Seawater pH and carbonate saturation are predicted to decrease dramatically by the end of the century. This process, designated ocean acidification (OA), threatens economically and ecologically important marine calcifiers, including the northern quahog (Mercenaria mercenaria). While many studies have demonstrated the adverse impacts of OA on bivalves, much less is known about mechanisms of resilience and adaptive strategies. Here, we examined clam responses to OA by evaluating cellular (hemocyte activities) and molecular (high-throughput proteomics, RNASeq) changes in hemolymph and extrapallial fluid (EPF—the site of biomineralization located between the mantle and the shell) in M. mercenaria continuously exposed to acidified (pH ~7.3; pCO2 ~2700 ppm) and normal conditions (pH ~8.1; pCO2 ~600 ppm) for one year. The extracellular pH of EPF and hemolymph (~7.5) was significantly higher than that of the external acidified seawater (~7.3). Under OA conditions, granulocytes (a sub-population of hemocytes important for biomineralization) were able to increase intracellular pH (by 54% in EPF and 79% in hemolymph) and calcium content (by 56% in hemolymph). The increased pH of EPF and hemolymph from clams exposed to high pCO2 was associated with the overexpression of genes (at both the mRNA and protein levels) related to biomineralization, acid–base balance, and calcium homeostasis, suggesting that clams can use corrective mechanisms to mitigate the negative impact of OA.
Continue reading ‘Proteomic and transcriptomic responses enable clams to correct the pH of calcifying fluids and sustain biomineralization in acidified environments’Posts Tagged 'mollusks'
Proteomic and transcriptomic responses enable clams to correct the pH of calcifying fluids and sustain biomineralization in acidified environments
Published 1 February 2023 Science ClosedTags: biological response, laboratory, molecular biology, mollusks, physiology
Combined effects of climate change stressors and predators with contrasting feeding-digestion strategies on a mussel species
Published 1 February 2023 Science ClosedTags: biological response, echinoderms, laboratory, mollusks, morphology, multiple factors, physiology, predation, temperature

Highlights
- Combined effects of climate change stressors and Predator Cues (PC) were evaluated.
- Ocean Acidification (OA), Warming (OW) and PC affected mussel traits.
- At the control temperature (15 °C), mussel byssal biogenesis increased with PC.
- PC affected mussel size, wet mass and calcification rate.
- The effects of starfish PC on some mussel traits were larger than those of snail PC.
Abstract
We investigated the combined effects of Ocean Warming (OW), Acidification (OA) and predator cues (Non-Consumptive Effects; NCEs) of two predators with contrasting feeding-digestion strategies on the mussel Perumytilus purpuratus. We considered starfish-NCEs (partially external digestion) and snail-NCEs (internal digestion). Mussels were exposed for 13 weeks to cross-factored OA (~500 and ~1400 μatm, pCO2) and OW (~15 and ~20 °C) conditions, in the presence/absence of NCEs from one or both predators. Mussels exposed to both NCEs exhibited smaller length and buoyant weight growth than those under control or snail-NCEs conditions. Mussels exposed to starfish-NCEs exhibited smaller wet mass than control mussels. OW and starfish-NCEs in isolation or combined with snail-NCEs increased mussel oxygen consumption. Byssal biogenesis was affected by the three-factors interaction. Clearance rates were affected by the OW × OA interaction. We suggest that mainly starfish-NCEs, in isolation or interacting with OA or/and OW, can threat mussel traits and the associated community.
Continue reading ‘Combined effects of climate change stressors and predators with contrasting feeding-digestion strategies on a mussel species’Clay-shielded estuarine gastropods are better protected against environmental acidification than unshielded individuals
Published 30 January 2023 Science ClosedTags: biological response, dissolution, laboratory, mollusks, North Pacific

Highlights
- Acidified estuaries compromise building and threaten dissolution of gastropods shells.
- Periostracum of Neripteron snails directs formation of an outer shell clay shield.
- Shield constructed of the mineral illite is tightly chemically-bonded to the periostracum.
- The more reflective unshielded shells showed a greater rate of dissolution.
- Ecological and evolutionary constraints on carbonate shell building predict outer protection.
