 and 0.02 m of reference stage, respectively. Analysis of additional unvented PTs revealed stage errors of up to ±1.2% when using barometric sensors within 15 km of the in-water sensor. Propagation of stage error to discharge using reference rating models resulted in cumulative discharge MAPEs of ±4.5% for vented and ±5.5% for unvented PTs. These findings highlight PTs as practical alternatives to reference instrumentation when deployed with standardized procedures, potentially expanding access to reliable streamflow data.

conterminous United States

PFAS remediation in a bioelectrochemical system inoculated with the west branch consortium (WBC-2)

Groundwater contamination by per - and polyfluoroalkyl substances (PFAS) poses a persistent environmental and public health concern. This study evaluates a two-chambered bioelectrochemical system (BES) inoculated with the West Branch Consortium (WBC-2) for PFAS remediation. Under an applied cathodic potential of −450 mV (versus Ag/AgCl), the BES with active WBC-2 achieved >99.0% perfluorooctanesulfonic acid (PFOS) removal within 21 days in deionized water with culture medium and > 98.9% removal of PFOS, perfluorooctanoic acid (PFOA), perfluorohexanoic acid (PFHxA), and perfluorohexanesulfonic acid (PFHxS) in contaminated groundwater after 102 days. Intermediate formation (e.g., PFOA, 6:2 fluorotelomer sulfonate (6:2 FTS), perfluoropropionic acid (PFPrA), perfluorobutanoic acid (PFBA)) and background-corrected fluoride release were consistent with PFOS transformation under anaerobic reducing conditions potentially involving defluorination. Following repeated PFOS spikes (100 μg/L on Days 0, 50, and 399), PFOA, PFPrA, and PFBA accumulated over 664 days. Despite being the dominant accumulated compound, PFOA accounted for <1.8% of the total spiked PFOS mass. Minimal PFOS transformation occurred in controls without active WBC-2, highlighting the importance of microbial metabolism. Biofilm analysis revealed dense colonization of rod-shaped bacteria on carbon fiber brushes. Enrichment of Bacillus , Agrobacterium , and other low-abundance taxa suggests selective adaptation to BES and PFAS conditions. These findings highlight BES driven by electrochemically stimulated microbial activity as a promising strategy for PFAS remediation.

Journal of Water Process Engineering

Systematic approach to prioritize wells for effective groundwater monitoring and management in the Arkansas Headwaters Basin, Colorado, USA

Study region The Arkansas Headwaters Basin, an intermountain basin in the Southern Rocky Mountains of North America. Study focus Our specific focus is choosing a set of wells to support a possible future regional groundwater-surface water model that would support water management. We present a three-step process using multiple criteria to score, predict, and choose prioritized wells that capture the full distribution of data including extremes. The three-step process provides accessible visualizations, fiscally efficient well prioritization, and screening useful for subsequent groundwater modeling. The novelty of the proposed methodology is the systematic approach integrating a scoring and a predictive approach to support a selection path. The systematic approach may be broadly adapted for other basins. New hydrological insights for the region Understanding regional hydrology hinges on efficient collection of hydrologic data that captures the relevant dynamics including extremes. The present study, a case study for a particular basin in the Southern Rocky Mountains, is the first use of a scripted (R software) strategy to select an economical and representative set of monitoring wells. Our findings suggest caution when using proximity as a proxy for correlation, because proximal wells in the same geologic formation and similar depths are not always correlated. In the Arkansas Headwaters Basin, subsurface geology may be less influential on groundwater elevations than broader hydrologic influences, such as regional drought.

Colorado

Decadal-scale characteristics of natural and anthropogenic dune morphology along North Carolina Barrier Islands (SE Atlantic Coast)

Dunes offer protection to coastal communities from storms, yet quantitative research characterizing human influences on dune systems across decades and 10–100 km spatial scales constitutes a known data gap. This study analyzed a multidecadal archive of dune crest elevations and positions to map mesoscale spatiotemporal trends in foredune morphology across the barrier island coastline of North Carolina, SE Atlantic coast. Results are summarized within four categories of coastline representing a range of coastal management intensities, integrating beach width and dune lateral accommodation space to assess the comparative influence of differing intensities of anthropogenic intervention on dune morphology. These categories account for the presence of structures, roads, and alongshore connectivity to anthropogenic sediment sources (beach nourishment). These categories show distinct dune and beach system characteristics. Dunes are largest in the areas with the longest legacy of coastal management. As categories decrease in development intensity, dune and beach system characteristics generally depart from the large, static dunes and narrow beaches consistent with hold-the-line management. Where small or transgressive dunes exist on heavily developed coasts, they coincide with relatively strong hydrodynamic forcing or sediment-limiting geology. There is evidence of indirect coastal management influence on undeveloped barrier islands with alongshore connectivity to developed barriers. While this analysis was conducted at the mesoscale, the results provide researchers with the framework to make insights at smaller scales than those demonstrated in this study.

