Posts Tagged 'mollusks'

Controls on boron isotope ratios in marine bivalve shells: insights from a controlled experiment across pH and temperature gradients

Documenting spatial and temporal patterns of ocean acidification and understanding the way marine organisms build carbonate skeletons is critical to assessing their potential vulnerability to present and future stressors. The boron isotopic composition (δ11Bc) of many marine carbonates provides insight into the pH at the site of calcification within biocalcifiers and, by extension, the pH of ambient seawater when the carbonate formed. The modification of seawater carbonate chemistry at the site of calcification by marine calcifiers and the utility of different taxa as paleo-pH proxy archives remains an area of active research. Despite the significance of marine bivalves to ecosystem function, high-resolution paleoclimatic studies, and the shellfish industry, their biocalcification mechanisms, controls on internal pH, and potential for reconstructing records of past seawater pH remain unclear. To address these gaps, a 20.5-week flowthrough tank experiment was conducted in which four species of commercially important bivalves from the northwest Atlantic Ocean were grown in tanks with controlled pHT (pH 7.4 to 8.0) and temperature conditions (6 to 12 °C). A total of 106 shell samples from 99 individuals of adult and juvenile Arctica islandica (ocean quahog), juvenile Mercenaria mercenaria (northern quahog or hard clam), juvenile Mya arenaria (soft-shell clam) and juvenile Placopecten magellanicus (Atlantic sea scallop) were analyzed from this controlled experiment to assess the seawater pH, temperature, and growth rate controls on shell δ11Bc.These four bivalve species, grown under identical, controlled conditions, showed differential responses to the same seawater temperature and pH, likely due to differences in how they regulate the pH of their internal fluids. Juvenile P. magellanicus and juvenile M. mercenaria demonstrated significant relationships (R≥0.60; p-value <0.006) between tank pHT and δ11Bc, suggesting potential utility as proxies for past ambient seawater pH. Conversely, the δ11Bc of juvenile A. islandica and juvenile M. arenaria did not yield a strong relationship with seawater pHT but instead yielded significant relationships with shell growth rate (linear extension), with a positive relationship for M. arenaria and a negative relationship for juvenile A. islandica. The δ11B results from the few (n=9) adult A. islandica shells measured show the most variability across the range of pH and temperatures (range of 16‰) and no significant relationship was found with seawater pH or growth rate. Despite rigorous oxidative cleaning of samples, the data suggest that adult A. islandica shells contain boron-rich organic phases resistant to traditional cleaning techniques. This suggests that the next step in the development of boron-based pH proxies in A. islandica requires additional research into robust cleaning and sampling methods of periostracum and other organics. Despite the need for further investigations to constrain growth rate effects and cleaning techniques in A. islandica and M. arenaria, there is potential for developing paleo-pH proxies from P. magellanicus and M. mercenaria to better understand spatial and temporal patterns of past, present and future ocean acidification.

Continue reading ‘Controls on boron isotope ratios in marine bivalve shells: insights from a controlled experiment across pH and temperature gradients’

Coupled ocean warming and acidification reduce shell integrity and bioenergetics in juvenile Mytilus coruscus

Under realistic climate change scenarios, marine bivalves face compounding stressors from concurrent ocean warming and acidification. Research has established the separate effects of these factors; however, the synergy driving physiological adaptation in mollusks has yet to be fully elucidated. We assessed the physiological responses of an ecologically significant mussel, Mytilus coruscus, to 2 mo exposure under varying environmental conditions (25°C/28°C and pH 7.7/8.1). Key metrics included shell properties, flesh weight, antioxidant defenses, bioenergetics, and gene expression. Compared to control groups, experimental groups showed reductions in shell hardness and compressive strength, >10% decrease in flesh weight, and 40-52% suppression of carbonic anhydrase and Ca2+-ATPase activities. Molecular analyses of the mantle tissue demonstrated compromised mitochondrial energy transduction (>40% reduction in ATP6 expression) alongside upregulated stress response markers (>2.1-fold COX3 increase). Notably, cellular energy allocation declined, accompanied by depletion of energy reserves (proteins, lipids, carbohydrates), indicating metabolic prioritization toward stress compensation. These findings elucidate how coupled stressors disrupt homeostasis through multilevel interactions, forcing energy trade-offs between defense mechanisms and growth processes, and confirm the tissue-specific vulnerability of the mantle and individual resilience of bivalves under multifactorial climate change.

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Ocean acidification does not affect the trophic transfer of Ag, Co, and Zn in the cuttlefish Sepia officinalis

Highlights

  • Trophic transfer of metallic radiotracer was assessed in cuttlefish under low pH condition.
  • High assimilation of Ag, Co and Zn in juvenile cuttlefish via diet.
  • Ocean acidification (pH 7.63) does not affect metal trophic transfer.
  • Digestive gland is main storage site for Ag and Co.
  • Zn displays broader tissue distribution.