Abstract
The effects of progressive global acidification on the shells of marine organisms is a topic of much current interest. Most studies on molluscan shell resistance to dissolution consider the carbonate mineral component, with less known about the protective role of the outer organic periostracum. Outer-shell resistance would seem especially important to gastropods living in carbonate-undersaturated and calcium-deficient estuarine waters that threaten shell dissolution and constrain CaCO3 production. We tested this prediction using gastropods from an acidified estuarine population (Neripteron violaceum) that form a clay shield outside the periostracum. Specifically, we aimed to show that the carbonate shell component lacks integrity, that the formation of the clay shield is directed by the organism, and that the clay shield functions to protect against shell dissolution. We found no evidence for any specific carbonate dissolution resistance strategy in the thin, predominantly aragonitic shells of these gastropods. Shield formation was directed by an ornamented periostracum which strongly bonded illite elements (e.g., Fe, Al and S), that become available through suspension in the water column. In unshielded individuals, CaCO3 erosion was initiated randomly across the shell (not age-related) and progressed rapidly when the periostracum was breached. A light reflectance technique showed qualitatively that shield consolidation is negatively-related to shell erosion. These findings support a conceptual framework for gastropod outer-shell responses to acidification that considers both environmental and evolutionary constraints on shell construction. We describe a novel strategy for shell protection against dissolution, highlighting the diversity of mechanisms available to gastropods facing extreme coastal acidification.
Continue reading ‘Clay-shielded estuarine gastropods are better protected against environmental acidification than unshielded individuals’Resistant calcification responses of Arctica islandica clams under ocean acidification conditions
Published 30 January 2023 Science ClosedTags: biological response, chemistry, laboratory, mollusks, morphology, North Atlantic
Highlights
- We cultured both juvenile and adult A. islandica collected from northern Norway under a range of pH
- Arctica islandica from Norway can maintain its shell growth even in aragonite undersaturated (Ω < 1) conditions.
- Our results show that shell growthresilience in acidified seawater is likely a multi-population adaptation in A. islandica.
Abstract
Ocean acidification (OA) directly impacts marine calcifying organisms including ecologically and commercially important shellfish species such as Arctica islandica (A. islandica). To test whether documented growth resilience of A. islandica to OA is a general response across ages and populations or a function of adaptation to local habitat, we cultured juvenile and adult clams collected from an environment with little pH variation under four pH levels (7.5, 7.7, 7.9, and 8.1) for three months and integrated our understanding with relevant literature. The average shell growth over the experiment among all (69) individuals was 57 ± 55 μm, and there were no statistically significant differences in growth among pH treatments, including the control treatment, despite the general growth rate differences between juveniles and adults. Our results show that A. islandica can maintain its shell growth even in aragonite undersaturated (Ω < 1) conditions (0.65 and 0.83 for pH 7.5 and 7.7 treatments, respectively), supporting the hypothesis that resistance to OA conditions is likely a generalized response across populations. Although the present results show A. islandica can maintain their shell growth under short-term OA, long-term impacts of OA on A. islandica shell growth and other physical parameters including shell density and microstructure are still needed to better assess the sustainability of A. islandica in a more acidified future and to provide guidance on managing this important shellfish stock.
Continue reading ‘Resistant calcification responses of Arctica islandica clams under ocean acidification conditions’Transgenerational transfer of the microbiome is altered by ocean acidification in oyster larvae
Published 10 January 2023 Science ClosedTags: biological response, BRcommunity, laboratory, molecular biology, mollusks, physiology, protists, reproduction
Ocean acidification will affect marine molluscs, however, transgenerational plasticity (TGP) can ameliorate some effects. Marine molluscs acquire members of their microbiome via the egg, yet we know little about how the microbiome can be influenced by transgenerational exposure to ocean acidification. We exposed adult Sydney Rock oysters (Saccostrea glomerata) from four genotypes to elevated and ambient PCO2 for nine weeks. Larvae were then raised in the same ambient and elevated PCO2 conditions. The relative abundance of bacteria in eggs and larvae were characterised using 16S RNA amplicon sequencing. Parental exposure to elevated PCO2 significantly altered the bacterial community composition of both eggs and larvae, but this was dependent on genotype. Parental exposure to elevated PCO2 caused five core Rhodobacteraceae ASVs to increase in relative abundance, and three Rhodobacteraceae ASVs to decrease in relative abundance. These findings show transfer of maternal microbiomes to larvae is altered by exposure to ocean acidification and this may play a role in TGP.
Continue reading ‘Transgenerational transfer of the microbiome is altered by ocean acidification in oyster larvae’Impacts of ocean acidification and warming on post-larval growth and metabolism in two populations of the great scallop (Pecten maximus L.)
