Limnology and Oceanography: Methods

OverviewThis dataset originates from a preliminary respirometry study on carabid beetles from the Elbe Estuary (Northern Germany), encompassing species from freshwater and saltmarsh habitats along a salinity gradient. The study was designed to establish and validate a workflow for measuring oxygen consumption, including chamber setup, sensor recording, drift correction, and calculation of absolute and mass-specific metabolic rates. Oxygen consumption was measured for five species (Carabus auratus, Carabus granulatus, Limodromus assimilis, Poecilus versicolor and Pterostichus niger) using sealed glass vials connected to an optical oxygen system. The dataset provides individual-level measurements and serves primarily as a methodological reference for future respirometry studies on ground-dwelling arthropods. The O2 consumption rates of carabid beetles showed interspecific differences and followed metabolic scaling theory, revealing an inverse relationship between body mass and mass-specific metabolic rates across species (Figure 3). O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=127 SRC="FIGDIR/small/720111v1_fig3.gif" ALT="Figure 3"> View larger version (17K): org.highwire.dtl.DTLVardef@f41f27org.highwire.dtl.DTLVardef@12939eeorg.highwire.dtl.DTLVardef@19a4630org.highwire.dtl.DTLVardef@17611ba_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 3:C_FLOATNO Oxygen consumption rates of Carabid species per (a) animal in [ml O2 h-1] and as (b) mass-specific consumption rate [ml O2 h-1 g-1]. Points represent mean oxygen consumption per individual (C. auratus: n = 2; L. assimilis: n = 6; P. versicolor: n = 7; P. niger: n = 6). C_FIG

The Mississippi River (MR) is the largest source of freshwater and nutrients to the Gulf of Mexico (GoM), strongly influencing stratification, primary production, and plankton organization. The interaction between buoyant plume waters and denser shelf waters in the northern Gulf of Mexico (nGoM) generates sharp density gradients that can promote fine-scale biological aggregation. We investigated how hydrographic structure associated with the MR plume controls the vertical distribution of plankton during May 2017 using an integrated instrumentation suite that included an in situ digital holographic imaging system (HOLOCAM) coupled with CTD and optical sensors. Phytoplankton thin layers were repeatedly detected at plume-edge stations within or immediately above a compressed pycnocline formed by bottom-trapped saline wedges. These layers were 1.2-3.5 m thick and exhibited chlorophyll-a concentrations up to threefold higher than background levels. The assemblage was dominated by chain-forming diatoms, particularly Chaetoceros debilis and C. socialis, whose local abundance maxima coincided with chlorophyll peaks. In contrast, copepods, appendicularians, and other zooplankton were broadly distributed throughout the upper water column and rarely aggregated within the layers. Redundancy analysis indicated that chlorophyll concentration and stratification intensity were primary drivers of community structure across stations. Satellite imagery revealed rapid short-term variability in plume extent, helping explain differences in stratification and thin layer development among sampling days. Our results demonstrate that salt-wedge dynamics at the plume-shelf interface constitute a key physical mechanism governing transient phytoplankton thin layer formation in the nGoM, while zooplankton responses remain weakly coupled at the temporal scales resolved here.

Understanding how warming alters estuarine plankton communities is essential for predicting future changes in biodiversity and ecosystem functioning. We conducted a four-week indoor mesocosm experiment using natural summer plankton from the Elbe River to examine the effects of warming (+2 {degrees}C and +4 {degrees}C) on abiotic conditions and responses of the plankton community. In this study, oxygen concentrations, primary producer biomass (chlorophyll a, microphytoplankton) and microzooplankton abundances declined sharply during the first 10 days across all treatments while mesozooplankton abundances increased. This suggests a strong top-down control by mesozooplankton on lower trophic levels across all temperature treatments. Primary producers biomass and oxygen concentrations recovered after an initial decline, however to lower levels compared to the onset of the experiment while micro- and mesozooplankton remained low during the second half of the experiment. Nutrient dynamics indicated progressive remineralization, with increasing ammonium, NOx, and silicate concentrations, while phosphate concentrations remained low throughout the experiment. Complementary DNA and RNA metabarcoding revealed similar community turnover over time in all treatments and temperature effects became only pronounced at the end of the experiment. Overall, warming effects were subtle relative to the strong internal trophic dynamics likely caused by the artificial mesocosm setup. Our findings of changes in plankton community dynamics indicate that biotic interactions, changes in trophic diversity and other environmental factors, i.e. oxygen concentrations are likely the drivers of this estuarine system rather than warming alone.