Abstract

Cephalopods are known to efficiently accumulate metals and may therefore play an important role in the trophic transfer of contaminants within marine food webs. However, the influence of environmental changes such as ocean acidification on trace element assimilation and retention in these organisms remains poorly understood. In the present study, the trophic transfer of three trace elements (Ag, Co, and Zn) was investigated in juvenile cuttlefish Sepia officinalis under two seawater pH conditions representative of present-day (pH 7.92) and near-future ocean acidification scenarios (pH 7.63). Using radiotracer techniques and a pulse-chase feeding experiment with radiolabelled shrimp, we quantified assimilation efficiencies, depuration kinetics, and tissue distribution of these elements following a single contaminated meal. Juvenile cuttlefish showed high assimilation efficiencies for all three trace elements: 94–100% for Ag and Co, and 77–78% for Zn. Depuration kinetics revealed element-specific retention patterns, with biological half-lives of several weeks to months for Ag and Zn, whereas Co was eliminated more rapidly. Tissue distribution showed a strong organotropism towards the digestive gland, which acted as the main storage compartment for Ag and Co, while Zn showed a wider distribution across tissues. No significant differences in assimilation efficiencies, depuration kinetics, or tissue distribution were observed between pH treatments. These results suggest that moderate ocean acidification scenarios projected for the coming century are unlikely to significantly affect trophic transfer and internal handling of trace elements in juvenile cuttlefish.

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Effects of ocean acidification on radular tooth material properties in Littorina littorea (Gastropoda, Mollusca)

Ocean acidification is known to affect calcified structures in marine organisms, yet its impact on non-calcified but functionally essential feeding tools remains poorly understood. The radula is a defining molluscan apomorphy, whose mechanical performance is critical for feeding and survival. Here we investigated the effects of reduced seawater pH on the radular teeth of the intertidal gastropod Littorina littorea. Individuals were maintained for seven weeks under acidified conditions (pH 7.5) or near-present-day conditions (pH 8.1) and compared with a field-collected control group. Radulae were analysed using scanning electron microscopy, confocal laser scanning microscopy, energy-dispersive X-ray spectroscopy, and nanoindentation.

Radulae from acid-treated individuals exhibited markedly increased tooth wear in the working zone despite largely preserved gross morphology. Wear was most pronounced at the cusps of central and lateral teeth and showed rounded profiles indicative of progressive abrasive wear. Acidic conditions caused pronounced changes in the outer tooth coating, including reduced silicon enrichment and substantial decreases in stiffness and hardness, while the inner tooth structure was only weakly affected. Confocal microscopy revealed treatment-specific autofluorescence patterns, suggesting pH-dependent alterations of the organic matrix. Differences between laboratory-maintained and field-collected individuals further indicate that feeding conditions influence radular tooth properties.

These results demonstrate that ocean acidification can impair radular function through material-level degradation of composite feeding structures, potentially reducing grazing efficiency and imposing sublethal fitness costs.

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The influence of localized water quality on Eastern oysters (Crassostrea virginica) and their internal microbiome under changing environmental conditions

Oysters are found ubiquitously in estuaries along the Georgia coast, where marsh morphology and large daily tidal fluctuations create dynamic and stressful conditions to which oysters may be locally adapted. Based on water quality data from the Sapelo Island National Estuarine Research Reserve, it is evident that changing climatic conditions are rapidly causing shifts in water quality that may be adversely affecting oyster health, especially as ocean acidification alters the carbonate buffering capacity, increasing the amplitude of daily pH variations. Importantly, the rate of change of conditions are not uniform within estuaries, varying on spatial and temporal scales. The symbiotic relationship between oysters and their internal microbiome has been increasingly analyzed as a metric for oyster health. As filter feeders, oysters continuously introduce microorganisms into their hemolymph. Core families of bacteria, including Mycoplasmataceae, have been identified to be associated with healthy oysters. The abundance of core groups, or of pathogenic genera like Vibrio, can be used as an indicator of oyster condition. Utilizing reciprocal transplant and common garden tank designs, we examined how changing variability in localized water quality conditions drive oyster health using physical and microbial indicators, including oyster growth, condition index, and shifts in microbial community dynamics. Our results suggest that low pH conditions are detrimental to oyster physiology, inducing stress, leading to a reduction in overall health and growth. Low pH causes a shift within the microbial composition, altering community dynamics, and increasing the abundance of stress-related bacteria, including Arcobacteraceae and Vibrionaceae. Drivers of oyster health and host-associated microbial dynamics are site- and scale-dependent and will need further research to fully understand which biotic or abiotic factors are most influential in oyster conditions amidst low pH conditions. Oysters are increasingly used in nature-based restoration efforts to support reef recovery and salt marsh expansion, making it critical to understand how relocation influences oyster health. Our results indicate that oyster condition is driven by destination rather than origin, with relocation success dependent on water quality at the transplant site.

