Gene losses, parallel evolution and heightened expression confer adaptations to dedicated cleaning behaviour.

BACKGROUND: Cleaning symbioses are captivating interspecific interactions in which a cleaner fish removes ectoparasites from its client, contributing to the health and diversity of natural fish communities and aquaculture systems. However, the genetic signatures underlying this specialized behaviour remain poorly explored. To shed light on this, we generated a high-quality chromosome-scale genome of the bluestreak cleaner wrasse Labroides dimidiatus, a dedicated cleaner with cleaning as primary feeding mechanism throughout its life. RESULTS: Compared with facultative and non-cleaner wrasses, L. dimidiatus was found with notable contractions in olfactory receptors implying their limited importance in dedicated cleaning. Instead, given its distinct tactile pre-conflict strategies, L. dimidiatus may rely more heavily on touch sensory perception, with heightened gene expression in the brain in anticipation of cleaning. Additionally, a reduction in NLR family CARD domain-containing protein 3 might enhance innate immunity of L. dimidiatus, probably assisting to reduce the impacts from parasite infections. In addition, convergent substitutions for a taste receptor and bone development genes across cleaners (L. dimidiatus and facultative cleaners) may provide them with evolved food discrimination abilities and jaw morphology that differentiate them from non-cleaners. Moreover, L. dimidiatus may exhibit specialized neural signal transductions for cleaning, as evidenced by positive selection in genes related to the glutamatergic synapse pathway. Interestingly, numerous glutamate receptors also demonstrated significantly higher expression in L. dimidiatus not engaged in cleaning, as compared to those involved in cleaning. Besides, apparent contractions in L. dimidiatus for protocadherins, which are responsible for neuronal development, may further promote specialized neural signal transductions in this species. CONCLUSIONS: This study reveals that L. dimidiatus harbours substantial losses in specific gene families, convergent evolutions across cleaners and a large-scale high gene expression in preparation for cleaning, allowing for adaptation to the dedicated cleaning behaviour.

Impacts of Deoxygenation and Hypoxia on Shark Embryos Anti-Predator Behavior and Oxidative Stress.

Climate change is leading to the loss of oxygen content in the oceans and endangering the survival of many marine species. Due to sea surface temperature warming and changing circulation, the ocean has become more stratified and is consequently losing its oxygen content. Oviparous elasmobranchs are particularly vulnerable as they lay their eggs in coastal and shallow areas, where they experience significant oscillations in oxygen levels. Here, we investigated the effects of deoxygenation (93% air saturation) and hypoxia (26% air saturation) during a short-term period (six days) on the anti-predator avoidance behavior and physiology (oxidative stress) of small-spotted catshark (Scyliorhinus canicula) embryos. Their survival rate decreased to 88% and 56% under deoxygenation and hypoxia, respectively. The tail beat rates were significantly enhanced in the embryos under hypoxia compared to those exposed to deoxygenation and control conditions, and the freeze response duration showed a significant opposite trend. Yet, at the physiological level, through the analyses of key biomarkers (SOD, CAT, GPx, and GST activities as well as HSP70, Ubiquitin, and MDA levels), we found no evidence of increased oxidative stress and cell damage under hypoxia. Thus, the present findings show that the projected end-of-the-century deoxygenation levels elicit neglectable biological effects on shark embryos. On the other hand, hypoxia causes a high embryo mortality rate. Additionally, hypoxia makes embryos more vulnerable to predators, because the increased tail beat frequency will enhance the release of chemical and physical cues that can be detected by predators. The shortening of the shark freeze response under hypoxia also makes the embryos more prone to predation.

Combined effects of climate change and BDE-209 dietary exposure on the behavioural response of the white seabream, Diplodus sargus.

