Limacina Helicina Antarctica Research Articles

A Promising Approach to Quantifying Pteropod Eggs Using Image Analysis and Machine Learning

A newly developed protocol to semi-automate egg counting in Southern Ocean shelled (thecosome) pteropods using image analysis software and machine learning algorithms was developed and tested for accuracy. Preserved thecosome pteropod (Limacina helicina antarctica) egg masses collected from two austral summer research voyages in East Antarctica were digitally photographed to develop a streamlined approach to enumerate eggs within egg masses using Fiji/ImageJ and the associated machine learning plugin known as Trainable Weka Segmentation. Results from this semi-automated approach were then used to compare with manual egg counts from eggs dissected from egg masses under stereomicroscope. A statistically significant correlation was observed between manual and semi-automated approaches (R2 = 0.92, p < 0.05). There was no significant difference between manual and automated protocols when egg counts were divided by the egg mass areas (mm2) (t(29.6) = 1.98, p = 0.06). However, the average time to conduct semi-automated counts (M = 7.4, SD = 1.2) was significantly less than that for the manual enumeration technique (M = 35.9, SD = 5.7; t(30) = 2.042, p < 0.05). This new approach is promising and, unlike manual enumeration, could allow specimens to remain intact for use in live culturing experiments. Despite some limitations that are discussed, this user-friendly and simplistic protocol can provide the basis for further future development, including the addition of macro scripts to improve reproducibility and through the association with other imaging platforms to enhance interoperability. Furthermore, egg counting using this technique may lead to a relatively unexplored monitoring tool to better understand the responses of a species highly sensitive to multiple stressors connected to climate change.

The use of swimmers from sediment traps to measure summer community structure of Southern Ocean pteropods

In the Southern Ocean, pteropods play an important role in biogeochemical cycling, and sediment traps are a valuable tool for investigating this role through the collection of passively sinking matter from productive surface waters to deep sea layers. Observations of ‘swimmers’ (e.g. organisms that actively swim into traps) can also prove valuable for studying zooplankton community structure. In this study, we used two separate sediment trap studies during the 2016–2017 summer to study pteropod population structure over time scales of 24 h and 28 days. In both studies, highest densities were measured for veliger-stage Limacina helicina antarctica (0.09–0.3 mm) relative to all species and age classes. Increases in shell diameters of veligers in all traps over time enabled the calculation of an intraseasonal potential growth rate of 0.0068 mm d−1. Swimmer flux rates ranged from 121 to 2674 ind. m−2 d−1 at 53 m depth, and the 24-h vertical flux study measured 960 ind. m−2 d−1 at 57 m depth and 6692 m−2 d−1 at 90 m depth. Among a suite of environmental and biological covariates tested, fluorescence and sinking particulate organic and inorganic carbon (POC and PIC) possessed the most predictive power to explain abundances of near-surface pteropod age class and species composition. Gymnosome abundances were largely influenced by increasing adult L. helicina antarctica counts. Changes to pteropod population and community dynamics in response to climate change will have cascading effects throughout Antarctic epipelagic food webs, and these results provide a regional snapshot of patterns in structure and sedimentation from an under-surveyed region of the Southern Ocean.

Particulate organic matter release below melting sea ice (Terra Nova Bay, Ross Sea, Antarctica): Possible relationships with zooplankton

