Posts Tagged 'laboratory'

Effects of ocean acidification on Lottia scutum settlement

The effects of ocean acidification on calcifying marine organisms are becoming more pronounced as atmospheric CO2 levels have increased due to anthropogenic carbon emissions (Etheridge et al., 1996). Studies on these effects have also increased over time. Ocean acidification (OA) has been shown to affect the feeding behavior and metabolic rates of larvae in a number of species (Vargas et al., 2013; Pan et al., 2015). Metabolic changes can significantly influence developmental rates, but little is still known about consequences of OA for non-feeding marine invertebrate larvae. In this study, we focus on the effects of OA conditions on the larval stage of Lottia scutum, a Pacific rocky intertidal limpet species that ranges from Alaska to southern California. Larvae were exposed to OA conditions (pH 7.3) at competency stage and monitored for settlement behavior and metamorphosis. Our results indicate that L. scutum larvae were able to successfully settle in OA and ambient seawater treatments. We did not find a negative effect of the specific OA conditions used in this study on the settlement of L. scutum. These findings provide insight into how environmental stress might affect early life stages, as well as how marine invertebrate larvae from regularly low pH environments fare in OA conditions.

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Molecular responses in an Antarctic bivalve and an ascidian to ocean acidification


  • The non-calcifying species Cnemidocarpa verrucosa sp. A showed a greater number of differentially expressed genes than the calcifying Aequiyoldia eightsii.
  • The Ocean Acidification caused an upregulation of genes involved in the immune system and antioxidant response in the ascidian Cnemidocarpa verrucosa sp. A.
  • The abundance of the key marine organisms (such as Cnemidocarpa verrucosa), could be affected by Ocean Acidification if pH predictions for polar regions come true.
  • Contrary to expected, Ocean Acidification could not affect the mollusk Aequiyoldia eightsii compared to the non-calcifying species.


Southern Ocean organisms are considered particularly vulnerable to Ocean acidification (OA), as they inhabit cold waters where calcite-aragonite saturation states are naturally low. It is also generally assumed that OA would affect calcifying animals more than non-calcifying animals. In this context, we aimed to study the impact of reduced pH on both types of species: the ascidian Cnemidocarpa verrucosa sp. A, and the bivalve Aequiyoldia eightsii, from an Antarctic fjord. We used gene expression profiling and enzyme activity to study the responses of these two Antarctic benthic species to OA. We report the results of an experiment lasting 66 days, comparing the molecular mechanisms underlying responses under two pCO2 treatments (ambient and elevated pCO2). We observed 224 up-regulated and 111 down-regulated genes (FC ≥ 2; p-value ≤ 0.05) in the ascidian. In particular, the decrease in pH caused an upregulation of genes involved in the immune system and antioxidant response. While fewer differentially expressed (DE) genes were observed in the infaunal bivalve, 34 genes were up-regulated, and 69 genes were downregulated (FC ≥ 2; p-value ≤ 0.05) in response to OA. We found downregulated genes involved in the oxidoreductase pathway (such as glucose dehydrogenase and trimethyl lysine dioxygenase), while the heat shock protein 70 was up-regulated. This work addresses the effect of OA in two common, widely distributed Antarctic species, showing striking results. Our major finding highlights the impact of OA on the non-calcifying species, results that differ from the general trend, in which one remarks the higher impact on calcifying species. Our result proposes a deep discussion about the potential effect on non-calcifying species, such as ascidians, a diverse and abundant group, that form extended three-dimensional clusters in the shallow waters and shelf areas along the Southern Ocean.

