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The positive effect of fully protected Marine Protected Areas (MPAs) on marine biodiversity, specifically on fishes, has been widely documented. In contrast, the potential of MPAs to mitigate the impact of adverse climatic conditions has seldom been investigated. Here, we assessed the effectiveness of MPAs, quantified as increasing fish biomass, across wide geographic and environmental gradients across the Mediterranean Sea. We performed underwater visual surveys within and outside MPAs to characterize fish assemblages in 52 rocky reef sites across an extent of over 3,300 km. We used the steep spatial temperature gradient across the Mediterranean as a 'space-for-time' substitution to infer climate-driven temporal changes. We found that, as expected, Mediterranean MPAs increased fish biomass. At the same time, higher seawater temperatures are associated with decreased fish biomass, changes in species composition, and shifts towards more thermophilic species. Importantly, we found that the rate of decrease in fish biomass with temperature was similar between protected and fished sites. Taken together, these results suggest that the capacity of MPAs to harbor higher fish biomass, compared to surrounding areas, is maintained across a broad temperature range. At the same time, MPAs will not be able to offset larger-scale biotic alterations associated with climate change. Policy implications: Our results suggest that sustained warming will likely reduce fish biomass in the Mediterranean Sea and shift community structure, requiring more conservative targets for fishery regulations. At the same time, protection from fishing will remain an important management tool even with future high-water temperatures, and MPAs are expected to continue to provide local-scale benefits to conservation and fisheries.

Relating environmental DNA (eDNA) signal strength to organism abundance requires a fundamental understanding of eDNA production. A number of studies have demonstrated that eDNA production may scale allometrically – that is, larger organisms tend to exhibit lower mass-specific eDNA production rates, likely due to allometric scaling in key processes related to eDNA production (e.g. surface area, excretion/egestion). While most previous studies have examined intra-specific allometry, physiological rates and organism surface area also scale allometrically across species. We therefore hypothesize that eDNA production will similarly exhibit inter-specific allometric scaling. To evaluate this hypothesis, we reanalyzed previously published eDNA data from Stoeckle et al. (2021) which compared metabarcoding read count to organism count and biomass data obtained from trawl surveys off the New Jersey coast. Using a Bayesian model we empirically estimated the value of the allometric scaling coefficient (‘b’) for Northwestern Atlantic bony fishes to be 0.77 (credible interval = 0.64 – 0.92), although our model failed to converge for Chondrichthyan species. We found that integrating allometry significantly improved correlations between organism abundance and metabarcoding read count relative to traditional metrics of abundance (density and biomass) for bony fishes. Although substantial unexplained variation remains in the relationship between read count and organism abundance, our study provides evidence that eDNA production may scale allometrically across species in some contexts. Future studies investigating the relationship between eDNA signal strength and metrics of fish abundance could potentially be improved by accounting for allometry; to this end, we developed an online tool that can facilitate the integration of allometry in eDNA/abundance relationships. This is a curated dataset obtained from: Stoeckle, M.Y., Adolf, J., Charlop-powers, Z., Dunton, K.J., Hinks, G., and Vanmorter, S.M. 2021. Trawl and eDNA assessment of marine fish diversity, seasonality, and relative abundance in coastal New Jersey, USA. ICES J. Mar. Sci. 78(1): 293–304. doi:10.1093/icesjms/fsaa225. We separated Chondrichtyan and Osteichthyan species into separate datasets, including only species which were detected using eDNA. Values of 0.001 were added to 'zeroes' for abundance data, due to the inclusion of a heteroscedastic residual error term in some of the models.

The mucus covers the fish's body, working as a protective barrier. Besides physical protection, mucus provides molecules that protect the fish from pathogens damaging 1,2. These include antimicrobial peptides secreted in the mucus, which play an essential role in defense against microbial pathogens since these belong to the innate immune system2,3. In this study, two adult Halobatrachus didactylus individuals were captured from the wild in Sesimbra. Then, mucus collection was performed by scraping the dorsal-lateral body of the fish with a sponge. Our objective was the identification of new peptides with bioactive potential in mucus samples by chromatography analysis. Size exclusion highperformance liquid chromatography (SE-HPLC) analysis performed on mucus samples from the two individuals revealed a similar profile with an intense highlight peak which resulted in a distribution of about 775 Dalton. With interest in that peak, the two mucus samples were pooled for fractionation by SEC. The resulting fraction was analyzed by liquid chromatography-tandem mass spectrometry (LCMS/MS) to identify the most probable peptide sequences. Identification from databases did not provide reliable results, indicating a lack of information on the matrix analyzed. We resorted to de novo sequencing with good results using PEAKS Studio software. Five identified peptides were selected according to their bioactivities predicted in silico. Furthermore, the five identified peptides were synthesized, and the molecular size was validated by SE-HPLC analysis. Overall, this chromatographic approach enabled the identification of promising peptides, which bioactivities will be evaluated in vitro in future work.