Timing and rates of release of particulate organic matter (POM) beneath the Antarctic sea ice during the melting season are relatively unknown. To shed light on this topic, we investigated: i) quantity and biochemical composition of POM released below annual sea ice in Terra Nova Bay (TNB, Ross Sea, Antarctica) through sediment traps deployed at 10- and 30-m depth; ii) the abundance and taxonomic composition of phytoplankton; iii) the abundance and biochemical composition of two zooplankton species, namely the pteropods Clione limacina antarctica and Limacina helicina antarctica. Overall, our results show that in late spring-early summer in TNB the melting of annual sea ice determines a rapid release of particles into the underlying water column, and that those particles are transported downward at the rate of hours to days. POM fluxes were dominated by lipids, which probably resulted from zooplankton grazing activity on sea-ice algae and faecal pellets that were released into the water column. This, after the initial injection of particles, which were likely associated with sinking sympagic microalgal biomass derived from the sea ice melting, enhanced the nutritional value of POM. POM released beneath the sea ice, in turn, had a possible effect on the biochemical composition of the two pteropod species. Our results confirm that melting of sea ice in spring-summer each year in Antarctic coastal waters represents a fundamental step in the transfer of energy towards the higher trophic levels. Evidence of a decline in Antarctic sea ice over the last five years, plausibly as a consequence of global warming, underlines a conspicuous decline in habitat availability for sea-ice algae and, as a consequence, a drop in the availability of food for pteropods and the higher trophic levels of the coastal Antarctic Ocean.

Feeding ecology and microbiome of the pteropod Limacina helicina antarctica

The pteropod (pelagic snail) Limacina helicina antarctica is a dominant grazer along the Western Antarctic Peninsula (WAP) and plays an important role in regional food web dynamics and biogeochemical cycling. For the first time, we examined the gut microbiome and feeding ecology of L. h. antarctica based on 16S and 18S rRNA gene sequences of gut contents in the WAP during austral summer. Eukaryotic gut contents of L. h. antarctica indicate that this species predominantly feeds on diatoms and dinoflagellates, supplementing its diet with ciliates and foraminifera. Mollicutes bacteria were a consistent component of the gut microbiome. Determining the gut microbiome and feeding ecology of L. h. antarctica aids in identifying the underlying mechanisms controlling pteropod abundance and distribution in a region of rapid environmental change.

Long-term observations of pteropod phenology along the Western Antarctic Peninsula

Shifts in phenology – annually occurring life history events – have been observed among many marine organisms due to global warming. We examined if phenological changes in the pteropod (pelagic snail) Limacina helicina antarctica have occurred along the Western Antarctic Peninsula, one of the most intensely warming regions on Earth, which would have important implications for regional food web dynamics. Pteropod shell diameters were analyzed from samples collected in the Palmer Antarctica Long-Term Ecological Research (PAL LTER) program year-round sediment trap from 2004 to 2018. There was considerable interannual variability in the time of appearance of a new pteropod cohort, which ranged from day of year 22–255, but no long-term, directional change. Mean L. h. antarctica growth rate for the time series was 0.009 mm day−1 and there was no significant long-term change in growth rate. This study represents the first in the Southern Ocean to illustrate that pteropods actively grow throughout the winter season. Sea ice was the dominant driver of pteropod phenology, with earlier sea ice retreat the year prior, lower winter sea surface temperature (SST) the year prior, and higher primary productivity in the same year leading to earlier pteropod time of appearance. Similarly, more open water with higher autumn SST, both the year prior, and elevated chlorophyll a the same year, promoted faster pteropod growth. These results indicate that while pteropods are responsive to considerable environmental variability, their phenology has remained relatively stable. The identified responses of pteropod phenology to environmental shifts are key for determining future effects of climate change on biogeochemical cycling and plankton trophic interactions in the region.

Effects of temperature and food concentration on pteropod metabolism along the Western Antarctic Peninsula