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Long-term preconditioning of the coral Pocillopora acuta does not restore performance in future ocean conditions

There is overwhelming evidence that tropical coral reefs are severely impacted by human induced climate change. Assessing the capability of reef-building corals to expand their tolerance limits to survive projected climate trajectories is critical for their protection and management. Acclimation mechanisms such as developmental plasticity may provide one means by which corals could cope with projected ocean warming and acidification. To assess the potential of preconditioning to enhance thermal tolerance in the coral Pocillopora acuta, colonies were kept under three different scenarios from settlement to 17 months old: present day (0.9 °C-weeks (Degree Heating Weeks), + 0.75 °C annual, 400 ppm pCO2) mid-century (2.5 °C-weeks, + 1.5 °C annual, 685 ppm pCO2) and end of century (5 °C-weeks, + 2 °C annual, 900 ppm pCO2) conditions. Colonies from the present-day scenario were subsequently introduced to the mid-century and end of century conditions for six weeks during summer thermal maxima to examine if preconditioned colonies (reared under these elevated conditions) had a higher physiological performance compared to naive individuals. Symbiodiniaceae density and chlorophyll a concentrations were significantly lower in mid-century and end of century preconditioned groups, and declines in symbiont density were observed over the six-week accumulated heat stress in all treatments. Maximum photosynthetic rate was significantly suppressed in mid-century and end of century preconditioned groups, while minimum saturating irradiances were highest for 2050 pre-exposed individuals with parents originating from specific populations. The results of this study indicate preconditioning to elevated temperature and pCO2 for 17 months did not enhance the physiological performance in P. acuta. However, variations in trait responses and effects on tolerance found among treatment groups provides evidence for differential capacity for phenotypic plasticity among populations which could have valuable applications for future restoration efforts.

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The effects of temperature and pH on egg release and germling development of the fucoid alga, Silvetia compressa

The rockweed, Silvetia compressa (Phaeophyceae, Fucales) is a fleshy, brown seaweed that occupies the middle intertidal zone on wave-exposed rocky shores from northern California to Baja California. Like other rockweeds in temperate latitudes worldwide, Silvetia is a canopy-forming foundation species that provides habitat and refuge from desiccation and thermal stress to a diverse community of intertidal organisms. Over the past several decades, Silvetia populations have exhibited declines driven likely by multiple anthropogenic disturbances, including alterations in environmental conditions associated with climate change, that are affecting both adults and early life history stages. Reproduction and development of Silvetia is particularly sensitive to abiotic stresses, thus, altered spawning, recruitment, and development are expected as global temperatures and carbon dioxide levels continue to rise and the ocean becomes more acidic. To examine the effects of temperature and pH on Silvetia early life history stages (egg and germling abundance, germination success, and germling development), I conducted a laboratory-controlled experiment. Collected reproductive tips of Silvetia were induced to release gametes in petri dishes (n=5 per treatment per month) with seawater conditions of four possible combinations of temperature (ambient 16℃ and a conservative future warm condition of 20℃) and pH (ambient 8.1 and a conservative future condition of 7.8) treatments. The number of eggs in petri dishes were quantified thirty minutes post-spawning and the number of germlings were counted a day later, with germination success. One-week post-spawning, the length of germlings remaining in petri dishes with maintained water conditions were measured. The effects of temperature and pH were variable among the different early life history stages of Silvetia with warming negatively impacting egg and germling counts and, potentially, germling length while future low pH conditions only reduced egg numbers under ambient temperature conditions. Despite documented peaks in reproductive output in Silvetia in winter, there were no clear temporal patterns for any measured parameter. Ocean warming may have a greater impact on Silvetia early life history stages than ocean acidification. Future impacts on early life history stages may result in continued declines of these ecosystem engineers which can have disproportionate effects on the ecosystem, including dramatic shifts in productivity, biodiversity, nutrient cycling, and ecosystem stability.

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Impact of dissolved CO2 on calcification in two large, benthic foraminiferal species

