Systematic review of the uncertainty of coral reef futures under climate change.

Climate change impact syntheses, such as those by the Intergovernmental Panel on Climate Change, consistently assert that limiting global warming to 1.5 °C is unlikely to safeguard most of the world's coral reefs. This prognosis is primarily based on a small subset of available models that apply similar 'excess heat' threshold methodologies. Our systematic review of 79 articles projecting coral reef responses to climate change revealed five main methods. 'Excess heat' models constituted one third (32%) of all studies but attracted a disproportionate share (68%) of citations in the field. Most methods relied on deterministic cause-and-effect rules rather than probabilistic relationships, impeding the field's ability to estimate uncertainty. To synthesize the available projections, we aimed to identify models with comparable outputs. However, divergent choices in model outputs and scenarios limited the analysis to a fraction of available studies. We found substantial discrepancies in the projected impacts, indicating that the subset of articles serving as a basis for climate change syntheses may project more severe consequences than other studies and methodologies. Drawing on insights from other fields, we propose methods to incorporate uncertainty into deterministic modeling approaches and propose a multi-model ensemble approach to generating probabilistic projections for coral reef futures.

Toward Best Practices for Controlling Mammalian Cell Culture Environments.

The characterization, control, and reporting of environmental conditions in mammalian cell cultures is fundamental to ensure physiological relevance and reproducibility in basic and preclinical biomedical research. The potential issue of environment instability in routine cell cultures in affecting biomedical experiments was identified many decades ago. Despite existing evidence showing variable environmental conditions can affect a suite of cellular responses and key experimental readouts, the underreporting of critical parameters affecting cell culture environments in published experiments remains a serious problem. Here, we outline the main sources of potential problems, improved guidelines for reporting, and deliver recommendations to facilitate improved culture-system based research. Addressing the lack of attention paid to culture environments is critical to improve the reproducibility and translation of preclinical research, but constitutes only an initial step towards enhancing the relevance of in vitro cell cultures towards in vivo physiology.

In situ monitoring reveals cellular environmental instabilities in human pluripotent stem cell culture.

Mammalian cell cultures are a keystone resource in biomedical research, but the results of published experiments often suffer from reproducibility challenges. This has led to a focus on the influence of cell culture conditions on cellular responses and reproducibility of experimental findings. Here, we perform frequent in situ monitoring of dissolved O2 and CO2 with optical sensor spots and contemporaneous evaluation of cell proliferation and medium pH in standard batch cultures of three widely used human somatic and pluripotent stem cell lines. We collate data from the literature to demonstrate that standard cell cultures consistently exhibit environmental instability, indicating that this may be a pervasive issue affecting experimental findings. Our results show that in vitro cell cultures consistently undergo large departures of environmental parameters during standard batch culture. These findings should catalyze further efforts to increase the relevance of experimental results to the in vivo physiology and enhance reproducibility.

Projecting coral responses to intensifying marine heatwaves under ocean acidification.

Over this century, coral reefs will run the gauntlet of climate change, as marine heatwaves (MHWs) become more intense and frequent, and ocean acidification (OA) progresses. However, we still lack a quantitative assessment of how, and to what degree, OA will moderate the responses of corals to MHWs as they intensify throughout this century. Here, we first projected future MHW intensities for tropical regions under three future greenhouse gas emissions scenario (representative concentration pathways, RCP2.6, RCP4.5 and RCP8.5) for the near-term (2021-2040), mid-century (2041-2060) and late-century (2081-2100). We then combined these MHW intensity projections with a global data set of 1,788 experiments to assess coral attribute performance and survival under the three emissions scenarios for the near-term, mid-century and late-century in the presence and absence of OA. Although warming and OA had predominately additive impacts on the coral responses, the contribution of OA in affecting most coral attributes was minor relative to the dominant role of intensifying MHWs. However, the addition of OA led to greater decreases in photosynthesis and survival under intermediate and unrestricted emissions scenario for the mid- and late-century than if intensifying MHWs were considered as the only driver. These results show that role of OA in modulating coral responses to intensifying MHWs depended on the focal coral attribute and extremity of the scenario examined. Specifically, intensifying MHWs and OA will cause increasing instances of coral bleaching and substantial declines in coral productivity, calcification and survival within the next two decades under the low and intermediate emissions scenario. These projections suggest that corals must rapidly adapt or acclimatize to projected ocean conditions to persist, which is far more likely under a low emissions scenario and with increasing efforts to manage reefs to enhance resilience.

