Kelp holdfast microclimates buffer invertebrate inhabitants from extreme temperatures.

Climate change is altering environmental conditions, with microclimates providing small-scale refuges within otherwise challenging environments. Durvillaea (southern bull kelp; rimurapa) is a genus of large intertidal fucoid algae, and some species harbour diverse invertebrate communities in their holdfasts. We hypothesised that animal-excavated Durvillaea holdfasts provide a thermal refuge for epibiont species, and tested this hypothesis using the exemplar species D. poha. Using a southern Aotearoa New Zealand population as a case-study, we found extreme temperatures outside the holdfast were 4.4 °C higher in summer and 6.9 °C lower in winter than inside the holdfast. A microclimate model of the holdfasts was built and used to forecast microclimates under 2100 conditions. Temperatures are predicted to increase by 2-3 °C, which may exceed the tolerances of D. poha. However, if D. poha or a similar congeneric persists, temperatures inside holdfasts will remain less extreme than the external environment. The thermal tolerances of two Durvillaea-associated invertebrates, the trochid gastropod Cantharidus antipodum and the amphipod Parawaldeckia kidderi, were also assessed; C. antipodum, but not P. kidderi, displayed metabolic depression at temperatures above and below those inside holdfasts, suggesting that they would be vulnerable outside the holdfast and with future warming. Microclimates, such as those within D. poha holdfasts or holdfasts of similar species, will therefore be important refuges for the survival of species both at the northern (retreating edge) and southern (expanding edge) limits of their distributions.

Assessing the utility of marine filter feeders for environmental DNA (eDNA) biodiversity monitoring.

Aquatic environmental DNA (eDNA) surveys are transforming how marine ecosystems are monitored. The time-consuming preprocessing step of active filtration, however, remains a bottleneck. Hence, new approaches that eliminate the need for active filtration are required. Filter-feeding invertebrates have been proven to collect eDNA, but side-by-side comparative studies to investigate the similarity between aquatic and filter-feeder eDNA signals are essential. Here, we investigated the differences among four eDNA sources (water; bivalve gill-tissue; sponges; and ethanol in which filter-feeding organisms were stored) along a vertically stratified transect in Doubtful Sound, New Zealand using three metabarcoding primer sets targeting fish and vertebrates. Combined, eDNA sources detected 59 vertebrates, while concurrent diver surveys observed eight fish species. There were no significant differences in alpha and beta diversity between water and sponge eDNA and both sources were highly correlated. Vertebrate eDNA was successfully extracted from the ethanol in which sponges were stored, although a reduced number of species were detected. Bivalve gill-tissue dissections, on the other hand, failed to reliably detect eDNA. Overall, our results show that vertebrate eDNA signals obtained from water samples and marine sponges are highly concordant. The strong similarity in eDNA signals demonstrates the potential of marine sponges as an additional tool for eDNA-based marine biodiversity surveys, by enabling the incorporation of larger sample numbers in eDNA surveys, reducing plastic waste, simplifying sample collection, and as a cost-efficient alternative. However, we note the importance to not detrimentally impact marine communities by, for example, nonlethal subsampling, specimen cloning, or using bycatch specimens.

The added costs of winter ocean warming for metabolism, arm regeneration and survival in the brittle star Ophionereis schayeri.

As the climate continues to change, it is not just the magnitude of these changes that is important - equally critical is the timing of these events. Conditions that may be well tolerated at one time can become detrimental if experienced at another, as a result of seasonal acclimation. Temperature is the most critical variable as it affects most aspects of an organism's physiology. To address this, we quantified arm regeneration and respiration in the Australian brittle star Ophionereis schayeri for 10 weeks in response to a +3°C warming (18.5°C, simulating a winter heatwave) compared with ambient winter temperature (15.5°C). The metabolic scaling rate (b=0.635 at 15.5°C and 0.746 at 18.5°C) with respect to size was similar to that of other echinoderms and was not affected by temperature. Elevated temperature resulted in up to a 3-fold increase in respiration and a doubling of regeneration growth; however, mortality was greater (up to 44.2% at 18.5°C), especially in the regenerating brittle stars. Metabolic rate of the brittle stars held at 18.5°C was much higher than expected (Q10≈23) and similar to that of O. schayeri tested in summer, which was near their estimated thermotolerance limits. The additional costs associated with the elevated metabolism and regeneration rates incurred by the unseasonably warm winter temperatures may lead to increased mortality and predation risk.