Rising atmospheric CO2 shifts the marine inorganic carbonate system and decreases seawater pH, a process often abbreviated to ‘ocean acidification’. Since acidification decreases the saturation state for crystalline calcium carbonate (e.g., calcite and aragonite), rising dissolved CO2 levels will either increase the energy demand for calcification or reduce the total amount of CaCO3 precipitated. Here we report growth of two large benthic photosymbiont-bearing foraminifera, Heterostegina depressa and Amphistegina lessonii, cultured at four different ocean acidification scenarios (400, 700, 1000 and 2200 ppm atmospheric pCO2). Using the alkalinity anomaly technique, we calculated the amount of calcium carbonate precipitated during the incubation and found that both species produced the most carbonate at intermediate CO2 levels. The chamber addition rates for each of the conditions were also determined and matched the changes in alkalinity. These results were complemented by micro-CT scanning of selected specimens to visualize the effect of CO2 on growth. The increased chamber addition rates at elevated CO2 concentrations suggest that both foraminifera species can take advantage of the increased availability of the inorganic carbon, despite a lower saturation state. This adds to the growing number of reports showing the variable response of foraminifera to elevated CO2 concentrations, which is likely a consequence of differences in calcification mechanisms.

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How does ocean acidification affect Zostera marina during a marine heatwave?


  • Under extreme conditions Z. marina grows in both leaf length and wet mass.
  • Increasing CO2 levels for Z. marina at high temperatures may stimulate growth.
  • Extremely high temperatures inhibit sucrose and starch synthesis in Z. marina.
  • Out of 223 identified differentially expressed genes 70 were upregulated.
  • Glycolysis and the TCA cycle controlling genes and metabolites were upregulated.


Extreme ocean events caused by global warming, such as marine heatwaves (MHWs) and ocean acidification (OA), are projected to intensify. A combination of extreme events may have severe consequences for marine ecosystemsZostera marina was selected to understand how seagrass adapts to OA in extremely hot conditions. By combining morphology, transcriptomics, and metabolomics under mesoscale experimental conditions, we systematically investigated the response characteristics of Z. marina. Extremely high temperatures had a pronounced effect on growth, and the combined effect of OA mitigated the inhibitory effect of MHW. Both transcriptomic and metabolomic results showed that Z. marina resisted OA and MHW by upregulating the TCA cycle, glycolysis, amino acid metabolism, and relevant genes, as well as by activating the antioxidant system. The results of this study serve to improve our understanding of dual effects of factors of climate change on seagrass and may be used to direct future management and conservation efforts.

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Brown seaweed Nemacystus decipiens intensifies the effects of ocean acidification on coral Montipora digitata

Photosynthetic marine macrophytes such as seaweeds have been proposed to provide habitat refugia for marine calcifiers against ocean acidification (OA) by increasing the local pH. However, the effectiveness of seaweed as a potential habitat refugia for marine calcifiers such as corals remains to be investigated. This study focused on the seaweed Nemacystus decipiens, which are widely farmed in the shallow reef lagoon of Okinawa coral reefs, Japan, and aimed to evaluate their response to high pCO2 and whether they can mitigate the effect of high pCO2 on the coral Montipora digitata. Corals were cultured with and without seaweed under control (300–400 μatm) or high pCO2 conditions (OA, 900–1000 μatm) for 2 weeks. Results showed that all photo-physiological parameters examined in the seaweed N. decipiens were not affected by high pCO2, suggesting that OA will not positively affect their productivity. The calcification rate of the coral M. digitata was found to decrease under OA and the effect was further exaggerated by the presence of seaweed. The present study suggests that farming seaweeds will not act as a potential habitat refugia for adjacent corals under future OA, but instead can exaggerate the negative effect of OA on coral calcification.

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Species-specific responses of macrophyte production to the increasing CO2 environment with potential ecosystem implications involved in the Baltic Sea

