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Limnology and Oceanography: Methods

Wiley

Preprints posted in the last 30 days, ranked by how well they match Limnology and Oceanography: Methods's content profile, based on 11 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

1
Characterization of the vertical distribution of plankton and the formation of thin layers in the northern Gulf of Mexico using digital holography

Vega, G. M.; Kerkar, A. U.; Nayak, A. R.; McFarland, M.; Lopes, R. M.

2026-03-31 ecology 10.64898/2026.03.29.714992 medRxiv
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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.

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Comparative food-web analysis of bluefin tuna spawning habitats in the eastern Indian Ocean and Gulf of Mexico

Stukel, M. R.; Landry, M. R.; Decima, M.; Fender, C. K.; Kranz, S. A.; Laiz-Carrion, R. L.; Malca, E.; QUINTANILLA, J. M.; Selph, K. E.; Swalethorp, R.; Yingling, N.

2026-03-20 ecology 10.64898/2026.03.18.711569 medRxiv
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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

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Influence of organs, body size and growth and domoic acid depuration in the king scallop, Pecten maximus.

Le Moan, E.; Hegaret, H.; Deleglise, M.; Ambroziak, M.; Vanmaldergem, J.; Derrien, A.; Terre-Terrillon, A.; Breton, F.; Fabioux, C.; Jean, F.; Flye-Sainte-Marie, J.

2026-03-25 ecology 10.64898/2026.03.23.708139 medRxiv
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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

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How size and shape affect the vertical velocity of cyanobacterial colonies

Sinzato, Y. Z.; Verspagen, J. M. H.; Uittenbogaard, R.; Visser, P. M.; Huisman, J.; Jalaal, M.

2026-03-27 ecology 10.64898/2026.03.25.714148 medRxiv
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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.

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Characterizing Industrial Pond Ecology Timeline in DISCOVR Cultivation Trials for Early Detection of Pond Crashes

Wilbourn, E. K.; Curtis, D.; Kolla, H.; Rai, P.; Lane, P.; McGowen, J.; Lane, T. W.; Poorey, K.

2026-04-03 ecology 10.64898/2026.03.31.715673 medRxiv
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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.

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On-site metabarcoding analysis of environmental DNA samples

Mauvisseau, Q.; Ewer, I.; Blumeris, I.; Iren Bongo, S.; Filipe Brito de Oliveira, L.; Gouvea, B.; Carolina Cei, A.; Ferreira Rodrigues, K.; de Arruda Francisco, J.; Sletteng Garvang, E.; Marena do Rego Henriques, V.; Hurtado Solano, S.; Kvalheim, L.; Kaylynne Lawrence, S.; Ramalho Maciel, B.; Isanda Masaki, H.; Fortunate Mashaphu, M.; Masimula, L.; Prudent Mokgokong, S.; Katrin Onshuus, E.; Lima Paiva, B.; Parker-Allie, F.; Du Plessis, M.; Puzicha, M.; Gabriel Da Silva Solano Reis, O.; Speelman, G.; Moritz Splitthof, W.; Stocco de Lima, A. C.; Strindberg, H.; Smoge Saevik, O.; Tafjord, N. J. D

2026-03-30 ecology 10.64898/2026.03.27.714757 medRxiv
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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.

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Overcoming software bottlenecks for scalable passive acoustic monitoring: insights from a global expert assessment

Malerba, M. E.; Perez-Granados, C.; Bell, K.; Palacios, M. M.; Bellisario, K. M.; Desjonqueres, C.; Marquez-Rodriguez, A.; Mendoza, I.; Meyer, C. F. J.; Ramesh, V.; Raick, X.; Rhinehart, T. A.; Wood, C. M.; Ziegenhorn, M. A.; Buscaino, G.; Campos-Cerqueira, M.; Duarte, M. H. L.; Gasc, A.; Hanf-Dressler, T.; Juanes, F.; do Nascimento, L. A.; Rountree, R. A.; Thomisch, K.; Toledo, L. F.; Toka, M.; Vieira, M.