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Ocean acidification and harmful algal blooms combine to suppress the growth and survival of North Atlantic bivalve larvae

While harmful algal blooms (HABs) and ocean acidification (OA) are environmental factors that can impair bivalves, the manner in which these two stressors may act and interact to impact bivalve larvae is poorly understood. This study exposed larvae of hard clams (Mercenaria mercenaria) and Eastern oysters (Crassostrea virginica) to a range of pCO2 levels found in estuaries (400–3,000 µatm) and three harmful algae, Alexandrium catenella, Dinophysis acuminata, and Margalefidinium polykrikoides, at densities found during HABs (500–7,000 cells mL-1), with one HAB species exposure per experiment. The combined OA and HAB treatment significantly reduced larval survival in all 21 experiments by 91 ± 4.6% (SE) compared to controls and reduced larval sizes in 92% of experiments by 40 ± 3.5%. Cultured M. polykrikoides had a stronger negative effect on larvae than cellular equivalent bloom populations. Densities of D. acuminata >750 cells mL-1 reduced larval survival and size (p < 0.01), but the addition of OA to D. acuminata did not suppress survival further. While the combined A. catenella and OA treatment reduced larval growth and survival at all densities (p < 0.01), A. catenella alone did not impact M. mercenaria survival or size at or below 1,000 cells mL-1 and did not impact C. virginica at any density. Oyster larvae were less impacted than hard clams by OA (33 vs. 67% of experiments) and by HABs (67 vs. 100% of experiments). Given the very low survival of bivalve larvae when exposed to combined HABs and OA in all experiments (<0.1–5%), bivalve restoration and conservation efforts should seek to avoid regions that experience these co-stressors.

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Research progress on the comprehensive response mechanisms of marine organisms to multiple environmental stressors

The ocean constitutes a vital component of the Earth’s ecosystem, serving as the breeding and habitat ground for marine organisms. Currently, the global marine ecosystem is confronting combined threats from multiple environmental stressors, such as seawater warming, acidification, hypoxia, and microplastic pollution. Research focusing solely on individual stressors can hardly reveal the authentic response patterns of marine organisms accurately. This paper presents a comprehensive review. It systematically integrates cutting-edge research findings from recent years. The review centers on two core themes. These themes are the interactive effects of multiple environmental stressors and the response mechanisms of marine organisms. Studies indicate significant species-specific differences in organism responses to combined stress. These differences exist across various organism groups. Additionally, the interactive effects of multiple environmental stressors often induce biological responses. These responses deviate from the predictions derived from single-factor studies. The research results presented herein can provide crucial theoretical support for the conservation of marine biological resources, the restoration of biodiversity, and the protection of the marine ecological environment. Meanwhile, they lay a foundation for the establishment of predictable marine stress-response relationship models.

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Climate change resilience and positive Scope for Growth in wild adult Sydney rock oysters, Saccostrea glomerata (Gould 1850)

Climate change resilience and positive Scope for Growth in wild adult Sydney rock oysters, Saccostrea glomerata (Gould 1850)

Oysters have ecological and economic importance worldwide as they provide ecosystem services and sustain profitable aquaculture industries. Calcifying bivalves including oysters have been found to be sensitive to ocean warming and acidification caused by anthropogenic climate change. This study tested whether adult wild Sydney rock oyster, Saccostrea glomerata, exposed to elevated pCO2 (331 μatm and 867 μatm) and temperature (24°C and 28°C) in an orthogonal design for five weeks, have resilience and can maintain sufficient scope for growth or are pushed into a suboptimal state. At the end of the exposure growth, condition index, clearance, ingestion and absorption efficiency and rates were measured and scope for growth calculated. Sydney rock oysters responded to elevated pCO2 and temperature with no change in overall growth or condition index, but significantly increased metabolic, clearance, ingestion, and absorption rates and positive Scope for Growth. Our results indicate that adult S. glomerata can cope with the moderate level of climate change stress predicted for 2100 through increased standard metabolic rate and increased energetic processes. If, however, food availability becomes limiting, and other environmental stressors interact with climate change stressors then resilience thresholds maybe breached for this economically, ecologically and indigenous significant and iconic oyster species.

Continue reading ‘Climate change resilience and positive Scope for Growth in wild adult Sydney rock oysters, Saccostrea glomerata (Gould 1850)’

Effects of ocean acidification on the growth, shell integrity, and vulnerability to thermal stress and predation in Pacific oysters (Magallana gigas), and bay mussels (Mytilus spp.)