Developing an effective marine eDNA monitoring: eDNA detection at pre-outbreak densities of corallivorous seastar (Acanthaster cf. solaris).

Outbreaks of the corallivorous Crown-of-Thorns Seastar (CoTS) Acanthaster cf. solaris contribute significantly to coral reef loss. Control of outbreaks is hampered because standard monitoring techniques do not detect outbreaks at early (low density) stages, thus preventing early intervention. We previously demonstrated that eDNA monitoring can detect CoTS at intermediate densities. Here, we test whether detection probability can be improved by (i) targeted site selection or collection at specific times and (ii) moving from an average eDNA copy number approach (based on the limit of quantification) to a presence/absence approach (based on the limit of detection). Using a dataset collected over three years and multiple reef sites, we demonstrated that adding water residence age, sea surface level and temperature into generalized linear models explained low amounts of variance of eDNA copy numbers. Site specific CoTS density, by contrast, was a significant predictor for eDNA copy numbers. Bayesian multi-scale occupancy modelling of the presence/absence data demonstrated that the probability of sample capture (θ) on most reefs with intermediate or high CoTS densities was >0.8. Thus, confirming CoTS presence on these reefs would only require 2-3 samples. Sample capture decreased with decreasing CoTS density. Collecting ten filters was sufficient to reliably (based on the lower 95 % Credibility Interval) detect CoTS below nominal outbreak levels (3 Ind. ha-1). Copy number-based estimates may be more relevant to quantify CoTS at higher densities. Although water residence age did contribute little to our models, sites with higher residence times may serve as sentinel sites accumulating eDNA. The approach based on presence or absence of eDNA facilitates eDNA monitoring to detect CoTS densities below outbreak thresholds and we continue to further develop this method for quantification.

Staying in place and moving in space: Contrasting larval thermal sensitivity explains distributional changes of sympatric sea urchin species to habitat warming.

For marine ectotherms, larval success, planktonic larval duration and dispersal trajectories are strongly influenced by temperature, and therefore, ocean warming and heatwaves have profound impacts on these sensitive stages. Warming, through increased poleward flow in regions with western boundary currents, such as the East Australia Current (EAC), provides opportunities for range extension as propagules track preferred conditions. Two sea urchin species, Centrostephanus rodgersii and Heliocidaris tuberculata, sympatric in the EAC warming hotspot, exhibit contrasting responses to warming. Over half a century, C. rodgersii has undergone marked poleward range extension, but the range of H. tuberculata has not changed. We constructed thermal performance curves (TPC) to determine if contrasting developmental thermal tolerance can explain this difference. The temperatures tested encompassed present-day distribution and forecast ocean warming/heatwave conditions. The broad and narrow thermal optimum (Topt) ranges for C. rodgersii and H. tuberculata larvae (7.2 and 4.7°C range, respectively) matched their realized (adult distribution) thermal niches. The cool and warm temperatures for 50% development to the feeding larva approximated temperatures at adult poleward range limits. Larval cool tolerances with respect to mean local temperature differed, 6.0 and 3.8°C respectively. Larval warm tolerances were similar for both species as are the adult warm range edges. The larvae of both species would be sensitive to heatwaves. Centrostephanus rodgersii has stayed in place and shifted in space, likely due to its broad cold-warm larval thermal tolerance and large thermal safety margins. Phenotypic plasticity of the planktonic stage of C. rodgersii facilitated its range extension. In contrast, larval cold intolerance of H. tuberculata explains its restricted range and will delay poleward extension as the region warms. In a warming ocean, we show that intrinsic thermal biology traits of the pelagic stage provide an integrative tool to explain species-specific variation in range shift patterns.