Macrophytes vary in their ability to utilize carbon in the form of HCO3 and/or CO2 for photosynthesis. Some functional groups that solely use CO2 for photosynthesis could receive competitive advantages from the predicted increase in CO2 compared to groups with efficient carbon acquisition strategies of HCO3. The aim of this study was to identify carbon use strategies in the common macrophytes (macroalgae, charophytes, seagrass, and other angiosperms) that represent a broad range of functional traits to CO2 concentrations in the northeastern Baltic Sea. Mechanistic assessment of the carbon physiology of macrophytes was used to predict productivity and competitive interactions between different functional groups under future climate. Carbon use strategies in macrophytes were determined by analysing the carbon isotopes (δ13C), pH drift experiments, and photosynthesis versus dissolved inorganic carbon. In addition, habitat mapping data was used to interpret the potential implications of the elevated CO2 to this coastal ecosystem. The results suggested that the primary productivity of macrophytes is often limited by carbon availability, and the increasing CO2 concentrations in the brackish Baltic Sea are expected to enhance photosynthetic production. While all species tested showed evidence of carbon concentrating mechanisms (CCMs), differential levels of CCM activity indicate varying levels of competitive fitness in a future high-CO2 environment. Overall, macrophytes which inhabit the shallowest and deepest parts of the vegetated zone are expected to experience physiological benefits under future CO2 conditions, while intermediate communities dominated by the perennial brown alga Fucus vesiculosus may experience loss of fitness. These fitness differences have implications for competitive interaction and species range under future climate.

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Pacific oysters do not compensate growth retardation following extreme acidification events

Ocean acidification caused by anthropogenic carbon dioxide emissions alters the growth of marine calcifiers. Although the immediate effects of acidification from global ocean models have been well studied on calcifiers, their recovery capacity over a wide range of pH has never been evaluated. This aspect is crucial because acidification events that arise in coastal areas can far exceed global ocean predictions. However, such acidification events could occur transiently, allowing for recovery periods during which the effects on growth would be compensated, maintained or amplified. Here we evaluated the recovery capacity of a model calcifier, the Pacific oyster Crassostrea gigas. We exposed juveniles to 15 pH conditions between 6.4 and 7.8 for 14 days. Oyster growth was retarded below pH 7.1 while shells were corroded at pH 6.5. We then placed the oysters under ambient pH > 7.8 for 42 days. Growth retardation persisted at pH levels below pH 7.1 even after the stress was removed. However, despite persistent retardation, growth has resumed rapidly suggesting that the oysters can recover from extreme acidification. Yet we found that the differences in individual weight between pH conditions below 7.1 increased over time, and thus the growth retardation cannot be compensated and may affect the fitness of the bivalves.

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Large-scale culturing of Neogloboquadrina pachyderma, its growth in, and tolerance of, variable environmental conditions

The planktic foraminifera Neogloboquadrina pachyderma is a calcifying marine protist and the dominant planktic foraminifera species in the polar oceans, making it a key species in marine polar ecosystems. The calcium carbonate shells of foraminifera are widely used in palaeoclimate studies because their chemical composition reflects the seawater conditions in which they grow. This species provides unique proxy data for past surface ocean hydrography, which can provide valuable insight to future climate scenarios. However, little is known about the response of N. pachyderma to variable and changing environmental conditions.Here, we present observations from large-scale culturing experiments where temperature, salinity and carbonate chemistry were altered independently. We observed overall low mortality, calcification of new chambers and addition of secondary calcite crust in all our treatments. In-culture asexual reproduction events also allowed us to monitor the variable growth of N. pachyderma’s offspring. Several specimens had extended periods of dormancy or inactivity after which they recovered. These observations suggest that N. pachyderma can tolerate, adapt to and calcify within a wide range of environmental conditions. This has implications for the species-level response to ocean warming and acidification, for future studies aiming to culture N. pachyderma and use in palaeoenvironmental reconstruction.

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Short-term exposure to independent and combined acidification and warming elicits differential responses from two tropical seagrass-associated invertebrate grazers

Ocean acidification and warming could affect animal physiology, key trophic interactions and ecosystem functioning in the long term. This study investigates the effects of four pH−temperature combination treatments simulating ocean acidification (OA), ocean warming (OW) and combined OA and OW conditions (FUTURE) relative to ambient present-day conditions (PRESENT) on the grazing of the juveniles of two seagrass-associated invertebrates namely the sea cucumber Stichopus cf. horrens and topshell Trochus maculatus over a 5-day exposure period. Diel and feeding activity of both species increased under OW and FUTURE to some extent, while the nighttime activity of Trochus but not Stichopus decreased under OA relative to PRESENT during the first 2 days. Fecal production of Stichopus did not differ among treatments, while the lowest fecal production of Trochus was observed under OA during the first 24 h of grazing. These responses suggest that Trochus may be initially more sensitive to OA compared with Stichopus. Interestingly, fecal production of Trochus in FUTURE was significantly higher than OA, suggesting that warming may ameliorate the negative effect of acidification. Diel activity, feeding and fecal production after 5 days did not differ among treatments for both species, suggesting acclimation to the acute changes in temperature and pH after a few days, although Stichopus acclimated rapidly than Trochus. The ability of the two juvenile invertebrate grazers to rapidly acclimate to increased temperature and lowered pH conditions after short-term exposure may favor their survival under projected changes in ocean conditions.