North Carolina

Assessing future hydrologic extremes using an integrated hydrology and river operations model in the Russian River watershed

Study region The Russian River watershed, situated in coastal, northern California, experiences hydrologic extremes, including periodic droughts and flooding. Water managers are working to maintain sustainable water supplies and environmental flows, while mitigating flood risks. Study focus This paper introduces an integrated hydrology and river operations model for the Russian River watershed. This model is distinct from models in previous studies because it represents surface-groundwater interactions and uses climate forcings to estimate dynamic water use demands that are superimposed onto both reservoir operations and water supply constraints. The model was used to examine three historical (1990–2015) and eight future (2016–2099) water use and climate change scenarios. New hydrological insights for the region The direct connection between streams and aquifers facilitated both annual aquifer replenishment by high winter streamflows and streamflow depletion by groundwater wells (19 % of pumped groundwater in alluvial aquifers from stream leakage) during critical low flow periods. Simulated streamflow changes included 59 % longer and 54 % more severe streamflow droughts, 26 % lower seasonal low streamflows, and up to 125 % higher peak streamflows, averaged over future climate and water use scenarios, suggesting increased future flood and water availability risks. Results showed the importance of reservoir operations for mitigating the impacts of increased hydroclimatic volatility, despite a decrease in reservoir reliability at Lake Mendocino, suggesting that reservoir management may be used to decrease future risks.

California

Hydrologic variability and groundwater age of springs in eastern Oregon and northern Nevada, USA

The ecological importance of springs in semiarid regions is far greater than their small size and sparse distribution, yet little is known about the hydrologic functioning of these systems. During 2016–22, 261 springs were visited in the volcanic terrane of eastern Oregon and northern Nevada. When conditions were suitable, measurements of discharge, water temperature, and specific conductance were made, and samples for the analysis of carbon-14, tritium, and water stable isotopes (WSI) were collected. A subset of 60 springs was revisited during different seasons in the same year and during the dry season in multiple years to evaluate variability in discharge, chemistry, and groundwater age. Specific conductance and WSI varied considerably among springs across the study area but were unexpectedly stable across seasons and years at individual springs. Seasonal and interannual variability in spring discharge was related to the residence time of the discharging groundwater. Springs discharging older groundwater (10 3 –10 4 years) had significantly less variability in their discharge compared to springs discharging younger groundwater (10 0 –10 1 years). Variability among springs discharging younger groundwater included cessation of late-summer discharge at 18 % of the repeat-visit springs. A logistic regression model predicted the age of discharging spring water with 89 % accuracy using only the spring latitude, longitude, elevation, and δ 2 H value. This study framework provides a simple, inexpensive, and robust method to provisionally assess the hydrologic behavior of springs having little or no prior information in understudied, semiarid regions across the globe.

Nevada, Oregon

Ringtail (Bassariscus astutus) survival in southwestern Oregon

The effective conservation and management of small carnivore populations requires understanding species’ life-history traits and identifying important vital rates that drive population trajectories. However, many of these species are rare or elusive and of state or federal conservation concern, and demographic information is often lacking and difficult to obtain. At the northern limit of their range in Oregon, USA, ringtail ( Bassariscus astutus ) occupy mid-elevation forests and are a species of conservation concern because of their limited distribution and suspected low density. We initiated a radio-telemetry study in 2020 to estimate monthly and annual survival of ringtail in southwest Oregon. We monitored 26 ringtail from November 2020 to October 2022 and estimated survival rates using a known-fate framework and Program MARK. Model-averaged monthly survival estimates ranged from lows of 0.963 (SE = 0.022, 95% CI = 0.887–0.988) in spring (Feb–May) of 2021 to highs of 0.980 (SE = 0.018, 95% CI = 0.889–0.997) in summer (Jun–Oct) of 2022. Model-averaged estimates of annual survival were 0.695 (SE = 0.176, 95% CI = 0.310–0.920) during 2020–2021 and 0.728 (SE = 0.168, 95% CI = 0.336–0.934) during 2021–2022. Predation was the leading cause of mortality, but notably, no mortalities could be attributed to avian predation. Survival rates were much higher within forested landscapes in Oregon than for populations in the southwestern United States, which may have implications for regional conservation and management strategies.

