Photoprotective mechanisms in Elysia species hosting Acetabularia chloroplasts shed light on host-donor compatibility in photosynthetic sea slugs.

Sacoglossa sea slugs have garnered attention due to their ability to retain intracellular functional chloroplasts from algae, while degrading other algal cell components. While protective mechanisms that limit oxidative damage under excessive light are well documented in plants and algae, the photoprotective strategies employed by these photosynthetic sea slugs remain unresolved. Species within the genus Elysia are known to retain chloroplasts from various algal sources, but the extent to which the metabolic processes from the donor algae can be sustained by the sea slugs is unclear. By comparing responses to high-light conditions through kinetic analyses, molecular techniques, and biochemical assays, this study shows significant differences between two photosynthetic Elysia species with chloroplasts derived from the green alga Acetabularia acetabulum. Notably, Elysia timida displayed remarkable tolerance to high-light stress and sophisticated photoprotective mechanisms such as an active xanthophyll cycle, efficient D1 protein recycling, accumulation of heat-shock proteins and α-tocopherol. In contrast, Elysia crispata exhibited absence or limitations in these photoprotective strategies. Our findings emphasize the intricate relationship between the host animal and the stolen chloroplasts, highlighting different capacities to protect the photosynthetic organelle from oxidative damage.

Thermal tolerance limits and physiological traits as indicators of Hediste diversicolor's acclimation capacity to global and local change drivers.

Global projections predict significant increases in ocean temperature and changes in ocean chemistry, including salinity variations by 2100. This has led to a substantial interest in the study of thermal ecophysiology, as temperature is a major factor shaping marine ectotherm communities. However, responses to temperature may be influenced by other factors such as salinity, highlighting the relevance of multiple stressor studies. In the present work, we experimentally evaluated the thermal tolerance of the marine ragworm Hediste diversicolor under predicted global change scenarios. Organisms were subjected to an experimental trial under control (24 °C), and two temperature treatment scenarios (ocean warming +3 °C - (27 °C) and heat wave +6 °C - (30 °C)), combined with salinity variations (20 and 30) in a full factorial design for 29 days. Environmental data from the field were collected during 2019 and 2020. At day 30 post exposure, upper thermal limits (Critical Thermal Maximum - CTMax), thermal safety margins (TSM) and acclimation capacity were measured. Higher acclimation temperatures led to higher thermal tolerance limits, confirming that H. diversicolor features some physiological plasticity, acclimation capacity and a positive thermal safety margin. This margin was greater considering in situ temperature data from 2019 than maximum temperatures for 2020 (CTMax > maximum habitat temperature-MHT). Moreover, smaller organisms displayed higher upper thermal limits suggesting that thermal tolerance is size dependent. Ragworms subjected to higher salinity also showed a higher CTMax than those acclimated to lower salinity. However, temperature and salinity showed an additive effect on CTMax, as no significant interaction was detected. We conclude that H. diversicolor can easily acclimate to increased water temperature, independently of salinity variations. Given the key role of ragworms in food webs in estuaries and coastal lagoons, substrate bioturbation and aquaculture, this information is relevant to support conservation actions, optimize culture protocols and identify thermal resistant strains.

Assessing the Trophic Impact of Bleaching: The Model Pair Berghia stephanieae/Exaiptasia diaphana.

Bleaching events associated with climate change are increasing worldwide, being a major threat to tropical coral reefs. Nonetheless, the indirect impacts promoted by the bleaching of organisms hosting photosynthetic endosymbionts, such as those impacting trophic interactions, have received considerably less attention by the scientific community. Bleaching significantly affects the nutritional quality of bleached organisms. The consequences promoted by such shifts remain largely overlooked, namely on specialized predators that have evolved to prey upon organisms hosting photosynthetic endosymbionts and benefit nutritionally, either directly or indirectly, from the available pool of photosynthates. In the present study, we advocate the use of the model predator-prey pair featuring the stenophagous nudibranch sea slug Berghia stephanieae that preys upon the photosymbiotic glass anemone Exaiptasia diaphana to study the impacts of bleaching on trophic interactions. These model organisms are already used in other research fields, and one may benefit from knowledge available on their physiology, omics, and culture protocols under controlled laboratory conditions. Moreover, B. stephanieae can thrive on either photosymbiotic or aposymbiotic (bleached) glass anemones, which can be easily maintained over long periods in the laboratory (unlike photosymbiotic corals). As such, one can investigate if and how nutritional shifts induced by bleaching impact highly specialized predators (stenophagous species), as well as if and how such effects cascade over consecutive generations. Overall, by using this model predator-prey pair one can start to truly unravel the trophic effects of bleaching events impacting coral reef communities, as well as their prevalence over time.