Using linear inverse ecosystem modeling as a data assimilation tool, we compare spawning grounds of Atlantic and Southern Bluefin Tuna (ABT and SBT, respectively) based on results from field campaigns in the Gulf of Mexico (GoM) and eastern Indian Ocean off northwest Australia (Argo Basin). Both regions are warm, stratified, low-nutrient waters dominated by cyanobacteria (Prochlorococcus). Despite these similarities, the Argo Basin is more productive, with [~]1.5X higher net primary production and nearly 2X higher production of top trophic levels in the model (tuna larvae, planktivorous fish, and predatory gelatinous zooplankton). Higher primary production in the Argo Basin is mainly driven by higher N2 fixation and storm mixing of new nutrients in the upper and lower euphotic zone, respectively. Increased ecosystem efficiency (secondary production of top trophic levels / primary production) results from differences in plankton food web organization. In the GoM, protistan zooplankton are the direct consumers of nearly all phytoplankton production. In contrast, higher rates of herbivory by crustaceans feeding on nanophytoplankton combines with a higher impact of appendicularians on cyanobacteria to convert plankton production into larval tuna prey more efficiently in the Argo Basin. Despite similarities in the proportions of phytoplankton production mediated by cyanobacteria and other picoplankton in both systems, food web pathways to larval tuna and other planktivorous fish are substantially shorter in the Argo Basin. Our results highlight the impact of distinct zooplankton ecological niches on ecosystem efficiency and suggest a need for better inclusion of plankton food-web structure in models simulating climate impacts on fisheries production. HIGHLIGHTSO_LIDeveloped food web models of tuna spawning habitat (Indian Ocean & Gulf of Mexico) C_LIO_LISpawning habitats in the Argo Basin and Gulf of Mexico (GoM) are both oligotrophic C_LIO_LIArgo Basin had higher net primary production in part as a result of nitrogen fixation C_LIO_LIArgo Basin had higher rates of direct herbivory by metazoan zooplankton C_LIO_LIThis resulted in greater ecosystem efficiency in the Argo Basin. C_LI

Since 1995, European fisheries of Pecten maximus have faced the presence of Pseudo-nitzschia species, which are able to produce the neurotoxin domoic acid responsible for Amnesic Shellfish Poisoning (ASP). As filter-feeders, scallops can accumulate and retain domoic acid much longer than most other bivalves, from months to years. When concentrations exceed the regulatory threshold, fisheries are closed leading to economic concern. Inter-individual variability increases the difficulty to predict the depuration dynamics. Quantifying the correlations between domoic acid depuration in P. maximus and individual physiological traits, particularly body size, could improve the understanding of contamination and depuration. We analysed toxin dynamics in organs and assessed the effects of body size and growth. This analysis was based on two datasets from an experimental and an in situ depuration monitoring of P. maximus exposed to a natural bloom of toxic P. australis. Results showed that the distribution of domoic acid shifted among organs between contamination and two months of depuration. Toxin concentrations correlated negatively with body size during contamination and after two months of depuration, but shifted to a positive correlation after 7 months of depuration. This suggested that smaller scallops both accumulate more domoic acid and depurate it more rapidly. Dilution by growth appeared to explain the inversion of the correlation between domoic acid and body size throughout depuration. These results yield useful information for modelling these mechanisms, thus providing valuable tools for scallop fishery management facing ASP. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=60 SRC="FIGDIR/small/708139v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@1fd317org.highwire.dtl.DTLVardef@15b9032org.highwire.dtl.DTLVardef@57dae8org.highwire.dtl.DTLVardef@1e4c7fc_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIExperimental and in situ datasets allowed to quantify DA proportion dynamics in organs in P. maximus C_LIO_LIDA concentration and body size are negatively correlated during contamination phase, but positively after a 7-month depuration C_LIO_LIConsidering dilution by growth is important for young scallops to assess DA depuration dynamics C_LIO_LIBoth depuration rate and dilution by growth need to be considered to model DA depuration over the whole scallop size range C_LI

