• Energy Research

Individuals of Astarte crenata, Ctenodiscus crispatus and Cistenides hyperborea were obtained from replicate SMBA (Scottish Marine Biological Association) box cores and 15 minute Agassiz trawls in the Barents Sea (stations B13, 74.3 degrees N, 30.0 degrees E; B16, 80.3 degrees N, 30.0 degrees E; JCR18006, RSS James Clark Ross) in July 2019. Individuals of the Aequiyoldia eightsi and Laternula elliptica were collected by SCUBA-assisted divers at Rothera Point, Adelaide Island, West Antarctic Peninsula (67.3 degrees S, 68.1 degrees W) in March-April 2019. Surficial sediment (less than 5 cm depth) for the Arctic species was collected using SMBA box cores at stations B13, B14 (76.3 degrees N, 30.3 degrees E) and B16 in July 2019 and, for the Antarctic species, by hand from the Hamble, UK (50.9 degrees N, 1.3 degrees W). Sediment was sieved (500 micrometer mesh) within a seawater bath to retain the fine fraction and remove macrofauna and debris. Species were distributed across 42 clear acrylic aquaria (internal LWH: 12 x 12 x 33 cm, 3 replicates treatment-1: species × location × climate scenario), designed to accommodate the appropriate density (Arctic, 2 ind. mesocosm-1; Antarctic, 1 ind. Mesocosm-1; size and burrowing requirements of each species (sediment depth: A. crenata, C. crispatus & C. hyperborea, 16 cm; A. eightsi, 12cm; L. elliptica, 19cm), and exposed to ambient (1 ± 0.5 degrees Celsius, around 400 ppm [CO2]) and indicative near-future (3 ± 0.5 degrees Celsius, around 550 ppm [CO2]) environmental conditions. Aquaria were randomly placed within one of two insulated seawater reservoirs (LWH: 1.2 x 1.2 x 0.8m, Tanks Direct, UK). All aquaria were overlain with seawater (salinity 33, 10 micrometer sand filtered, UV sterilized) to a combined sediment and water depth of 31 cm and maintained in the dark. After acclimation to ambient mesocosm conditions (21 days, 09-29/09/2019), the water temperature and atmospheric CO2 of the aquaria in the future treatment was gradually increased at 0.5 degrees Celsius and 50 ppm increments every 7 days to reach the desired near-future climate conditions. During both the acclimation and experimental period (92 days, 21/10/2019 - 21/01/2020), C. crispatus and C. hyperborea were fed ad libitum once a week with commercially available fish food (Aquarian Tropical Flake; 0.03 g), and A. crenata, A. eightsi and L. elliptica were fed ad libitum three times per week with 100 ml of precultured phytoplankton (33:33:33 mix, Isochrysis sp., Tetraselmis sp., and Phaeodactylum sp.; Algal Culture Laboratory, National Oceanography Centre Southampton). To avoid accumulation of excess food and associated nutrients, partial seawater changes (weekly 50 percentage seawater exchange) were undertaken. Temperature, pH, salinity and total alkalinity were periodically measured across all experimental mesocosms (T,pH,S: weekly, AT: week 2, week 6, week 11). Total alkalinity analysis followed the standard HCl titration protocol of the Carbonate Facility at the University of Southampton. From which, DIC, [pCO2], [calcite], [aragonite], [HCO3] and [CO3] were calculated using the CO2calc software. CO2(ppm), H2O(ppt), H2O(C), Cell_Temperature(C), Cell_Pressure(kPa), CO2_Absorption and H2O_Absorption were measured continuously in the ambient air and future climate regime gas mixture from the start (21/10/2019) to end (21/01/2020) of the experimental period. All measurements were calculated using an Infra-red gas analyzer LiCOR LI-820 model and recorded using the interface software on a standard laptop. Behavioural activity was quantified using measurements of movement and burial behaviour at the sediment surface. Individuals (morphology, +/- 0.01 mm; blotted wet weight, +/- 0.001 g) were placed in temperature-acclimatised viewing trays containing sediment (depth 5 cm) overlain with sea water (depth 3 cm) and viewed (less than or equal to 60 minutes) with a benchtop video camera (Logitech C920 HD Pro Webcam, 1080p). The time taken for each individual to initiate movement (response time, s) and to complete burial (burial time, s) was recorded (SkyStudioPro time-lapse software; 3 frame s-1, autosave every 300 frames) and analysed frame by frame (VLC Media Player). Faunal mediated sediment particle reworking metrics were estimated at the end of the experimental