Published 29 December 2022 Science ClosedTags: biological response, field, molecular biology, mollusks, morphology, mortality, multiple factors, North Atlantic, physiology, reproduction, temperature
Ocean acidification and warming are key stressors for many marine organisms. Some organisms display physiological acclimatisation or plasticity, but this may vary across species ranges, especially if populations are adapted to local climatic conditions. Understanding how acclimatisation potential varies among populations is therefore important in predicting species responses to climate change. We carried out a common garden experiment to investigate how different populations of the economically important great scallop (Pecten maximus) from France and Norway responded to variation in temperature and pCO2 concentration. After acclimation, post-larval scallops (spat) were reared for 31 days at one of two temperatures (13°C and 19°C) under either ambient or elevated pCO2 (pH 8.0 and pH 7.7). We combined measures of proteomic, metabolic, and phenotypic traits to produce an integrative picture of how physiological plasticity varies between the populations. The proteome of French spat showed significant sensitivity to environmental variation, with 12 metabolic, structural and stress-response proteins responding to temperature and/or pCO2. Principal component analysis revealed seven energy metabolism proteins in French spat that were consistent with countering ROS stress under elevated temperature. Oxygen uptake in French spat did not change under elevated temperature, but increased under elevated pCO2. In contrast, Norwegian spat reduced oxygen uptake under both elevated temperature and pCO2. Metabolic plasticity seemingly allowed French scallops to maintain greater energy availability for growth than Norwegian spat. However, increased physiological plasticity and growth in French spat may come at a cost, as French (but not Norwegian) spat showed reduced survival under elevated temperature.
Continue reading ‘Impacts of ocean acidification and warming on post-larval growth and metabolism in two populations of the great scallop (Pecten maximus L.)’Wild oyster population resistance to ocean acidification adversely affected by bacterial infection
Published 22 December 2022 Science ClosedTags: biological response, laboratory, molecular biology, mollusks, multiple factors, North Pacific, pathogens, physiology, prokaryotes

The carbon dioxide induced ocean acidification (OA) process is well known to have profound effects on physiology, survival and immune responses in marine organisms, and particularly calcifiers including edible oysters. At the same time, some wild populations could develop a complex and sophisticated immune system to cope with multiple biotic and abiotic stresses, such as bacterial infections and OA, over the long period of coevolution with the environment. However, it is unclear how immunological responses and the underlying mechanisms are altered under the combined effect of OA and bacterial infection, especially in the ecologically and economically important edible oysters. Here, we collected the wild population of oyster species Crassostrea hongkongensis (the Hong Kong oyster) from their native estuarine area and carried out a bacterial challenge with the worldwide pervasive pathogen of human foodborne disease, Vibrio parahaemolyticus, to investigate the host immune responses and molecular mechanisms under the high-CO2 and low pH-driven OA conditions. The wild population had a high immune resistance to OA, but the resistance is compromised under the combined effect of OA and bacterial infection both in vivo or in vitro. We classified all transcriptomic genes based on expression profiles and functional pathways and identified the specifically switched on and off genes and pathways under combined effect. These genes and pathways were mainly involved in multiple immunological processes including pathogen recognition, immune signal transduction and effectors. This work would help understand how the immunological function and mechanism response to bacterial infection in wild populations and predict the dynamic distribution of human health-related pathogens to reduce the risk of foodborne disease under the future climate change scenario.
Continue reading ‘Wild oyster population resistance to ocean acidification adversely affected by bacterial infection’Observed and projected impacts of coastal warming, acidification, and deoxygenation on Pacific oyster (Crassostrea gigas) farming: a case study in the Hinase Area, Okayama Prefecture and Shizugawa Bay, Miyagi Prefecture, Japan
Published 20 December 2022 Science ClosedTags: biological response, chemistry, field, fisheries, modeling, mollusks, morphology, multiple factors, North Pacific, oxygen, regionalmodeling, reproduction, temperature
Coastal warming, acidification, and deoxygenation are progressing, primarily due to the increase in anthropogenic CO2. Coastal acidification has been reported to have effects that are expected to become more severe as acidification progresses, including inhibiting formation of the shells of calcifying organisms such as shellfish. However, compared to water temperature, an indicator of coastal warming, spatiotemporal variations in acidification and deoxygenation indicators such as pH, aragonite saturation state (Ωarag), and dissolved oxygen in coastal areas of Japan have not been observed and projected. Moreover, many species of shellfish are important fisheries resources, including Pacific oyster (Crassostrea gigas). Therefore, there is concern regarding the future combined impacts of coastal warming, acidification, and deoxygenation on Pacific oyster farming, necessitating evaluation of current and future impacts to facilitate mitigation measures. We deployed continuous monitoring systems for coastal warming, acidification, and deoxygenation in the Hinase area of Okayama Prefecture and Shizugawa Bay in Miyagi Prefecture, Japan. In Hinase, the Ωarag value was often lower than the critical level of acidification for Pacific oyster larvae, although no impact of acidification on larvae was identified by microscopy examination. Oyster larvae are anticipated to be affected more seriously by the combined impacts of coastal warming and acidification, with lower pH and Ωarag values and a prolonged spawning period, which may shorten the oyster shipping period and lower the quality of oysters. No significant future impact of surface-water deoxygenation on Pacific oysters was identified. To minimize the impacts of coastal warming and acidification on Pacific oyster and related local industries, cutting CO2 emissions is mandatory, but adaptation measures such as regulation of freshwater and organic matter inflow from rivers and changes in the form of oyster farming practiced locally might also be required.