Mid-Trophic Level (MTL) organisms--including krill, forage fish, and mesopelagic fish-- are abundant in the California Current System (CCS) and play an essential role in transferring energy and biomass from primary producers to top predators. However, their spatiotemporal distribution and variability remain poorly understood, particularly with respect to vertical structure across epipelagic and mesopelagic habitats and coastal-offshore gradients. This lack of understanding emerges from both the complexity of MTL interactions with a heterogeneous environment and the challenges associated with sampling these organisms at high spatial and temporal resolution. To address this gap, we analyze 11 years of fisheries acoustic observations in the CCS (2006-2016) to characterize the spatiotemporal dynamics of MTLs as inferred from acoustic backscatter. Acoustic observations at 38 and 120 kHz, collected during day and night across depth strata from 15 to 495 m, reveal consistent cross-shore, seasonal, and latitudinal patterns in the backscatter of acoustically defined zooplankton, epipelagic fish, and mesopelagic fish communities. These patterns include: (1) weaker cross-shore gradients in mesopelagic relative to epipelagic communities; (2) a temporal succession among communities associated with seasonal upwelling; and (3) a multimodal latitudinal distribution with distinct coastal backscatter peaks. We further investigate relationships between acoustic backscatter and co-located environmental variables from in situ, remote sensing, and reanalysis products to elucidate plausible mechanisms underlying MTL dynamics. HighlightsO_LIFisheries acoustics resolve variability in mid-trophic communities C_LIO_LIEleven years of backscatter reveal consistent patterns in the California Current C_LIO_LIEpipelagic backscatter declines faster from the coast to offshore than mesopelagic C_LIO_LISeasonal changes in community composition are linked to upwelling dynamics C_LIO_LIBackscatter exhibits multimodal latitudinal distributions with distinct peaks C_LI

We developed a model to predict surface water temperature across U.S. lakes using satellite remote sensing and in situ observations to enhance cyanobacterial harmful algal bloom (cyanoHAB) forecasting. The study focused on Sentinel-3 Ocean and Land Colour Instrument (OLCI) sensor resolved lakes. We developed random forest models using both Landsat-derived and in-situ-measured surface water temperature. Landsat models offered broad spatial and temporal coverage of all OLCI resolved lakes, but they were sensitive to cloud cover and required filtering to minimize error. In contrast, the in situ model represented fewer OLCI resolved lakes, but yielded lower mean absolute error and bias. The models predicted lake surface temperature across the entire calendar year, with best performance (RMSEapplied=1.11; biasapplied=0.01; MAEapplied=0.77) from the in situ model. This approach allowed for the continuous prediction of lake surface temperatures from 1.1 to 31.6 {degrees}C for unfrozen, open-water conditions critical for improving the accuracy of cyanoHAB forecasting. A key strength of this study was the use of an extensive dataset and model validation against in situ observations, which improved predictive accuracy throughout the year across all seasons. The predictive model offers a water resource tool for management, ecosystem protection, and public health.

Cyanobacterial colonies often exploit their buoyancy and large size to float upwards rapidly and form dense surface blooms, which can degrade water quality, threaten ecosystems and public health, and impose substantial economic costs. Yet, how the morphology of cyanobacterial colonies controls their vertical velocity remains poorly understood. We conducted detailed three-dimensional morphological characterization of colonies of the cyanobacterium Microcystis in lake samples at the single-colony level and performed controlled flotation experiments in stratified flows. Using particle tracking in a vertical density gradient, we separately quantified the contributions of colony shape and buoyant density at the level of individual colonies. Our results show that the shape factor in Stokes law varies systematically with colony size. Consequently, the vertical velocity of colonies does not scale with the square of colony size but only with a power of 1.13, as larger colonies have a more irregular shape and therefore experience enhanced drag. We therefore correct the commonly used Stokes law to account for the size-dependent change in the shape factor. Interestingly, implementation of this power law relationship in a vertical migration model shows widespread chaotic dynamics in the migration trajectories of Microcystis colonies. These results highlight the importance of morphological plasticity in cyanobacterial colonies and can inform predictive models and hydrodynamic control strategies for toxic blooms. Our methodology to simultaneously determine the density, shape factor and velocity is broadly applicable to suspended aggregates with complex shapes in freshwater and marine systems.