Decabromodiphenyl-ether (BDE-209) is a persistent organic pollutant ubiquitously found in marine environments worldwide. Even though this emerging chemical contaminant is described as highly toxic, bioaccumulative and biomagnifiable, limited studies have addressed the ecotoxicological implications associated with its exposure in non-target marine organisms, particularly from a behavioural standpoint. Alongside, seawater acidification and warming have been intensifying their impacts on marine ecosystems over the years, compromising species welfare and survival. BDE-209 exposure as well as seawater acidification and warming are known to affect fish behaviour, but information regarding their interactive effects is not available. In this study, long-term effects of BDE-209 contamination, seawater acidification and warming were studied on different behavioural traits of Diplodus sargus juveniles. Our results showed that D. sargus exhibited a marked sensitivity in all the behaviour responses after dietary exposure to BDE-209. Fish exposed to BDE-209 alone revealed lower awareness of a risky situation, increased activity, less time spent within the shoal, and reversed lateralization when compared to fish from the Control treatment. However, when acidification and/or warming were added to the equation, behavioural patterns were overall altered. Fish exposed to acidification alone exhibited increased anxiety, being less active, spending more time within the shoal, while presenting a reversed lateralization. Finally, fish exposed to warming alone were more anxious and spent more time within the shoal compared to those of the Control treatment. These novel findings not only confirm the neurotoxicological attributes of brominated flame retardants (like BDE-209), but also highlight the relevance of accounting for the effects of abiotic variables (e.g. pH and seawater temperature) when investigating the impacts of environmental contaminants on marine life.

Camouflage and Exploratory Avoidance of Newborn Cuttlefish under Warming and Acidification.

Ocean warming and acidification have been shown to elicit deleterious effects on cephalopod mollusks, especially during early ontogeny, albeit effects on behavior remain largely unexplored. This study aimed to evaluate, for the first time, the effect of end-of-the-century projected levels of ocean warming (W; + 3 °C) and acidification (A; 980 µatm pCO2) on Sepia officinalis hatchlings' exploratory behavior and ability to camouflage in different substrate complexities (sand and black and white gravel). Cuttlefish were recorded in open field tests, from which mobility and exploratory avoidance behavior data were obtained. Latency to camouflage was registered remotely, and pixel intensity of body planes and background gravel were extracted from photographs. Hatching success was lowered under A and W combined (AW; 72.7%) compared to control conditions (C; 98.8%). Motion-related behaviors were not affected by the treatments. AW delayed camouflage response in the gravel substrate compared to W alone. Moreover, cuttlefish exhibited a higher contrast and consequently a stronger disruptive pattern under W, with no changes in background matching. These findings suggest that, although climate change may elicit relevant physiological challenges to cuttlefish, camouflage and mobility of these mollusks are not undermined under the ocean of tomorrow.

Effects of Hypoxia on Coral Photobiology and Oxidative Stress.

Global ocean oxygen (O2) content is decreasing as climate change drives declines in oxygen solubility, strengthened stratification of seawater masses, increased biological oxygen consumption and coastal eutrophication. Studies on the biological effects of nocturnal decreased oxygen concentrations (hypoxia) on coral reefs are very scarce. Coral reefs are fundamental for supporting one quarter of all marine species and essential for around 275 million people worldwide. This study investigates acute physiological and photobiological responses of a scleractinian coral (Acropora spp.) to overnight hypoxic conditions (<2 mg/L of O2). Bleaching was not detected, and visual and physical aspects of corals remained unchanged under hypoxic conditions. Most photobiological-related parameters also did not show significant changes between treatments. In addition to this, no significant differences between treatments were observed in the pigment composition. However, hypoxic conditions induced a significant decrease in coral de-epoxidation state of the xanthophyll cycle pigments and increase in DNA damage. Although the present findings suggest that Acropora spp. is resilient to some extent to short-term daily oxygen oscillations, long-term exposure to hypoxia, as predicted to occur with climate change, may still have deleterious effects on corals.

Access to Cleaning Services Alters Fish Physiology Under Parasite Infection and Ocean Acidification.

Cleaning symbioses are key mutualistic interactions where cleaners remove ectoparasites and tissues from client fishes. Such interactions elicit beneficial effects on clients' ecophysiology, with cascading effects on fish diversity and abundance. Ocean acidification (OA), resulting from increasing CO2 concentrations, can affect the behavior of cleaner fishes making them less motivated to inspect their clients. This is especially important as gnathiid fish ectoparasites are tolerant to ocean acidification. Here, we investigated how access to cleaning services, performed by the cleaner wrasse Labroides dimidiatus, affect individual client's (damselfish, Pomacentrus amboinensis) aerobic metabolism in response to both experimental parasite infection and OA. Access to cleaning services was modulated using a long-term removal experiment where cleaner wrasses were consistently removed from patch reefs around Lizard Island (Australia) for 17 years or left undisturbed. Only damselfish with access to cleaning stations had a negative metabolic response to parasite infection (maximum metabolic rate-MO2Max; and both factorial and absolute aerobic scope). Moreover, after an acclimation period of 10 days to high CO2 (∼1,000 µatm CO2), the fish showed a decrease in factorial aerobic scope, being the lowest in fish without the access to cleaners. We propose that stronger positive selection for parasite tolerance might be present in reef fishes without the access to cleaners, but this might come at a cost, as readiness to deal with parasites can impact their response to other stressors, such as OA.

