Posts Tagged 'laboratory'

Effects of salinity and pH of seawater on the reproduction of the sea urchin Paracentrotus lividus

Fertilization and early development are usually the most vulnerable stages in the life of marine animals, and the biological processes during this period are highly sensitive to the environment. In nature, sea urchin gametes are shed in seawater, where they undergo external fertilization and embryonic development. In a laboratory, it is possible to follow the exact morphological and biochemical changes taking place in the fertilized eggs and the developing embryos. Thus, observation of successful fertilization and the subsequent embryonic development of sea urchin eggs can be used as a convenient biosensor to assess the quality of the marine environment. In this paper, we have examined how salinity and pH changes affect the normal fertilization process and the following development of Paracentrotus lividus. The results of our studies using confocal microscopy, scanning and transmission electron microscopy, and time-lapse Ca2+ image recording indicated that both dilution and acidification of seawater have subtle but detrimental effects on many aspects of the fertilization process. They include Ca2+ signaling and coordinated actin cytoskeletal changes, leading to a significantly reduced rate of successful fertilization and, eventually, to abnormal or delayed embryonic development.

Continue reading ‘Effects of salinity and pH of seawater on the reproduction of the sea urchin Paracentrotus lividus’

The role of aspartic acid in reducing coral calcification under ocean acidification conditions

Biomolecules play key roles in regulating the precipitation of CaCO3 biominerals but their response to ocean acidification is poorly understood. We analysed the skeletal intracrystalline amino acids of massive, tropical Porites spp. corals cultured over different seawater pCO2. We find that concentrations of total amino acids, aspartic acid/asparagine (Asx), glutamic acid/glutamine and alanine are positively correlated with seawater pCO2 and inversely correlated with seawater pH. Almost all variance in calcification rates between corals can be explained by changes in the skeletal total amino acid, Asx, serine and alanine concentrations combined with the calcification media pH (a likely indicator of the dissolved inorganic carbon available to support calcification). We show that aspartic acid inhibits aragonite precipitation from seawater in vitro, at the pH, saturation state and approximate aspartic acid concentrations inferred to occur at the coral calcification site. Reducing seawater saturation state and increasing [aspartic acid], as occurs in some corals at high pCO2, both serve to increase the degree of inhibition, indicating that biomolecules may contribute to reduced coral calcification rates under ocean acidification.

Continue reading ‘The role of aspartic acid in reducing coral calcification under ocean acidification conditions’

Fluctuating seawater pCO2/pH induces opposing interactions with copper toxicity for two intertidal invertebrates

Highlights

• Fluctuating pCO2/pH doubled copper-induced antioxidant activity and DNA damage in mussels.

• In contrast, fluctuating conditions mitigated some of the effects of copper for ragworms.

• These effects were associated with the differing acid-base physiology of the two species.

• Physiology was as important as changing copper chemistry in determining overall toxicity.

Abstract

Global ocean pCO2 is increasing as a result of anthropogenic CO2 emissions, driving a decline in seawater pH. However, coastal waters already undergo fluctuations in pCO2/pH conditions over far shorter timescales, with values regularly exceeding those predicted for the open ocean by the year 2100. The speciation of copper, and therefore its potential toxicity, is affected by changing seawater pH, yet little is known concerning how present-day natural fluctuations in seawater pH affect copper toxicity to marine biota. Here, we test the hypothesis that a fluctuating seawater pCO2/pH regime will alter the responses of the mussel Mytilus edulis and the ragworm Alitta virens to sub-lethal copper, compared to a static seawater pCO2/pH scenario. Mussels and worms were exposed to 0.1 and 0.25 μM copper respectively, concentrations determined to produce comparable toxicity responses in these species, for two weeks under a fluctuating 12-hour pCO2/pH cycle (pH 8.14–7.53, pCO2 445–1747 μatm) or a static pH 8.14 (pCO2 432 μatm) treatment. Mussels underwent a haemolymph acidosis of 0.1–0.2 pH units in the fluctuating treatments, alongside two-fold increases in the superoxide dismutase activity and DNA damage induced by copper, compared to those induced by copper under static pH conditions. Conversely, ragworms experienced an alkalosis of 0.3 pH units under fluctuating pH/pCO2, driven by a two-fold increase in coelomic fluid bicarbonate. This mitigated the copper-induced oxidative stress to slightly reduce both antioxidant activity and DNA damage, relative to the static pH + copper treatment. These opposing responses suggest that differences in species acid-base physiology were more important in determining toxicity responses than the pH-induced speciation change. With variability in seawater chemistry predicted to increase as climate change progresses, understanding how fluctuating conditions interact with the toxicity of pH-sensitive contaminants will become more crucial in predicting their risk to coastal biota.