2026-04-01 ecology 10.64898/2026.03.30.715176 medRxiv
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Passive acoustic monitoring (PAM) enables non-invasive sampling of wildlife across broad spatial, temporal and taxonomic scales. Its ongoing and widespread use has generated unprecedented volumes of acoustic data, shifting the primary bottleneck from data collection to the storage, processing, integration, and interpretation of PAM outputs. Although many software tools exist to address these challenges, differences in their design, scope, and usability often create fragmented and complex analytical workflows. To identify the key barriers and opportunities shaping the implementation of PAM surveys, we conducted a structured expert solicitation involving 30 international practitioners working across terrestrial and aquatic ecosystems. Experts identified and ranked their most critical pain points in current PAM workflows, spanning data storage, processing, and interpretation. The top challenge identified related to accurate species identification using deep learning and artificial intelligence (AI) models, especially in noisy soundscapes or for underrepresented taxa. Eight additional priority challenges included workflow fragmentation, limited availability of user-friendly analytical and visualisation tools, uneven access to software, manual validation bottlenecks, computational constraints, and difficulties in data handling, standardisation, and sharing. Participants also proposed practical mitigation strategies for these priority challenges, supported by step-by-step guidance to help overcome key barriers. Together, these insights provide a roadmap toward more scalable, open-access, and collaborative software systems, which are increasingly essential to realise the full potential of PAM in global biodiversity monitoring.

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ExocubeBio: an in-situ fluidic platform for microbial exposure on the International Space Station

Burr, D. J.; Nitsche, R.; Ravaro, E.; Wipf, S.; Ganga, P. L.; Balsamo, M.; Pellari, S. S.; Caltavituro, F.; Gisi, M.; de Almeida, R. C.; Manieri, P.; Sgambati, A.; Moratto, C.; Nürnberg, D. J.; Kish, A.; Elsaesser, A.

2026-03-25 microbiology 10.64898/2026.03.25.714121 medRxiv
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Space-based platforms currently represent the most accurate means to experimentally assess the influence of the space environment on biological systems. However, performing such experiments remains technically challenging and requires highly specialized instrumentation. This study describes the current development and hardware qualification of ExocubeBio, a miniaturized experimental platform for in-situ biological space exposure. This experiment is scheduled for installation on the exterior of the International Space Station in 2027, as part of Exobio, the European Space Agencys new generation exobiology exposure facility. ExocubeBio will expose live microbial samples to the low Earth orbit environment, and combine autonomous in-situ optical density and fluorescence measurements, with the capacity to return preserved samples to Earth. Achieving these experimental goals requires a specialized, robust and reliable hardware system. The ExocubeBio hardware testing described here includes assessment of material biocompatibility and durability, functional validation of the miniaturized fluidic system, and optimization of the integrated optical subsystem for optical density and fluorescence measurements. These results demonstrate that the ExocubeBio experimental hardware components can each execute their core functional and operational requirements; subsystems allow for sample exposure, in-situ measurements of microbial cultures, and the chemical preservation of samples for post-flight analysis. As ExocubeBio transitions from hardware development to mission readiness, the results presented here validate the overall design and engineering approaches utilized. By combining the strengths of in-situ monitoring and sample return, ExocubeBio represents a significant advancement in space-based experimentation, and will provide new insights into microbial responses to the space environment.

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Coral restoration alters reef soundscapes but machine learning and manual analyses suggest different recovery rates

Croasdale, E. M.; Saponari, L.; Dale, C.; Shah, N.; Williams, B.; Lamont, T. A. C.