Oregon

Per- and polyfluoroalkyl substances (PFAS) mass flux and mass balance at an aqueous film-forming foam release site in semiarid eastern New Mexico, USA

Passive flux meters (PFMs) directly measure groundwater chemistry mass flux and Darcy flux, providing insight into contaminant source-zone architecture and transport properties. This study uses PFMs to characterize PFAS flux in groundwater at a semiarid site with a thick (greater than 90-m) unsaturated zone where groundwater has been contaminated with per - and polyfluoroalkyl substances (PFAS) related to the use of aqueous film-forming foam (AFFF) for fire training and fire suppression. PFAS mass discharge (PFAS mass flux integrated over a control plane) in groundwater downgradient from several PFAS release areas is calculated using PFM results. In groundwater downgradient from fire-training areas, total PFAS mass discharge (summed across 14 compounds) was estimated to be between 6.0 and 31 g per day in 2020 and between 5.9 and 23 g per day in 2021. Site-specific documentation, generic information on AFFF properties, and literature values of PFAS concentration in AFFF are used to estimate site-specific PFAS-application rates at fire-training areas. These PFAS-application rates are compared to groundwater PFAS-discharge rates. Results suggest that transformation processes (exact pathways unknown) have led to increased discharge of measured PFAS in groundwater relative to initial AFFF formulations. The mass balance approach has broad applicability as a high-level approach that can provide insight into PFAS transport at AFFF sites.

New Mexico

Evaluating the effects of a multi-modal deterrent on upstream passage by invasive carp and native species

Invasive carp, including Silver Carp ( Hypophthalmichthys molitrix ) and Grass Carp ( Ctenopharyngodon idella ), are increasing their range within the Mississippi River Basin, threatening native fishes and ecosystem function. We conducted a 3-year field test of a multi-modal deterrent that uses sound, bubbles, and light to test its ability to impede upstream passage by invasive carp. The test was performed at a lock and dam on the Cumberland River, Kentucky, USA, using acoustic telemetry to track the movements of Silver Carp, Grass Carp, and several native fish species. When the deterrent was operating, the estimated probability of upstream lock passage was 53% lower (95% CI: 41%–63%) for Silver Carp but 74% higher (95% CI: 3%–190%) for Grass Carp compared to times when the deterrent was not operating. Paddlefish ( Polyodon spathula ) passage was reduced by 43% (95% CI: 71% reduction to 9% increase), although this was not statistically significant. We detected no significant effects on Freshwater Drum ( Aplodinotus grunniens ) or Smallmouth Buffalo ( Ictiobus bubalus ). However, sample sizes were small for these native species, limiting our ability to identify effects. Water temperature, tailwater elevation, and vessel lockages affected passage rates, although this varied among species.

Kentucky

Linking stream-reach nitrogen loads and groundwater “reachsheds” to inform wastewater-nitrogen management actions, Cape Cod, Massachusetts

Study Region Cape Cod, Massachusetts, U.S.A. Study Focus Anthropogenic nitrogen (N) is a key factor in degrading groundwater and surface-water quality, particularly in coastal New England where onsite wastewater systems are prevalent. This study evaluated whether direct N-load measurements in streams on Cape Cod, Massachusetts, coupled with flow-path information from groundwater-flow models, can effectively identify potential land areas where nitrogen mitigation could substantially reduce loads to receiving waters. Nitrogen fluxes were measured along 63 stream reaches during winter and summer and paired with simulated groundwater recharge areas to identify and rank potential areas for reduction of nonpoint-source N inputs. New Hydrologic Insights for the Region Reach-scale nitrate-N loads ranged from −39.1–1182 kg-N/yr per 100 m of stream, indicating spatially variable groundwater inputs across seasons. “Reachsheds” — areas contributing groundwater recharge to specific stream reaches — were delineated using a regional groundwater-flow model. Strong correlations were found between observed N loads and land-use characteristics, especially the number of septic systems and total N inputs from the sum of considered sources. Observed N loads were moderately correlated with recharge area size and wastewater flow estimates. Correlating reach-specific groundwater N loads with land use and parcel-scale nitrogen-yield data identified reachsheds with the highest potential for N load reduction. This approach enables targeted implementation of restoration efforts to optimize nutrient management and support regional load reduction.