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Proteomic and transcriptomic analysis of large yellow croaker (Larimichthys crocea) during early development under hypoxia and acidification stress

In recent years, aquatic ecosystems have been exposed to various stressors such as hypoxia and acidification, which has become an issue of significant concern. Many studies in fish have investigated the regulatory mechanisms of the response to hypoxia and acidification stress at the molecular level. However, molecular studies on hypoxia acidification dual stress conditions in rhubarb fish are less. For this study, the juvenile large yellow croaker was used as the study object. Four experimental groups were established, including the control group (normal group, N107; DO = 7.0 mg/L, pH = 8.1), hypoxia group (H107; DO = 3.5 mg/L, pH = 8.1), acidification group (A107; DO = 7.0 mg/L, pH = 7.3), and hypoxia–acidification group (dual stress group, D107; DO = 3.5 mg/L, pH = 7.3). Study of its response mechanism under hypoxia acidification conditions by transcriptome and proteome analysis. The present study revealed that the number of quantifiable proteins was 6303. Five pairwise comparisons between experimental groups demonstrated that a total of 265 DEGs/DEPs showed associations between the transcriptome and proteome at the level of quantitative and differential expression. Comparative proteomic and transcriptomic analyses were performed to identify differentially expressed genes/proteins in juvenile Larimichthys crocea under hypoxia and acidification stress. The GO term enrichment analysis showed that hypoxia had a greater effect on the organism. The KEGG pathway enrichment analysis showed that pathways associated with the extracellular matrix ECM–receptor interaction and protein digestion and absorption pathways were notably affected by hypoxia and acidification stress. Among these, the protein digestion and absorption pathway was significantly affected in all five pairwise comparisons between experimental groups. The ECM–receptor interaction pathway was significantly enriched under dual stress, indicating that dual stress had a greater detrimental effect on fish growth than single stressors. The study provides valuable insights into the potential combined effects of decreased pH and DO in Sciaenidae and elucidates the mechanism underlying the response of L. crocea to simultaneous hypoxia–acidification stress during early development.

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Examining the reproductive success of bull kelp (Nereocystis luetkeana, Phaeophyceae, Laminariales) in climate change conditions

Climate change is affecting marine ecosystems in many ways, including raising temperatures and leading to ocean acidification. From 2014 to 2016, an extensive marine heat wave extended along the west coast of North America and had devastating effects on numerous species, including bull kelp (Nereocystis luetkeana). Bull kelp is an important foundation species in coastal ecosystems and can be affected by marine heat waves and ocean acidification; however, the impacts have not been investigated on sensitive early life stages. To determine the effects of changing temperatures and carbonate levels on Northern California’s bull kelp populations, we collected sporophylls from mature bull kelp individuals in Point Arena, CA. At the Bodega Marine Laboratory, we released spores from field-collected bull kelp, and cultured microscopic gametophytes in a common garden experiment with a fully factorial design crossing modern conditions (11.63 ± 0.54°C and pH 7.93 ± 0.26) with observed extreme climate conditions (15.56 ± 0.83°C and 7.64 ± 0.32 pH). Our results indicated that both increased temperature and decreased pH influenced growth and egg production of bull kelp microscopic stages. Increased temperature resulted in decreased gametophyte survival and offspring production. In contrast, decreased pH had less of an effect but resulted in increased gametophyte survival and offspring production. Additionally, increased temperature significantly impacted reproductive timing by causing female gametophytes to produce offspring earlier than under ambient temperature conditions. Our findings can inform better predictions of the impacts of climate change on coastal ecosystems and provide key insights into environmental dynamics regulating the bull kelp lifecycle.