Continue reading ‘Fluctuating seawater pCO2/pH induces opposing interactions with copper toxicity for two intertidal invertebrates’

Coccolithophore community response to ocean acidification and warming in the Eastern Mediterranean Sea: results from a mesocosm experiment

Mesocosm experiments have been fundamental to investigate the effects of elevated CO2 and ocean acidification (OA) on planktic communities. However, few of these experiments have been conducted using naturally nutrient-limited waters and/or considering the combined effects of OA and ocean warming (OW). Coccolithophores are a group of calcifying phytoplankton that can reach high abundances in the Mediterranean Sea, and whose responses to OA are modulated by temperature and nutrients. We present the results of the first land-based mesocosm experiment testing the effects of combined OA and OW on an oligotrophic Eastern Mediterranean coccolithophore community. Coccolithophore cell abundance drastically decreased under OW and combined OA and OW (greenhouse, GH) conditions. Emiliania huxleyi calcite mass decreased consistently only in the GH treatment; moreover, anomalous calcifications (i.e. coccolith malformations) were particularly common in the perturbed treatments, especially under OA. Overall, these data suggest that the projected increase in sea surface temperatures, including marine heatwaves, will cause rapid changes in Eastern Mediterranean coccolithophore communities, and that these effects will be exacerbated by OA.

Continue reading ‘Coccolithophore community response to ocean acidification and warming in the Eastern Mediterranean Sea: results from a mesocosm experiment’

Epiphytes provide micro-scale refuge from ocean acidification

Highlights

• OA induced bleaching and reduced metabolism in non-epiphytized coralline.

• Epiphytized corallines were less susceptible to the detrimental effects of OA.

• Epiphytized corallines had thicker diffusive boundary layer than non-epiphytized.

• Non-calcifying epiphytes provide small scale refuge from OA.

• Epiphytic refugia may protect corallines under future OA conditions.

Abstract

Coralline algae, a major calcifying component of coastal shallow water communities, have been shown to be one of the more vulnerable taxonomic groups to ocean acidification (OA). Under OA, the interaction between corallines and epiphytes was previously described as both positive and negative. We hypothesized that the photosynthetic activity and the complex structure of non-calcifying epiphytic algae that grow on corallines ameliorate the chemical microenvironmental conditions around them, providing protection from OA. Using mesocosm and microsensor experiments, we showed that the widespread coralline Ellisolandia elongata is less susceptible to the detrimental effects of OA when covered with non-calcifying epiphytic algae, and its diffusive boundary layer is thicker than when not covered by epiphytes. By modifying the microenvironmental carbonate chemistry, epiphytes, facilitated by OA, create micro-scale shield (and refuge) with more basic conditions that may allow the persistence of corallines associated with them during acidified conditions. Such ecological refugia could also assist corallines under near-future anthropogenic OA conditions.