Ocean Acidification Alleviates Dwarf Eelgrass (Zostera noltii) Lipid Landscape Remodeling under Warming Stress.

Coastal seagrass meadows provide a variety of essential ecological and economic services, including nursery grounds, sediment stabilization, nutrient cycling, coastal protection, and blue carbon sequestration. However, these ecosystems are highly threatened by ongoing climatic change. This study was aimed to understand how the dwarf eelgrass Zostera noltii leaf lipid landscapes are altered under predicted ocean warming (+4 °C) and hypercapnic (ΔpH 0.4) conditions. Warming and hypercapnic conditions were found to induce a severe reduction in the leaf total fatty acid, though the combined treatment substantially alleviated this depletion. The lipid discrimination revealed a significant increase in the relative monogalactosyldiacylglycerol (MGDG) content in both hypercapnic and warming conditions, allied to plastidial membrane stabilization mechanisms. Hypercapnia also promoted enhanced phosphatidylglycerol (PG) leaf contents, a mechanism often associated with thylakoid reinvigoration. In addition to changing the proportion of storage, galacto- and phospholipids, the tested treatments also impacted the FA composition of all lipid classes, with warming exposure leading to decreases in polyunsaturated fatty acids (PUFAs); however, the combination of both stress conditions alleviated this effect. The observed galactolipid and phospholipid PUFA decreases are compatible with a homeoviscous adaptation, allowing for the maintenance of membrane stability by counteracting excessive membrane fluidity. Neutral lipid contents were substantially increased under warming conditions, especially in C18 fatty acids (C18), impairing their use as substrates for fatty acylated derivatives essential for maintaining the osmotic balance of cells. An analysis of the phospholipid and galactolipid fatty acid profiles as a whole revealed a higher degree of discrimination, highlighting the higher impact of warming and the proposed stress alleviation effect induced by increased water-dissolved CO2 availability. Still, it is essential to remember that the pace at which the ocean is warming can overcome the ameliorative capacity induced by higher CO2 availability, leaving seagrasses under severe heat stress beyond their lipid remodeling capacity.

Cuttlefish Buoyancy in Response to Food Availability and Ocean Acidification.

Carbon dioxide concentration in the atmosphere is expected to continue rising by 2100, leading to a decrease in ocean pH in a process known as ocean acidification (OA). OA can have a direct impact on calcifying organisms, including on the cuttlebone of the common cuttlefish Sepia officinalis. Moreover, nutritional status has also been shown to affect the cuttlebone structure and potentially affect buoyancy. Here, we aimed to understand the combined effects of OA (980 µatm CO2) and food availability (fed vs. non-fed) on the buoyancy of cuttlefish newborns and respective cuttlebone weight/area ratio (as a proxy for calcification). Our results indicate that while OA elicited negative effects on hatching success, it did not negatively affect the cuttlebone weight/area ratio of the hatchlings-OA led to an increase in cuttlebone weight/area ratio of fed newborns (but not in unfed individuals). The proportion of "floating" (linked to buoyancy control loss) newborns was greatest under starvation, regardless of the CO2 treatment, and was associated with a drop in cuttlebone weight/area ratio. Besides showing that cuttlefish buoyancy is unequivocally affected by starvation, here, we also highlight the importance of nutritional condition to assess calcifying organisms' responses to ocean acidification.

The past, present and future of cleaner fish cognitive performance as a function of CO2 levels.

Ocean acidification is one of the many consequences of climate change. Various studies suggest that marine organisms' behaviour will be impaired under high CO2. Here, we show that the cognitive performance of the cleaner wrasse, Labroides dimidiatus, has not suffered from the increase of CO2 from pre-industrial levels to today, and that the standing variation in CO2 tolerance offers potential for adaptation to at least 750 µatm. We acclimated cleaners over 30 days to five levels of pCO2, from pre-industrial to high future CO2 scenarios, before testing them in an ecologically relevant task-the ability to learn to prioritize an ephemeral food source over a permanent one. Fish learning abilities remained stable from pre-industrial to present-day pCO2. While performance was reduced under mid (750 µatm) and high CO2 (980 µatm) scenarios, under the former 36% of cleaners still solved the task. The presence of tolerant individuals reveals potential for adaptation, as long as selection pressure on cognitive performance is strong. However, the apparent absence of high CO2 tolerant fish, and potentially synergistic effects between various climate change stressors, renders the probability of further adaptation unlikely.