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Seasonal production dynamics of high latitude seaweeds in a changing ocean: implications for bottom-up effects on temperate coastal food webs

As the oceans absorb excess heat and CO2 from the atmosphere, marine primary producers face significant changes to their abiotic environments and their biotic interactions with other species. Understanding the bottom-up consequences of these effects on marine food webs is essential to informing adaptive management plans that can sustain ecosystem and cultural services. In response to this need, this dissertation provides an in-depth consideration of the effects of global change on foundational macroalgal (seaweed) species in a poorly studied, yet highly productive region of our world’s oceans. To explore how seaweeds within seasonally dynamic giant kelp forest ecosystems will respond to ocean warming and acidification, I employ a variety of methods: year-round environmental monitoring using an in situ sensor array, monthly subtidal community surveys, and a series of manipulative experiments. I find that a complementary phenology of macroalgal production currently characterizes these communities, providing complex habitat and a nutritionally diverse energy supply to support higher trophic levels throughout the year. I also find that future ocean warming and acidification will lead to substantial shifts in the phenology, quantity and quality of macroalgal production in these systems. My results suggest that the giant kelp Macrocystis pyrifera may be relatively resilient to the effects of global change in future winter and summer seasons at high latitudes. In contrast, the calcifying coralline algae Bossiella orbigniana and Crusticorallina spp. and the understory kelps Hedophyllum nigripes and Neoagarum fimbriatum will experience a suite of negative impacts, especially in future winter conditions. The resulting indirect effects on macroalgal-supported coastal food webs will be profound, with projected reductions in habitat and seasonal food supply on rocky reefs. Coming at a time of heightened interest in seaweed production potential at high latitudes, this dissertation provides a comprehensive evaluation of the future of these foundational organisms in a changing environment.

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The response of coral skeletal nano structure and hardness to ocean acidification conditions

Ocean acidification typically reduces coral calcification rates and can fundamentally alter skeletal morphology. We use atomic force microscopy (AFM) and microindentation to determine how seawater pCO2 affects skeletal structure and Vickers hardness in a Porites lutea coral. At 400 µatm, the skeletal fasciculi are composed of tightly packed bundles of acicular crystals composed of quadrilateral nanograins, approximately 80–300 nm in dimensions. We interpret high adhesion at the nanograin edges as an organic coating. At 750 µatm the crystals are less regular in width and orientation and composed of either smaller/more rounded nanograins than observed at 400 µatm or of larger areas with little variation in adhesion. Coral aragonite may form via ion-by-ion attachment to the existing skeleton or via conversion of amorphous calcium carbonate precursors. Changes in nanoparticle morphology could reflect variations in the sizes of nanoparticles produced by each crystallization pathway or in the contributions of each pathway to biomineralization. We observe no significant variation in Vickers hardness between skeletons cultured at different seawater pCO2. Either the nanograin size does not affect skeletal hardness or the effect is offset by other changes in the skeleton, e.g. increases in skeletal organic material as reported in previous studies.

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Development of optical fibre pH sensors for marine microenvironments

The oceans absorb approximately one-third of the CO2 emission into the atmosphere causing a decline in seawater pH, a process known as ocean acidification (OA). This decline in pH reflects changes to the seawater carbonate system and is expected to have an impact on the marine environment and ecosystems. pH changes along coastal ecosystems are highly variable and knowledge of these marine environments can be used to enhance our understanding of OA impacts on marine organisms.

The micro-to-centimetre thick layer directly surrounding many aquatic organisms is known as the diffusion boundary layer (DBL). The DBL creates a region which reduces the exposure of calcifying species to OA conditions. The pH within the DBL is dependent on light-controlled metabolic activities and exhibits different pH behaviour to bulk seawater. The challenge of detecting in situ pH variations and attributing OA effects highlights a need for a fresh research approach and innovative analyses.