Integrating environmental variability to broaden the research on coral responses to future ocean conditions.

Our understanding of the response of reef-building corals to changes in their physical environment is largely based on laboratory experiments, analysis of long-term field data, and model projections. Experimental data provide unique insights into how organisms respond to variation of environmental drivers. However, an assessment of how well experimental conditions cover the breadth of environmental conditions and variability where corals live successfully is missing. Here, we compiled and analyzed a globally distributed dataset of in-situ seasonal and diurnal variability of key environmental drivers (temperature, pCO2 , and O2 ) critical for the growth and livelihood of reef-building corals. Using a meta-analysis approach, we compared the variability of environmental conditions assayed in coral experimental studies to current and projected conditions in their natural habitats. We found that annual temperature profiles projected for the end of the 21st century were characterized by distributional shifts in temperatures with warmer winters and longer warm periods in the summer, not just peak temperatures. Furthermore, short-term hourly fluctuations of temperature and pCO2 may regularly expose corals to conditions beyond the projected average increases for the end of the 21st century. Coral reef sites varied in the degree of coupling between temperature, pCO2 , and dissolved O2 , which warrants site-specific, differentiated experimental approaches depending on the local hydrography and influence of biological processes on the carbonate system and O2 availability. Our analysis highlights that a large portion of the natural environmental variability at short and long timescales is underexplored in experimental designs, which may provide a path to extend our understanding on the response of corals to global climate change.

Temperature transcends partner specificity in the symbiosis establishment of a cnidarian.

Coral reef research has predominantly focused on the effect of temperature on the breakdown of coral-dinoflagellate symbioses. However, less is known about how increasing temperature affects the establishment of new coral-dinoflagellate associations. Inter-partner specificity and environment-dependent colonization are two constraints proposed to limit the acquisition of more heat tolerant symbionts. Here, we investigated the symbiotic dynamics of various photosymbionts in different host genotypes under "optimal" and elevated temperature conditions. To do this, we inoculated symbiont-free polyps of the sea anemone Exaiptasia pallida originating from Hawaii (H2), North Carolina (CC7), and the Red Sea (RS) with the same mixture of native symbiont strains (Breviolum minutum, Symbiodinium linucheae, S. microadriaticum, and a Breviolum type from the Red Sea) at 25 and 32 °C, and assessed their ITS2 composition, colonization rates, and PSII photochemical efficiency (Fv/Fm). Symbiont communities across thermal conditions differed significantly for all hosts, suggesting that temperature rather than partner specificity had a stronger effect on symbiosis establishment. Overall, we detected higher abundances of more heat resistant Symbiodiniaceae types in the 32 °C treatments. Our data further showed that PSII photophysiology under elevated temperature improved with thermal pre-exposure (i.e., higher Fv/Fm), yet, this effect depended on host genotype and was influenced by active feeding as photochemical efficiency dropped in response to food deprivation. These findings highlight the role of temperature and partner fidelity in the establishment and performance of symbiosis and demonstrate the importance of heterotrophy for symbiotic cnidarians to endure and recover from stress.

Unfamiliar partnerships limit cnidarian holobiont acclimation to warming.