Neurobiological and behavioural responses of cleaning mutualisms to ocean warming and acidification.

Cleaning interactions are textbook examples of mutualisms. On coral reefs, most fishes engage in cooperative interactions with cleaners fishes, where they benefit from ectoparasite reduction and ultimately stress relief. Furthermore, such interactions elicit beneficial effects on clients' ecophysiology. However, the potential effects of future ocean warming (OW) and acidification (OA) on these charismatic associations are unknown. Here we show that a 45-day acclimation period to OW (+3 °C) and OA (980 µatm pCO2) decreased interactions between cleaner wrasses (Labroides dimidiatus) and clients (Naso elegans). Cleaners also invested more in the interactions by providing tactile stimulation under OA. Although this form of investment is typically used by cleaners to prolong interactions and reconcile after cheating, interaction time and client jolt rate (a correlate of dishonesty) were not affected by any stressor. In both partners, the dopaminergic (in all brain regions) and serotoninergic (forebrain) systems were significantly altered by these stressors. On the other hand, in cleaners, the interaction with warming ameliorated dopaminergic and serotonergic responses to OA. Dopamine and serotonin correlated positively with motivation to interact and cleaners interaction investment (tactile stimulation). We advocate that such neurobiological changes associated with cleaning behaviour may affect the maintenance of community structures on coral reefs.

Ocean warming and acidification may challenge the riverward migration of glass eels.

The dramatic decline of European eel ( Anguilla anguilla) populations over recent decades has attracted considerable attention and concern. Furthermore, little is known about the sensitivity of the early stages of eels to projected future environmental change. Here, we investigated, for the first time, the potential combined effects of ocean warming (OW; Δ + 4°C; 18°C) and acidification (OA; Δ - 0.4 pH units) on the survival and migratory behaviour of A. anguilla glass eels, namely their preference towards riverine cues (freshwater and geosmin). Recently arrived individuals were exposed to isolated and combined OW and OA conditions for 100 days, adjusting for the salinity gradients associated with upstream migration. A two-choice test was used to investigate migratory activity and shifts in preference towards freshwater environments. While OW decreased survival and increased migratory activity, OA appears to hinder migratory response, reducing the preference for riverine cues. Our results suggest that future conditions could potentially favour an early settlement of glass eels, reducing the proportion of fully migratory individuals. Further research into the effects of climate change on eel migration and habitat selection is needed to implement efficient conservation plans for this critically endangered species.

Body size and season influence elemental composition of tissues in ocean sunfish Mola mola juveniles.

The effects of body size and season on the elemental composition of ocean sunfish Mola mola were assessed for the first time. A total of 57 by-caught juvenile specimens measuring between 31.8 and 120.3 cm were sampled in spring and autumn. Concentrations of trace elements (three essential - Co, Cu, Zn, and three non-essential - As, Cd, Pb) were determined in five body tissues [gills, gelatin (subcutaneous white gelatinous layer), liver, white muscle and red muscle]. Elemental composition of M. mola tissues was found to vary with both body size and season. When an effect of size was verified, the most common trend was a decrease in elemental levels with increasing fish size, most likely deriving from the fast growth rate of this fish (i.e. dilution effect) and the occurrence of an ontogenetic shift in dietary preferences. Differently, Zn levels increased with fish size in both gills and red muscle, potentially deriving from a greater physiological need in those tissues as fish grow. Seasonal differences in trace element levels were observed for approximately half of the studied cases (tissue/trace element) with greater elemental concentrations being mostly found in autumn. Such pattern was most obvious in liver tissue, presumably resulting from a greater dietary elemental uptake in the end of summer/autumn. A general absence of seasonal differences was observed in the gills, white muscle and red muscle. Interestingly, a trend of enhanced concentrations in spring was observed for gelatin, potentially deriving from past long-term differences in exposure to trace elements.

Bioaccumulation and ecotoxicological responses of juvenile white seabream (Diplodus sargus) exposed to triclosan, warming and acidification.