Continue reading ‘Epiphytes provide micro-scale refuge from ocean acidification’

Physiological and ecotoxicological interactions of copper and ocean acidification in the polychaete worms Hediste diversicolor and Alitta virens

For coastal aquatic habitats the change in seawater pH occurring as a result of ocean acidification has the potential to alter the speciation and toxicity of the many contaminants that remain in high concentrations in coastal systems. Of particular concern are metals, such as copper, whose speciation is pH sensitive within the OA range. A meta-analysis of studies to date investigating OA-contaminant interactions using marine invertebrates reveals that 72% of the 44 studies conducted have indeed focused on metals such as copper, with only a few studies looking at polycyclic aromatic hydrocarbons (PAH) and pharmaceuticals. No clear trends in the pH-effect size on contaminant toxicity for either species or contaminant group were present however, suggesting species specific physiological responses may influence this interaction as well as contaminant chemistry. A relatively understudied group were the polychaetes, a key functional group for many coastal sediments. Sediments act as a sink for contaminants where they can accumulate to high concentrations. Hence there is high potential for polychaetes to experience elevated metal exposures under reduced seawater pH as OA progresses. To address this knowledge gap, the responses of two common coastal polychaete, Alitta virens and Hediste diversicolor, were studied under three different experimental scenarios (both water-borne and sediment based) focusing on the physiological and toxicological responses under combined exposures to ocean acidification and copper. Water-borne exposures of Alitta virens to 0.25 μM copper under ambient seawater (pH 8.10) showed a significant increase in DNA damage, along with a rise in both SOD activity and lipid peroxidation. However, when exposed to copper under OA conditions (pH 7.70) there was no further increase in DNA damage and a significant decrease in SOD activity was observed alongside a fall in lipid peroxidation suggesting that OA looks to buffer the toxicity responses to this species. This is in contrast to previous studies using mussels and sea urchins, where copper toxicity responses were significantly higher when exposed under OA conditions. To assess whether local adaptations to high levels of copper contamination influences this OA-copper interaction, a population comparison using a metal resistance population of the harbour ragworm, Hediste diversicolor and a nearby non-resistant population was then conducted. Exposures were run using copper concentrations that elicit comparable toxicity responses, using 0.50 uM copper for the resistant population, compared to 0.25 uM for the non-resistant population, reflecting the two-fold differences in LC50 values for these population. These experiments reveal a significant increase by 19.70% in metabolic rate effect size (the combined stressor when compared to the control) in the resistant population compared to a decrease by 24.02% the non-resistant population, along with differences in ammonia excretion rate and the O:N ratio, thus revealing an energetic cost of this genetic resistance when faced with the combined stressors of OA and copper. These data are in line with the emerging energy limited tolerance to stressors’ hypothesis which states that tolerance to stress can be energy limited, with bioenergetics playing a central role in the tolerance to environmental stress. Finally, a more environmentally realistic exposure scenario was conducted using Alitta virens to test the influence of sediment and tidal cycles on worm acid-base and oxidative stress responses. Field measurements of sediment pH revealed that the pHNBS range over a tidal cycle varies from 6.97 to 7.87, indicating that polychaetes are already experiencing pH’s lower than the predictions for near future open oceans. In aquarium exposures, with overlying water of pHNBS 8.10, sediment pHNBS remained within the range of 7.45 to 7.31, when the overlying water was manipulated to OA conditions (pHNBS 7.70) sediment pHNBS was within the same range as the ambient treatment. The lack of change in sediment pH, despite a 0.40 unit drop in seawater pH, removed any comparative differences in physiological and toxicity end points in the worms between treatments. Tidal emersion induced a slight reduction in sediment pH, with a significant copper effect on sediment pH causing a further decrease in pH levels. Interestingly emersion resulted in a significant OA-copper interaction for coelomic fluid bicarbonate, which increased over the emersion period, however, there was no emersion driven acidosis within coelomic fluid. Overall this work further points to contaminant-OA interactions being species specific driven, in part driven by animal physiology. It also highlights the importance of environmentally relevant exposures with sediment dwelling organisms experiencing lower pH levels than the overlying seawater which could potentially affect metal speciation and could lead to OA-contaminant interactions occurring very differently in this environment. These are important considerations for ecotoxicology studies in the face of global ocean changes.