Ocean drifters from oil-on-water exercise in North Sea (Frigg oil field) June 2019. Described in more detail in Brekke, C., Espeseth, M. M., Dagestad, K.-F., Röhrs, J., Hole, L. R., & Reigber, A. (2021). Integrated analysis of multisensor datasets and oil drift simulations - a free-floating oil experiment in the open ocean. Journal of Geophysical Research: Oceans, 126, e2020JC016499. https://doi.org/10.1029/2020JC016499 Work is funded by grant no. 237906 (CIRFA) of the Norwegian Research Council. Work is funded by grant no. 237906 (CIRFA) of the Norwegian Research Council

The monitoring of the fish community in Doñana wetlands was initiated in 2004 as part of the Monitoring Program of Natural Resources and Processes. The aim was to obtain a temporal and continuous series of data in the abundance and distribution of fish species to analyze the evolution of their numbers and estimates biodiversity values. Data were recorded annually between 2004-2019 by more than 2 members of the monitoring team which performed samplings in different locations twice per year in winter-spring and summer seasons when the study sites are flooded. The fishes were sampled at the 139 stations classified according to their location (on either aeolian sands or marshland). Funnel traps were used as a sampling method. Between 5-9 funnel traps were randomly distributed (until 50 cm of depth) in each location, depending of the flooded area and depth. The traps were left for 24 hours and emptied the content into white sorting pans. Individuals were counted and identified until the maximun taxonomic level in the field and realease. During samplings, it was identified 15 families. The most abundances were Poecilidae and Cyprinidae. Data recorded during the surveys included species identification, number of individuals, sex and life stage (pupa, larvae, inmature, mature) of the organisms when possible, as well as the time and georreferenced data of the observation. Between 2004-2007 data was registered in Excel file and since 2008 data was recorded in CyberTracker sequence). The protocol used has been supervised by researchers and the data have been validated by the members who performed the sampling. 1. Don_fish_ev_20221222: eventID, intitutionCode, institutionID, datasetName, eventDate, year, month, day, country, stateProvince, location, localityID, locality, decimalLatitude, decimalLongitude, habitat, sampleSizeUnit, sampleSizeEffort, DynamicPropiertiesEvent, eventRemarks, recordeBy.-- 2. Don_fish_occ_20221222: eventID, occurrenceID, individualCount, sex, lifeStage, kingdom, phylum, order, family, genus, specificEpithet, scientificName.-- 3. Don_fish_mof_20221222: OccurrenceID, measurementID, measurementType, measurementValue, measurementUnit, measurementMethod. Dataset are structured following well-established data formats. Three files are provided. The first file (Don_fish_ev_20221222) contains the information of each event (eventID, event date, geographical coordinates, sample effort, etc…); the second file (Don_fish_occ_20221222) contains the information of the occurrences of fish species recorded in each station, taxonomic classification; and the third file (Don_fish_mof_20221222) provide information of the biometric variable (weight) of fish sample in each occurrence. We acknowledge financial support from National Parks Autonomous Agency (OAPN) between 2002-2007; Singular Scientific and Technical Infrastructures from the Spanish Science and Innovation Ministry (ICTS-MICINN); Ministry of Agriculture, Livestock, Fisheries and Sustainable Development from the Regional Government of Andalusia (CAGPDES-JA) since 2007; and Doñana Biological Station from the Spanish National Research Council (EBD-CSIC) since all the study period (2005). Peer reviewed