Triclosan (TCS) is a synthetic microbial compound widely used in the formulation of various personal care products. Its frequent detection in marine ecosystems, along with its physical and chemical properties, suggest that TCS can be highly persistent, being easily bioaccumulated by biota and, therefore, eliciting various toxicological responses. Yet, TCS's mechanisms of bioaccumulation and toxicity still deserve further research, particularly focusing on the interactive effects with climate change-related stressors (e.g. warming and acidification), as both TCS chemical behaviour and marine species metabolism/physiology can be strongly influenced by the surrounding abiotic conditions. Hence, the aim of this study was to assess TCS bioaccumulation and ecotoxicological effects (i.e. animal fitness indexes, antioxidant activity, protein chaperoning and degradation, neurotoxicity and endocrine disruption) in three tissues (i.e. brain, liver and muscle) of juvenile Diplodus sargus exposed to the interactive effects of TCS dietary exposure (15.9 µg kg-1 dw), seawater warming (ΔTºC = +5 °C) and acidification (ΔpCO2 âˆ¼ +1000 µatm, equivalent to ΔpH = -0.4 units). Muscle was the primary organ of TCS bioaccumulation, and climate change stressors, particularly warming, significantly reduced TCS bioaccumulation in all fish tissues. Furthermore, the negative ecotoxicological responses elicited by TCS were significantly altered by the co-exposure to acidification and/or warming, through either the enhancement (e.g. vitellogenin content) or counteraction/inhibition (e.g. heat shock proteins HSP70/HSC70 content) of molecular biomarker responses, with the combination of TCS plus acidification resulting in more severe alterations. Thus, the distinct patterns of TCS tissue bioaccumulation and ecotoxicological responses induced by the different scenarios emphasized the need to further understand the interactive effects between pollutants and abiotic conditions, as such knowledge enables a better estimation and mitigation of the toxicological impacts of climate change in marine ecosystems.

Distinct Bleaching Resilience of Photosynthetic Plastid-Bearing Mollusks Under Thermal Stress and High CO2 Conditions.

The impact of temperature on photo-symbiotic relationships has been highly studied in the tropical reef-forming corals but overlooked in less charismatic groups such as solar-powered sacoglossan sea slugs. These organisms display one of the most puzzling symbiotic features observed in the animal kingdom, i.e., their mollusk-plastid association, which enables them to retain photosynthetic active chloroplasts (i.e., kleptoplasts) retrieved from their algae feed sources. Here we analyze the impact of thermal stress (+4°C) and high pCO2 conditions (ΔpH = 0.4) in survival, photophysiology (i.e., bleaching, photosynthetic efficiency, and metabolism) and stress defense mechanisms (i.e., heat shock and antioxidant response) of solar-powered sacoglossan sea slugs, from tropical (Elysia crispata) and temperate (E. viridis) environments. High temperature was the main factor affecting the survival of both species, while pH only affected the survival of the temperate model. The photobiology of E. viridis remained stable under the combined scenario, while photoinhibition was observed for E. crispata under high temperature and high pCO2. In fact, bleaching was observed within all tropical specimens exposed to warming (but not in the temperate ones), which constitutes the first report where the incidence of bleaching in tropical animals hosting photosynthetic symbionts, other than corals, occurs. Yet, the expulsion of kleptoplasts by the tropical sea slug, allied with metabolic depression, constituted a physiological response that did not imply signs of vulnerability (i.e., mortality) in the host itself. Although the temperate species revealed greater heat shock and antioxidant enzyme response to environmental stress, we argue that the tropical (stenotherm) sea slug species may display a greater scope for acclimatization than the temperate (eurytherm) sea slug. E. crispata may exhibit increased capacity for phenotypic plasticity by increasing fitness in a much narrower thermal niche (minimizing maintenance costs), which ultimately may allow to face severe environmental conditions more effectively than its temperate generalist counterpart (E. viridis).

Effects of acute waterborne exposure to harmful algal toxin domoic acid on foraging and swimming behaviours of fish early stages.

Domoic acid (DA) is a neurotoxin naturally produced by Pseudo-nitzschia diatoms that may be transferred through the marine food web and cause mass mortality events at higher trophic levels. Yet, the effects of the dissolved marine toxin on foraging responses and swimming performances of fish early stages are poorly known. Here we evaluated the effects of short-term exposure (24 h) to a single dose of domoic acid (136 µg DA L-1) on larvae (15-20 days post-hatch) of three commercially important fish species (the sea breams Diplodus sargus and Sparus aurata and the meagre Argyrosomus regius). Although DA exposure did not elicit significant effects on larval survival (p > 0.05) and swimming performance (p > 0.05), the toxin significantly affected the fish capture success (p < 0.001). Our findings suggest that toxigenic Pseudo-nitzschia blooms may compromise fish early stages, in particular larvae feeding behaviours, leading to complications in the development and increasing fish vulnerability and mortality.