Combined effect of marine heatwaves and light intensity on the cellular stress response and photophysiology of the leather coral Sarcophyton cf. glaucum.

Marine heatwaves (MHW) are threatening tropical coral reef ecosystems, leading to mass bleaching events worldwide. The combination of heat stress with high irradiance is known to shape the health and redox status of corals, but research is biased toward scleractinian corals, while much less is known on tropical symbiotic soft corals. Here, we evaluated the cellular stress response and the photophysiological performance of the soft coral Sarcophyton cf. glaucum, popularly termed as leather coral, under different global change scenarios. Corals were exposed to different light intensities (high light, low light, ∼662 and 253 µmol photons m-2 s-1) for 30 days (time-point 1) and a subsequent MHW simulation was carried out for 10 days (control 26 vs 32 °C) (time-point 2). Subsequently, corals were returned to control temperature and allowed to recover for 30 days (time-point 3). Photophysiological performance (maximum quantum yield of photosystem II (Fv/Fm), a measure of photosynthetic activity; dark-level fluorescence (F0), as a proxy of chlorophyll a content (Chl a); and zooxanthellae density) and stress biomarkers (total protein, antioxidants, lipid peroxidation, ubiquitin, and heat shock protein 70) were assessed in corals at these three time-points. Corals were especially sensitive to the combination of heat and high light stress, experiencing a decrease in their photosynthetic efficiency under these conditions. Heat stress resulted in bleaching via zooxanthellae loss while high light stress led to pigment (Chl a) loss. This species' antioxidant defenses, and protein degradation were particularly enhanced under heat stress. A recovery was clear for molecular parameters after 30 days of recovery, whereby photophysiological performance required more time to return to basal levels. We conclude that soft corals distributed along intertidal areas, where the light intensity is high, could be especially vulnerable to marine heatwave events, highlighting the need to direct conservation efforts toward these organisms.

Cellular stress response and acclimation capacity of the ditch shrimp Palaemon varians to extreme weather events - How plastic can a plastic species be?

Species from shallow marine environments are particularly vulnerable to extreme weather events (heatwaves and extreme rainfall) that can promote abrupt environmental shifts, namely in temperature and salinity (respectively). To assess how these shifts impact species' cellular stress responses (CSR), ditch shrimps Palaemon varians were exposed to a chronic (28 days) thermohaline stress experiment. Three levels of temperature (20, 23 and 26 °C) and two levels of salinity (20 and 40) were tested in a full factorial experiment, and shrimps sampled at the 7th, 14th, 21st and 28th day of exposure. Survival, wet weight (as proxy for growth), and cellular stress biomarkers associated with oxidative stress (LPO - Lipid Peroxidation, GST - Glutathione-S-Transferase, SOD - Superoxide Dismutase, TAC - Total Antioxidant Capacity and CAT - Catalase) and protein denaturation (UBI - Ubiquitin and HSP-70 - Heat Shock Protein 70 kDa) were analysed in shrimps' muscle at each sampling day. Temperature and time of exposure significantly affected biomarker levels, with shrimps exposed to 20 and 26 °C revealing more pronounced differences. No interactions were detected between temperature and salinity, suggesting that these factors display additive effects on shrimps' CSR. Antioxidant agents (CAT and TAC) increased under elevated temperature, while protein denaturation markers (UBI and HSP-70) were mostly affected by time of exposure, decreasing at 28 days. Total protein reserves increased throughout time and no effects on wet weight were observed. A negative correlation between wet weight and HSP-70 was detected, suggesting that HSP-70 levels are dependent on organism size. Peak survival (~73 %) was found under 20 °C and salinity 40 and lower survival (~30-40 %) was associated with higher temperatures (23 and 26 °C) and lower salinity (20). We conclude that P. varians displays some level of acclimation capacity but differences in survival may indicate effects on osmoregulation processes and the need for longer timeframes to fully acclimate to heat and hyposaline stress.