2026-04-02 ecology 10.64898/2026.03.31.710564 medRxiv
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Coral restoration is recognised as a critical tool to mitigate pantropical degradation of reef ecosystems. Robust monitoring of restoration progress is crucial for projects to evaluate their success, improve practice, and share knowledge. However, traditional visual surveys often fail to capture the full impact of coral restoration on reef function. Therefore, we employed Passive Acoustic Monitoring (PAM) to assess whether the soundscape of a coral restoration site in the Seychelles differs from adjacent healthy and degraded reference reefs. We applied two methods of soundscape analysis: manual detection of unidentified fish sounds; and machine learning-based Uniform Manifold Approximation and Projection analysis. Results were approach-specific: the manual approach highlighted similarities in fish calls between the restoration site and the healthy reference reef, while the machine learning approach extracted broader soundscape patterns, clustering the restoration site alongside the degraded reference reef. Although this is a single-site study, these findings suggest that a) coral restoration alters reef soundscapes, though recovery time may be taxon-specific, and b) multiple metrics are needed to bridge single-taxon and broad soundscape scales. This study contributes to the evolving field of soundscape ecology in coral reef ecosystems, highlighting the utility of PAM in monitoring changes to reef function through coral restoration.

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Phytoplankton size structure and biogeochemical responses to nutrient enrichment in an oligotrophic coral reef

Suarez-Caballero, J. L.; Nakamura, T.

2026-04-08 ecology 10.64898/2026.04.06.716629 medRxiv
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Tropical coral reef ecosystems worldwide are being impacted by combined pressures of climate change and human activities that introduce large quantities of nutrients and sediments into coastal areas. In this context, phytoplankton represent a critical link between dissolved inorganic nutrients and coral reef food webs, yet their role in these ecosystems remains understudied. We investigated ecological responses of the summer phytoplankton community of Shiraho Reef (Ishigaki Island, Okinawa, Japan) to nutrient enrichment using field-based microcosm experiments under natural light and temperature conditions in September 2022 and 2023. Treatments included single and combined additions of nitrogen, phosphorus, and silicon. Chlorophyll a (Chl a) concentrations increased after three days under combined nutrient conditions, whereas single-nutrient additions produced limited responses, indicating a strong co-limitation by nitrogen and phosphorus in the reef. Analysis of size-fractionated Chl a revealed shifts from picophytoplankton that typically dominate tropical oligotrophic ecosystems toward larger groups supported by enhanced nutrient availability. Our results show short-term impacts of nutrient enrichment events on phytoplankton size structure and biogeochemical cycling in coral reefs, and highlight the importance of pelagic processes in coral reef carbon dynamics under nutrient-enrichment.

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Real-time, in situ fluorescence and optical density measurements of liquid cultures in simulated microgravity

Lantin, S.; Bansal, M.; Alper, H.; Lee, J. A.

2026-03-25 microbiology 10.64898/2026.03.23.713711 medRxiv
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As human space exploration expands to the Moon, Mars, and beyond, there is a growing need to study the effects of altered gravity on the microbial systems that we will bring with us for life support. Because spaceflight experiment opportunities are rare and resource-intensive, most space biology experiments are conducted using ground-based simulators. The most common microgravity simulator for microbial experiments, the rotating wall vessel, can approximate the low-shear and low-turbulence conditions that characterize microgravity. However, current designs do not allow for real-time measurement of growth or metabolic activity during rotation: experiments require destructive sampling or disruption of the microgravity simulation conditions. Here, we describe the development of an in situ spectroscopy system compatible with the Cell Spinpod rotating wall vessel, which enables measurement of both optical absorbance and fluorescence with high temporal resolution, producing growth curves similar to those from an off-the-shelf plate reader. These results are validated using two common microbial hosts: Escherichia coli and Saccharomyces cerevisiae. The Spinpod Optical System has the potential to diversify the types of microbiology experiments possible in simulated microgravity, allowing the measurement of not only growth curve parameters but also metabolic activity, gene expression, or community dynamics. It thus has the potential to improve the quality of experiments seeking to characterize microbial responses to spaceflight conditions.

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The influence of pH on the growth and on the formation of nutrient-stress induced scum-forming blooms in cyanobacterial cultures

Dervaux, J.; Brunet, P.