Differential behavioural responses to venlafaxine exposure route, warming and acidification in juvenile fish (Argyrosomus regius).

Antidepressants, such as venlafaxine (VFX), which are considered emerging environmental pollutants, are increasingly more present in the marine environment, and recent evidence suggest that they might have adverse effects on fish behaviour. Furthermore, altered environmental conditions associated to climate change (e.g. warming and acidification) can also have a determinant role on fish behaviour, fitness and survival. Yet, the underlying interactions between these environmental stressors (pharmaceuticals exposure and climate change) are still far from being fully understood. The aim of this study was to assess behavioural responses (in juvenile meagre (Argyrosomus regius) exposed to VFX via water ([VFX] ~20µgL-1) and via dietary sources ([VFX] ~160µgkg-1 dry weight), as well as to increased temperature (ΔT°C=+5°C) and high CO2 levels (ΔpCO2 ~1000µatm; equivalent to ΔpH=-0.4units). Overall, VFX bioaccumulation in fish plasma was enhanced under the combination of warming and acidification. VFX triggered fish exploration, whereas fish activity and shoal cohesion were reduced. Acidification alone decreased fish exploration and shoal cohesion, and reversed fish preference to turn leftwards compared to control conditions. Such alterations were further enhanced by VFX exposure. The combination of warming and acidification also reduced shoal cohesion and loss of lateralization, regardless of VFX exposure. The distinct behaviour observed when VFX contamination, acidification and warming acted alone or in combination highlighted the need to consider the likely interactive effects of seawater warming and acidification in future research regarding the toxicological aspects of chemical contaminants.

Seagrass ecophysiological performance under ocean warming and acidification.

Seagrasses play an essential ecological role within coastal habitats and their worldwide population decline has been linked to different types of anthropogenic forces. We investigated, for the first time, the combined effects of future ocean warming and acidification on fundamental biological processes of Zostera noltii, including shoot density, leaf coloration, photophysiology (electron transport rate, ETR; maximum PSII quantum yield, Fv/Fm) and photosynthetic pigments. Shoot density was severely affected under warming conditions, with a concomitant increase in the frequency of brownish colored leaves (seagrass die-off). Warming was responsible for a significant decrease in ETR and Fv/Fm (particularly under control pH conditions), while promoting the highest ETR variability (among experimental treatments). Warming also elicited a significant increase in pheophytin and carotenoid levels, alongside an increase in carotenoid/chlorophyll ratio and De-Epoxidation State (DES). Acidification significantly affected photosynthetic pigments content (antheraxanthin, ß-carotene, violaxanthin and zeaxanthin), with a significant decrease being recorded under the warming scenario. No significant interaction between ocean acidification and warming was observed. Our findings suggest that future ocean warming will be a foremost determinant stressor influencing Z. noltii survival and physiological performance. Additionally, acidification conditions to occur in the future will be unable to counteract deleterious effects posed by ocean warming.

Seahorses under a changing ocean: the impact of warming and acidification on the behaviour and physiology of a poor-swimming bony-armoured fish.

Seahorses are currently facing great challenges in the wild, including habitat degradation and overexploitation, and how they will endure additional stress from rapid climate change has yet to be determined. Unlike most fishes, the poor swimming skills of seahorses, along with the ecological and biological constraints of their unique lifestyle, place great weight on their physiological ability to cope with climate changes. In the present study, we evaluate the effects of ocean warming (+4°C) and acidification (ΔpH = -0.5 units) on the physiological and behavioural ecology of adult temperate seahorses, Hippocampus guttulatus. Adult seahorses were found to be relatively well prepared to face future changes in ocean temperature, but not the combined effect of warming and acidification. Seahorse metabolism increased normally with warming, and behavioural and feeding responses were not significantly affected. However, during hypercapnia the seahorses exhibited signs of lethargy (i.e. reduced activity levels) combined with a reduction of feeding and ventilation rates. Nonetheless, metabolic rates were not significantly affected. Future ocean changes, particularly ocean acidification, may further threaten seahorse conservation, turning these charismatic fishes into important flagship species for global climate change issues.