Sex-specific thermal tolerance limits in the ditch shrimp Palaemon varians: Eco-evolutionary implications under a warming ocean.

As global temperatures continue to rise due to global change, marine heatwaves are also becoming more frequent and intense, impacting marine biodiversity patterns worldwide. Organisms inhabiting shallow water environments, such as the commercially relevant ditch shrimp Palaemon varians, are expected to be the most affected by rising temperatures. Thus, addressing species' thermal ecology and climate extinction-risk is crucial to foster climate-smart conservation strategies for shallow water ecosystems. Here, we estimated sex-specific upper thermal tolerance limits for P. varians via the Critical Thermal Maximum method (CTmax), using loss of equilibrium as endpoint. We further calculated thermal safety margins for males and females and tested for correlations between upper thermal limits and shrimps' body size. To determine sex-biased variation in P. varians' traits (CTmax, weight and length), we compared trait variation between females and males through the coefficient of variation ratio (lnCVR). Females displayed an average CTmax value 1.8% lower than males (CTmaxfemales = 37.0 °C vs CTmaxmales = 37.7 °C). This finding may be related to the larger body size exhibited by females (156% heavier and 39% larger than males), as both length and weight had a significant effect on CTmax. The high energetic investment of females in offspring may also contribute to the differences recorded in thermal tolerance. Overall, organisms with a smaller body-size displayed a greater tolerance to elevated temperature, thus suggesting that smaller individuals may be positively selected in warmer environments. This selection may result in a reduction of size-at-maturity and shifts in sex ratio, given the sexual dimorphism in body size of shrimps. The thermal safety margin of P. varians was narrow (∼2.2 °C for males and ∼1.5 °C for females), revealing the vulnerability of this species to ocean warming and heatwaves.

Tolerant Larvae and Sensitive Juveniles: Integrating Metabolomics and Whole-Organism Responses to Define Life-Stage Specific Sensitivity to Ocean Acidification in the American Lobster.

Bentho-pelagic life cycles are the dominant reproductive strategy in marine invertebrates, providing great dispersal ability, access to different resources, and the opportunity to settle in suitable habitats upon the trigger of environmental cues at key developmental moments. However, free-dispersing larvae can be highly sensitive to environmental changes. Among these, the magnitude and the occurrence of elevated carbon dioxide (CO2) concentrations in oceanic habitats is predicted to exacerbate over the next decades, particularly in coastal areas, reaching levels beyond those historically experienced by most marine organisms. Here, we aimed to determine the sensitivity to elevated pCO2 of successive life stages of a marine invertebrate species with a bentho-pelagic life cycle, exposed continuously during its early ontogeny, whilst providing in-depth insights on their metabolic responses. We selected, as an ideal study species, the American lobster Homarus americanus, and investigated life history traits, whole-organism physiology, and metabolomic fingerprints from larval stage I to juvenile stage V exposed to different pCO2 levels. Current and future ocean acidification scenarios were tested, as well as extreme high pCO2/low pH conditions that are predicted to occur in coastal benthic habitats and with leakages from underwater carbon capture storage (CCS) sites. Larvae demonstrated greater tolerance to elevated pCO2, showing no significant changes in survival, developmental time, morphology, and mineralisation, although they underwent intense metabolomic reprogramming. Conversely, juveniles showed the inverse pattern, with a reduction in survival and an increase in development time at the highest pCO2 levels tested, with no indication of metabolomic reprogramming. Metabolomic sensitivity to elevated pCO2 increased until metamorphosis (between larval and juvenile stages) and decreased afterward, suggesting this transition as a metabolic keystone for marine invertebrates with complex life cycles.