The ocean is absorbing approximately one third of the anthropogenic carbon dioxide (CO₂) emissions to the atmosphere. As a result, the pH of the ocean is declining steadily, a phenomenon known as ocean acidification (OA). This decline in pH and the associated reductions in calcium carbonate saturation states of the water can have widespread consequences for marine life, particularly to calcifying organisms. In this thesis, I aim to understand the effects of OA on the growth, shell integrity, and susceptibility to secondary stressors like heatwaves or predation, of two important shellfish species in British Columbia, Pacific oysters (Magallana gigas) and bay mussels (Mytilus spp.). I also aim to identify potential tipping points beyond which the biological responses of these shellfish to OA rapidly become more pronounced. I reared oysters and mussels in experimental mesocosms, in four pCO₂ treatments for eight-weeks to determine growth. I subsequently exposed these OA-acclimated animals to a secondary stressor by simulating heatwave conditions to assess thermal tolerance, and by introducing a predatory sea star to assess vulnerability to predation. Finally, shell condition was visually assessed, and shells were mechanically crushed to determine integrity. I found that OA decreased the growth of both oysters and mussels. No tipping point was observed for oyster growth, but reduced growth only emerged at the highest levels of OA in mussels. Sensitivity to atmospheric warming was not increased after exposure to acidic conditions for either species, although oysters had a considerably higher thermal tolerance than mussels. Mussel vulnerability to predation did increase, although the relationship was complex and depended on predator size. OA negatively affected shell strength, and possible tipping points emerged for this response metric in both species. Overall, OA was shown to negatively affect both species, but patterns of effect and the presence of potential tipping points depended on the species and the response metric. Understanding how these ecologically and commercially important bivalves are responding to OA is important for understanding how changing ocean chemistry will affect marine ecosystems, and to inform aquaculture managers on mitigation strategies.

Continue reading ‘Effects of ocean acidification on the growth, shell integrity, and vulnerability to thermal stress and predation in Pacific oysters (Magallana gigas), and bay mussels (Mytilus spp.)’

Synergistic effects of ocean acidification and thermal stress on shell biomineralization and parasitism in the white clam Leukoma asperrima (Bivalvia: Veneridae)

Ocean acidification (OA) and global warming are fundamentally altering the biomineralization processes of calcifying marine organisms. This study evaluates shell malformations and parasitism in the white clam Leukoma asperrima at Bique Beach, Panama, from December 2024 to November 2025. Environmental parameters (pH, temperature) were monitored monthly across two sampling stations (n=1100). Results indicate that 13.6% of the population exhibited shell malformations, and 6.3% were parasitized by the pea crab Pinnotheres pisum. A strong positive correlation was found between pH and healthy individuals (r=0.97, p<0.001), whereas critical pH levels (min. 5.75) were associated with increased shell fragility and dissolution. Despite thermal tolerance observed up to 35.7°C, the synergistic effect of OA and local stressors compromises the structural integrity of L. asperrima, threatening the sustainability of this socio-economic resource in the Tropical Eastern Pacific.

Continue reading ‘Synergistic effects of ocean acidification and thermal stress on shell biomineralization and parasitism in the white clam Leukoma asperrima (Bivalvia: Veneridae)’

Coexpression among eastern oyster host and microbiome genes suggests coordinated regulation of calcifying fluid chemistry

Significance

Oysters and many marine animals build shells by controlling the chemistry of extracellular fluids where minerals form, yet whether microbes in these fluids influence calcification remains unclear. We show that oysters maintain favorable conditions for mineral formation by regulating the carbonate chemistry of the shell-forming fluid, and that resident microbes respond to these changes by expressing nitrogen- and sulfur-cycling genes capable of altering pH, alkalinity, and carbonate availability. Many of these microbial transcripts were tightly correlated with oyster immune and signaling genes, suggesting that host and microbiome processes may be linked within the calcifying environment. These findings point to a host–microbiome interaction in the regulation of calcifying-fluid chemistry that directly links microbial activity to the carbonate chemistry underlying biomineralization.

Abstract

Marine animals that build shells, such as oysters, carefully regulate the chemistry of their internal calcifying fluids, but the molecular mechanisms behind this control, as well as whether microbes play a role in calcification, are poorly understood. To better understand oysters’ molecular mechanisms and the role of their calcifying-fluid microbes, we conducted experiments that simulated a tidal cycle, measured calcifying fluid pH and total dissolved inorganic carbon, and characterized host and microbial gene expression via transcriptomics. These experiments showed that calcifying fluid pH remained relatively stable throughout tidal pH fluctuations, with corresponding increases in oyster transcripts for ion transport and acid–base regulation. These data provide direct evidence that tidal fluctuations drive rapid changes in oyster calcifying fluid chemistry. Most surprisingly, increases in microbial transcripts related to nitrogen and sulfur cycling correlated to higher calcifying fluid DIC, and coexpression network analysis revealed patterns of gene expression that linked oyster immune and neural pathways to microbial redox processes, providing molecular evidence of potential host modulation of microbial metabolism. Together, these results reveal that oysters actively regulate their calcifying fluid pH over short timescales, and the endemic microbiome metabolic responses can yield metabolites that influence calcifying fluid pH, alkalinity, and ultimately calcification. These data offer a perspective on oyster physiological capacity and, most importantly, the potential role of microbes in oyster calcification. In light of ongoing changes in ocean pH and temperature, oysters provide a model for studying animal–microbial responses to environmental acidification and how their interactions may shape biomineralization.