Continue reading ‘Physiological and ecotoxicological interactions of copper and ocean acidification in the polychaete worms Hediste diversicolor and Alitta virens’

Nutrient availability modulates the effects of climate change on growth and photosynthesis of marine macroalga Pyropia haitanensis (Bangiales, Rhodophyta)

The present research investigated the effect of pCO2 levels (C), seawater temperature (T), and nutrient availability (N) on the growth and physiochemical changes in Pyropia haitanensis. With nutrient enrichment, the interaction of higher pCO2 increased relative growth rates (RGR) by 105.9% when temperature increased (22 °C) compared with the control (lower T, lower C, and lower N: LTLCLN). The higher pCO2 decreased the Pm rates at the lower temperature (18 °C), yet displayed no interaction with higher T or N levels. The higher N increased dark respiration rate (Rd) at 18 °C. At 22 °C, higher pCO2 significantly enhanced the maximum ratio of (quantum yields (Fv/Fo) and the maximum quantum yield (ψpo), while it sharply decreased the absorption of photons per active reaction center (ABS/RC) and dissipation of energy fluxes (per RC) (DIo/RC). Higher temperature obviously reduced the Fv/Fo and ψpo under ambient CO2 level. The higher pCO2 significantly increased the phycoerythrin (PE) and phycocyanin (PC) contents, while higher temperature decreased the PE contents with elevated CO2 and declined the PC content regardless of CO2 condition. At lower nutrient condition, higher pCO2 increased Chl a content. Soluble carbohydrates (SC) and soluble protein (SP) content almost was unchanged among all treatments. Our findings indicate that nutrient availability may regulate photosynthetic mechanism to offset the negative effect of future ocean warming on P. haitanensis, thereby sustaining or increasing the biomass yield of the algae.

Continue reading ‘Nutrient availability modulates the effects of climate change on growth and photosynthesis of marine macroalga Pyropia haitanensis (Bangiales, Rhodophyta)’

Comparative metabolome analysis provides new insights into increased larval mortality under seawater acidification in the sea urchin Strongylocentrotus intermedius

Highlights

• Seawater acidification’s effects on Strongylocentrotus intermedius larvae were investigated.

• A comparative metabolomics analysis was performed.

• Significantly differentially expressed metabolites were identified and annotated.

• Metabolic pathways related to mortality induced by seawater acidification were observed.

Abstract

Mortality and metabolic responses of four-armed larvae of Strongylocentrotus intermedius under CO2-induced seawater acidification were investigated. Gametes of S. intermedius were fertilized and developed to the four-armed larval stage in either current natural seawater pH levels (as Control; pH = 7.99 ± 0.01) or laboratory-controlled acidified conditions (OA1: ΔpH = −0.3 units; OA2: ΔpH = −0.4 units; OA3: ΔpH = −0.5 units) according to the predictions of the Intergovernmental Panel on Climate Change (IPCC). The degrees of spicule exposure and asymmetry and mortality of four-armed larvae of S. intermedius were observed; each had a significant linearly increasing trend as the seawater pH level decreased. Comparative metabolome analysis identified a total of 87 significantly differentially expressed metabolites (SDMs, UP: 57, DOWN: 30) in OA-treated groups compared with the control group. Twenty-three SDMs, including carnitine, lysophosphatidylcholine (LPC) 18:3, lysophosphatidyl ethanolamine (LPE) 16:1, glutathione (GSH) and L-ascorbate, exhibited a linear increasing trend with decreasing seawater pH. Nine SDMs exhibited a linear decreasing trend as the seawater pH declined, including hypoxanthine, guanine and thymidine. Among all SDMs, we further mined 48 potential metabolite biomarkers responding to seawater acidification in four-armed larvae of S. intermedius. These potential metabolite biomarkers were mainly enriched in five pathways: glycerophospholipid metabolism, glutathione metabolism, purine metabolism, pyrimidine metabolism and the tricarboxylic acid cycle (TCA cycle). Our results will enrich our knowledge of the molecular mechanisms employed by sea urchins in response to CO2-induced seawater acidification.

Continue reading ‘Comparative metabolome analysis provides new insights into increased larval mortality under seawater acidification in the sea urchin Strongylocentrotus intermedius’

Elevated temperature and CO2 have positive effects on the growth and survival of larval Australasian snapper

Highlights

• Larval Snapper were positively affected by projected end of century temperature and pCO2 from fertilization to 16 days post-hatching.