Enhancing the resilience of corals to rising temperatures is now a matter of urgency, leading to growing efforts to explore the use of heat tolerant symbiont species to improve their thermal resilience. The notion that adaptive traits can be retained by transferring the symbionts alone, however, challenges the holobiont concept, a fundamental paradigm in coral research. Holobiont traits are products of a specific community (holobiont) and all its co-evolutionary and local adaptations, which might limit the retention or transference of holobiont traits by exchanging only one partner. Here we evaluate how interchanging partners affect the short- and long-term performance of holobionts under heat stress using clonal lineages of the cnidarian model system Aiptasia (host and Symbiodiniaceae strains) originating from distinct thermal environments. Our results show that holobionts from more thermally variable environments have higher plasticity to heat stress, but this resilience could not be transferred to other host genotypes through the exchange of symbionts. Importantly, our findings highlight the role of the host in determining holobiont productivity in response to thermal stress and indicate that local adaptations of holobionts will likely limit the efficacy of interchanging unfamiliar compartments to enhance thermal tolerance.

Additive impacts of deoxygenation and acidification threaten marine biota.

Deoxygenation in coastal and open-ocean ecosystems rarely exists in isolation but occurs concomitantly with acidification. Here, we first combine meta-data of experimental assessments from across the globe to investigate the potential interactive impacts of deoxygenation and acidification on a broad range of marine taxa. We then characterize the differing degrees of deoxygenation and acidification tested in our dataset using a ratio between the partial pressure of oxygen and carbon dioxide (pO2 /pCO2 ) to assess how biological processes change under an extensive, yet diverse range of pO2 and pCO2 conditions. The dataset comprised 375 experimental comparisons and revealed predominantly additive but variable effects (91.7%, additive; 6.0%, synergistic; and 2.3%, antagonistic) of the dual stressors, yielding negative impacts across almost all responses examined. Our data indicate that the pO2 /pCO2 -ratio offers a simplified metric to characterize the extremity of the concurrent stressors and shows that more severe impacts occurred when ratios represented more extreme deoxygenation and acidification conditions. Importantly, our analysis highlights the need to assess the concurrent impacts of deoxygenation and acidification on marine taxa and that assessments considering the impact of O2 depletion alone will likely underestimate the impacts of deoxygenation events and their ecosystem-wide consequences.

A framework for experimental scenarios of global change in marine systems using coral reefs as a case study.

Understanding the consequences of rising CO2 and warming on marine ecosystems is a pressing issue in ecology. Manipulative experiments that assess responses of biota to future ocean warming and acidification conditions form a necessary basis for expectations on how marine taxa may respond. Although designing experiments in the context of local variability is most appropriate, local temperature and CO2 characteristics are often unknown as such measures necessitate significant resources, and even less is known about local future scenarios. To help address these issues, we summarize current uncertainties in CO2 emission trajectories and climate sensitivity, examine region-specific changes in the ocean, and present a straightforward global framework to guide experimental designs. We advocate for the inclusion of multiple plausible future scenarios of predicted levels of ocean warming and acidification in forthcoming experimental research. Growing a robust experimental base is crucial to understanding the prospect form and function of marine ecosystems in the Anthropocene.

Defining CO2 and O2 syndromes of marine biomes in the Anthropocene.

Research efforts have intensified to foresee the prospects for marine biomes under climate change and anthropogenic drivers over varying temporal and spatial scales. Parallel with these efforts is the utilization of terminology, such as 'ocean acidification' (OA) and 'ocean deoxygenation' (OD), that can foster rapid comprehension of complex processes driving carbon dioxide (CO2 ) and oxygen (O2 ) concentrations in the global ocean and thus, are now widely used in discussions within and beyond academia. However, common usage of the terms 'acidification' and 'deoxygenation' alone are subjective and, without adequate contextualization, have the potential to mislead inferences over drivers that may ultimately shape the future state of marine ecosystems. Here we clarify the usage of the terms OA and OD as global, climate change-driven processes and discuss the various attributes of elevated CO2 and reduced O2 syndromes common to coastal ecosystems. We support the use of the existing terms 'coastal acidification' and 'coastal deoxygenation' because they help differentiate the sometimes rapid and extreme nature of CO2 and O2 syndromes in coastal ecosystems from the global, climate change-driven processes of OA and OD. Given the complexity and breadth of the processes involved in altering CO2 and O2 concentrations across marine ecosystems, we provide a workflow to enable contextualization and clarification of the usage of existing terms and highlight the close link between these two gases across spatial and temporal scales in the ocean. These distinctions are crucial to guide effective communication of research within the scientific community and guide policymakers responsible for intervening on the drivers to secure desirable future ocean states.