2026-04-09 plant biology 10.64898/2026.04.07.716915 medRxiv
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The growth of cultures and formation of mucilage blooms in reaction to salt stress of cyanobacterial cultures are investigated with a focus on the influence of pH. In non-buffered medium, cultures show their pH increasing from 6.5 just after inoculation, up to 11 during the exponential phase. We record the time-evolution of concentration and pH, with different initial OD0. In a second set of experiments, we extract the doubling time of the unbuffered cultures in comparison with those inoculated in pH-buffered BG11 media at four different pH from 6.3 to 10.5 : in the most acid media, all cultures die or grow very slowly. At pH = 10.5, we obtain the fastest growth for all four strains, allowing to qualify these cyanobacteria as being alkaliphiles, though for all strains with comparable initial OD0, the doubling time is shorter for unbuffered cultures. Following a previous study [31]), we finally investigate the influence of pH on mucilage formation and biomass uplift induced by salt stress, involving EPS floculation by cations. Our results show that operating in buffered media significantly influences the mucilage formation, though the observed regimes cannot be simply correlated to the pH value.

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Integrating conventional tagging and acoustic telemetry improves estimates of post-release survival in a highly targeted reef fish

Hyman, A. C.; Collins, A.; Ramsay, C.; Allen, M. S.; Wilms, S.; Barbieri, L.; Frazer, T. K.

2026-03-20 ecology 10.64898/2026.03.16.711647 medRxiv
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Accurate estimation of post-release survival is fundamental to fisheries stock assessment and effective management. Conventional tag-return studies and acoustic telemetry are commonly used to estimate this probability, yet each approach has limitations when applied independently. Using gag (Mycteroperca microlepis) as a case study, we integrated data from a large-scale conventional tagging program and an acoustic telemetry experiment within a discrete-time statistical modeling framework that links relative recapture risk with telemetry-derived fate. This approach enabled estimation of post-release survival across a broad gradient of capture depths representative of recreational fishing conditions. Estimated survival was high in shallow waters ({approx}97%) but declined with increasing capture depth, consistent with depth-related barotrauma. Applying model predictions to depth distributions from the recreational fishery yielded annual and monthly post-release survival probabilities. Annual estimates were consistent with values assumed in recent stock assessments, while monthly values highlighted seasonal patterns potentially relevant for management. This integrated framework advances post-release survival estimation by combining the extensive sample sizes and environmental coverage characteristic of conventional tagging data with the direct fate observations provided by acoustic telemetry, and offers a transferable approach for other highly targeted fisheries.

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Impact of Regularization Methods and Outlier Removal on Unsupervised Sample Classification

Heckman, C. A.

2026-04-10 bioinformatics 10.64898/2026.04.07.716815 medRxiv
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BackgroundHigh-content assays (HCAs) have problems distinguishing biologically significant effects from the incidental effects of non-repeatable technical factors. Non-repeatable results are attributed to variations in the cell culture environment and the numerous, heterogeneous descriptors evaluated. The aim here was to determine whether preprocessing operations impacted the reproducibility of class assignments of experimental data. MethodsBatch effects that could affect reproducibility, i.e., signal/noise ratio, instrumental conditions, and segmentation, were controlled variables. The remaining batch effects, variations in materials, personnel, and culture environment could not be controlled. Descriptors values were measured directly from images. Exploratory factor analysis was used to solve the identifiable and interpretable feature, factor 4. In each of five trials, one sample was treated with the same chemical mixture (EXP) and another with the solvent vehicle alone (CON). ResultsRepeated CON and EXP samples showed significant differences among factor 4 means in data regularized within each trial. The mean of Trial 3 CON differed significantly from all other CON samples. These differences disappeared upon regularization to comprehensive databases. Among repeated EXPs, the Trial 2 mean differed from three other EXPs, but regularization to comprehensive databases had little effect. However, classification patterns were unchanged after regularization to any comprehensive database derived by the same protocol. After regularization to datasets derived by two different protocols, the classification pattern differed but only reflected elevation of differences that had been marginal to statistical significance. Outlier removal was deleterious. Even with the most sparing definition of outliers, over 3% of a single samples contents were removed from most trials. Elimination based on the overall within-trial distributions caused type I and type II errors. ConclusionsNon-repeatable factor 4 means in repeated trials had negligible influence on classification outcomes, so repeatability may not be a good indicator of assay quality. Irreducible batch effects, combined with small sample sizes and skewed distributions of descriptors values, may account for non-repeatability. As the current results are based on real-world data, they suggest that non-repeatability is an uncorrectable feature of these assays. Classification patterns are not affected by several irreducible technical factors, namely materials, personnel, and non-repeatable environmental variables.