The objective of this research is to develop optical fibre pH sensors capable of continuous pH measurement, and suitable for measuring pH variation in marine microenvironments. The development, characterisation, and applications of optical fibre pH sensors are described. The pH sensing components consist of pH-sensitive indicators immobilised in an optimised sol-gel matrix, minimising indicator leaching without the need for a covalent bond. This research explores two approaches, absorbance-based and fluorescence-based pH sensors.

The absorbance-based sensor applied meta-cresol purple (mCP) as the pH-sensitive indicator. The pH sensor has a usable lifetime of 7 days and a dynamic range of pH 7.4 to 9.7. This self-referencing pH sensor was utilised for real-time pH measurements within the DBL of the seaweed Ulva sp., and successfully used to monitor metabolic activity for 100 hours, achieving a precision of 0.02 pH units. This sensor conformed to the GOA-ON Weather quality guideline and demonstrated its capability to identify short-term variation in biological and environmental studies.

The fluorescence pH sensor utilises a time-domain dual-lifetime referencing scheme (t-DLR). The fluorescence pH sensing materials required the synthesis of pH-sensitive iminocoumarin and the encapsulation of pH-inert reference Ru(dpp)3 in polyacrylonitrile (PAN). The pH is determined from the ratio of the combined excitation intensity to the emission intensity of the reference indicator. This approach allows the signal to be referenced internally, independent of fluorescence dye concentration and variations in excitation light intensity. The t-DLR instrumentation used commercial electronic and optical components, integrated with custom-made electronic circuits. The pH sensor has a dynamic range of pH 7.8 to 9.3 and a precision of 0.02 pH units. The pH sensor was insensitive to changes in salinity and had negligible dye leaching and minimal photobleaching.

This work accomplished the development of mCP-based and dual-layer t-DLR fluorescent-based optical fibre pH sensors. This highlights the versatility of optical fibre pH sensors and the potential for a wider range of applications.

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A bone morphogenetic protein regulates the shell formation of Crassostrea gigas under ocean acidification

Bone morphogenetic proteins (BMPs) are key factors controlling osteoblast differentiation, which have been proved to be involved in the hard tissue formation of marine mollusks. In the present study, a member of BMPs gene (CgBMP7) was identified from Pacific oyster Crassostrea gigas (C. gigas) with the aim to understand its possible role in the regulation of shell formation under ocean acidification (OA) conditions. The open reading frame (ORF) of CgBMP7 was of 1254 bp encoding a polypeptide of 417 amino acids. The deduced amino acid sequence of CgBMP7 was comprised of one signal peptide, one prodomain and one TGF-β domain, which shared 21.69%-61.10% identities with those from other species. The mRNA transcript of CgBMP7 was ubiquitously expressed in all the tested tissues of adult oysters with a higher expression level in mantle, notably highest in the middle fold of the three folds of mantle. The expression level of bone morphogenetic protein type I receptor (CgBMPR1B) mRNA was also highest in the MF and up-regulated dramatically post recombinant BMP7 protein (rCgBMP7) stimulation. After the blockage of BMPR1B with inhibitor LDN193189 (LDN), the mRNA expression level and phosphorylation level of CgSmad1/5/8 in mantle were decreased, and the mRNA expression level of CgCaM and Cgengrailed-1 were down-regulated significantly. During the oysters were exposed to acidified seawater for weeks, the expression levels of CgBMP7, CgBMPR1B and CgSmad1/5/8 in the middle fold of mantle decreased significantly (p < 0.01) at the 4th week, and CgCaM and Cgengrailed-1 also exhibited the same variable expression patterns as CgBMP7. In addition, the growth of shell in the treatment group (pH 7.8) was slower than that in the control group (pH 8.1). These results collectively indicated that BMP7 was able to trigger the BMPR-Smad signaling pathway and involved in controlling the formation of oyster calcified shell under OA conditions.