Massachusetts

Surface-wave relocation and characterization of the 2024–2025 dyking episode along the Fentale–Dofen segment of the Ethiopian rift

From late-December 2024 to mid-March 2025, a 50-km-long dyke intrusion triggered over 300 earthquakes (magnitude 4 to 5.9) between Fentale and Dofen volcanoes along the Northern Main Ethiopian Rift. Dyke intrusions periodically occur along the Fentale–Dofen magmatic segment and are an expression of ongoing rift extension. Preliminary analyses using interferometric synthetic aperture radar revealed extensive ground deformation (up to 60 cm), which closely matched the temporal and spatial evolution of surface manifestations and earthquake locations from global catalogues. While global catalogues are critical for real-time monitoring, the precision of locations in remote and or sparsely instrumented regions can be low. In this investigation, we present surface-wave relocation results of the dyking episode that began near Fentale volcano in December 2024. We estimate relative locations using differential traveltimes measured from regional-to-teleseismic distance surface-wave observations of earthquakes reported by the U.S. Geological Survey. Relative relocations reduce the initial region of diffuse seismicity to a 50-km-long narrow band bounding the strike of surface manifestations and the zone of maximum surface deformation. We demonstrate the precision of surface-wave relocations over incremental time periods, capturing the progression of dyking from seismic onset through seismic migration and caldera subsidence. Results showcase the utility of surface-wave relocations in the characterization of dyking episodes and provide complementary insights into the current understanding of the Fentale–Dofen volcanic plumbing system.

Geophysical Journal International

Slow slip detectability in seafloor pressure records offshore Alaska

In subduction zones worldwide, seafloor pressure data are used to observe tectonic deformation, particularly from megathrust earthquakes and slow slip events (SSEs). However, such measurements are also sensitive to oceanographic circulation-generated pressures over a range of frequencies that conflate with tectonic signals of interest. Using seafloor pressure and temperature data from the Alaska Amphibious Community Seismic Experiment, and sea surface height data from satellite altimetry, we evaluate the efficacy of various seasonal and oceanographic pressure signal proxy corrections and conduct synthetic tests to determine their impact on the timing and amplitude prediction of ramp-like signals typical of SSEs. We find that subtracting out the first mode of the complex empirical orthogonal functions of the pressure records on either the shelf or slope yields signal root-mean-square error (RMS) reductions up to 73% or 80%, respectively. Additional correction with proxies that exploit the depth-dependent spatial coherence of pressure records provides cumulative variance reductions up to 83% and 93%, respectively. Our detectability tests show that the timing and amplitude of synthetic SSE-like ramps can be well constrained for ramp amplitudes ≥4 cm on the shelf and ≥2 cm on the slope, using a fully automated detector. The principal limits on detectability are residual abrupt changes in pressure that occur as part of the transition to and from summer to winter conditions but are not adequately characterized by our seasonal corrections, as well as the inability to properly account for instrumental drift, which is not readily separated from the seasonal signal.

Alaska

Deaf, deafblind, and hard of hearing university student experiences with earthquake early warning in the United States: Evaluating language planning and technology access

The growing literature on deaf and hard of hearing (DHH+) populations and disasters demonstrates that emergency communication (including alerts) is not reaching global DHH + individuals with dangerous impacts for morbidity and mortality. This is the first research study in the U.S. to qualitatively explore the experiences of DHH + persons with earthquake early warning (EEW) through group-based dialogue sessions. The study investigates eight DHH + university students'past earthquake experiences, access to EEW alerts, and perceptions of ShakeAlert Ⓡ , an EEW system for detecting earthquakes and alerting residents of California, Oregon, and Washington. Findings highlight key gaps in disaster alert usability within four thematic areas: lack of messaging in participants' language(s), unclear alert messaging, deficient message delivery mechanisms for deafblind persons, and insufficient access to earthquake information and training that leads to dependence on informal information networks. Weaknesses identified in these four themes reduce DHH + trust in EEW systems and compromise the capacity of alert recipients to take swift protective action or to mentally prepare before shaking starts. The study also underscores structural factors such as insufficient linguistic representation in disaster language planning and technology design, which ignores the linguistic and sensory access needs of DHH + individuals. Building on disaster language planning frameworks, we recommend involving DHH + populations to co-develop EEW alerts. By centering DHH + perspectives, this research contributes to ongoing efforts to ensure that EEW systems reach everyone.