Chelating ion exchange resins are widely used to eliminate metal interferences in the analysis of ammonium (NH4+) in soil extraction solutions. However, their potential to co-adsorb NH4+ remains underexplored. Here, synthetic metal ion solutions containing 6-30 mg L-1 NH4+ and the metal cations Ca2+, Mg2+, Cu2+, Mn2+, and Zn2+ were treated with Amberlite IRC-748 resin. The resin efficiently removed Ca2+ (-42.2%), Mg2+ (-21.1%), Cu2+ (-99.9%), Mn2+ (-56.9%), and Zn2+ (-93.6%). However, NH4+ losses of 2.2-5.6% were observed, indicating concentration-dependent co-adsorption. While these losses may be acceptable for concentration measurements via routine assays such as photometric analysis, they may still affect the accuracy of high-precision N analyses that rely on quantitative NH4+ recovery. This highlights a methodological caveat for resin-treated samples, especially in low-NH4+ environments. We therefore recommend including recovery assessments and correction factors when using chelating resins to improve accuracy in NH4+ quantification.

Seamounts are prominent deep-ocean features that strongly influence geological processes, ocean circulation, and benthic biodiversity. Despite their importance, most seamounts remain unmapped and poorly characterized, particularly in the southeast Pacific Ocean, a region recognized for high marine endemism and ecological isolation. In this study, we present a quantitative habitat characterization of a previously undocumented seamount, informally named Solito Seamount, located between the Nazca-Desventuradas Marine Park and the Juan Fernandez Archipelago. High-resolution multibeam bathymetry and backscatter intensity data were integrated with in situ observations from two remotely operated vehicle (ROV) dives (SO643 and SO645) to investigate how geomorphology and substrate distribution influence benthic community patterns. An automated and hierarchical quantitative mapping framework incorporating objective terrain analysis and multivariate statistical techniques, including principal component analysis and clustering, was applied to delineate five distinct megahabitat types: flat, basal slope, valley, ridge slope, and ridge crest. ROV video transects traversing these megahabitats revealed five associated substrate type forming macrohabitats: bedrock, bedrock with sediment veneer, sediment-rock transition, sediment, and coral rubble. Outputs were used to investigate how environmental heterogeneity structures megafaunal assemblages of Solito Seamount. Multivariate analysis revealed a combined effect of megahabitat type and substrate type on benthic megafaunal assemblages across the depth gradient. These compositional dissimilarities were primarily driven by habitat-forming taxa. In the deeper dive (SO643), a broad suite of taxa contributed to dissimilarities, and assemblages were primarily organised by megahabitat. The ridge crest hosted a distinct reef-building scleractinian community, whereas the ridge slope hosted mixed antipatharian, gorgonian and actiniarian assemblages. In contrast, the shallower dive exhibited simpler patterns with few taxa driving dissimilarities. Substrate effects were most pronounced with coral rubble forming a distinct habitat characterised by sponges (Stelletta sp.). Pronounced biological differences between dives may also represent depth-dependent structuring resulting from differences in oxygen regimes associated with water masses, underscoring the role of oceanographic forcing. This study provides the first quantitative habitat map of this previously undocumented seamount, delivering essential baseline information for this largely unexplored region of the southeast Pacific. The integrated multi-scale geophysical and biological approach presented here offers a robust framework for advancing seamount ecosystem understanding and supporting future biodiversity assessments and conservation planning.

For sustainable algal biomass cultivation, we need substantial improvement in annualized productivity by reducing the frequency of crop failure and improved growth in open raceway pond systems. In this study, high-performing strains were identified and optimized for biomass productivity. We utilized next-generation sequencing methods to quantify the ecological features of open raceway systems cultivated at in Arizona. We utilized data from several months of cultivation runs to construct a rich time-series of the ecology dynamics using amplicon sequencing and used custom anomaly detection, "PondSentry", for the early prediction of pond crashes. PondSentry uses tensor decomposition of higher-order joint moments to detect incipient anomalies in multivariate data and displays significant improvements from standard knowledge-based anomaly detection methods. The PondSentry strategy identifies signs of deteriorating pond health at an average of three days before an actual crash event, with rank order of the ecological features plausible for crop failures driven by organisms such as Amoeboaphelidium occidentale FD01. These findings are independently confirmed with PCR and microscopy studies at an Arizona cultivation site. PondSentrys time-series-based anomaly detection of crashes provides a suitable monitoring strategy for eukaryotic crash agents in unialgal culture. The early warnings can be used to time interventions or harvests to prevent biomass loss. The PondSentry strategy strengthens the role of data science and data-driven methods in algal cultivation and can increase the feasibility of algal-biomass based products.