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The reproductive capacities of the calanoid copepods Parvocalanus crassirostis and Acartia pacifica under different pH and temperature conditions

The increasing atmospheric CO2 concentrations and warming of marine waters have encouraged experiments on multi-stressor interactions in marine organisms. We conducted a multigenerational experiment to assess reproductive capacities regarding egg production in calanoid copepods Parvocalanus crassirostis and Acartia pacifica under different pH and temperature conditions. The experimental set-up allowed assessing the tandem effect of warming and acidification on the number of eggs produced by healthy copepod pairs under two pH conditions of 8.20 and 7.50 (hard selection) as well as with a gradual reduction of 0.05 pH units at each generation (soft selection). The results are quite interesting, with very diverse performance across temperatures. The number of eggs produced under hard selection was higher at pH 8.20 compared to pH 7.50 for both species, with the maximum number of eggs produced at 24–28 °C, whereas under soft selection, there was no significant difference in the egg production rate at 24–28 °C across generations and there was an improvement in the number of eggs produced at 8–16 °C. The results provide evidence that in a future ocean scenario of lower pH and higher temperature, the two species, and possibly the copepod population at large, might not decrease. Copepod populations might be resilient, and the transcriptomic evidence of adaptation to increased temperature and lower pH is a ray of hope. We believe further studies are needed to provide more robust datasets to underpin the hypothesis of adaptation to climate change.

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Transcriptomic responses in the nervous system and correlated behavioural changes of a cephalopod exposed to ocean acidification

The nervous system is central to coordinating behavioural responses to environmental change, likely including ocean acidification (OA). However, a clear understanding of neurobiological responses to OA is lacking, especially for marine invertebrates. We evaluated the transcriptomic response of the central nervous system (CNS) and eyes of the two-toned pygmy squid ( Idiosepius pygmaeus ) to OA conditions, using a de novo transcriptome assembly created with long read PacBio ISO-sequencing data. We then correlated patterns of gene expression with CO treatment levels and OA-affected behaviours in the same individuals. OA induced transcriptomic responses within the nervous system related to various different types of neurotransmission, neuroplasticity, immune function and oxidative stress. These molecular changes may contribute to OA-induced behavioural changes, as suggested by correlations between gene expression profiles, CO treatment and OA-affected behaviours. This study provides the first molecular insights into the neurobiological effects of OA on a cephalopod and correlates molecular changes with whole animal behavioural responses, helping to bridge the gap in our knowledge between environmental change and animal responses.

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Short-term exposure to combined condition of low salinity and pH affects ROS-mediated stress in disk abalone (Haliotis discus hannai)

Climate change due to global warming can alter the salinity and pH in aquatic ecosystems. Low salinity (LS) and ocean acidification (OA) are stressors involved in osmotic regulation and can alter the antioxidant capacity of the body. In this study, we observed Na+/K+-ATPase (NKA) expression and activity in disk abalone gill tissue and changes in hemolymph osmolarity in relation to osmotic regulation over a short period (5 days). To confirm the degree of oxidative stress caused by changes in salinity and pH, changes in H2O2 levels, reactive oxygen species (ROS) levels, antioxidant enzyme (superoxide dismutase [SOD] and catalase [CAT]) expression, and caspase-7 expression were investigated at the molecular level. The degree of DNA damage was evaluated using the comet assay. mRNA expression, activity of gill NKA, and osmolarity of the hemolymph were significantly decreased in the LS group. Nonetheless, no noteworthy distinction was observed in mRNA expression or NKA activity between the control group and OA group. Hemolymph H2O2 levels and mRNA expression of SOD, CAT, and caspase-7 were significantly higher under the LS + OA condition than under single conditions of LS and OA. Further, caspase-7 mRNA expression and DNA damage increased with increasing exposure time. The group exposed to LS + OA showed the highest levels of caspase-7 expression and DNA damage. These results indicate that a combination of low salinity and pH induces more stress than a single condition does. Unmanageable ROS-mediated stress caused by environmental changes can lead to cell death and DNA damage.

Continue reading ‘Short-term exposure to combined condition of low salinity and pH affects ROS-mediated stress in disk abalone (Haliotis discus hannai)’

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