Summer Is Coming! Tackling Ocean Warming in Atlantic Salmon Cage Farming.

Atlantic salmon (Salmo salar) cage farming has traditionally been located at higher latitudes where cold seawater temperatures favor this practice. However, these regions can be impacted by ocean warming and heat waves that push seawater temperature beyond the thermo-tolerance limits of this species. As more mass mortality events are reported every year due to abnormal sea temperatures, the Atlantic salmon cage aquaculture industry acknowledges the need to adapt to a changing ocean. This paper reviews adult Atlantic salmon thermal tolerance limits, as well as the deleterious eco-physiological consequences of heat stress, with emphasis on how it negatively affects sea cage aquaculture production cycles. Biotechnological solutions targeting the phenotypic plasticity of Atlantic salmon and its genetic diversity, particularly that of its southernmost populations at the limit of its natural zoogeographic distribution, are discussed. Some of these solutions include selective breeding programs, which may play a key role in this quest for a more thermo-tolerant strain of Atlantic salmon that may help the cage aquaculture industry to adapt to climate uncertainties more rapidly, without compromising profitability. Omics technologies and precision breeding, along with cryopreservation breakthroughs, are also part of the available toolbox that includes other solutions that can allow cage farmers to continue to produce Atlantic salmon in the warmer waters of the oceans of tomorrow.

Salinity shapes the stress responses and energy reserves of marine polychaetes exposed to warming: From molecular to functional phenotypes.

Estuarine systems are critical transition zones influenced by sea, land and freshwater. An array of human activities impacts these areas leading to multiple-stressor interactions. Temperature and salinity are among the most relevant drivers in estuaries, shaping species growth, reproduction and distribution. However, few studies provide an overview of cellular rewiring processes under multiple-stressor environments. Here, we tested how salinity could shape the response of ragworms Hediste diversicolor, an important bioindicator and commercial species, to elevated temperature. We exposed polychaetes to three temperatures for a month, simulating control, ocean warming and heatwave conditions (24, 27 and 30 °C, respectively) combined with two salinities (20 and 30). We quantified whole-organism performance (wet weight gain and survival), along with cellular stress response (CSR) and energy reserves of worms after 14 and 28 days of exposure. Significant three-way interactions between temperature, salinity and exposure time show the non-linearity of molecular responses. Worms at a salinity of 20 were more sensitive to warming than worms exposed to a salinity of 30. The combination of high temperature and low salinity can act synergistically to induce oxidative stress and macromolecular damage in worm tissues. This finding was supported by an induction of the CSR, with a concomitant decrease of energy reserves, pointing towards a metabolic compensation strategy. However, under a higher salinity (30), the need for a CSR upon thermal challenge was reduced and energy content increased with temperature, which suggests that environmental conditions were within the optimum range. Heatwaves striking low-salinity areas of estuaries can therefore negatively impact the cellular physiology of H. diversicolor, with greater metabolic costs. However, extreme stress levels were not reached as worms incremented wet weight and survival was high under all conditions tested. Our findings are important for the optimization of ragworm aquaculture and adaptive conservation strategies of estuarine systems.

Conserved fatty acid profiles and lipid metabolic pathways in a tropical reef fish exposed to ocean warming - An adaptation mechanism of tolerant species?