Microplastic ingestion induces asymmetry and oxidative stress in larvae of the sea urchin Pseudechinus huttoni.

Determining the effects of microplastic (MP) ingestion by marine organisms, especially during the sensitive larval stages, is an important step in understanding wider ecosystem responses. We investigated the ingestion, retention (1-5 µm), and short-term exposure effects (1-4 µm) of spherical MPs by larvae of the sea urchin Pseudechinus huttoni. Larvae ingested MPs in a dose-dependent manner and successfully egested particles after a short retention period. Survival was not significantly affected by exposure to MPs over the 10-day experimental period, however, a teratogenic response in terms of delayed development resulted in an increase of larval arm asymmetry. Additionally, MP exposure resulted in oxidative damage to lipids and proteins in larval body tissue despite a significant upregulation of antioxidant defences. The findings indicate MP exposure may impair cellular function, leading to negative consequences for an organism's fitness and survival.

Cross-generational response of a tropical sea urchin to global change and a selection event in a 43-month mesocosm study.

Long-term experimental investigations of transgenerational plasticity (TGP) and transgenerational acclimatization to global change are sparse in marine invertebrates. Here, we test the effect of ocean warming and acidification over a 25-month period of Echinometra sp. A sea urchins whose parents were acclimatized at ambient or one of two near-future (projected mid and end of the 21st century) climate scenarios for 18 months. Several parameters linked to performance exhibited strong effects of future ocean conditions at 9 months of age. The Ambient-Ambient group (A-A, both F0 and F1 at ambient conditions) was significantly larger (21%) and faster in righting response (31%) compared to other groups. A second set of contrasts revealed near-future scenarios caused significant negative parental carryover effects. Respiration at 9 months was depressed by 59% when parents were from near-future climate conditions, and righting response was slowed by 28%. At 10 months, a selective pathogenic mortality event led to significantly higher survival rates of A-A urchins. Differences in size and respiration measured prior to the mortality were absent after the event, while a negative parental effect on righting (29% reduction) remained. The capacity to spawn at the end of the experiment was higher in individuals with ambient parents (50%) compared to other groups (21%) suggesting persistent parental effects. Obtaining different results at different points in time illustrates the importance of longer term and multigeneration studies to investigate effects of climate change. Given some animals in all groups survived the pathogenic event and that effects on physiology (but not behavior) among groups were eliminated after the mortality, we suggest that similar events could constitute selective sweeps, allowing genetic adaptation. However, given the observed negative parental effects and reduced potential for population replenishment, it remains to be determined if selection would be sufficiently rapid to rescue this species from climate change effects.

Reduced seawater pH alters marine biofilms with impacts for marine polychaete larval settlement.

Ocean acidification (OA) can negatively affect early-life stages of marine organisms, with the key processes of larval settlement and metamorphosis potentially vulnerable to reduced seawater pH. Settlement success depends strongly on suitable substrates and environmental cues, with marine biofilms as key settlement inducers for a range of marine invertebrate larvae. This study experimentally investigated (1) how seawater pH determines growth and community composition of marine biofilms, and (2) whether marine biofilms developed under different pH conditions can alter settlement success in the New Zealand serpulid polychaete Galeolaria hystrix. Biofilms were developed under six pH(T) treatments (spanning from 7.0 to 8.1 [ambient]) in a flow-through system for up to 14 months. Biofilms of different ages (7, 10 and 14 months) were used to assay successful settlement of competent G. hystrix larvae reared under ambient conditions. Biofilm microbiomes were characterized through amplicon sequencing of the small subunit ribosomal rRNA gene (16S and 18S). Biofilm community composition was stable over time within each pH treatment and biofilm age did not affect larval settlement selectivity. Seawater pH treatment strongly influenced biofilm community composition, as well as subsequent settlement success when biofilms were presented to competent Galeolaria larvae. Exposure to biofilms incubated under OA-treatments caused a decrease in larval settlement of up to 40% compared to the ambient treatments. We observed a decrease in settlement on biofilms relative to ambient pH for slides incubated at pH 7.9 and 7.7. This trend was reversed at pH 7.4, resulting in high settlement, comparable to ambient biofilms. Settlement decreased on biofilms from pH 7.2, and no settlement was observed on biofilms from pH 7.0. For the first time, we show that long-term incubation of marine biofilms under a wide range of reduced seawater pH treatments can alter marine biofilms in such a way that settlement success in marine invertebrates can be compromised.