period by establishing the redistribution of optically distinct particulate tracers (luminophores: 30g aquaria-1, fluorescent green, 125 - 250µm silica sand,; Glass Pebbles Ltd., UK). Luminophores were evenly distributed across the sediment surface immediately after the partial seawater change. After 12 days, images (RGB colour, JPEG compression) of the vertical redistribution of luminophores were taken from each side of each aquarium using a digital SLR camera. Extracted profile data was obtained using a custom-made semi-automated macro that runs in ImageJ (version 1.47s, released 3rd June, 2013), a java-based public domain program developed at the US National Institutes of Health (http://rsb.info.nih.gov/ij/index.html). The ventilatory behaviour of the infauna (bioirrigation) was estimated from absolute changes in the concentration (10mM, 1.0289g NaBr dissolved per 1l seawater mesocosm-1) of the inert tracer sodium bromide (Delta [Br-], mg L-1; negative values indicate increased infaunal ventilatory activity) over an 8 h period on day 12, determined using a Tecator flow injection auto-analyser (FIA Star 5010 series). Accumulated water column concentrations (delta micromol L-1) of NH4-N, NOx-N (i.e. NO3-N + NO-N) and PO4-P were determined at the start and end of the experimental period from standardized samples (taken from the centre of each mesocosm at around 5cm depth, 0.45 micrometer NALGENE filtered) following standard protocols using a Lachat Quikchem 8500 flow-injection auto-analyser. Sediment was analysed using a Malvern Mastersizer 2000 He-Ne LASER diffraction sizer for sediment particle size at the University of Cambridge following a standard protocol (provided). Sediment organic material content (percentage) was analysed at the University of Cambridge following a standard protocol (provided). Full descriptions and additional information provided in Williams et al. [submitted for publication] in main manuscript and supplementary information. Behaviour: Logitech C920 HD Pro Webcam, 1080p Behaviour: SkyStudioPro time-lapse software V 1.1.0.30 Behaviour: VLC Media Player V 3.0.12 Sediment particle reworking: Canon 400D digital SLR camera Sediment particle reworking: ImageJ (version 1.47s) Bioirrigation: Tecator flow injection auto-analyser (FIA Star 5010 series) Nutrients: Lachat Quikchem 8500 flow-injection auto-analyser Total Alkalinity: Apollo SciTech Alkalinity Titrator AS-ALK2 Salinity: WTW (TM) TetraCom (TM) 325 Standard temperature-conductivity combination electrode Temperature: NBS scale, Mettler-Toledo InLab Expert Pro temperature-pH combination electrode pH: NBS scale, Mettler-Toledo InLab Expert Pro temperature-pH combination electrode pCO2, CO3, HCO3, Ωcalcite, Ωaragonite, TCO2: Calculated using the CO2calc software [version 4.0.9] CO2(ppm), H2O(ppt), H2O(C), Cell_Temperature(C), Cell_Pressure(kPa), CO2_Absorption and H2O_Absorption : Infra-red gas analyzer LiCOR Li-820 CO2(ppm), H2O(ppt), H2O(C), Cell_Temperature(C), Cell_Pressure(kPa), CO2_Absorption and H2O_Absorption: ACER laptop CO2(ppm), H2O(ppt), H2O(C), Cell_Temperature(C), Cell_Pressure(kPa), CO2_Absorption and H2O_Absorption: LiCOR Windows software Sediment: Malvern Mastersizer 2000 He-Ne LASER diffraction sizer Measurements were collected during the 92 day lab-based experiment carried out on replicate macrofaunal mesocosms of Astarte crenata, Ctenodiscus crispatus, Cistenides hyperborea from the Western Barents Sea and Aequiyoldia eightsi and Laternula elliptica from Rothera Point, Antarctic Peninsula. Macrofauna were collected on cruise JR18006 in the Western Barents Sea and by hand during scuba dives in the cove adjacent to Rothera research station. Experimental measurements include carbonate chemistry indices measured in water samples; carbon dioxide concentration levels; benthic invertebrate response rate, burial rate, sediment particle reworking, bioirrigation, and associated nutrient concentrations; benthic invertebrate body size and individual biomass; sediment organic material content, and sediment particle size. Supported by the INSPIRE Doctoral Training Partnership programme (grant NE/S007210/1) and 'The Changing Arctic Ocean Seafloor (ChAOS) - how changing sea ice conditions impact biological communities, biogeochemical processes and ecosystems' project (NE/N015894/1 and NE/P006426/1, 2017/2021), Natural Environment Research Council (NERC) in the UK. Standard protocols were followed and data entry double checked by independent person. Missing values for behavioural activity (response rate, burial rate) relate to the individual not completing the associated activity within the 60 minute timeframe. Missing biomass data is due to mortalities. Original unedited raw videos and processed, stitched together timelapses are provided. Original unedited raw fSPI images and processed, stitched together fSPI profiles are provided. 