Climate warming is causing rapid spatial expansion of ocean warm pools from equatorial latitudes towards the subtropics. Sedentary coral reef inhabitants in affected areas will thus be trapped in high temperature regimes, which may become the "new normal". In this study, we used clownfish Amphiprion ocellaris as model organism to study reef fish mechanisms of thermal adaptation and determine how high temperature affects multiple lipid aspects that influence physiology and thermal tolerance. We exposed juvenile fish to two different experimental conditions, implemented over 28 days: average tropical water temperatures (26 °C, control) or average warm pool temperatures (30 °C). We then performed several analyses on fish muscle and liver tissues: i) total lipid content (%), ii) lipid peroxides, iii) fatty acid profiles, iv) lipid metabolic pathways, and v) weight as body condition metric. Results showed that lipid storage capacity in A. ocellaris was not affected by elevated temperature, even in the presence of lipid peroxides in both tissues assessed. Additionally, fatty acid profiles were unresponsive to elevated temperature, and lipid metabolic networks were consequently well conserved. Consistent with these results, we did not observe changes in fish weight at elevated temperature. There were, however, differences in fatty acid profiles between tissue types and over time. Liver showed enhanced α-linolenic and linoleic acid metabolism, which is an important pathway in stress response signaling and modulation on environmental changes. Temporal oscillations in fatty acid profiles are most likely related to intrinsic factors such as growth, which leads to the mobilization of energetic reserves between different tissues throughout time according to organism needs. Based on these results, we propose that the stability of fatty acid profiles and lipid metabolic pathways may be an important thermal adaptation feature of fish exposed to warming environments.

Calcium homeostasis and stable fatty acid composition underpin heatwave tolerance of the keystone polychaete Hediste diversicolor.

Extreme weather events, such as heatwaves, are becoming increasingly frequent, long-lasting and severe as global climate change continues, shaping marine biodiversity patterns worldwide. Increased risk of overheating and mortality across major taxa have been recurrently observed, jeopardizing the sustainability of ecosystem services. Molecular responses of species, which scale up to physiological and population responses, are determinant processes that modulate species sensitivity or tolerance to extreme weather events. Here, by integrating proteomic, fatty acid profiling and physiological approaches, we show that the tolerance of the intertidal ragworm Hediste diversicolor, a keystone species in estuarine ecosystems and an emergent blue bio-resource, to long-lasting heatwaves (24  vs 30 °C for 30 days) is shaped by calcium homeostasis, immune function and stability of fatty acid profiles. These features potentially enabled H. diversicolor to increase its thermal tolerance limit by 0.81 °C under the heatwave scenario and maintain survival. No growth trade-offs were detected, as wet weight remained stable across conditions. Biological variation of physiological parameters was lower when compared to molecular measures. Proteins showed an overall elevated coefficient of variation, although decreasing molecular variance under the heatwave scenario was observed for both proteins and fatty acids. This finding is consistent with the phenomenon of physiological canalization in extreme environments and contradicts the theory that novel conditions increase trait variation. Our results show that keystone highly valued marine polychaetes are tolerant to heatwaves, confirming the potential of H. diversicolor as a blue bio-resource and opening new avenues for sustainable marine aquaculture development.

Different sensitivity to heatwaves across the life cycle of fish reflects phenotypic adaptation to environmental niche.

Predicting responses of marine organisms to global change requires eco-physiological assessments across the complex life cycles of species. Here, we experimentally tested the vulnerability of a demersal temperate fish (Sparus aurata) to long-lasting heatwaves, on larval, juvenile and adult life-stages. Fish were exposed to simulated coastal (18 °C), estuarine (24 °C) summer temperatures, and heatwave conditions (30 °C) and their physiological responses were assessed based on cellular stress response biomarkers (heat shock protein 70 kDa, ubiquitin, antioxidant enzymes, lipid peroxidation) and phenotypic measures (histopathology, condition and mortality). Life-stage vulnerability can be ranked as larvae > adults > juveniles, based on mortality, tissue pathology and the capacity to employ cellular stress responses, reflecting the different environmental niches of each life stage. While larvae lacked acclimation capacity, which resulted in damage to tissues and elevated mortality, juveniles coped well with elevated temperature. The rapid induction of cytoprotective proteins maintained the integrity of vital organs in juveniles, suggesting adaptive phenotypic plasticity in coastal and estuarine waters. Adults displayed lower plasticity to heatwaves as they transition to deeper habitats for maturation, showing tissue damage in brain, liver and muscle. Life cycle closure of sea breams in coastal habitats will therefore be determined by larval and adult stages.