Global variability in seawater Mg:Ca and Sr:Ca ratios in the modern ocean.

Seawater Mg:Ca and Sr:Ca ratios are biogeochemical parameters reflecting the Earth-ocean-atmosphere dynamic exchange of elements. The ratios' dependence on the environment and organisms' biology facilitates their application in marine sciences. Here, we present a measured single-laboratory dataset, combined with previous data, to test the assumption of limited seawater Mg:Ca and Sr:Ca variability across marine environments globally. High variability was found in open-ocean upwelling and polar regions, shelves/neritic and river-influenced areas, where seawater Mg:Ca and Sr:Ca ratios range from ∼4.40 to 6.40 mmol:mol and ∼6.95 to 9.80 mmol:mol, respectively. Open-ocean seawater Mg:Ca is semiconservative (∼4.90 to 5.30 mol:mol), while Sr:Ca is more variable and nonconservative (∼7.70 to 8.80 mmol:mol); both ratios are nonconservative in coastal seas. Further, the Ca, Mg, and Sr elemental fluxes are connected to large total alkalinity deviations from International Association for the Physical Sciences of the Oceans (IAPSO) standard values. Because there is significant modern seawater Mg:Ca and Sr:Ca ratios variability across marine environments we cannot absolutely assume that fossil archives using taxa-specific proxies reflect true global seawater chemistry but rather taxa- and process-specific ecosystem variations, reflecting regional conditions. This variability could reconcile secular seawater Mg:Ca and Sr:Ca ratio reconstructions using different taxa and techniques by assuming an error of 1 to 1.50 mol:mol, and 1 to 1.90 mmol:mol, respectively. The modern ratios' variability is similar to the reconstructed rise over 20 Ma (Neogene Period), nurturing the question of seminonconservative behavior of Ca, Mg, and Sr over modern Earth geological history with an overlooked environmental effect.

Sea urchin larvae show resilience to ocean acidification at the time of settlement and metamorphosis.

Extensive research has shown that the early life stages of marine organisms are sensitive to ocean acidification (OA). Less is known, however, on whether larval settlement and metamorphosis may be affected, or by which mechanisms. These are key processes in the life cycle of most marine benthic organisms, since they mark the transition between the free swimming larval stage to benthic life. We investigated whether OA could affect the larval settlement success of the sea urchin Evechinus chloroticus, a key coastal species with ecological, economic and cultural importance in New Zealand. We performed four settlement experiments to test whether reduced seawater pH (ranging from 8.1 to 7.0, at an interval of ~0.2 pH units) alters larval settlement and metamorphosis success. Our results show that settlement success was not significantly reduced when the larvae were exposed to a range of reduced seawater pH treatments (8.1-7.0) at time of settlement (on direct effects). Similarly, when presented with crustose coralline algae (CCA) pre-conditioned in seawater pH of either pH 8.1 or 7.7 for 28 days, larval settlement success remained unaltered (on indirect effects). We conclude that competent larvae in this species are resilient to OA at time of settlement. Further research on a range of taxa that vary in settlement selectivity and behaviour is needed in order to fully understand the effects of OA on the life cycle of marine invertebrates and the consequences it might have for future coastal marine ecosystems.