Pteropods (pelagic snails) are abundant zooplankton in the Southern Ocean where they are important grazers of phytoplankton, prey for higher trophic levels, and sensitive to environmental change. The Western Antarctic Peninsula (WAP) is a highly dynamic and productive region that has undergone rapid warming, but little is known about how environmental changes there will affect pteropod physiology. In this study, the effects of warming seawater temperatures and shifting food availability on Limacina helicina antarctica metabolism (respiration and excretion) were determined by conducting shipboard experiments that exposed pteropods to a range of temperatures and phytoplankton (food) concentrations. Highest respiration (up to 69 μmol O2 gDW−1 h−1) and usually highest excretion rates occurred under higher temperature with more limited metabolic response to food concentration, indicating these factors do not always have an additive effect on pteropod metabolism. The proportion of dissolved organic matter (DOM) to total organic and inorganic dissolved constituents was high and was also significantly affected by shifts in temperature and food. Dissolved organic carbon, nitrogen, and phosphorus (DOC, DON, and DOP) were on average 27, 51, and 11.5% of the total C, N, and P metabolized, respectively. The proportion of total N excreted as DON and the proportion of total P excreted as DOP were significantly affected by a combination of shifting temperature and food concentrations. There were no effects of temperature or food on DOC excretion (mean 8.79 μmol C gDW−1 h−1; range 0.44 to 44) as a proportion of total C metabolized. Metabolic O2:N ratio ranged from 2 to 9 and decreased significantly with increasing temperature and food, indicating a shift toward increased protein catabolism. Metabolic ratios of C, N, and P were all below the canonical Redfield ratio, which has implications for phytoplankton nutrient uptake and bacterial production. Respiration rates at ambient conditions of other WAP pteropods, and excretion rates for Clio pyramidata, were also measured, with respiration rates ranging from 24.39 (Spongiobranchaea australis) to 28.86 (L. h. antarctica) μmol O2 gDW−1 h−1. Finally, a CO2 perturbation experiment measuring L. h. antarctica metabolism under pre-industrial and elevated dissolved pCO2 conditions showed no significant change in mean L. h. antarctica respiration or excretion rates with higher pCO2. These insights into the metabolic response of pteropods to ocean variability increase our understanding of the role of zooplankton in biogeochemical cycles and help predict future responses to climate change.

Zooplankton diel vertical migration during Antarctic summer

Zooplankton diel vertical migration (DVM) during summer in the polar oceans is presumed to be dampened due to near continuous daylight. We analyzed zooplankton diel vertical distribution patterns in a wide range of taxa along the Western Antarctic Peninsula (WAP) to assess if DVM occurs, and if so, what environmental controls modulate DVM in the austral summer. Zooplankton were collected during January and February in paired day-night, depth-stratified tows through the mesopelagic zone along the WAP from 2009-2017, as well as in day and night epipelagic net tows from 1993-2017. The copepod Metridia gerlachei, salp Salpa thompsoni, pteropod Limacina helicina antarctica, and ostracods consistently conducted DVM between the mesopelagic and epipelagic zones. Migration distance for M. gerlachei and ostracods decreased as photoperiod increased from 17 to 22 h daylight. The copepods Calanoides acutus and Rhincalanus gigas, as well as euphausiids Thysanoessa macrura and Euphausia crystallorophias, conducted shallow (mostly within the epipelagic zone) DVMs into the upper 50 m at night. Rhincalanus gigas, T. macrura, and L. h. antarctica DVM behavior was modulated by chlorophyll a concentration, mixed layer depth, and depth of the subsurface chlorophyll a maximum, respectively. Carnivorous and detritivorous taxa – including the calanoid copepod Paraeuchaeta antarctica, ostracods, chaetognaths, and Tomopteris spp. polychaetes – as well as seasonally migrating copepods, were most abundant in the mesopelagic zone regardless of the diel cycle. Paraeuchaeta antarctica underwent reverse DVM within the top 100 m. The impacts of Antarctic zooplankton summer DVM and the resident mesopelagic assemblage on carbon export should be better quantified.

Changes in Genome-Wide Methylation and Gene Expression in Response to Future pCO2 Extremes in the Antarctic Pteropod Limacina helicina antarctica