The prevalence of nitrogen limitation and nitrogen-phosphorus co-limitation (henceforth referred to as nitrogen-related limitation) in Norwegian lakes and their relationships with atmospheric nitrogen deposition, climate, dissolved organic matter (DOM), and catchment characteristics were assessed across space and time. Routine monitoring data from 1,529 lakes in the national Vannmiljo database were analyzed for two multi-year periods (1995-2009 and 2010-2025). Limitation was inferred using the molar NO--N/TP ratio as an indicator of dissolved inorganic nitrogen availability. Nitrogen-related limitation was widespread in both periods and exhibited strong regional structure, with highest prevalence in northern regions and lowest prevalence in southwestern Norway. Overall prevalence increased from 31% to 38% between periods, with significant increases in western regions. Regional-scale models identified climate, forest cover, DOM, agriculture, and atmospheric nitrogen deposition as predictors of limitation probability, whereas study period per se and bog/peatland cover were not significant. At the local scale, atmospheric nitrogen deposition and DOM were the only consistent predictors, with substantially lower explanatory power than at the regional scale. These results indicate that large-scale environmental gradients play a major role in shaping nutrient stoichiometry in Norwegian lakes. Because the monitoring dataset primarily represents lakes affected by human activities, the findings are particularly relevant for water management. The widespread occurrence of nitrogen-related limitation suggests that nitrogen availability may influence phytoplankton growth in many systems and that dual-nutrient management strategies addressing both nitrogen and phosphorus may be required in many regions.

Benthic remineralization of organic matter is key to carbon and nutrient cycling, influencing both long-term carbon storage in the sediments and the release of nutrients that support primary production in the water column. With its multiple forms and ages of sea ice, Nares Strait in the Canadian Arctic offers a unique opportunity to address the knowledge gap of variability of benthic remineralization rates along a natural sea ice gradient. Here, we incubated sediment cores in different locations in Nares Strait characterised by different sea ice conditions ranging from first-year ice to multi-year ice, to measure oxygen and nutrient fluxes. To identify potential drivers, we measured environmental variables, identified macrofauna and calculated a suite of taxonomic and functional diversity indices. Our analyses showed that benthic fluxes varied significantly between the northern and southern regions of Nares Strait. The presence of deposit feeders and sea ice cover (number of days since ice-free) were the main drivers in benthic fluxes, explaining 22.6% and 13.9% of the benthic flux variation, respectively. Overall, functional diversity was a better predictor of benthic fluxes than taxonomic diversity, indicating its primary importance in controlling benthic ecosystems functioning. Our results reveal that, from a benthic biogeochemical point of view, Nares Strait seems to be dissected into two main sub-regions: (i) a permanently and highly sea ice-covered area north of Kennedy Channel, resembling deeper regions of the Arctic Ocean and (ii) a seasonally ice-covered area between the North Water Polynya and Kane Basin, where benthic fluxes values are equivalent to those reported in similar continental Arctic shelves. Consequently, the rapid functional shifts resulting from the ongoing decline in sea ice could enhance benthic remineralisation rates if deposit feeder were to become dominant in certain areas, reducing the role of the region and by extension, the Arctic, as a carbon sink.

Environmental DNA (eDNA) methods offer a powerful tool for monitoring aquatic species, yet field applications remain challenged by uncertainty in DNA transport, mixing, and detection, particularly in flowing or tidally influenced systems. One approach to improve confidence in eDNA surveys is the use of controlled DNA sources (positive controls), but questions remain regarding how the biological condition of the source influences eDNA release and detectability. This study evaluated differences in eDNA concentrations emitted from live versus dead fish in a controlled, shallow, well-mixed channel. Using a fixed point-sampling design, we measured eDNA concentrations over time and modeled the effects of treatment, sampling time, temperature, and water velocity. Dead fish consistently released significantly higher concentrations of eDNA than live fish, while eDNA concentrations declined over time in both treatments. Water temperature and velocity did not significantly influence detection, and the rate of eDNA decline was similar between live and dead treatments. These findings highlight the importance of source condition and site-specific mixing dynamics when interpreting positive control experiments and underscore the value of site characterization when designing eDNA sampling protocols.