Ocean acidification affects microbial community and invertebrate settlement on biofilms.

Increased atmospheric CO2 is driving ocean acidification (OA), and potential changes in marine ecosystems. Research shows that both planktonic and benthic communities are affected, but how these changes are linked remains unresolved. Here we show experimentally that decreasing seawater pH (from pH 8.1 to 7.8 and 7.4) leads to reduced biofilm formation and lower primary producer biomass within biofilms. These changes occurred concurrently with a re-arrangement of the biofilm microbial communities. Changes suggest a potential shift from autotrophic to heterotrophic dominated biofilms in response to reduced pH. In a complimentary experiment, biofilms reared under reduced pH resulted in altered larval settlement for a model species (Galeolaria hystrix). These findings show that there is a potential cascade of impacts arising from OA effects on biofilms that may drive important community shifts through altered settlement patterns of benthic species.

Little evidence of adaptation potential to ocean acidification in sea urchins living in "Future Ocean" conditions at a CO2 vent.

Ocean acidification (OA) can be detrimental to calcifying marine organisms, with stunting of invertebrate larval development one of the most consistent responses. Effects are usually measured by short-term, within-generation exposure, an approach that does not consider the potential for adaptation. We examined the genetic response to OA of larvae of the tropical sea urchin Echinometra sp. C. raised on coral reefs that were either influenced by CO2 vents (pH ~ 7.9, future OA condition) or nonvent control reefs (pH 8.2). We assembled a high quality de novo transcriptome of Echinometra embryos (8 hr) and pluteus larvae (48 hr) and identified 68,056 SNPs. We tested for outlier SNPs and functional enrichment in embryos and larvae raised from adults from the control or vent sites. Generally, highest F ST values in embryos were observed between sites (intrinsic adaptation, most representative of the gene pool in the spawned populations). This comparison also had the highest number of outlier loci (40). In the other comparisons, classical adaptation (comparing larvae with adults from the control transplanted to either the control or vent conditions) and reverse adaptation (larvae from the vent site returned to the vent or explanted at the control), we only observed modest numbers of outlier SNPs (6-19) and only enrichment in two functional pathways. Most of the outliers detected were silent substitutions without adaptive potential. We conclude that there is little evidence of realized adaptation potential during early development, while some potential (albeit relatively low) exists in the intrinsic gene pool after more than one generation of exposure.

Beyond Biodiversity: Can Environmental DNA (eDNA) Cut It as a Population Genetics Tool?

Population genetic data underpin many studies of behavioral, ecological, and evolutionary processes in wild populations and contribute to effective conservation management. However, collecting genetic samples can be challenging when working with endangered, invasive, or cryptic species. Environmental DNA (eDNA) offers a way to sample genetic material non-invasively without requiring visual observation. While eDNA has been trialed extensively as a biodiversity and biosecurity monitoring tool with a strong taxonomic focus, it has yet to be fully explored as a means for obtaining population genetic information. Here, we review current research that employs eDNA approaches for the study of populations. We outline challenges facing eDNA-based population genetic methodologies, and suggest avenues of research for future developments. We advocate that with further optimizations, this emergent field holds great potential as part of the population genetics toolkit.

Species-level biodiversity assessment using marine environmental DNA metabarcoding requires protocol optimization and standardization.