California

Expression of corticoid-regulatory genes in the gills of Atlantic salmon (Salmo salar) parr and smolt and during salinity acclimation

In teleost fishes, cortisol is the major corticoid and has both glucocorticoid and mineralocorticoid actions. However, how fish tissues discriminate between these distinct corticosteroid actions is unclear. In mammals, the major factors responsible for intracellular corticosteroid regulation are glucocorticoid receptors ( gr s) and the mineralocorticoid receptor ( mr ), but their role in osmoregulation of fish is unclear. 11β-hydroxysteroid dehydrogenases ( hsd11b s) control the levels of intracellular corticosteroids by converting from bioactive forms to inert forms. To investigate how Atlantic salmon ( Salmo salar ) respond to cortisol in different physiological or environmental conditions, we performed comparisons of parr and smolt, and osmotic challenge experiments to examine the physiological responses and gill transcript levels of genes underlying cortisol-signalling, including gr1 , gr2 , mr , hsd11b2 and hsd11b3 . Because cortisol may interact with growth hormone and prolactin during salinity changes, transcript levels encoding growth hormone receptors ( ghr1 , ghr2 ) and the prolactin receptor ( prlr ) were also examined. Hsd11b2 transcript levels in seawater-acclimated fish were consistently lower compared to fish acclimated to fresh water. After transfer to seawater, prlr transcript levels in fish significantly decreased and transcript levels of ghr1 , ghr2 and hsd11b3 showed no change or were slightly higher than those of freshwater control groups. Gr1 , gr2 and mr transcript levels were slightly but consistently higher in fish acclimated to fresh water relative to seawater. Our results indicate that changes in corticosteroid receptor and hsd11b2 transcript levels in the gills may be important mechanisms that regulate corticoid signals to achieve ion homeostasis in Atlantic salmon.

Journal of Fish Biology

Egg thiamine concentrations reflect dietary differences between four hatchery strains, including two morphotypes, of Lake Trout in Lake Ontario

Objective Test whether dietary differences among hatchery strains of Lake Trout Salvelinus namaycush were associated with variation in egg thiamine concentrations and assess the potential for using strain-specific traits to mitigate thiamine deficiency complex (TDC) in Lake Ontario restoration efforts. Methods Stomach content analysis and stable isotope mixing models were used to compare diets among four hatchery strains of Lake Trout, representing lean and humper morphotypes. Egg thiamine concentrations were measured to test whether dietary differences correlated with susceptibility to TDC. Results Significant differences in diet and egg thiamine concentrations were observed among Lake Trout strains. A humper-like morphotype strain exhibited elevated thiamine levels, which were associated with reduced consumption of prey known to degrade thiamine levels when consumed, Alewife Alosa pseudoharengus and Rainbow Smelt Osmerus mordax , and increased reliance on Round Goby Neogobius melanostomus or sculpin species (Slimy Sculpin Cottus cognatus and Deepwater Sculpin Myoxocephalus thompsonii ). These findings support the link between the consumption of Alewife and Rainbow Smelt and thiamine deficiency. Conclusions Our findings support a link between strain-specific dietary preferences and susceptibility to TDC in Lake Trout. Preferential stocking of strains less prone to TDC, such as the strain derived from a humper morph from Lake Superior, could enhance restoration success in Lake Ontario. This strategy may be especially effective given the continued dominance of Alewife in the lake's prey base and the current management priorities supporting fisheries for Pacific salmon Oncorhynchus spp.