Molecular assessment of wild populations in the marine realm: Importance of taxonomic, seasonal and habitat patterns in environmental monitoring.

Scientists are currently faced with the challenge of assessing the effects of anthropogenic stressors on aquatic ecosystems. Cellular stress response (CSR) biomarkers are ubiquitous and phylogenetically conserved among metazoans and have been successfully applied in environmental monitoring but they can also vary according to natural biotic and abiotic factors. The reported variability may thus limit the wide application of biomarkers in monitoring, imposing the need to identify variability levels in the field. Our aim was to carry out a comprehensive in situ assessment of the CSR (heat shock protein 70 kDa, ubiquitin, antioxidant enzymes) and oxidative damage (lipid peroxidation) in wild populations across marine taxa by collecting fish, crustaceans, mollusks and cnidarians during two different seasons (spring and summer) and two habitat types (coast and estuary). CSR end-point patterns were different between taxa with mollusks having higher biomarker levels, followed by the cnidarians, while fish and crustaceans showed lower biomarker levels. The PCA showed clear clusters related to mobility/sessile traits with sessile organisms showing greater levels (>2-fold) of CSR proteins and oxidative damage. Mean intraspecific variability in the CSR measured by the coefficient of variation (% CV) (including data from all seasons and sites) was elevated (35-94%). Overall, there was a seasonal differentiation in biomarker patterns across taxonomic groups, especially evident in fish and cnidarians. A differentiation in biomarker patterns between habitat types was also observed and associated with phenotypic plasticity or local adaptation. Overall, specimens collected in the estuary had lower biomarker levels when compared to specimens collected in the coast. This work highlights the importance of assessing baseline biomarker levels across taxa, seasons and habitats prior to applying biomarker analyses in environmental monitoring. Selecting bioindicator species, defining sampling strategies, and identifying confounding factors are crucial preliminary steps that ensure the success of biomarkers as powerful tools in biomonitoring.

Molecular Plasticity under Ocean Warming: Proteomics and Fitness Data Provides Clues for a Better Understanding of the Thermal Tolerance in Fish.

Ocean warming is known to alter the performance and fitness of marine organisms albeit the proteome underpinnings of species thermal tolerance are still largely unknown. In this 1-month experiment we assessed the vulnerability of the gilt-head sea bream Sparus aurata, taken here as a biological model for some key fisheries species, to ocean warming (control 18°C, nursery ground temperature 24°C and heat wave 30°C). Survival was impaired after 28 days, mainly at 30°C although fishes' condition was unaltered. Muscle proteome modulation was assessed at 14 and 21 days, showing that protein expression profiles were similar between fish exposed to 18 and 24°C, differing from fish exposed to 30°C. Fish subjected to 24°C showed an enhanced glycolytic potential and decreased glycogenolysis mainly at 14 days of exposure. Fish subjected to 30°C also showed enhanced glycolytic potential and up-regulated proteins related to gene expression, cellular stress response (CSR), and homeostasis (mostly cytoskeletal dynamics, acid-base balance, chaperoning). However, inflammatory processes were elicited at 21 days along with a down-regulation of the tricarboxylic acid cycle. Thus, juvenile fish seem able to acclimate to 24°C but possibly not to 30°C, which is the predicted temperature for estuaries during heat waves by the year 2100. This may be related with increasing constraints on organism physiology associated with metabolic scope available for performance and fitness at higher temperatures. Consequently, recruitment of commercial sea breams may be in jeopardy, highlighting the need for improved management plans for fish stocks.

Seasonal changes in stress biomarkers of an exotic coastal species - Chaetopleura angulata (Polyplacophora) - Implications for biomonitoring.