Continue reading ‘Coexpression among eastern oyster host and microbiome genes suggests coordinated regulation of calcifying fluid chemistry’

Unravelling marine benthic functioning shifts under ocean acidification

Ocean acidification (OA) driven by increasing atmospheric CO2 is altering marine biodiversity. However, impacts of OA on ecosystem functioning at the community level, including calcification, primary production and nutrient uptake, remain largely unknown. Here, we conducted community transplant experiments at natural CO2 vents to assess how declining pH affects marine community species composition, biomass, and key ecosystem processes over time. Our results indicate that community shifts caused by declining pH lead to decreased biomass and calcification rates, while photosynthesis and nutrient uptake rates increased. By leveraging OA field model systems and in situ measurements of ecosystem functioning, this study provides critical insights into how OA-induced biodiversity loss reshapes the structure and functioning of temperate marine coastal ecosystems.

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Intracellular acid-base regulation mediates a trade-off between shell and somatic growth in a clam under ocean acidification

Highlights

  • Clams actively regulate intracellular pH against ocean acidification via CAc
  • RNAi confirms CAc’s essential role in H+ efflux, measured by in vivo SIET.
  • A CAc-sAC-NKA network forms a conserved regulatory pathway for acid-base balance.
  • DEB model shows this pH defense sustains shell linear growth despite metabolic costs.

SUMMARY

Ocean acidification (OA) is predicted to threaten marine bivalves, casting them as passive victims of changing carbonate chemistry. Contributing to a revised understanding, we identified a conserved mechanism for acid-base regulation that supports intracellular resilience. Using the Manila clam Ruditapes philippinarum as a model, this study demonstrated that intracellular pH (pHi) homeostasis under elevated pCO2 was maintained through cytosolic carbonic anhydrase (CAc)-mediated H+ efflux. A causal link was established by combining in vivo scanning ion-selective electrode technique (SIET) with RNA interference (RNAi), where RpCAc knockdown suppressed H+ efflux and compromised pHi. A coordinated regulatory network involving CAc, soluble adenylyl cyclase (sAC), and Na+/K+-ATPase (NKA) was synergistically upregulated, suggesting an evolved adaptive pathway. Dynamic Energy Budget (DEB) modeling, calibrated with experimental data, revealed that this cellular compensation carries a high energetic cost, leading to a significant reallocation of resources: shell growth was maintained, but somatic growth was severely suppressed. These results elucidate a conserved cytoprotective mechanism that enables short-term tolerance of OA at a substantial somatic cost, redefining resilience to include energetic trade-offs.

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Investigation of the adaptive mechanisms to ocean acidification in Patella species from CO2 vent systems of the Mediterranean Sea