North American Journal of Fisheries Management

High resolution identification and quantification of diffuse deep groundwater discharge in mountain rivers using continuous boat-mounted helium measurements

Discharge of deeply sourced groundwater to streams is difficult to locate and quantify, particularly where both discrete and diffuse discharge points exist, but diffuse discharge is one of the primary controls on solute budgets in mountainous watersheds. The noble gas helium is a unique identifier of deep groundwater discharge because groundwater with long residence times is commonly enriched in helium. In this study, a portable mass spectrometer was used to measure longitudinal variation in dissolved helium concentrations in two mountainous rivers at high spatial resolution not feasible with traditional sampling techniques. Helium profiles were then simulated using a mass-balance model to quantify longitudinal variation in groundwater discharge to the receiving rivers. Results indicate helium concentrations were enriched by multiple orders of magnitude above atmospheric equilibrium in both rivers and that this persisted for up to 18 km below observed pulse inputs in the Colorado River. Helium mass-balance models match observed longitudinal patterns with the exception of sharp initial increases in helium observed in the rivers. Increased longitudinal groundwater discharge rates correspond to mapped geologic structures in both watersheds that likely transport deep geothermal water. Models show variable sensitivity to spatial assignment of input variables representing the groundwater source, illustrating the importance of collecting data from discrete groundwater discharges where possible. The methodology shows promise for field experiments designed to assess air–water exchange rates and to quantify total groundwater discharge from a combination of discrete and diffuse sources.

Colorado, Utah

Hydrologic response of groundwater and streamflow to natural and anthropogenic drivers of change in headwaters of the upper Colorado River basin during recent wet (1982–1999) and drought (2000–2022) conditions

Study region: Headwaters of the upper Colorado River basin (UCOL), USA Study focus: Surface-water and groundwater numerical models incorporating water-use information were used to investigate changes in climate, water use, and simulated hydrologic responses of snow processes, evapotranspiration, groundwater, and streamflow during recent wet (1982–1999) and drought (2000–2022) periods in the headwater subregions of the upper Colorado River basin. New hydrologic insights for the region: Decreases in average streamflow between wet and drought periods ranged from 20 % in the Colorado River headwaters subregion to 23 % in the Gunnison River headwaters subregion. Like streamflow, average surface runoff was statistically less during the drought than the wet period, with decreases from 24–31 % in the headwaters. On a volume basis, runoff decreases were greater than streamflow decreases in both the Colorado River and Gunnison River headwaters. Although the amount of water-year groundwater discharge to streams remained nearly the same between the wet and drought periods, groundwater as a percentage of streamflow increased between the wet and drought periods, highlighting the importance of groundwater in sustaining streamflow during drought conditions. Multiple linear regression analyses revealed that snowmelt-only models were better than the best precipitation and temperature models at explaining streamflow variability from all headwater subregions for both the wet and drought periods.

Arizona, Colorado, New Mexico, Utah, Wyoming

Manipulation of the Symbiodiniaceae microbiome confers multigenerational impacts on symbioses and reproductive ecology of its Exaiptasia diaphana host

Symbiodiniaceae-associated microbiota strongly influence cnidarian symbioses. We systematically reduced the bacterial and fungal communities associated with Symbiodiniaceae to study potential effects on the cnidarian holobiont Exaiptasia diaphana (Aiptasia). Clonal anemones were inoculated with xenic Breviolum minutum (SSB01) and microbiome-manipulated cultures after antibacterial or antifungal treatment. The asexual reproduction of pedal laceration allowed for three generations of clonal aposymbiotic Aiptasia to be utilised in this study, from the initial adult generation (G0), to the first (G1), and second (G2) generation. We inoculated small and large G1 Aiptasia with SSB01 algae and monitored onset of symbiosis, rate of algal proliferation, and holobiont characteristics. Sequencing the 16S and 18S rRNA gene regions identified significant differences in the bacterial and fungal communities of the G0 and G1 generations, alongside differences between the size classes of small and large G1 anemones. The microbiome of larger G1 individuals was distinct to the smaller G1 anemones, suggesting a microbiome maturation process. Control Breviolum minutum cultures exhibited a significantly greater proliferation rate in large G1 anemones when compared to antibacterial or antifungal treated cultures, whereas the opposite trend was documented in the small G1 anemones. Although no differences were observed between algal photochemical parameters, or the growth and polyp activity of G1 juveniles, we observed a significant influence in the production of G2 clones between treatments. Overall, we provide strong ecological implications of manipulating Symbiodiniaceae microbiome, not for the algae themselves, but for the maturation of the host Aiptasia, as well as for the cnidarian holobiont over multiple generations.

The ISME Journal