3 replicate water samples were taken for each independent treatment level in the experiments. Standards and blanks were utilised when analysing total alkalinity as per the HCl protocol. Standard protocols were followed and data entry double checked by independent person. LiCOR instruments were calibrated 3 months prior to the start of the experimental period.

Experiments were conducted between 2006 and 2015 with individuals hand collected by SCUBA divers in the austral summer, from 6 - 15m depth, near Rothera Research Station, Adelaide Island (67° 34' 2'' S, 68° 08' 0'' W). Common marine ectotherms were selected from different phyla to represent a range of functional groups, for which aquarium husbandry is well established and long term temperature limits have been estimated under normoxia. All animals remained submerged throughout the transfer from the sea to the flow-through aquarium system at the station. To control for size-dependent effects on survival we selected individuals of a similar size within each species group at the start of the experiment, and only studied fully reproductive adults. The 3.0 °C temperature-oxygen experiment was conducted in flow through aquaria at Rothera Research Station. In Rothera the tanks had a constant exchange of seawater that was balanced to allow temperature and oxygen treatments to be maintained while preventing any build-up of metabolic waste. In all experiments, seawater chemistry was monitored every 2-3 days using Nutrafin aquarium test kits. Ammonia, nitrite and nitrates were maintained well below 0.4, 0.2 and 5 mg L-1 to prevent toxicity from metabolic by-products. A small piece of one tentacle was removed from each anemone and preserved in 96% ethanol. DNA was extracted from each tentacle using the DNeasy Blood and Tissue kit (Qiagen) according to manufacturer's instructions. The cytochrome oxidase subunit I gene (COI) mitochondrial region was amplified using 1-2 µl extracted DNA and MyTaq DNA polymerase mix (30 µl reactions; Bioline UK (now Meridien Biosceince), with 10nmol each of the universal COI primers for invertebrates (LCO 1490 5'-GGTCAACAAATCATAAAGATATTGG-3'; HCO 2198 5''''''-TAAACTTCAGGGTGACCAAAAAATCA-3') (Folmer et al. 1994). PCR conditions were: 94 °C for 5 minutes, 5 cycles of 94 °C for 1 min, 45 °C for 1.5 min, 72 °C for 1.5 min, followed by 30 cycles of 94 °C for 1 min, 50 °C for 1 min, 72 °C for 1 min and a final elongation stage of 5 min at 72 °C. The COI fragments were bi-directionally sequenced by Source Bioscience (Cambridge, UK). The species identity of each individual was analysed using Blast sequence similarity searching of INSDC (International Nucleotide Sequence Database Collaboration) (https://www.insdc.org/). Temperatures were raised at the same rate in all experiments (0.3 ± 0.1 °C d-1) until the incubation temperature was reached and subsequently monitored daily. Animals were held at temperatures of 6.0 and 8.0 °C, temperatures that were 0 to 4 °C above the long-term limits calculated from experiments with different rates of warming. Control animals were kept at 0.0 °C in the main holding aquarium. Acute thermal limits: Individuals that survived beyond the duration of incubations at both 6.0 and 8.0 °C were then tested to see if acute thermal limits were elevated due to acclimation to these elevated incubation temperatures. These data were compared with acute thermal limits conducted on control individuals that had been kept in the aquarium at 0.0 °C. For the acute temperature ramping trials individuals were transferred to plastic jacketed tanks (Engineering Design and Plastics Ltd), whose jackets were filled with 25 % v/v ethanol in water solution, that was heated or cooled by LTD20G thermocirculators (Grant Instruments). Temperatures were increased at 1.0 ± 0.1 °C d-1 until