The continuous increase in anthropogenic carbon dioxide (CO2) emissions into the atmosphere is one of the main factors contributing to ocean acidification (OA). In fact, CO2 is partially absorbed by the oceans, where it alters carbonate chemistry and seawater pH, which is expected to decrease from the current level of 8.1 to 7.7 by 2100. OA exerts harmful impacts primarily on calcifying organisms, as it affects the availability of carbonates, which makes their calcareous structures thinner and more fragile. Moreover, several studies have described the detrimental effects of OA across many marine taxa, affecting important physiological and metabolic mechanisms. On the other hand, research conducted at CO2 vent systems, which are volcanic naturally acidified sites, showed that several organisms can survive under acidified conditions through specific tolerance and/or adaptive strategies. Among these organisms, the gastropod Patella spp. is one of the few calcifiers capable of inhabiting naturally acidified sites, such as the Castello Aragonese vent systems at Ischia Island and the San Giorgio vent systems at Sicily Island. Nonetheless, the complex mechanisms that allow survival and potential adaptation of these organisms to natural OA conditions need to be understood. Therefore, this PhD study aimed at investigating the potential molecular, physiological, metabolic, genetic, and epigenetic mechanisms that enable these organisms to tolerate and survive under OA conditions through a stepwise approach. Specifically, this PhD research attempted to answer the following questions: • Does OA entail a stressful condition in resident populations of Patella spp. living at reduced pH conditions, thereby affecting their overall well-being and health status? • Are there specific physiological, metabolic, and biochemical mechanisms that contribute in defining tolerance to OA? • Are limpets genetically adapted to OA? • Is DNA methylation contributing to promote tolerance to OA in limpets? • What is the role of environmental conditions in shaping the response to OA? The first chapter of this thesis considered three Patella species (P. caerulea, P. rustica, and P. ulyssiponensis) collected from the CO2 vent systems of the Castello Aragonese (Ischia Island). This vent system exhibits a natural acidification gradient ranging from ambient pH (N1: pH = 8.1), to intermediate pH (N2: pH = 7.7), and to extremely low pH (N3: pH < 7.4). Resident populations were collected along the gradient and at San Pietro, an additional ambient pH site (pH = 8.1), located at a distance of 4 km from the Castello vent. In addition, a 30-day in situ transplant experiment was conducted using P. caerulea, in order to evaluate the short-term responses induced by OA. Morphometric traits, such as shell length, height, width, and soft-tissue weight, were measured, and a set of biomarkers related to antioxidant system, energy metabolism, neurotoxicity, and biomineralization was applied. For resident populations, P. caerulea showed increased size and energy reserves at the extremely acidified site, likely related to a shift from erect calcified algae to biofilm, or to reduced competition and/or predatory pressure under acidified conditions. Biochemical endpoints measured in both P. caerulea and P. ulyssiponensis were not modified by OA. Conversely, P. rustica exhibited significant modulation of nearly all biochemical parameters, likely due to its different position on the rocky shore, which makes this species more exposed to tidal fluctuations and therefore to an additional source of disturbance, besides OA. Short-term exposure of P. caerulea to OA resulted in a decrease in protein content and an increase in glycogen content at the extremely acidified site, with the induction of superoxide dismutase and glutathione-S-transferase activities at intermediate pH, suggesting the activation of compensatory mechanisms to cope with reduced pH conditions. Overall, results revealed a distinct response to OA of the three species of Patella. Moreover, the increased size and energy-related endpoints observed in P. caerulea and P. rustica highlighted the need to further investigate energy metabolism aspects, in order to better understand the trade-offs between compensatory mechanisms and the energetic cost underlying tolerance to OA. Based on these evidences, the second chapter focused exclusively on P. caerulea, with the aim of deeply investigating metabolic and physiological stress-responses, comparing resident populations of the Castello Aragonese vent systems and transplanted organisms, similarly to the first chapter. Respiration and ammonia excretion rates were measured four times across the year. Additionally, untargeted metabolomics analyses were performed to investigate metabolic pathways potentially involved in response to OA. Only during summer, OA increased respiration rate in limpets from the most acidified site, while simultaneously reduced excretion rates, likely to allocate more energy resources to face the increasing temperature, besides OA. Furthermore, both resident and transplanted populations up-regulated carnitine metabolism, suggesting that OA induced an increase of energy production through β-oxidation and subsequent Krebs cycles. Moreover, several metabolites involved in osmoregulation, oxidative stress, and nucleic acid mechanisms were increased. Overall, results seem to confirm the presence of negative effects and of an energetic cost underpinning tolerance to OA. The third and final chapter investigated the potential influence of phenotypic plasticity, genetic adaptation, and DNA methylation in tolerance to OA in adult and juvenile populations of P. caerulea collected from two CO2 vent systems of the Mediterranean Sea. Adult and juvenile specimens were sampled along the acidification gradient of the Castello Aragonese vent systems of Ischia Island (San Pietro/N1: pH = 8.1; N2: pH = 7.7; N3: pH < 7.4) and from the San Giorgio vent systems of Sicily Island (Patti: pH = 8.1; San Giorgio: pH = 7.8). Following genomic DNA extractions from foot tissue and individual libraries preparation with the NEB Next® Enzymatic Methyl-seq Kit, samples were sequenced on the Illumina NovaSeq 6000 sequencer. Data processing and analyses were conducted on Euler platform mainly using biscuit tool, which enabled to simultaneously extract genomic and epigenomic information from DNA methylation sequencing. Population genomics and epigenomics analyses revealed divergent patterns between the Ischia and Sicily populations. Populations from the Ischia vent revealed marked signs of genomic differentiation, particularly in adults from the intermediate and extremely low pH sites, while reduced differences in DNA methylation levels were detected, especially in adults. These findings suggest a strong signature of purifying selection acting on standing genetic variation, through a within-generation response, likely driven by the more pronounced pH fluctuations occurring at these sites. Conversely, no genomic differentiation was observed between the Sicily populations, but greater differences in DNA methylation were detected between acidified and non-acidified sites at both adult and juvenile stages. These results revealed that this epigenetic mechanism, rather than genomic changes, may play a key role in the response to the milder pH variations of this vent and potentially enhance organisms’ tolerance to OA. In conclusion, this PhD project investigated tolerance to OA in limpets through a holistic approach that, for the first time, integrated morphological, physiological, metabolic, biochemical, genetic, and epigenetic analyses. Overall, findings revealed that Patella spp. has the ability to survive under acidified conditions even though with a physiological and metabolic cost, which could be partially compensated by more favorable environmental conditions. This study further highlights the importance of conducting research in naturally acidified environments, since it allows to formulate more realistic hypotheses about the ability of marine organisms to persist in future changing oceans.