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Status of Round Goby Invasion Fronts in New York and Quebec: Implications for Lake Champlain

George, S. D.; Diebboll, H. L.; Pearson, S. H.; Goldsmit, J.; Drouin, A.; Vachon, N.; Cote, G.; Daudelin, S.; Bartron, M. L.; Modley, M. D.; Littrell, K. A.; Getchell, R. G.; Fiorentino, R. J.; Sadekoski, T. R.; Finkelstein, J. S.; Darling, M. J.; Parent, G. J.; Atkins, L. M.

2026-03-25 ecology 10.64898/2026.03.23.712452 medRxiv
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Invasive round goby Neogobius melanostomus have advanced eastward through the state of New York and provinces of Ontario and Quebec over the past two decades and are approaching Lake Champlain, one of the largest lakes in North America. This manuscript describes international efforts to monitor round goby populations during 2021-2025 on (a) the southern approach to Lake Champlain via the Hudson River and Champlain Canal, and (b) the northern approach to Lake Champlain via the Saint Lawrence River and Richelieu River. Monitoring utilized environmental DNA (eDNA), backpack electrofishing, beach seining, benthic trawling, and viral hemorrhagic septicemia virus (VHSV) testing. In the Champlain Canal, round goby were captured as far north as the downstream side of the C1 dam (97 kilometers [km] from Lake Champlain) while eDNA detections occurred as far north as the upstream side of the C2 dam (90 km from Lake Champlain). In the Richelieu River, round goby were captured as far south as Saint-Marc-sur-Richelieu (82 km from Lake Champlain) while the southern-most eDNA detections occurred near the Canadian side of the international border (4 km from Lake Champlain). Water temperature influenced habitat usage of round goby in the Champlain Canal, with catch rates in near-shore areas declining at < 10 {degrees}C. All VHSV test results were non-detections at the mouth of the Richelieu River, while one positive and two inconclusive results occurred along the Champlain Canal. Together, these data have informed multiple mitigation measures and have implications for management of aquatic invasive species across North America.

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A comprehensive reference database to support untargeted metabolomics in Pseuudomonas putida

Ross, D. H.; Chang, C.; Vasquez, J.; Overstreet, R.; Schultz, K.; Metz, T.; Bade, J.

2026-03-24 bioinformatics 10.64898/2026.03.20.713193 medRxiv
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Pseudomonas putida strain KT2440 is a crucial model organism for synthetic biology and bioengineering applications, yet there currently exists no comprehensive metabolomics database comparable to those available for other model organisms. This gap hinders the use of untargeted metabolomics for exploratory analyses in this system. We developed the P. putida metabolome reference database (PPMDB v1) to address this limitation by consolidating metabolite information from multiple sources and expanding coverage through computational predictions. The database was constructed by curating metabolites from BioCyc, BiGG, and other literature sources, then computationally expanding this collection using BioTransformer environmental transformation predictions to generate additional predicted metabolites. We enhanced the databases utility for molecular annotation in metabolomics studies by incorporating analytical properties including collision cross-sections, tandem mass spectra, and gas-phase infrared spectra. These analytical properties were gathered from existing measurement data or predicted using computational tools. We further augmented the database through inclusion of reaction information and pathway annotations, facilitating biological interpretation of metabolomics data. This publicly available resource fills a critical gap in P. putida research infrastructure, supporting metabolite annotation and biological interpretation in untargeted metabolomics studies and enabling in-depth exploratory analyses of this important synthetic biology platform at the molecular level. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=110 SRC="FIGDIR/small/713193v1_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@c8828forg.highwire.dtl.DTLVardef@1f3a5c5org.highwire.dtl.DTLVardef@1084535org.highwire.dtl.DTLVardef@1f7ca4a_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Sublethal Behavioural and Neurotoxic Effects of Wastewater Effluent Exposure in a Freshwater Crustacean

Sandoval Herrera, N.; Johansson Kvarnström, E.; Lovin, L.; Fick, J.; McCallum, E. S.