the last animal was no longer responding to the stimuli detailed above. Acclimatisation in field animals was tested for in only one species, the common starfish O. validus. To test for field acclimatisation the assessment of CTmax of freshly collected O. validus was repeated between 2006 and 2015 in both summer and winter. There was also a test of individuals from Scott Base (77° 50' 5'' S, 166° 46' 0'' E) to allow for regional differences and compare with a site with even less annual and seasonal variation than Rothera. As above, temperatures were increased at 1.0 ± 0.1 °C d-1 until the last animal was no longer responding to the stimuli. For analysis field temperature from the Rothera Time Series was measured by CTD (conductivity, temperature, depth) casts at 15m depth and was averaged for the month of animal collection. Further details on methodology are in the associated manuscript. This data assesses the ability of 8 species, from 7 classes representing a range of functional groups, to survive, for 100 to 303 days, at temperatures 0 to 4 degrees Celsius above previously calculated long-term temperature limits. Survivors were then tested for acclimation responses to acute warming. Acclimatisation in the field was tested in the seastar Odontaster validus collected in different years, seasons and locations within Antarctica. Finally, we tested the importance of oxygen limitation in controlling survival duration by incubating 7 species under normoxia (20%) and mild hyperoxia (30%). This study was funded by Natural Environment Research Council core funding to the British Antarctic Survey and Spitfire DTP funding to R.E.S. All data have been quality checked by the data creators.

The data contains three supporting datasets: 1. Mid-intertidal data 2. Vertical transect data 3. GPS coordinates for all sites

AbstractMarine encrusting communities play vital roles in benthic ecosystems and have major economic implications with regards to biofouling. However, their ability to persist under projected warming scenarios remains poorly understood and is difficult to study under realistic conditions. Here, using heated settlement panel technologies, we show that after 18 months Antarctic encrusting communities do not acclimate to either +1 °C or +2 °C above ambient temperatures. There is significant up-regulation of the cellular stress response in warmed animals, their upper lethal temperatures decline with increasing ambient temperature and population genetic analyses show little evidence of differential survival of genotypes with treatment. By contrast, biofilm bacterial communities show no significant differences in community structure with temperature. Thus, metazoan and bacterial responses differ dramatically, suggesting that ecosystem responses to future climate change are likely to be far more complex than previously anticipated.

The biodiversity of marine and coastal habitats is experiencing unprecedented change. While there are well-known drivers of these changes, such as overexploitation, climate change and pollution, there are also relatively unknown emerging issues that are poorly understood or recognized that have potentially positive or negative impacts on marine and coastal ecosystems. In this inaugural Marine and Coastal Horizon Scan, we brought together 30 scientists, policymakers and practitioners with transdisciplinary expertise in marine and coastal systems to identify new issues that are likely to have a significant impact on the functioning and conservation of marine and coastal biodiversity over the next 5-10 years. Based on a modified Delphi voting process, the final 15 issues presented were distilled from a list of 75 submitted by participants at the start of the process. These issues are grouped into three categories: ecosystem impacts, for example the impact of wildfires and the effect of poleward migration on equatorial biodiversity; resource exploitation, including an increase in the trade of fish swim bladders and increased exploitation of marine collagens; and new technologies, such as soft robotics and new biodegradable products. Our early identification of these issues and their potential impacts on marine and coastal biodiversity will support scientists, conservationists, resource managers and policymakers to address the challenges facing marine ecosystems.