Continue reading ‘Observed and projected impacts of coastal warming, acidification, and deoxygenation on Pacific oyster (Crassostrea gigas) farming: a case study in the Hinase Area, Okayama Prefecture and Shizugawa Bay, Miyagi Prefecture, Japan’Size matters: physiological sensitivity of the scallop Argopecten purpuratus to seasonal cooling and deoxygenation upwelling-driven events
Published 20 December 2022 Science ClosedTags: biological response, field, fisheries, mollusks, multiple factors, oxygen, socio-economy, South Pacific, temperature
Environment imposes physiological constraints which are life-stage specific as growth-maintenance and/or growth-reproduction energetic requirements are size and volume-dependent. The scallop Argopecten purpuratus, one of the most important bivalve species subjected to fishery and aquaculture along the Humboldt Current System, inhabits spaces affected by continuous changes in temperature, pH, oxygen, and food availability driven by remote and local oceanographic processes. Specifically, in Chile, this species is mainly cultured in central-north Chile where is permanently affected by upwelling events of dissimilar intensity and duration which generate local conditions of acidification, deoxygenation, and cooling with different magnitudes. However, to date, it remains unknown how this economic valuable resource is physiologically affected throughout its life cycle by the continuous environmental changes driven by upwelling events of different intensities and duration along the year. Here, for the first time, A. purpuratus life-stage physiological sensitivity was assessed at a seasonal scale through a year-field experiment where growth, calcification, and survivorship were evaluated. Our study shows how seasonal differences in the upwelling phenology (here measured as changes in temperature, dissolved oxygen, pH, and primary productivity, but also as the number, duration, and intensity of cooling and de-oxygenation events) notably impacted the A. purpuratus physiological performance from juvenile to adult life-stages. This was especially noticeable during the spring season which showed the most intense cooling and deoxygenation events driven by stronger favorable-upwelling winds and the lowest growth and gross calcification rates (the highest decalcification rates) where adult stages showed the lowest performance. On the other hand, A. purpuratus survivorship was not significantly affected by upwelling intensity which would be providing evidence of the high physiological flexibility and well-locally adapted is this species to fluctuating and occasional stressful environmental conditions. Our results are significantly relevant in the climate change context as some upwelling systems are at risk to change shortly (i.e., an upwelling intensification in frequency and intensity) as a consequence of changes in the atmospheric pressures that modulate favourable-upwelling winds. These changes may certainly increase the climate related-risks of the entire socio-ecological systems related to the fishery and aquaculture of A. purpuratus along the Humboldt Current System.
Continue reading ‘Size matters: physiological sensitivity of the scallop Argopecten purpuratus to seasonal cooling and deoxygenation upwelling-driven events’Shelled pteropod abundance and distribution across the Mediterranean Sea during spring
Published 16 December 2022 Science ClosedTags: abundance, biological response, BRcommunity, chemistry, community composition, field, Mediterranean, mollusks, otherprocess, zooplankton
Highlights
- First estimate of pteropod distribution across the Mediterranean Sea in spring.
- Highest abundance recorded in the oligotrophic Eastern Mediterranean basin.
- Temperature, aragonite saturation, oxygen and salinity main drivers of distribution.
- Pteropods and planktic foraminifera are inversely distributed in the Med Sea.
Abstract
Thecosome pteropods are a dominant group of calcifying pelagic molluscs and an important component of the food web. In this study, we characterise spring pteropod distribution throughout the Mediterranean Sea, an understudied region for this common group of marine calcifying organisms. This semi-enclosed sea is rapidly changing under climatic and anthropogenic forcings. The presence of surface water biogeochemical gradients from the Atlantic Ocean/Gibraltar Strait to the Eastern Mediterranean Sea allowed us to investigate pteropod distribution and their ecological preferences. In the ultra-oligotrophic Eastern Mediterranean Sea, we found the mean upper 200 m pteropod standing stock of 2.13 ind. m-3 was approximately 5x greater than the Western basin (mean 0.42 ind. m-3). Where standing stocks were high, pteropods appeared largely in the same family grouping belonging to Limacinidae. Temperature, O2 concentration, salinity, and aragonite saturation (Ωar) explain 96% of the observed variations in the community structure at the time of sampling, suggesting that pteropods might show a preference for environmental conditions with a lower energetic physiological demand. We also document that pteropods and planktonic foraminifera have an opposite geographical distribution in the Mediterranean Sea. Our findings indicate that in specific pelagic ultra-oligotrophic conditions, such as the Eastern Mediterranean Sea, different feeding strategies could play an important role in regulating calcifying zooplankton distribution.