Knowledge on baseline values of stress biomarkers in natural conditions is urgent due to the need of reference values for monitoring purposes. Here we assessed the cellular stress response of the chiton Chaetopleura angulata in situ. Biomarkers commonly used in environmental monitoring (heat shock protein 70kDa, total ubiquitin, catalase, glutathione-S-transferase, superoxide-dismutase, lipid peroxidation) were analyzed in the digestive system, gills and muscle of C. angulata, under spring and summer conditions in order to assess seasonal tissue-specific responses. Season had an effect on all targeted organs, especially affecting the digestive system which displayed clear seasonal clusters. The respective Integrated Biomarker Response (IBR) showed a 7.2-fold seasonal difference. Muscle and gills showed similar IBRs between seasons making them appropriate organs to monitor chemical pollution as they were less responsive to seasonal variation. The most stable biomarkers in these organs were ubiquitin and superoxide-dismutase thus being reliable for monitoring purposes.

Comparing biomarker responses during thermal acclimation: A lethal vs non-lethal approach in a tropical reef clownfish.

Knowledge of thermal stress biology for most tropical fish species in reef ecosystems under climate change is still quite limited. Thus, the objective of this study was to measure the time-course changes of thermal stress biomarkers in the commercially exploited coral reef fish Amphiprion ocellaris, during a laboratory simulated event of increased temperature. Heat shock protein 70kDa (Hsp70) and total ubiquitin (Ub) were determined in the muscle (lethal method) and in the fin (non-lethal alternative method) under two temperature treatments (control - 26°C and elevated temperature - 30°C) throughout one month with weekly samplings. Results suggest that biomarker basal levels are tissue-specific and influence the degree of response under temperature exposure. Responses were highly inducible in the muscle but not in fin tissue, indicating that the latter is not reliable for monitoring purposes. Thermal stress was observed in the muscle after one week of exposure (both biomarkers increased significantly) and Ub levels then decreased, suggesting the animals were able to acclimate by maintaining high levels of Hsp70 and through an effective protein turnover. In addition, the results show that mortality rates did not differ between treatments. This indicates that A. ocellaris is capable of displaying a plastic response to elevated temperature by adjusting the protein quality control system to protect cell functions, without decreasing survival. Thus, this coral reef fish species presents a significant acclimation potential under ocean warming scenarios of +4°C. Monitoring of thermal stress through a non-lethal method, fin-clipping, although desirable proved to be inadequate for this species.

Physiological, cellular and biochemical thermal stress response of intertidal shrimps with different vertical distributions: Palaemon elegans and Palaemon serratus.

The ability to cope with high temperature variations is a critical factor in intertidal communities. Two species of intertidal rocky shore shrimps (Palaemon sp.) with different vertical distributions were collected from the Portuguese coast in order to test if they were differentially sensitive to thermal stress. Three distinct levels of biological organization (organismal, biochemical, and cellular) were surveyed. The shrimp were exposed to a constant rate of temperature increase of 1°C x h(-1), starting at 20°C until reaching the CTMax (critical thermal maximum). During heat stress, two biomarkers of protein damage were quantified in the muscle via enzyme-linked immunosorbent assays: heat shock proteins HSP70 (hsp70/hsc70) and total ubiquitin. Muscle histopathological alterations caused by temperature were also evaluated. CTMax values were not significantly different between the congeners (P. elegans 33.4 ± 0.5 °C; P. serratus 33.0 ± 0.5 °C). Biomarker levels did not increase along the temperature trial, but P. elegans (higher intertidal) showed higher amounts of HSP70 and total ubiquitin than P. serratus (lower intertidal). HSP70 and total ubiquitin levels showed a positive significant correlation in both species, suggesting that their association is important in thermal tolerance. Histopathological observations of muscle tissue in P. serratus showed no gross alterations due to temperature but did show localized atrophy of muscle fibers at CTMax. In P. elegans, alterations occurred at a larger scale, showing multiple foci of atrophic muscular fascicles caused by necrotic or autolytic processes. In conclusion, Palaemon congeners displayed different responses to stress at a cellular level, with P. elegans having greater biomarker levels and histopathological alterations.

Synergy of environmental variables alters the thermal window and heat shock response: an experimental test with the crab Pachygrapsus marmoratus.