• Elevated temperature increased the size of larvae, however high pCO2 had no effect.

• High pCO2 significantly increased survival at 16 days post-hatch, but elevated temperature had no effect.

• Some species and populations of marine fish exhibit positive effects from projected environmental change.

Abstract

Rising water temperature and increased uptake of CO2 by the ocean are predicted to have widespread impacts on marine species. However, the effects are likely to vary, depending on a species’ sensitivity and the geographical location of the population. Here, we investigated the potential effects of elevated temperature and pCO2 on larval growth and survival in a New Zealand population of the Australasian snapper, Chrysophyrs auratus. Eggs and larvae were reared in a fully cross-factored experiment (18 °C and 22 °C/pCO2 440 and 1040 μatm) to 16 days post hatch (dph). Morphologies at 1 dph and 16 dph were significantly affected by temperature, but not CO2. At 1dph, larvae at 22 °C were longer (7%) and had larger muscle depth at vent (14%), but had reduced yolk (65%) and oil globule size (16%). Reduced yolk reserves in recently hatched larvae suggests higher metabolic demands in warmer water. At 16 dph, larvae at elevated temperature were longer (12%) and muscle depth at vent was larger (64%). Conversely, survival was primarily affected by CO2 rather than temperature. Survivorship at 1 dph and 16 dph was 24% and 54% higher, respectively, under elevated CO2 compared with ambient conditions. Elevated temperature increased survival (24%) at 1 dph, but not at 16 dph. These results suggest that projected climate change scenarios may have an overall positive effect on early life history growth and survival in this population of C. auratus. This could benefit recruitment success, but needs to be weighed against negative effects of elevated CO2 on metabolic rates and swimming performance observed in other studies on the same population.

Continue reading ‘Elevated temperature and CO2 have positive effects on the growth and survival of larval Australasian snapper’

Investigating the effects of climate co-stressors on surf smelt energy demands

Surf smelt (Hypomesus pretiosus) are ecologically and economically important to the Pacific Northwest. They play a critical role in the food web and support numerous commercially important species and are an economically important baitfish. Surf smelt interact closely with the nearshore environment, utilizing approximately 10% of Puget Sound coastlines for spawning throughout the year. Surf smelt spawn at high tide and adhere fertilized eggs to beach sediment, causing their embryos to be exposed to air and seawater throughout embryonic development. Because of this unique life history, surf smelt may be susceptible to anthropogenic stressors including coastal development and climate change. However, very few studies have attempted to test the tolerance of surf smelt to climate change, including elevated temperature and ocean acidification. The purpose of this study was to examine the interactive effects of climate co-stressors ocean acidification and seawater warming on the energy demands of developing surf smelt. Surf smelt embryos and larvae were collected and placed into experimental basins under three temperature treatments (12°C, 15°C, and 18°C) and two total carbon treatments (ambient and elevated) for a period of 14 days for the embryos, and 4 days for the larvae. Increased temperature significantly decreased yolk size in developing surf smelt embryos and larvae. During this time, embryo yolk sacs in the high temperature treatment were on average 10.2% smaller than embryo yolk sacs in ambient temperature water. Larval yolk and oil globules mirrored this trend with larvae in the high temperature treatment having on average 32.5% smaller yolk sacs and 20.0% smaller oil globules compared to larvae in ambient temperature. While no effect of acidification as a singular stressor was observed, the interaction with temperature significantly increased surf smelt embryo heart rates by 5% above ambient conditions. These results indicate that near-future climate change scenarios are going to impact the energy demands of developing surf smelt, a result that may have a variety of potential impacts including altered hatch times, larval deformities, and increased mortality, all of which will increase interannual variability in adult recruitment. The results of this study highlight the need to increase focus on studying surf smelt in the context of ecological and climate change research.

Continue reading ‘Investigating the effects of climate co-stressors on surf smelt energy demands’


Subscribe to the RSS feed

Powered by FeedBurner

Follow AnneMarin on Twitter

Blog Stats

  • 1,365,201 hits

OA-ICC HIGHLIGHTS

Ocean acidification in the IPCC AR5 WG II

OUP book