25 individuals were sequenced from the 5 sets of sampling/experimental work detailed above. All analyses were performed by Novogene and comprised the following stages and application of software packages. Raw reads were quality controlled for error rate using Phred and GC content using the Illumina CASAVA v1.8 software. Reads were cleaned with the removal of Illumina kit adapter sequences and adapter contamination, where the level of uncertain nucleotides N > 10% and where low quality bases as defined by the Phred Q20 score constituted more than 50% of a read. De novo transcriptome assembly was performed using Trinity version r20140413p1 with parameters min_kmer_cov=2, min_glue=2, others were set to default, using the modules Inchworm, Chrysalis and Butterfly (Grabherr et al., 2011). Hierarchical clustering was performed using the Corset program in Trinity to remove read redundancy. The longest transcripts from each cluster were selected as unigenes. Annotation of the unigenes was performed using seven databases (NR, NT, KO, SwissProt, Pfam, Go and KOG). Blast searching against NT was performed using NBCI blast 2.2.28+ with an e-value threshold of 1e-5 (Altschul et al., 1997). Diamond v0.8.22 (Buchfink et al., 2015) was used to blast search the unigenes against NR, SwissProt and KOG. The e-value threshold for NR and SwissProt was 1e-5 and 1e-3 for KOG. Pfam (Finn et al., 2008) was screened using the hmmscan package in HMMER v3.1b1 with an e-value threshold of 0.01. GO annotation was based on the protein annotation results from NR and Pfam using Blast2GO vb2g4pipe_2.5 (Goetz et al., 2008; Young et al., 2010) with an e-value threshold of 1e-6. KEGG mapping was performed using KAAS (KEGG Automated Annotation Server) v.r140224 with an e-value threshold of 1e-8 (Mao et al., 2005; Moriya et al., 2007; Kanehisa et al., 2008). GO enrichment was performed using GOSeqtopGO vGOSeq 1.32.0, topGO-2.32.0) with a corrected p value of <0.05. KEGG enrichment was performed using KOBAS v3.0 with a corrected p value of <0.05. The de novo transcriptome was used as a reference assembly and the reads from each library mapped back to the transcriptome and quantified using Bowtie2 vbowtie2-2.2.2.2 and RSEM vRSEM-v1.3.0 (Li et al., 2011) with output referenced as FPKM (Fragments Per Kilobase of transcript sequence per Million base pairs sequenced). The threshold for expression was set at FPKM >3.0. Differential expression between the different sets of samples was calculated using DEGseq v1.12.0 (Wang et al., 2010) with normalization via TMM and FDR calculated using BH (Benjamini and Hochberg, 1995) with output threshold of log2fold change >1 and adjusted p value <0.005. Protein gene identifiers were extracted from the SwissProt annotations for the MI v. MO differential expression analysis for analysis via the STRING v11 program (https://string-db.org/) to visualize protein-protein interactions (Szklarczyk et al., 2019). Transcriptomic analyses were undertaken on both in situ collected and experimentally warmed blue mussels (Mytilus edulis) from Greenland. M. edulis were collected from the Godthabsfjorden near Nuuk, Greenland (64.45555 -51.14416) at the following locations and dates: Inner fjord (64.45941, -50.31030) on 11/06/2018; outer fjord (64.19666, -51.69) on 13/06/2018, and sub-tidal (64.19666, -51.69) on 13/06/2018 (outer fjord at 20-40cm below the lowest low water mark). The in situ collected inner and outer fjord intertidal animals with outer fjord subtidal animals used as controls were collected at 27 °C, 19 °C and 3 °C, respectively. Some of the outer fjord subtidal M. edulis were experimentally warmed to 22 °C and 32 °C for one hour to mimic high aerial exposure temperatures in the inner and outer fjord intertidal, respectively. RNA-Seq was performed on 5 animals for each treatment, with all subsequent bioinformatics analyses performed by Novogene, China. This work was supported by the Carlsberg Foundation, the Independent Research Fund Denmark (Danmarks Frie Forskningsfond) (DFF-International Postdoc; case no. 7027-00060B), a Marie Sklodowska-Curie Individual Fellowship (IF) under contract number 797387 and Aage V. Jensens Fond (Aage V. Jensens Foundation) and NERC-UKRI core funding to the British Antarctic Survey.