Continue reading ‘Shelled pteropod abundance and distribution across the Mediterranean Sea during spring’Bathyal octopus, Muusoctopus leioderma, living in a world of acid: first recordings of routine metabolic rate and critical oxygen partial pressures of a deep water species under elevated pCO2
Published 14 December 2022 Science ClosedTags: biological response, laboratory, mollusks, North Pacific, physiology
Elevated atmospheric CO2 as a result of human activity is dissolving into the world’s oceans, driving a drop in pH, and making them more acidic. Here we present the first data on the impacts of ocean acidification on a bathyal species of octopus Muusoctopus leioderma. A recent discovery of a shallow living population in the Salish Sea, Washington United States allowed collection via SCUBA and maintenance in the lab. We exposed individual Muusoctopus leioderma to elevated CO2 pressure (pCO2) for 1 day and 7 days, measuring their routine metabolic rate (RMR), critical partial pressure (Pcrit), and oxygen supply capacity (α). At the time of this writing, we believe this is the first aerobic metabolic data recorded for a member of Muusoctopus. Our results showed that there was no change in either RMR, Pcrit or α at 1800 µatm compared to the 1,000 µatm of the habitat where this population was collected. The ability to maintain aerobic physiology at these relatively high levels is discussed and considered against phylogeny and life history.
Continue reading ‘Bathyal octopus, Muusoctopus leioderma, living in a world of acid: first recordings of routine metabolic rate and critical oxygen partial pressures of a deep water species under elevated pCO2’Differential gene expression analysis in the scallop Argopecten purpuratus exposed to altered pH and temperature conditions in an upwelling-influenced farming area
Published 13 December 2022 Science ClosedTags: biological response, fisheries, laboratory, mitigation, molecular biology, mollusks, multiple factors, physiology, South Pacific, temperature

Increased carbon dioxide in the atmosphere and its absorption across the ocean surface will alter natural variations in pH and temperature levels, occurring in coastal upwelling ecosystems. The scallop Argopecten purpuratus, one of the most economically important species farmed in northern Chile, has been shown to be vulnerable to these environmental drivers. However, the regulatory responses at the gene-level of scallops to these climate stressors remain almost unknown. Consequently, we used an orthogonal experimental design and RNAseq approach to analyze the acute effects of variability in pH and temperature on gene expression in the muscle tissue of A. purpuratus. In respect to control conditions (pH ~ 8.0/ 14 °C), the influence of low pH (~ 7.7) and temperature (14 °C) induced the activation of several genes associated with apoptotic signaling pathways and protein localization to plasma membrane. Elevated temperature (18 °C) and pH (~8.0) conditions increased the expression of transcripts associated with the activation of muscle contraction, regulation, and sarcomere organization effects on muscle tissue. In scallops exposed to low pH and elevated temperature, the genes expressed were differentially associated with the oxidation-reduction process, signal translation, and positive regulation of GTPase activity. These results indicated that the differentially expressed genes under the experimental conditions tested are mainly related to the mitigation of cellular damage and homeostasis control. Our results add knowledge about the function of the adductor muscle in response to stressors in scallops. Furthermore, these results could help in the identification of molecular biomarkers of stress necessary to be integrated into the aquaculture programs for the mitigation of climate change.
Continue reading ‘Differential gene expression analysis in the scallop Argopecten purpuratus exposed to altered pH and temperature conditions in an upwelling-influenced farming area’Promoting pinto abalone (Haliotis kamtschatkana) recovery in the Salish Sea: the effects of fluctuating temperature and elevated CO2 on survival, growth, and radula morphology
Published 12 December 2022 Science ClosedTags: biological response, fisheries, laboratory, mitigation, mollusks, morphology, mortality, multiple factors, North Pacific, temperature
Overharvesting of pinto abalone (Haliotis kamtschatkana) in the Salish Sea between 1959 and 1994 caused severe population declines. This led to the Washington Department of Fish and Wildlife classifying pinto abalone as a “species of concern.” The Puget Sound Restoration Fund (PSRF) is committed to help pinto abalone recover by outplanting juveniles at specific sites around the Salish Sea. Survival of outplanted individuals is different at each site, but it is not clear why. Differences in water chemistry parameters, such as temperature and pH, could explain the differences in survival, either through differences in the mean conditions or through short term exposure to more extreme conditions. Future ocean warming and acidification could make fluctuations in water chemistry parameters more severe. The goal of my thesis was to simulate in lab the outplanting of abalone post-sets in fluctuating temperature and elevated CO2 conditions. I utilized an ocean acidification system to create atmospheres that affect seawater pH. I hypothesized that temperature fluctuations and high dissolved CO2 (low pH) will negatively affect survival, growth, and shell and radula morphology. Fluctuating temperatures yielded lower survival and greater growth, determined by mean shell length, compared to constant temperature. High CO2 yielded comparable survival and smaller growth than low CO2. Traditional morphological analysis of the radula found that fluctuating temperatures caused the radula to grow in a more compact manner, with smaller teeth formed closer together. Geometric morphological analysis found that radula tooth orientation was not affected by any of the treatments. This is the first study to find any effects of water chemistry on abalone radula morphology. Overall, the presence of a single stressor was detrimental to pinto abalone post-sets. However, the combination of stressors performed similarly to the absence of stressors. This indicates that fluctuating temperature can mitigate the negative effects of high CO2, possibly by increasing metabolic rate. In support of pinto abalone recovery efforts, PSRF can utilize my findings to evaluate water chemistry parameters at their outplant sites. I recommend that pinto abalone be outplanted in areas around the Salish Sea that are characterized by near constant temperatures, around 10°C, and low dissolved CO2 (high pH around 8.2). Due to yearly, seasonal, and weekly changes in water chemistry conditions, constant conditions do not exist. Outplant sites with the smallest fluctuations in water chemistry parameters should be used. In addition, ocean warming and acidification are expected to occur in concert. My findings indicate pinto abalone post-sets should be able to survive and grow under future climate scenarios, when outplanted into both temperatures that fluctuate on weekly scales, from 10°C up to 14°C, and acidification within 0.2 pH units when these conditions occur together, not separately.