Symbiodinium mitigate the combined effects of hypoxia and acidification on a noncalcifying cnidarian.

Anthropogenic nutrient inputs enhance microbial respiration within many coastal ecosystems, driving concurrent hypoxia and acidification. During photosynthesis, Symbiodinium spp., the microalgal endosymbionts of cnidarians and other marine phyla, produce O2 and assimilate CO2 and thus potentially mitigate the exposure of the host to these stresses. However, such a role for Symbiodinium remains untested for noncalcifying cnidarians. We therefore contrasted the fitness of symbiotic and aposymbiotic polyps of a model host jellyfish (Cassiopea sp.) under reduced O2 (~2.09 mg/L) and pH (~ 7.63) scenarios in a full-factorial experiment. Host fitness was characterized as asexual reproduction and their ability to regulate internal pH and Symbiodinium performance characterized by maximum photochemical efficiency, chla content and cell density. Acidification alone resulted in 58% more asexual reproduction of symbiotic polyps than aposymbiotic polyps (and enhanced Symbiodinium cell density) suggesting Cassiopea sp. fitness was enhanced by CO2 -stimulated Symbiodinium photosynthetic activity. Indeed, greater CO2 drawdown (elevated pH) was observed within host tissues of symbiotic polyps under acidification regardless of O2 conditions. Hypoxia alone produced 22% fewer polyps than ambient conditions regardless of acidification and symbiont status, suggesting Symbiodinium photosynthetic activity did not mitigate its effects. Combined hypoxia and acidification, however, produced similar numbers of symbiotic polyps compared with aposymbiotic kept under ambient conditions, demonstrating that the presence of Symbiodinium was key for mitigating the combined effects of hypoxia and acidification on asexual reproduction. We hypothesize that this mitigation occurred because of reduced photorespiration under elevated CO2 conditions where increased net O2 production ameliorates oxygen debt. We show that Symbiodinium play an important role in facilitating enhanced fitness of Cassiopea sp. polyps, and perhaps also other noncalcifying cnidarian hosts, to the ubiquitous effects of ocean acidification. Importantly we highlight that symbiotic, noncalcifying cnidarians may be particularly advantaged in productive coastal waters that are subject to simultaneous hypoxia and acidification.

Surviving but not thriving: inconsistent responses of zooxanthellate jellyfish polyps to ocean warming and future UV-B scenarios.

Complex changes to UV radiation at the Earth's surface are occurring concurrently with ocean warming. Despite few empirical tests, jellyfish are hypothesised to be increasing in some parts of the world because they are robust to environmental stressors. Here we examine the effects of UV-B and ocean warming projections on zooxanthellate jellyfish polyps. We exposed Cassiopea sp. polyps to three levels of UV-B (future-low (1.43 Wm(2)), current (1.60 Wm(2)), future-high (1.77 Wm(2))) and two levels of temperature (current-day (25 °C) and future (28 °C)) over 6 weeks. The intensity of UV-B was varied throughout the day to mimic diel variation in UV-B irradiance. Polyp survival, asexual reproduction and YII were measured. In the current and future-high UV-B treatments, more polyps were produced in 25 °C than 28 °C. This pattern, however, was reversed under future-low UV-B conditions, where more polyps were produced at 28 °C. YII was highest under current summer conditions and future conditions of low UV-B and increased temperature. YII, however, was reduced under high UV-B conditions but was further reduced with warming. Our results suggest that although Cassiopea polyps may survive elevated UV-B and warming conditions, they are unlikely to thrive. If, however, UV-B radiation decreases then ocean warming may facilitate increases in Cassiopea populations.