The intertidal zone is an extremely variable habitat, imposing stressful conditions on its inhabiting communities. Tolerance towards extremes of temperature, salinity and pH are crucial in these habitats. Despite the vast literature on stress tolerance, few studies have focused on the synergistic effects of several variables on thermal tolerance and HSP70 (heat shock protein 70 kDa) levels. In this work, the crabs were exposed to three experimental conditions 1) thermal ramp at standard pH (8) and saline conditions (35‰) (named T), 2) thermal ramp at standard pH (8) and hyposaline conditions (15‰) (named T plus HypoS), and 3) thermal ramp at lower pH (7) and standard saline conditions (35‰) (named T plus A). Two physiological parameters (Critical Thermal Maximum - CTMax, and osmolality) and a stress biomarker (HSP70) were chosen for this analysis. These parameters were measured in all of the aforementioned conditions. CTMax for each set of conditions was reached by exposing the organisms to a rate of temperature increase of 1 °C h(-1) until loss of equilibrium. Haemolymph samples were taken every 2 °C to quantify HSP70 and osmolality. Results showed that CTMax did not differ between crabs solely exposed to T stress and crabs exposed to T plus HypoS stress. However, HSP70 production was impaired in T plus HypoS stress. When crabs were exposed to T plus A stress, they showed a significantly higher CTMax, suggesting that short-term exposure to acidified conditions may alter the thermal window of this species. Nevertheless, in T plus A conditions HSP70 production was impaired as well. Regarding osmolality it decreased according to temperature increase in all tested stress conditions. This study showed that the heat stress response is altered by the synergistic effect of variables. Physiological end-points (i.e. CTMax) may vary and the expression of stress proteins such as HSP70 may be impaired.

Effect of temperature in multiple biomarkers of oxidative stress in coastal shrimp.

Various studies in captivity and in the wild have pointed to the effect of season, and temperature in particular, in the levels of the oxidative stress biomarkers currently used for environmental quality assessment. However, knowledge on how temperature affects the oxidative stress response is unavailable for most species. This study investigated the effect of increasing temperature on lipid peroxidation, catalase activity, superoxide dismutase and glutathione-S-transferase in the shrimps, Palaemon elegans and Palaemon serratus. It was concluded that increasing temperatures significantly affect all the biomarkers tested in both species, with the exception of superoxide dismutase in P. serratus which was not affected by temperature. The oxidative stress response was more intense in P. elegans, than in P. serratus, producing higher peaks of all biomarkers at temperatures between 22°C and 26°C, followed by low levels at higher temperatures. It was concluded that monitoring of ecosystems using oxidative stress biomarkers should take into account the species and thermal history of the organisms. Sampling should be avoided during heat waves and immediately after heat waves.

Effect of increasing temperature in the differential activity of oxidative stress biomarkers in various tissues of the Rock goby, Gobius paganellus.

Oxidative stress biomarkers have been widely used in the development of ecological indices and in the assessment of exposure of aquatic organisms to contaminants from agricultural, industrial and urban pollution. However, temperature is known to also have a significant effect on oxidative stress biomarkers. This way, temperature is a confounding factor that may result in difficulties in the interpretation of oxidative stress biomarkers response patterns. Since climate change is expected to result in more frequent and intense heat wave events it is pertinent to investigate the effect of increasing temperature in the oxidative stress response of common aquatic organisms. It is also important to assess the differential response of different body tissues, given that they are differently exposed to temperature depending on their location and physiological function. This study investigates the effect of increasing temperature (20 °C-34 °C) in the response of multiple biomarkers of oxidative stress: lipid peroxidation, glutathione-S-transferase, superoxide dismutase and catalase activities, in the muscle, liver and gills of a common coastal fish, the Rock goby, Gobius paganellus. The response of the oxidative stress biomarkers analysed were always higher in the gills than in the other tissues. Muscle generally presented the lower levels of any of the biomarkers tested when compared to other tissues. Nevertheless, muscle tissue always responded significantly to temperature, as did the liver, while the gills were unresponsive in terms of lipid peroxidation and glutathione-S-transferase. Unresponsive tissues to temperature may be particularly interesting as indicators of pollution, given that temperature will not be a confounding variable in their oxidative stress response.