Continue reading ‘Promoting pinto abalone (Haliotis kamtschatkana) recovery in the Salish Sea: the effects of fluctuating temperature and elevated CO2 on survival, growth, and radula morphology’Biological responses of the predatory blue crab and its hard clam prey to ocean acidification and low salinity
Published 9 December 2022 Science ClosedTags: biological response, BRcommunity, crustaceans, growth, laboratory, mesocosms, mollusks, morphology, mortality, multiple factors, performance, predation, salinity
How ocean acidification (OA) interacts with other stressors is understudied, particularly for predators and prey. We assessed long-term exposure to decreased pH and low salinity on (1) juvenile blue crab Callinectes sapidus claw pinch force, (2) juvenile hard clam Mercenaria mercenaria survival, growth, and shell structure, and (3) blue crab and hard clam interactions in filmed mesocosm trials. In 2018 and 2019, we held crabs and clams from the Chesapeake Bay, USA, in crossed pH (low: 7.0, high: 8.0) and salinity (low: 15, high: 30) treatments for 11 and 10 wk, respectively. Afterwards, we assessed crab claw pinch force and clam survival, growth, shell structure, and ridge rugosity. Claw pinch force increased with size in both years but weakened in low pH. Clam growth was negative, indicative of shell dissolution, in low pH in both years compared to the control. Growth was also negative in the 2019 high-pH/low-salinity treatment. Clam survival in both years was lowest in the low-pH/low-salinity treatment and highest in the high-pH/high-salinity treatment. Shell damage and ridge rugosity (indicative of deterioration) were intensified under low pH and negatively correlated with clam survival. Overall, clams were more severely affected by both stressors than crabs. In the filmed predator-prey interactions, pH did not substantially alter crab behavior, but crabs spent more time eating and burying in high-salinity treatments and more time moving in low-salinity treatments. Given the complex effects of pH and salinity on blue crabs and hard clams, projections about climate change on predator-prey interactions will be difficult and must consider multiple stressors.
Continue reading ‘Biological responses of the predatory blue crab and its hard clam prey to ocean acidification and low salinity’Molecular features associated with resilience to ocean acidification in the northern quahog, Mercenaria mercenaria
Published 6 December 2022 Science ClosedTags: biological response, laboratory, molecular biology, mollusks, North Atlantic, reproduction
The increasing concentration of CO2 in the atmosphere and resulting flux into the oceans will further exacerbate acidification already threatening coastal marine ecosystems. The subsequent alterations in carbonate chemistry can have deleterious impacts on many economically and ecologically important species including the northern quahog (Mercenaria mercenaria). The accelerated pace of these changes requires an understanding of how or if species and populations will be able to acclimate or adapt to such swift environmental alterations. Thus far, studies have primarily focused on the physiological effects of ocean acidification (OA) on M. mercenaria, including reductions in growth and survival. However, the molecular mechanisms of resilience to OA in this species remains unclear. Clam gametes were fertilized under normal pCO2 and reared under acidified (pH ~ 7.5, pCO2 ~ 1200 ppm) or control (pH ~ 7.9, pCO2 ~ 600 ppm) conditions before sampled at 2 days (larvae), 32 days (postsets), 5 and 10 months (juveniles) and submitted to RNA and DNA sequencing to evaluate alterations in gene expression and genetic variations. Results showed significant shift in gene expression profiles among clams reared in acidified conditions as compared to their respective controls. At 10 months of exposure, significant shifts in allele frequency of single nucleotide polymorphisms (SNPs) were identified. Both approaches highlighted genes coding for proteins related to shell formation, bicarbonate transport, cytoskeleton, immunity/stress, and metabolism, illustrating the role these pathways play in resilience to OA.