Reduced salinity increases susceptibility of zooxanthellate jellyfish to herbicide toxicity during a simulated rainfall event.

Accurately predicting how marine biota are likely to respond to changing ocean conditions requires accurate simulation of interacting stressors, exposure regimes and recovery periods. Jellyfish populations have increased in some parts of the world and, despite few direct empirical tests, are hypothesised to be increasing because they are robust to a range of environmental stressors. Here, we investigated the effects of contaminated runoff on a zooxanthellate jellyfish by exposing juvenile Cassiopea sp. medusae to a photosystem II (PSII) herbicide, atrazine and reduced salinity conditions that occur following rainfall. Four levels of atrazine (0ngL(-1), 10ngL(-1), 2µgL(-1), 20µgL(-1)) and three levels of salinity (35 ppt, 25 ppt, 17 ppt) were varied, mimicking the timeline of light, moderate and heavy rainfall events. Normal conditions were then slowly re-established over four days to mimic the recovery of the ecosystem post-rain and the experiment continued for a further 7 days to observe potential recovery of the medusae. Pulse-amplitude modulated (PAM) chlorophyll fluorescence, growth and bell contraction rates of medusae were measured. Medusae exposed to the combination of high atrazine and lowest salinity died. After 3 days of exposure, bell contraction rates were reduced by 88% and medusae were 16% smaller in the lowest salinity treatments. By Day 5 of the experiment, all medusae that survived the initial pulse event began to recover quickly. Although atrazine decreased YII under normal salinity conditions, YII was further reduced when medusae were exposed to both low salinity and atrazine simultaneously. Atrazine breakdown products were more concentrated in jellyfish tissues than atrazine at the end of the experiment, suggesting that although bioaccumulation occurred, atrazine was metabolised. Our results suggest that reduced salinity may increase the susceptibility of medusae to herbicide exposure during heavy rainfall events.

Irukandji jellyfish polyps exhibit tolerance to interacting climate change stressors.

Increasing ocean temperatures and strengthening boundary currents have caused the poleward migration of many marine species. Cubozoan jellyfish known to cause Irukandji syndrome have historically been confined to tropical waters but may be expanding into subtropical regions. Here, we examine the interactive effects of warming and acidification on the population dynamics of polyps of an Irukandji jellyfish, Alatina nr mordens, and the formation of statoliths in newly metamorphosed medusae, to determine if this jellyfish could tolerate future conditions predicted for southeast Queensland (SEQ), Australia. Two experiments, examining the orthogonal factors of temperature and pH, were undertaken. Experiment 1 mimicked the current, ca. 2050 and ca. 2100 summer temperature and pH conditions predicted for SEQ using A1F1 scenarios (temperature: 25, 27, 29 °C; pH: 7.9, 7.8, 7.6) and Experiment 2 mimicked current and future winter conditions (18 and 22 °C, pH 7.9, 7.8, 7.6). All polyps in Experiment 1 survived and budded. Fewer polyps budded in the lower pH treatments; however, patterns varied slightly among temperature treatments. Statoliths at pH 7.6 were 24% narrower than those at pH 7.8 and 7.9. Most polyps survived the winter conditions mimicked by Experiment 2 but only polyps in the 22 °C, pH 7.9 treatment increased significantly. The current absence of A. nr mordens medusae in SEQ, despite the polyps' ability to tolerate the current temperature and pH conditions, suggests that ecological, rather than abiotic factors currently limit their distribution. Observations that budding was lower under low pH treatments suggest that rates of asexual reproduction will likely be much slower in the future. We consider that A. nr mordens polyps are likely to tolerate future conditions but are unlikely to thrive in the long term. However, if polyps can overcome potential ecological boundaries and acidification proceeds slowly A. nr mordens could expand polewards in the short term.