In April of 2007, field surveys of fish assemblages were conducted at 31 locations. Each sampling location was a 200-500 m stretch encompassing all types of geomorphological forms in the river channel (e.g. riffles, runs, and pools). Two commonly utilized electrofishing protocols were applied to maximize the capture of fish. Specifically, for shallow water locations, fish specimens were collected by means of backpack electrofishing. The crew sampled the reach in an upstream direction with two passes. For non-wadable areas, boat electrofishing was applied by moving the boat slowly in a downstream direction. All collected fish were identified to species level, measured, and weighed. Most individuals were released to the sampling sites, and a subset of fish species was fixed in 7% formalin for final preservation.Several environmental variables were recorded in situ after fish collection. Water temperature (℃), pH, dissolved oxygen (mgL-1), and conductivity (µScm-1) were measured by using a multi-parametric probe (WTW Multi 340i). Water depth (m) and channel width (m) were calculated using a depth sounder, and a Leica CRF900 rangefinder camera, respectively. An altimeter was used to measure altitude (m). Current velocity (ms-1) was determined using a flowmeter device. The percentage of substrate particle size was visually estimated and divided into four types: sand, silt, cobble, and boulder. File description:fish_abundance_data.csv31 sites (rows) × 62 species (columns) community matrix fish_trait_data.csv62 species (rows) × 9 functional traits (columns) matrix site_environment_data.csv31 sites (rows) × 12 environmental variables (columns) matrix fish_model_data.csvoccupancy, total mean abundance, niche position, niche breadth, trait vector 1, trait vector 2, trait vector 3, trait vector 4 and one phylogenetic vector for each fish species

In situ primary productivity based on 14C assimilation and nutrients from samples collected by many research projects and on numerous cruises globally, 1985-2008.

We combine consistently dated benthic carbon isotopic records distributed over the entire Atlantic Ocean with numerical simulations performed by a glacial configuration of the Norwegian Earth System Model with active ocean biogeochemistry, in order to interpret the observed Cibicides δ13C changes at the stadial-interstadial transition corresponding to the end of Heinrich Stadial 4 (HS4) in terms of ocean circulation and remineralization changes. We show that the marked increase in Cibicides δ13C observed at the end of HS4 between ~2000 and 4200 m in the Atlantic can be explained by changes in nutrient concentrations as simulated by the model in response to the halting of freshwater input in the high latitude glacial North Atlantic. Our model results show that this Cibicides δ13C signal is associated with changes in the ratio of southern-sourced (SSW) versus northern-sourced (NSW) water masses at the core sites, whereby SSW is replaced by NSW as a consequence of the resumption of deep water formation in the northern North Atlantic and Nordic Seas after the freshwater input is halted. Our results further suggest that the contribution of ocean circulation changes to this signal increases from ~40 % at 2000 m to ~80 % at 4000 m. Below ~4200 m, the model shows little ocean circulation change but an increase in remineralization across the transition marking the end of HS4. The simulated lower remineralization during stadials than interstadials is particularly pronounced in deep subantarctic sites, in agreement with the decrease in the export production of carbon to the deep Southern Ocean during stadials found in previous studies.

These data are fish surveys from May to December 2021 following the construction of the clusters of artificial reefs. There are three clusters, each with 9 artificial reefs all of which are spaced the same distance and each differs per cluster (1 meter, 3 meters, and 5 meters). Reefs were constructed in less than 4 meters of water in the Bight of Old Robinson, on Abaco, The Bahamas.

Reference datasets (Nov2022 update) for Mitohelper (https://github.com/aomlomics/mitohelper) Mitohelper is a repository built to facilitate experimental design, alignment visualization, and reference sequence analysis in fish eDNA studies. Refer to our paper and Mitohelper's wiki for database construction pipeline. I. Reference database files in tab-separated format, containing gene, taxonomy, and sequence information: mitofish.all.Nov2022.tsv (776,210 records) mitofish.12S.Nov2022.tsv (44,560 records) mitofish.12S.Nov2022_NR.fasta (fasta file of 12S rRNA gene records) mitofish.COI.Nov2022.tsv (314,143 records) II. De-replicated QIIME 2-compatible 12S/12S+16S+18S rRNA reference datasets: 12S-seqs-derep-uniq.qza 12S-tax-derep-uniq.qza 12S-16S-18S-seqs.qza 12S-16S-18S-tax.qza If you use Mitohelper, please cite: Jean Lim, S, Thompson, LR. Mitohelper: A mitochondrial reference sequence analysis tool for fish eDNA studies. Environmental DNA. 2021; 00: 1– 10. https://doi.org/10.1002/edn3.187 Major update: The 12S rRNA gene sequence dataset is now filtered to only contain mitochondrial genomes annotated with 12S rRNA gene sequences. Sequences of the 12S rRNA gene are now extracted from complete mitochondrial genomes to construct a more gene-specific 12S rRNA dataset. 12S rRNA gene sequences in mitohelper's dataset are available for download as mitofish.12S.Nov2022_NR.fasta