Continue reading ‘Molecular features associated with resilience to ocean acidification in the northern quahog, Mercenaria mercenaria’Ocean acidification affects the bioenergetics of marine mussels as revealed by high-coverage quantitative metabolomics
Published 25 November 2022 Science ClosedTags: biological response, laboratory, molecular biology, mollusks, North Pacific, performance, physiology

Highlights
- The metabolic response of mussels to acidification was evaluated.
- Acidification decreased energy storage and increased energy demands.
- Acidification affected amino acid metabolism and biosynthesis.
- Carry-over effects of acidification on cellular energy allocation were observed.
Abstract
Ocean acidification has become a major ecological and environmental problem in the world, whereas the impact mechanism of ocean acidification in marine bivalves is not fully understood. Cellular energy allocation (CEA) approach and high-coverage metabolomic techniques were used to investigate the acidification effects on the energy metabolism of mussels. The thick shell mussels Mytilus coruscus were exposed to seawater pH 8.1 (control) and pH 7.7 (acidification) for 14 days and allowed to recover at pH 8.1 for 7 days. The levels of carbohydrates, lipids and proteins significantly decreased in the digestive glands of the mussels exposed to acidification. The 14-day acidification exposure increased the energy demands of mussels, resulting in increased electron transport system (ETS) activity and decreased cellular energy allocation (CEA). Significant carry-over effects were observed on all cellular energy parameters except the concentration of carbohydrates and cellular energy demand (Ec) after 7 days of recovery. Metabolomic analysis showed that acidification affected the phenylalanine, tyrosine and tryptophan biosynthesis, taurine and hypotaurine metabolism, and glycine, serine and threonine metabolism. Correlation analysis showed that mussel cell energy parameters (carbohydrates, lipids, proteins, CEA) were negatively/positively correlated with certain differentially abundant metabolites. Overall, the integrated biochemical and metabolomics analyses demonstrated the negative effects of acidification on energy metabolism at the cellular level and implicated the alteration of biosynthesis and metabolism of amino acids as a mechanism of metabolic perturbation caused by acidification in mussels.
Continue reading ‘Ocean acidification affects the bioenergetics of marine mussels as revealed by high-coverage quantitative metabolomics’Sex and gametogenesis stage are strong drivers of gene expression in Mytilus edulis exposed to environmentally relevant plasticiser levels and pH 7.7
Published 23 November 2022 Science ClosedTags: biological response, laboratory, molecular biology, mollusks, morphology, multiple factors, physiology, reproduction, toxicants
Plastic pollution and changes in oceanic pH are both pressing environmental issues. Little emphasis, however, has been placed on the influence of sex and gametogenesis stage when investigating the effects of such stressors. Here, we examined histology and molecular biomarkers of blue mussels Mytilus edulis exposed for 7 days to a pH 7.7 scenario (− 0.4 units) in combination with environmentally relevant concentrations (0, 0.5 and 50 µg/L) of the endocrine disrupting plasticiser di-2-ethylhexyl phthalate (DEHP). Through a factorial design, we investigated the gametogenesis cycle and sex-related expression of genes involved in pH homeostasis, stress response and oestrogen receptor-like pathways after the exposure to the two environmental stressors. As expected, we found sex-related differences in the proportion of developing, mature and spawning gonads in histological sections. Male gonads also showed higher levels of the acid–base regulator CA2, but females had a higher expression of stress response-related genes (i.e. sod, cat, hsp70). We found a significant effect of DEHP on stress response-related gene expression that was dependent on the gametogenesis stage, but there was only a trend towards downregulation of CA2 in response to pH 7.7. In addition, differences in gene expression between males and females were most pronounced in experimental conditions containing DEHP and/or acidified pH but never the control, indicating that it is important to consider sex and gametogenesis stage when studying the response of mussels to diverse stressors.
Continue reading ‘Sex and gametogenesis stage are strong drivers of gene expression in Mytilus edulis exposed to environmentally relevant plasticiser levels and pH 7.7′Calmodulin regulates the calcium homeostasis in mantle of Crassostrea gigas under ocean acidification
Published 18 November 2022 Science ClosedTags: biological response, laboratory, molecular biology, mollusks, physiology
The biosynthesis of shell is a complicated calcification process in the marine bivalve, which can be severely impacted by ocean acidification (OA). Calmodulin (CaM) is a pivotal calcium regulator and thought to be crucial for calcification. In the present study, a CaM (designated CgCaM) with calcium-binding activity was identified from the Pacific oyster Crassostrea gigas with the objective to understand its possible role in the regulation of calcium homeostasis under acidification treatment. The open reading frame (ORF) of CgCaM was of 474 bp encoding a 17.5 kDa protein with four continuous EF-hand domains. CgCaM shared high similarity with CaMs from other invertebrates and vertebrates. The mRNA transcript of CgCaM was constitutively expressed in all detected tissues with the higher expression level in mantle, especially highest in the middle fold of the three folds of mantle. CgCaM was found to be mainly distributed in the mantle epithelium. When the oysters were exposed to acidified seawater, the expression level of CgCaM in the middle fold of mantle and the content of Ca2+ in this fold both decreased significantly. These results collectively suggested that CgCaM was involved in the regulation of calcium homeostasis in the middle fold of mantle under acidification treatment.