2026-04-01 ecology 10.64898/2026.03.30.715298 medRxiv
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The increasing discharge of treated wastewater effluent poses a growing threat to freshwater ecosystems. Although wastewater treatment plants reduce chemical pollution, they do not fully remove many biologically active compounds. Behavioural responses in aquatic organisms provide sensitive and ecologically relevant indicators of sublethal contaminant exposure, offering insight into underlying physiological disruption and potential ecological consequences. Here, we examined the behavioural and neurotoxic effects of a seven-day experimental exposure to treated wastewater effluent in the noble crayfish (Astacus astacus). We quantified four ecologically important behaviours: (1) shelter use, a key antipredator strategy, (2) food seeking, (3) the ability to detect and respond to wastewater-associated olfactory cues, and (4) locomotor activity was assessed across all behavioural contexts. Cholinesterase (ChE) activity was measured as a biomarker of neurotoxicity. Exposure to wastewater effluent significantly altered crayfish behaviour. Exposed individuals exhibited higher locomotor activity compared to controls, exposed crayfish avoided areas containing wastewater cues, spending less time near the effluent source. Similarly, ChE activity was significantly reduced in exposed crayfish, indicating neurotoxic effects. The concurrence of ChE inhibition and behavioural modification suggests that effluent-derived contaminants may interfere with neural signalling pathways underlying crayfish locomotion and habitat selection. Overall, our results demonstrate that short-term exposure to treated wastewater effluent can induce both neurochemical disruption and ecologically relevant behavioural changes in A. astacus. Such alterations may increase vulnerability to predation and influence population dynamics in effluent-receiving waters, highlighting the importance of integrating behavioural endpoints with mechanistic biomarkers in assessing sublethal impacts of wastewater contamination.

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Robustness and management performance of MSY reference points derived from the hockey-stick stock-recruitment model under structural uncertainty

Ichinokawa, M.; Okamura, H.

2026-03-30 ecology 10.64898/2026.03.27.714336 medRxiv
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The hockey-stick (HS) stock recruitment relationship (SRR) has been widely used as an empirical alternative to conventional SRRs such as the Beverton-Holt (BH) and Ricker (RI) models. However, the management performance and risks associated with estimating maximum-sustainable-yield (MSY) reference points (RPs) based on HS remain insufficiently understood. This study first defines deterministic and stochastic MSY RPs under the HS model and provides an overview of their properties. We then conduct simulation experiments to investigate the bias and management consequences that arise when MSY RPs are estimated from the HS model (HS-derived MSY RPs) rather than from the true SRR (e.g., BH) across a range of biological and stochastic parameters, with particular focus on scenarios with insufficient data contrast. Our results show that HS-derived MSY RPs tend to exhibit higher bias but lower variance than MSY RPs derived from the true SRR. Management strategy evaluation simulations further reveal that management procedures combining HS-derived MSY RPs with adaptive model learning and some precautionary measures gradually reduce this bias and achieve average spawning biomass and yield that are comparable to those obtained under management based on the true BH SRR. We also show that the management effectiveness of the precautionary measures depends on life-history traits and recruitment variability. These findings indicate that although HS-derived MSY RPs may be biased and require cautious use, combining them with appropriate precautionary measures allows management to remain robust while limiting variability and yield losses. This broadens the range of management options that are available for supporting sustainable fisheries management.

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Eco-Friendly Antifouling Solutions: Hazard Assessment of Synthetic Derivatives of Natural Compounds

Leite, J. P.; Lima, E.; Pereira, D.; Cidade, H.; Correia-da-Silva, M.; Ruivo, R.; Santos, M.