O_LIEnvironmental DNA (eDNA) metabarcoding of water samples is increasingly used to detect fish species in streams. Several studies have concluded that it can outperform traditional inventory methods and recommend using it at large scales for fish-based ecological assessments. However, there is no standard protocol that can guarantee sufficient detection rates and repeatability, despite companies offering an extensive range of analyses. C_LIO_LIWe compared eDNA metabarcoding performed by four companies. Following their guidelines, samples were collected in a small tropical stream in the Maroni River (French Guiana) that hosts a species-rich fish community. We compared their inventories to each other and to a list of species captured during an extensive fish inventory performed immediately after sampling eDNA, as well as to current data on the species distributions. C_LIO_LIThe number of species detected by eDNA metabarcoding ranged from 5 to 48 among the companies, but these inventories contained many inaccuracies. All companies combined, 63 species were detected, of which 10 (16%) had never been reported in the Maroni River. The extensive inventory identified 50 species in the local fish community, of which 16-46 were not detected by eDNA metabarcoding (i.e. false negative detection rate of 32%-92% among the companies). C_LIO_LIReanalysis of raw sequencing data decreased differences among companies greatly, highlighting the importance of using a comprehensive and accurate DNA barcode database to assign species. Dissimilarity indices, calculated to compare the local fish community (based on presence/absence or fish catches) to eDNA detection, revealed large differences regardless of the company. C_LIO_LISummary and applications. The large percentage of species not detected by eDNA metabarcoding of water samples could strongly bias fish-diversity inventories in streams that host species-rich communities. This issue is not well documented in the literature, and we recommend that similar studies in the future focus on other stream contexts. The large differences between commercial eDNA inventories and the local fish community challenge the use of eDNA metabarcoding for fish-based ecological assessments of streams. The variable performance of eDNA companies indicates the need for a standard protocol and access to a comprehensive DNA database before beginning large-scale eDNA programmes. C_LI Highlights- eDNA metabarcoding of water samples is widely used to detect species in streams - Detection performances of 4 private companies were compared to an exhaustive fish inventory - The number of undetected species varies from 32 to 92% depending on the company - Such discrepancies challenge the use of eDNA for fish-based ecological assessments

Nutrient availability, ecosystem productivity, and consumer assemblages are intricately linked through complex interactions and feedbacks. Nutrients influence the diversity and functional roles of consumers via shifts in resource quality and quantity, and consumers can alter ecosystem production and nutrient availability. However, our understanding of how characteristics of consumers respond to and influence concomitant shifts in nutrient availability and production is limited. We quantified the response of well-studied consumer assemblages (benthic invertebrates and zooplankton) to realistic nutrient loads that altered gross primary production (GPP) and ecosystem respiration (ER). We fertilized 14 outdoor experimental ponds for 2 months and monitored total water column carbon (TC), nitrogen (TN), and phosphorus (TP), GPP, ER, and net ecosystem production (NEP) weekly. Then, we evaluated how fertilization and the variation in nutrients and metabolism caused by fertilization were related to shifts in consumer assemblages. Fertilization increased water column TN and TP and reduced TC:TP ratios, TN:TP ratios, and rates of GPP and ER. However, consumer assemblages were more tightly linked to variation in nutrient availability and production across ponds than to fertilization. Greater declines in benthic diversity occurred in ponds with higher average TN:TP ratios during the experiment. Consistent with predicted effects of cladocerans on nutrient availability, shifts in cladoceran abundances were positively associated with average water column TN:TP ratios during the experiment. Finally, elevated GPP and ER were associated with greater increases in the abundance of benthic invertebrate predators, suggesting the possibility of top-down control. Our study highlights the critical role of consumer-mediated processes in the interaction between nutrient availability and production. Manuscript HighlightsO_LIFertilization reduced pond gross primary production and ecosystem respiration rates. C_LIO_LIInvertebrate predator abundance was inversely related to gross primary production. C_LIO_LIShifts in consumer assemblages were tightly linked to nutrients and production. C_LI

Environmental DNA metabarcoding is a powerful monitoring tool for assessing aquatic biodiversity, as well as the sustainability and impacts of fisheries and aquaculture. However, conventional laboratory workflows remain time-consuming and dependent on dedicated infrastructures. Here, we present a field trial of a fully portable, off-grid eDNA metabarcoding pipeline that enables end-to-end analysis within a few days using compact equipment, including a BentoLab workstation and an Oxford Nanopore Technologies (ONT) MinION sequencer. The workflow was implemented during two international training courses in Norway and Brazil, where students and early career researchers collected environmental samples, extracted and amplified DNA, prepared DNA libraries, and sequenced on-site before performing bioinformatics and statistical analyses. In the case study detailed here, seven eDNA samples collected and analysed on-site in the Oslofjord allowed detection of 16 fish and elasmobranch species. Although overall diversity was lower than in earlier studies using Illumina-based sequencing, our protocol reliably detected key species and demonstrates that portable eDNA metabarcoding is feasible for rapid ecological assessment, surveillance of high-risk regions and/or deployment in remote or resourcelZllimited settings.