DNA extraction from environmental samples (environmental DNA; eDNA) for metabarcoding-based biodiversity studies is gaining popularity as a noninvasive, time-efficient, and cost-effective monitoring tool. The potential benefits are promising for marine conservation, as the marine biome is frequently under-surveyed due to its inaccessibility and the consequent high costs involved. With increasing numbers of eDNA-related publications have come a wide array of capture and extraction methods. Without visual species confirmation, inconsistent use of laboratory protocols hinders comparability between studies because the efficiency of target DNA isolation may vary. We determined an optimal protocol (capture and extraction) for marine eDNA research based on total DNA yield measurements by comparing commonly employed methods of seawater filtering and DNA isolation. We compared metabarcoding results of both targeted (small taxonomic group with species-level assignment) and universal (broad taxonomic group with genus/family-level assignment) approaches obtained from replicates treated with the optimal and a low-performance capture and extraction protocol to determine the impact of protocol choice and DNA yield on biodiversity detection. Filtration through cellulose-nitrate membranes and extraction with Qiagen's DNeasy Blood & Tissue Kit outperformed other combinations of capture and extraction methods, showing a ninefold improvement in DNA yield over the poorest performing methods. Use of optimized protocols resulted in a significant increase in OTU and species richness for targeted metabarcoding assays. However, changing protocols made little difference to the OTU and taxon richness obtained using universal metabarcoding assays. Our results demonstrate an increased risk of false-negative species detection for targeted eDNA approaches when protocols with poor DNA isolation efficacy are employed. Appropriate optimization is therefore essential for eDNA monitoring to remain a powerful, efficient, and relatively cheap method for biodiversity assessments. For seawater, we advocate filtration through cellulose-nitrate membranes and extraction with Qiagen's DNeasy Blood & Tissue Kit or phenol-chloroform-isoamyl for successful implementation of eDNA multi-marker metabarcoding surveys.

Environmental DNA (eDNA) metabarcoding reveals strong discrimination among diverse marine habitats connected by water movement.

While in recent years environmental DNA (eDNA) metabarcoding surveys have shown great promise as an alternative monitoring method, the integration into existing marine monitoring programs may be confounded by the dispersal of the eDNA signal. Currents and tidal influences could transport eDNA over great distances, inducing false-positive species detection, leading to inaccurate biodiversity assessments and, ultimately, mismanagement of marine environments. In this study, we determined the ability of eDNA metabarcoding surveys to distinguish localized signals obtained from four marine habitats within a small spatial scale (<5 km) subject to significant tidal and along-shore water flow. Our eDNA metabarcoding survey detected 86 genera, within 77 families and across 11 phyla using three established metabarcoding assays targeting fish (16S rRNA gene), crustacean (16S rRNA gene) and eukaryotic (cytochrome oxidase subunit 1) diversity. Ordination and cluster analyses for both taxonomic and OTU data sets show distinct eDNA signals between the sampled habitats, suggesting dispersal of eDNA among habitats was limited. Individual taxa with strong habitat preferences displayed localized eDNA signals in accordance with their respective habitat, whereas taxa known to be less habitat-specific generated more ubiquitous signals. Our data add to evidence that eDNA metabarcoding surveys in marine environments detect a broad range of taxa that are spatially discrete. Our work also highlights that refinement of assay choice is essential to realize the full potential of eDNA metabarcoding surveys in marine biodiversity monitoring programs.

Hematological Analysis of the Ascidian Botrylloides leachii (Savigny, 1816) During Whole-Body Regeneration.

Whole-body regeneration (WBR)-the formation of an entire adult from only a small fragment of its own tissue-is extremely rare among chordates. Exceptionally, in the colonial ascidian Botrylloides leachii (Savigny, 1816) a fully functional adult is formed from their common vascular system after ablation of all adults from the colony in just 10 d, thanks to their high blastogenetic potential. While previous studies have identified key genetic markers and morphological changes, no study has yet focused on the hematological aspects of regeneration despite the major involvement of the remaining vascular system and the contained hemocytes in this process. To dissect this process, we analyzed colony blood flow patterns using time-lapse microscopy to obtain a quantitative description of the velocity, reversal pattern, and average distance traveled by hemocytes. We also observed that flows present during regeneration are powered by temporally and spatially synchronized contractions of the terminal ampullae. In addition, we revised previous studies of B. leachii hematology as well as asexual development using histological sectioning and compared the role played by hemocytes during WBR. We found that regeneration starts with a rapid healing response characterized by hemocyte aggregation and infiltration of immunocytes, followed by increased activity of hemoblasts, recruitment of macrophage-like cells for clearing the tissues of debris, and their subsequent disappearance from the circulation concomitant with the maturation of a single regenerated adult. Overall, we provide a detailed account of the hematological properties of regenerating B. leachii colonies, providing novel lines of inquiry toward the decipherment of regeneration in chordates.