2026-04-01 ecology 10.64898/2026.03.31.715569 medRxiv
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The accumulation of microorganisms and macroorganisms on aquatic surfaces poses economic and ecological challenges, particularly in maritime transport. Traditional antifouling methods, such as biocidal coatings containing toxic compounds like tributyltin (TBT) and copper, are effective but harmful to the environment. This study investigates eco-friendly antifouling alternatives, focusing on nature-inspired compounds (NIAFs) GBA 26 (GBA) and DPC345DHC (DH345), derived from polyphenols and flavonoids, respectively. The ecotoxicity of these compounds was evaluated using standardized assays with various species, including embryos of Danio rerio (zebrafish) (OECD TG 236), the algae Raphidocelis subcapitata (OECD TG 201), and the bacteria Vibrio fischeri (ISO 11348-2), along with nuclear receptor transactivation assays in Mytilus galloprovincialis (Mediterranean mussel). Gallic acid derivative GBA and 24h-transformation products showed low toxicity in zebrafish embryos, while dihydrochalcone DH345 inflicted developmental toxicity in zebrafish at 1 mg/L and above. Comparatively, tralopyril, a commercial biocide, exhibited significant toxicity at lower concentrations. Transcriptomic analysis of zebrafish embryos treated with GBA revealed selective gene modulation related to stress response, ion transport, and protein synthesis. Both, GBA and DH345, were shown to inhibit algae growth at 0.1 mg/L. Vibrio fischeri assay showed no toxic effects for any of the tested compounds. Nuclear receptor transactivation assays conducted with GBA revealed no activation of PPAR or PXR receptors. These findings suggest GBA and DH345 as potential eco-friendly antifouling agents with lower environmental risks than established antifoulants such as tralopyril. However, further research is needed to evaluate their potential long-term ecological impacts, particularly chronic toxicity across various organisms. This study advances the pursuit of sustainable antifouling solutions that prioritize environmental protection.

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Alkaline phosphatase activity supports heterotrophic carbon acquisition in a coastal time series site and a representative marine bacterium

Sachdev, E.; Adams, J. C.; Lanpher, K. B.; Perry, S.; Tostado, C.; Bowman, J. S.; Ingall, E. D.; Diaz, J. M.

2026-03-25 microbiology 10.64898/2026.03.24.713987 medRxiv
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Phosphorus is a vital nutrient required for the functioning of living organisms. In aquatic environments, dissolved inorganic phosphate is considered its most bioavailable form. However, phosphate can be scarce, which has the potential to limit microbial metabolism and ecosystem functioning. To overcome phosphate scarcity, microbes produce alkaline phosphatase (AP) to access dissolved organic phosphorus (DOP). Here, we conducted a year-long study of alkaline phosphatase activity (APA) at the Ellen Browning Scripps Memorial Pier, a nutrient-rich coastal site. APA was observed throughout the year despite phosphate-replete conditions, suggesting that the role of APs in microbial nutrition is not completely understood. We tested the hypothesis that APA may promote acquisition of organic carbon liberated from DOP hydrolysis by growing the heterotrophic marine bacterium Ruegeria pomeroyi on three DOP compounds as sole carbon sources and assessing APA. Controlling for carbon concentration, all DOP sources supported growth, but at lower levels than glucose, with the highest growth observed on glucose-6-phosphate (G6P), followed by adenosine monophosphate (AMP) and adenosine triphosphate (ATP). Moreover, cell-specific APA was significantly enhanced in carbon-deplete conditions and during growth on G6P, relative to cultures grown on replete glucose or nucleotides. These findings suggest alkaline phosphatases (APs) are part of a generic carbon stress response and likely play a role in acquiring certain forms of organic carbon by R. pomeroyi, with implications for other taxa. Overall, this study helps advance the current state of knowledge regarding microbial phosphorus cycling and carbon utilization in aquatic environments.