Continue reading ‘Calmodulin regulates the calcium homeostasis in mantle of Crassostrea gigas under ocean acidification’Intestinal microbiota perturbations in the gastropod Trochus niloticus concurrently exposed to ocean acidification and environmentally relevant concentrations of sulfamethoxazole
Published 15 November 2022 Science ClosedTags: biological response, molecular biology, mollusks, multiple factors, physiology, toxicants

Highlights
- Exposure to OA leads to the microbiota dysbiosis in the intestine of T. niloticus.
- Exposure to SMX barely affected the intestinal microbiota of T. niloticus.
- Exposure to SMX accelerated spread of sulfonamide ARGs.
Abstract
Ocean acidification (OA) and antibiotic pollution pose severe threats to the fitness of keystone species in marine ecosystems. However, the combined effects of OA and antibiotic pollution on the intestinal microbiota of marine organisms are still not well known. In this study, we exposed the herbivorous gastropod Trochus niloticus, a keystone species to maintains the stability of coral reef ecosystems, to acidic seawater (pH 7.6) and/or sulfamethoxazole (SMX, 100 ng/L, 1000 ng/L) for 28 days and determined their impacts on (1) the accumulation of SMX in the intestine of T. niloticus; (2) the characteristics of the intestinal microbiota in T. niloticus; (3) the relative abundances of sulfonamide resistance genes (i.e., sul1 and sul2) and intI1 in the intestinal microbiota of T. niloticus. Our results show that OA exposure leads to dramatic microbiota dysbiosis in the intestine of T. niloticus, including changes in bacterial community diversity and structure, decreased abundances of dominant species, existences of characteristic taxa, and altered functional predictions. In addition, SMX exposure at environmentally relevant concentrations had little effect on the intestinal microbiota of T. niloticus, whether in isolation or in combination with OA. However, after exposure to the higher SMX concentration (1000 ng/L), the accumulation of SMX in the intestine of T. niloticus could induce an increase in the copies of sul2 in the intestinal microbiota. These results suggest that the intestinal health of T. niloticus might be affected by OA and SMX, which might lead to fitness loss of the keystone species in coral reef ecosystems.
Continue reading ‘Intestinal microbiota perturbations in the gastropod Trochus niloticus concurrently exposed to ocean acidification and environmentally relevant concentrations of sulfamethoxazole’Impacts of seawater pH buffering on the larval microbiome and carry-over effects on later-life disease susceptibility in Pacific oysters
Published 11 November 2022 Science ClosedTags: abundance, biological response, fisheries, laboratory, molecular biology, mollusks, mortality, multiple factors, otherprocess, pathogens, reproduction, temperature
Ocean acidification upwelling events and the resulting lowered aragonite saturation state of seawater have been linked to high mortality of marine bivalve larvae in hatcheries. Major oyster seed producers along North America’s west coast have mitigated impacts via seawater pH buffering (e.g., addition of soda ash). However, little consideration has been given to whether such practice may impact the larval microbiome, with potential carry-over effects on immune competency and disease susceptibility in later-life stages. To investigate possible impacts, Pacific oysters (Crassostrea gigas) were reared under soda ash pH buffered or ambient pH seawater conditions for the first 24 h of development. Both treatment groups were then reared under ambient pH conditions for the remainder of the developmental period. Larval microbiome, immune status (via gene expression), growth, and survival were assessed throughout the developmental period. Juveniles and adults arising from the larval run were then subjected to laboratory-based disease challenges to investigate carry-over effects. Larvae reared under buffered conditions showed an altered microbiome, which was still evident in juvenile animals. Moreover, reduced survival was observed in both juveniles and adults of the buffered group under a simulated marine heatwave and Vibrio exposure compared with those reared under ambient conditions. Results suggest that soda ash pH buffering during early development may compromise later-life stages under stressor conditions, and illustrate the importance of a long-view approach with regard to hatchery husbandry practices and climate change mitigation.
Continue reading ‘Impacts of seawater pH buffering on the larval microbiome and carry-over effects on later-life disease susceptibility in Pacific oysters’