Maternal antioxidant provisioning mitigates pollutant-induced oxidative damage in embryos of the temperate sea urchin Evechinus chloroticus.

One mechanism of pollution resistance in marine populations is through transgenerational plasticity, whereby offspring capacity to resist pollution reflects parental exposure history. Our study aimed to establish correlations between oxidative stress biomarkers and key reproductive fitness parameters in the temperate sea urchin Evechinus chloroticus following exposure to dietary polycyclic aromatic hydrocarbons (PAHs). PAH-exposed adults exhibited total gonad tissue concentrations of PAHs in excess of 4 and 5 times baseline levels, for females and males respectively. Antioxidant enzymes were upregulated and oxidative lipid and protein damage to gonad tissues occurred. In addition, early stage offspring reflected maternal antioxidant status, with progeny derived from exposed females demonstrating significantly higher baselines than those derived from control females. Maternal exposure history enhanced the capacity of embryos to minimise oxidative damage to lipids and proteins following exposure to additional PAHs, but provided less of an advantage in protection against oxidative DNA damage. Abnormal embryonic development was largely independent of oxidative damage, remaining high in all embryo populations regardless of parental PAH-history. Overall, results document evidence for maternal transfer of antioxidant potential in E. chloroticus, but imply that a short-term inherited resilience against oxidative stress may not necessarily translate to a fitness or survival gain.

Ocean acidification has little effect on developmental thermal windows of echinoderms from Antarctica to the tropics.

As the ocean warms, thermal tolerance of developmental stages may be a key driver of changes in the geographical distributions and abundance of marine invertebrates. Additional stressors such as ocean acidification may influence developmental thermal windows and are therefore important considerations for predicting distributions of species under climate change scenarios. The effects of reduced seawater pH on the thermal windows of fertilization, embryology and larval morphology were examined using five echinoderm species: two polar (Sterechinus neumayeri and Odontaster validus), two temperate (Fellaster zelandiae and Patiriella regularis) and one tropical (Arachnoides placenta). Responses were examined across 12-13 temperatures ranging from -1.1 °C to 5.7 °C (S. neumayeri), -0.5 °C to 10.7 °C (O. validus), 5.8 °C to 27 °C (F. zelandiae), 6.0 °C to 27.1 °C (P. regularis) and 13.9 °C to 34.8 °C (A. placenta) under present-day and near-future (2100+) ocean acidification conditions (-0.3 pH units) and for three important early developmental stages 1) fertilization, 2) embryo (prehatching) and 3) larval development. Thermal windows for fertilization were broad and were not influenced by a pH decrease. Embryological development was less thermotolerant. For O. validus, P. regularis and A. placenta, low pH reduced normal development, albeit with no effect on thermal windows. Larval development in all five species was affected by both temperature and pH; however, thermal tolerance was not reduced by pH. Results of this study suggest that in terms of fertilization and development, temperature will remain as the most important factor influencing species' latitudinal distributions as the ocean continues to warm and decrease in pH, and that there is little evidence of a synergistic effect of temperature and ocean acidification on the thermal control of species ranges.

In situ developmental responses of tropical sea urchin larvae to ocean acidification conditions at naturally elevated pCO2 vent sites.