Continue reading ‘Investigation of the adaptive mechanisms to ocean acidification in Patella species from CO2 vent systems of the Mediterranean Sea’

pCO2-induced seawater acidification influencing cadmium toxicity on antioxidant defenses responses in juvenile Manila clam Ruditapes philippinarum

Highlights

  • GSH system and SOD-CAT act as complementary lines of antioxidant defense
  • SA alters Cd effects on antioxidants depending on metal concentration
  • High SA overrides Cd effects on antioxidant defenses in Manila clams
  • Combined high SA and Cd exposure overwhelms antioxidant capacity
  • Candidate biomarkers for monitoring SA or Cd stress are proposed

Abstract

Ocean acidification is known to interact with heavy metals, impacting physiological processes of marine organisms. This study investigated antioxidant defenses of juvenile Manila clam Ruditapes philippinarum exposed to cadmium (Cd) across ambient-relevant to high concentrations, under pCO2-induced seawater acidification (SA) scenarios corresponding to IPCC ocean pH projections. Results revealed that clam’s antioxidant system, encompassing GSH defense system and SOD-CAT defense lines, collectively combated oxidative stress dependent on specific stressors and their stress levels. GSH system is vital for detoxification and maintaining redox balance, while SOD and CAT are essential for scavenging ROS. Cd exposure notably activated GSH redox cycle, and SA markedly inhibited the antioxidants associated with this cycle. SOD and CAT exhibited distinct regulatory pathways with asynchronous responses to SA and Cd co-exposure. SA conditions modulate Cd-induced antioxidant response dependent on metal concentrations. Antioxidant biomarkers responded more prominently to SA and Cd interactions than to individual exposure, particularly, high SA effects could override Cd effects on antioxidant responses. Although SA effects did not directly induce lipid peroxidation, elevated MDA levels under Cd exposure occurred only under SA conditions, indicating insufficient antioxidant defense against lipid peroxidation under excess co-exposure. GSH and SOD were more sensitive to SA exposure, whereas MDA and GST were sensitive to Cd exposure, suggesting their potential as biomarkers for assessing SA or Cd-induced oxidative stress, respectively. These findings provide insights into interplay between metal toxicity and ocean acidification on antioxidant defenses in bivalves, shedding light on their strategies to combat metal pollution amidst global ocean change.

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Metabolic rate measurements of two benthic invertebrates under simulated climate change conditions

Background

Climate change is profoundly altering marine ecosystems through ocean warming and acidification. These stressors are especially pronounced in the Mediterranean Sea, a climate change hotspot projected to warm faster than the global average. Increased temperatures and reduced pH directly affect metabolic processes in marine invertebrates by elevating respiration rates up to species-specific thermal limits, beyond which physiological performance declines. Ocean acidification further disrupts metabolic processes by increasing energetic maintenance costs. Sessile and sedentary marine invertebrates, such as sponges and benthic gastropods, are particularly exposed to such environmental shifts due to their limited ability to escape unfavorable conditions, making physiological plasticity and local adaptation crucial for persistence.

New information

This manuscript presents a dataset of oxygen consumption rates and wet weight measurements for two low-mobility marine species, the gastropod Hexaplex trunculus and the sponge Chondrilla nucula. Using a common garden experiment, individuals from North and South Aegean populations were exposed for three months to simulated climate change conditions combining increased temperature and reduced pH. The dataset documents respiration measurements obtained using metabolic chambers after three months of exposure, allowing comparisons across species, geographic origin, and experimental treatments.The dataset accounts for intraspecific variation in these responses, providing insight into potential adaptive differences among geographically distinct populations. These data provide a resource for future analyses of metabolic responses of marine invertebrates to combined warming and acidification conditions.

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Resilient adults but vulnerable larvae: demographic pathways of chiton decline under ocean acidification

Highlights

  • Natural CO₂ seep systems showed reduced intertidal chiton abundance.
  • Adult chitons showed resilience to acidification in field and lab experiments.
  • Larval survival and recruitment were strongly impaired under acidified seawater.
  • Population declines are linked to early life-stage vulnerability.
  • Loss of chitons may reduce grazing and bulldozing, reshaping intertidal communities.

Abstract

Ocean acidification (OA) is a major threat to marine calcifiers; however, the sensitivity across taxa and life stages remains elusive. In this study, we combined field surveys of natural CO₂ seeps with laboratory exposure, transplantation, and larval settlement experiments to assess the effect of OA on chitons, a group of calcifying grazers and bulldozers that play critical roles in the structure of rocky intertidal ecosystems. Field surveys revealed approximately 98.6% reduction in chiton (Acanthopleura loochooanaLiolophura japonica, and Acanthochitona rubrolineata) abundance at acidified habitats (pH 7.6), despite greater microalgal food availability and no detectable increase in predator abundance. Laboratory CO₂-exposure experiments showed no direct effect of OA on adult A. loochooana survival, which is consistent with the presence of protective structural features in the valves that confer resistance to dissolution. Transplant experiments revealed no evidence of increased adult A. loochooana mortality in the acidified habitats (pH 7.6). In contrast, larvae showed pronounced sensitivity to OA, with acidified seawater (pH 7.6) reducing larval settlement by approximately 81.5% compared to control conditions (pH 8.1); early life stages were the most vulnerable. These findings suggest that OA-associated decline in chiton abundance is mainly mediated by impaired recruitment rather than by direct adult mortality, predation, or food limitation. Given the role of chitons as grazers and bulldozers, their loss could substantially change intertidal community dynamics by decreasing grazing pressure and disturbing algal and microbial assemblages. Our findings underscore the criticality of considering life-stage vulnerability and ecological function when evaluating the ecosystem-level consequences of OA.