Climate change is causing increased coastal freshening in Antarctica, leading to reduced salinity. For Antarctica’s endemic echinoderms, adapted to the stable polar environment, the impact of rapid reductions in coastal salinity on physiology and behaviour is currently unknown. Six common Antarctic echinoderms (the sea urchin Sterechinus neumayeri ; the sea star Odontaster validus ; the brittle star Ophionotus victoriae ; and three sea cucumbers Cucumaria georgiana , Echinopsolus charcoti and Heterocucumis steineni ), were directly transferred from ambient salinity (34.5‰) to a range of salinity dilutions (29–9‰) for 24 h. All species showed reduced activity and the establishment of a temporary osmotic gradient between coelomic fluid and external seawater. Most species exhibited a depression in oxygen consumption across tolerated salinities; however, at very low salinities that later resulted in mortality, oxygen consumption increased to levels comparable to those at ambient. Low salinity tolerance varied substantially between species, with O. victoriae being the least tolerant (24 h LC 50 (lethal for 50% of animals) = 19.9‰) while E. charcoti and C. georgiana demonstrated the greatest tolerance (24 h LC 50 = 11.5‰). These findings demonstrate the species-specific response of Antarctica’s endemic echinoderms to short-term hypoosmotic salinity events, providing valuable insight into this phylum’s ability to respond to an underreported impact of climate change.

Cet article présente les résultats de notre cinquième exercice annuel d'analyse de l'horizon, qui vise à identifier les sujets qui peuvent de plus en plus affecter la conservation de la diversité biologique, mais qui n'ont pas encore été largement pris en compte. Une équipe de scanners d'horizon professionnels, de chercheurs, de praticiens et d'un journaliste a identifié 15 sujets qui ont été identifiés via un processus itératif de type Delphi. Les 15 sujets comprennent un krach financier induit par le marché du carbone, l'expansion géographique rapide de la culture des macroalgues, le contrôle génétique des espèces invasives, la thérapie probiotique pour les amphibiens et une maladie fongique émergente des serpents. Este documento presenta el resultado de nuestro quinto ejercicio anual de exploración del horizonte, que tiene como objetivo identificar temas que pueden afectar cada vez más a la protección de la diversidad biológica, pero que aún no se han considerado ampliamente. Un equipo de escáneres profesionales de horizontes, investigadores, profesionales y un periodista identificaron 15 temas que se identificaron a través de un proceso iterativo similar a Delphi. Los 15 temas incluyen un colapso financiero inducido por el mercado de carbono, la rápida expansión geográfica del cultivo de macroalgas, el control genético de especies invasoras, la terapia probiótica para anfibios y una enfermedad fúngica emergente de serpientes. This paper presents the output of our fifth annual horizon-scanning exercise, which aims to identify topics that increasingly may affect conservation of biological diversity, but have yet to be widely considered. A team of professional horizon scanners, researchers, practitioners, and a journalist identified 15 topics which were identified via an iterative, Delphi-like process. The 15 topics include a carbon market induced financial crash, rapid geographic expansion of macroalgal cultivation, genetic control of invasive species, probiotic therapy for amphibians, and an emerging snake fungal disease. تعرض هذه الورقة مخرجات تمريننا السنوي الخامس لمسح الآفاق، والذي يهدف إلى تحديد الموضوعات التي قد تؤثر بشكل متزايد على الحفاظ على التنوع البيولوجي، ولكن لم يتم النظر فيها على نطاق واسع بعد. حدد فريق من ماسحات الأفق المهنية والباحثين والممارسين وصحفي 15 موضوعًا تم تحديدها من خلال عملية تكرارية تشبه عملية دلفي. تشمل الموضوعات الخمسة عشر الانهيار المالي الناجم عن سوق الكربون، والتوسع الجغرافي السريع لزراعة الطحالب الكبيرة، والتحكم الوراثي في الأنواع الغازية، والعلاج بالبروبيوتيك للبرمائيات، ومرض فطريات الثعابين الناشئة.