Laboratory experiments suggest that calcifying developmental stages of marine invertebrates may be the most ocean acidification (OA)-sensitive life-history stage and represent a life-history bottleneck. To better extrapolate laboratory findings to future OA conditions, developmental responses in sea urchin embryos/larvae were compared under ecologically relevant in situ exposures on vent-elevated pCO2 and ambient pCO2 coral reefs in Papua New Guinea. Echinometra embryos/larvae were reared in meshed chambers moored in arrays on either venting reefs or adjacent non-vent reefs. After 24 and 48 h, larval development and morphology were quantified. Compared with controls (mean pH(T) = 7.89-7.92), larvae developing in elevated pCO2 vent conditions (pH(T) = 7.50-7.72) displayed a significant reduction in size and increased abnormality, with a significant correlation of seawater pH with both larval size and larval asymmetry across all experiments. Reciprocal transplants (embryos from vent adults transplanted to control conditions, and vice versa) were also undertaken to identify if adult acclimatization can translate resilience to offspring (i.e. transgenerational processes). Embryos originating from vent adults were, however, no more tolerant to reduced pH. Sea temperature and chlorophyll-a concentrations (i.e. larval nutrition) did not contribute to difference in larval size, but abnormality was correlated with chlorophyll levels. This study is the first to examine the response of marine larvae to OA scenarios in the natural environment where, importantly, we found that stunted and abnormal development observed in situ are consistent with laboratory observations reported in sea urchins, in both the direction and magnitude of the response.

Dietary pollutants induce oxidative stress, altering maternal antioxidant provisioning and reproductive output in the temperate sea urchin Evechinus chloroticus.

Evidence is growing to suggest that the capacity to withstand oxidative stress may play an important role in shaping life-history trade-offs, although little is known on the relationship in broadcast spawning marine invertebrates. In this group, variation in gamete quantity and quality are important drivers of offspring survival and successful recruitment. Therefore the provisioning of eggs with antioxidants may be an important driver of life history strategies because they play a critical role in preventing damage from reactive oxygen species to macromolecules. In this study, a suite of oxidative stress biomarkers was measured in the gonads and eggs of the sea urchin Evechinus chloroticus exposed to polycyclic aromatic hydrocarbons (PAHs). Links between oxidative stress markers and core components of fitness including fecundity, gamete quality and maternal transfer of antioxidants were assessed. Experimental induction of oxidative stress was achieved via exposure to a mix of four PAHs over a 21-day period. In PAH exposed individuals, we observed a significant upregulation of the antioxidant defence and detoxification enzymes SOD, CAT, GR, GPx and GST, as well as a greater pool of the non-enzymatic antioxidant glutathione in gonad tissue and eggs. In contrast, glutathione redox status was not affected by PAH exposure, with the percentage of reduced glutathione remaining at approximately 80% in both gonad tissue and released eggs. PAH-exposed adults experienced greater than three- and five-fold increases in oxidative protein and lipid damage, respectively, in gonad tissue. In contrast, eggs maintained low levels of damage, not differing from baseline levels found in eggs released from PAH-naïve mothers. PAH exposure also resulted in a 2-fold reduction in fecundity of reproductively mature females but no significant alteration to egg diameter. Although PAH-exposed females released fewer eggs, successful fertilisation of those eggs was slightly enhanced with average rates ranging from 90-99% in comparison to 76-90% in control eggs. Early-stage offspring reflected maternal antioxidant status with populations derived from PAH-exposed mothers demonstrating significantly higher antioxidant levels than those derived from PAH-naïve mothers. This maternally inherited protection enhanced the capacity of embryos to minimise oxidative damage to lipids and proteins during early development but, despite this, did not reduce the proportion of morphological abnormalities in the population. Overall, these findings indicate that when faced with short-term contaminant stress E. chloroticus has the capacity to trade high reproductive output during a spawning event for a greater antioxidant investment in eggs. However, this production of potentially more resilient offspring did not translate to a fitness gain, at least for the early larval stages in the present experimental conditions.