The use of field water for laboratory rearing of mosquitoes could offer a better representation of the natural aquatic environment than laboratory tap or deionised water. For logistical reasons, such water may be stored in the laboratory environment for an extended period, but its stability is poorly documented. This study evaluated the influence of laboratory storage conditions on the kinetics of physicochemical parameters of breeding water collected from a field habitat. To capture within-habitat variability, water was collected from multiple spatial points from a breeding site and transferred into plastic containers for storage under laboratory conditions. Water physicochemical parameters were measured in the field to establish baseline readings, while laboratory measurements were done at 2-3-day intervals over 2 months to evaluate temporal changes. A linear mixed-effects model was fitted to evaluate the determinants of changes in physicochemical parameters under laboratory storage. Most parameters exhibited high stability; however, water temperature increased significantly by an average of [~]1.5 (p= 0.046) relative to the field. Water pH demonstrated a long-term rise over the 2-month storage period with a transient, significant dip of 0.71 units after a week of storage (p< 0.001). Overall, LMM analyses revealed that ambient relative humidity was the strongest statistical predictor of change in all water parameters except pH (p< 0.05). Ambient temperature correlated positively with water temperature and ammonium nitrogen (NH4-N) (p<0.002), and negatively with dissolved oxygen (p< 0.002). These results indicate that stored field water is highly sensitive to the laboratory microclimate. Specifically, water temperature, pH, and NH4-N serve as candidate indicators for storage-related physicochemical drift. We recommend the rigorous standardisation of insectary humidity and temperature, and monitoring of water parameters, which are likely relevant for bioassay reproducibility.

With metabolic disease on the rise across the globe, the devices that can provide precise and reliable estimates of energy expenditure and macronutrient oxidation can play a critical role in the development and evaluation of therapeutic regimes and wearable technologies that can be used outside of the laboratory. Whereas, metabolic carts can provide short-term (minutes to hours) metabolic measurements, whole-room calorimeters enable long-duration (hours to days) metabolic assessment, providing insights into how metabolism changes in response to meals, activity, sleep, etc. Obtaining accurate metabolic measurement via whole-room calorimetry, however, requires rigorous methods for calibration and quality assurance. To date most room calorimeters have been tuned to assess energy expenditure over long periods of time, i.e. 24-hours. Here we present novel calibration and signal processing techniques and recommendations that aim to improve the utility of metabolic chambers for use over different measurement epochs. This work serves as both a transparent description of our hardware, validation procedures, and data processing approaches.

Ocean warming is currently leading to distributional shifts of species and an alternation of coastal communities. Vulnerable species that are most sensitive to ocean warming are able to use several acclimation mechanisms, with one of the fastest being a shift in and shuffling of their partnerships with symbiotic microorganisms. Assessing symbiosis-focused mechanisms of acclimation and adaptation in response to ocean warming is a technical challenge due to the difficulty of accurately simulating the de novo formation of coastal communities. Here, we use the Kiel Outdoor Benthocosm facility to assess which sponges species are experimentally recruited and whether they exhibit symbiosis-focused mechanisms of acclimation following selection to ocean warming. We observed one sponge species (Haliclona sp.) and found that this sponge exhibited significant shifts in the membership and composition of its associated microbiome in response to ocean warming, with much of this being attributed to the rare microbiota. Moreover, Haliclona sp. maintained the diversity and dominance of its microbiome members. Four bacteria taxa were differentially abundant at elevated temperatures, with two being a Francisella sp. that is a suspected pathogen and an uncultured Francisellaceae that is most closely related to sulfur-oxidizing endosymbionts. Changes to the Haliclona sp. microbiome are largely consistent with a limited acclimation response, which could indicate that this sponge may use microbial symbionts as part of a mechanism to acclimate and adapt to a warmer future ocean.