Continue reading ‘Resilient adults but vulnerable larvae: demographic pathways of chiton decline under ocean acidification’

Experimental exposure to climate change scenarios imposed alterations on the morphological traits of sessile and low-motility marine invertebrates

Background

Over the past 50 years, the oceans have absorbed over 90% of global warming heat, leading to warming, acidification and declining oxygen levels that are disrupting marine ecosystems and altering species distributions and productivity. The vulnerability of marine organisms to these changes depends on their biological traits, habitat conditions and adaptive capacity, influencing their growth, behaviour and overall population health. Micro-computed tomography (micro-CT) has been previously used for studying the morphological traits of marine invertebrates, which provide important insights into species functionality and responses to climate change and ocean acidification. Micro-CT enables non-destructive, high-resolution 3D analysis of internal and external structures, allowing precise measurement of traits such as density, porosity and morphology that are valuable for climate change research.

New information

The present manuscript describes micro-CT imaging datasets generated to investigate the effects of climate change on the morphological structure of two benthic marine invertebrates: the low-motility gastropod Hexaplex trunculus (Linnaeus, 1758) and the sessile sponge Chondrilla nucula Schmidt, 1862. Both species are considered particularly vulnerable to environmental stressors. To date, no study has investigated the effects of ocean warming and acidification on sponges using micro-CT technology. Using a common garden experimental design, individuals from geographically distinct populations exposed to different natural environmental regimes were subjected to combined warming and acidification scenarios to assess their morphological responses and adaptive capacity.

Continue reading ‘Experimental exposure to climate change scenarios imposed alterations on the morphological traits of sessile and low-motility marine invertebrates’

Ocean acidification disrupts the biomineralization process in the oyster Crassostrea virginica via intracellular calcium signaling dysregulation

Calcium is a key component in the shell and skeleton structure, serving as a second messenger for regulating biomineralization across many species. Ocean acidification (OA) is well-studied for causing shell dissolution in marine bivalve species by disordering calcium deposition. However, the regulatory pathway of calcification affected by OA remains unclear. This study assessed eastern oyster (Crassostrea virginica) to determine how calcium signaling responds to elevated pCO2 and influences shell formation. Under elevated pCO2, increased calcium influx was found in mantle epithelial cells, followed by the upregulation of calmodulin, a primary sensor of intracellular calcium. Expression levels of shell matrix proteins (SMPs), representing shell construction conditions, were significantly upregulated in the CO2-induced mantle cells. Larval C. virginica exhibited developmental stage-dependent alterations in calcium signaling and SMPs disarrangement stimulated by pCO2. Pharmaceutical blockage of the calcium binding on calmodulin induced abnormal expression of downstream genes and shell matrix changes consistent with those caused by elevated pCO2. Restored SMPs expressions in CO2-treated mantle cells were achieved by rescuing the level of calcineurin, a downstream effector of calmodulin. These findings suggest that shell deformities under OA are primarily caused by the disruption of the calcium-calmodulin signaling pathway in mantle epithelial cells.

Continue reading ‘Ocean acidification disrupts the biomineralization process in the oyster Crassostrea virginica via intracellular calcium signaling dysregulation’

Influence of ocean warming and acidification on juveniles of the true giant clam, Tridacna gigas, and its microalgal symbionts

Uncontrolled carbon dioxide emissions from human activities contribute to ocean warming and acidification. These alterations in ocean chemistry threaten marine organisms, such as the true giant clam, Tridacna gigas, which is already imperiled due to overharvesting and habitat destruction. To gain an understanding of the physiological and molecular responses of T. gigas and its symbiotic dinoflagellates to ocean warming and acidification, we subjected juvenile individuals to different treatments simulating predicted seawater pH (7.6 and 8.0) and temperature (28°C, 30°C, 32°C and 34°C) levels for the next century. Juvenile giant clams were able to tolerate sustained exposure to temperatures of up to 32°C and pH as low as 7.6, while exposure to higher temperature (34°C), regardless of pH level, resulted in total mortality after a week. However, symbiosis was compromised even in the sublethal treatments, as indicated by the decrease in Symbiodiniaceae density and changes in symbiont gene expression. Symbionts significantly upregulated genes involved in splicing, translation, fatty acid metabolism, and DNA repair, which may constitute an adaptive response, while downregulating genes involved in photosynthesis and transmembrane transport, suggests impaired transfer of photosynthates to the host. These findings demonstrate the vulnerability of the juvenile T. gigas holobiont to heat stress, highlighting the critical importance of continued conservation and management alongside efforts to mitigate global changes in ocean conditions to safeguard this iconic marine bivalve.

Continue reading ‘Influence of ocean warming and acidification on juveniles of the true giant clam, Tridacna gigas, and its microalgal symbionts’

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