Epigenetic processes such as variation in DNA methylation may promote phenotypic plasticity and the rapid acclimatization of species to environmental change. The extent to which an organism may mount an epigenetic response to current and future climate extremes may influence its capacity to acclimatize or adapt to global change on ecological rather than evolutionary time scales. The thecosome pteropod Limacina helicina antarctica is an abundant macrozooplankton endemic to the Southern Ocean and is considered a bellwether of ocean acidification as it is highly sensitive to variation in carbonate chemistry. In this study, we quantified variation in DNA methylation and gene expression over time across different ocean acidification regimes. We exposed L. helicina antarctica to pCO2 levels mimicking present-day norms in the coastal Southern Ocean of 255 atm pCO2, present-day extremes of 530 atm pCO2, and projected extremes of 918 atm pCO2 for up to 7 days before measuring global DNA methylation and sequencing transcriptomes in animals from each treatment across time. L. helicina antarctica significantly reduced DNA methylation by 29–56% after 1 day of exposure to 918 atm pCO2 before DNA methylation returned to control levels after 6 days. In addition, L. helicina antarctica exposed to 918 atm pCO2 exhibited drastically more differential expression compared to cultures replicating present-day pCO2 extremes. Differentially expressed transcripts were predominantly downregulated. Furthermore, downregulated genes were enriched with signatures of gene body methylation. These findings support the potential role of DNA methylation in regulating transcriptomic responses by L. helicina antarctica to future ocean acidification and in situ variation in pCO2 experienced seasonally or during vertical migration. More broadly, L. helicina antarctica was capable of mounting a substantial epigenetic response to ocean acidification despite little evidence of metabolic recovery or transcriptional homeostasis in this species at future pCO2 levels.

Combined stress of ocean acidification and warming influence survival and drives differential gene expression patterns in the Antarctic pteropod, Limacina helicina antarctica.

The ecologically important thecosome pteropods in the Limacina spp. complex have recently been the focus of studies examining the impacts global change factors – e.g., ocean acidification (OA) and ocean warming (OW) – on their performance and physiology. This focus is driven by conservation concerns where the health of pteropod populations is threatened by the high susceptibility of their shells to dissolution in low aragonite saturation states associated with OA and how coupling of these stressors may push pteropods past the limits of physiological plasticity. In this manipulation experiment, we describe changes in the transcriptome of the Antarctic pteropod, Limacina helicina antarctica, to these combined stressors. The conditions used in the laboratory treatments met or exceeded those projected for the Southern Ocean by the year 2100. We made two general observations regarding the outcome of the data: (1) Temperature was more influential than pH in terms of changing patterns of gene expression, and (2) these Antarctic pteropods appeared to have a significant degree of transcriptomic plasticity to respond to acute abiotic stress in the laboratory. In general, differential gene expression was observed amongst the treatments; here, for example, transcripts associated with maintaining protein structure and cell proliferation were up-regulated. To disentangle the effects of OA and OW, we used a weighted gene co-expression network analysis to explore patterns of change in the transcriptome. This approach identified gene networks associated with OW that were enriched for transcripts proposed to be involved in increasing membrane fluidity at warmer temperatures. Together these data provide evidence that L.h.antarctica has a limited capacity to acclimate to the combined conditions of OA and OW used in this study. This reduced scope of acclimation argues for continued study of how adaptation to polar aquatic environments may limit the plasticity of present-day populations in responding to future environmental change.

Characteristics of swimming shelled Antarctic pteropods (Limacina helicina antarctica) at intermediate Reynolds number regime

Shelled Antarctic pteropods (aquatic snails nicknamed ``sea butterflies'') swim with a pair of parapodia (or ``wings'') in a high efficiency propulsion regime when the Reynolds number based on the flapping exceeds 35.

Seasonal transcriptomes of the Antarctic pteropod, Limacina helicina antarctica

High latitude seas will be among the first marine systems to be impacted by ocean acidification (OA). Previous research studying the effects of OA on the pteropod, Limacina helicina antarctica, has led this species to be identified as a sentinel organism for OA in polar oceans. Here, we present gene expression data on L. h. antarctica, collected in situ during the seasonal transition from early spring to early summer. Our findings suggest that after over-wintering under seasonal sea ice, pteropods progress toward full maturity in the early summer when food becomes increasingly available. This progression is highlighted by a dramatic shift in gene expression that supports the development of cytoskeletal structures, membrane ion transportation, and metabolically important enzymes associated with glycolysis. In addition, we observed signs of defense of genomic integrity and maturation as evidenced by an up-regulation of genes involved in DNA replication, DNA repair, and gametogenesis. These data contribute to a broader understanding of the life-cycle dynamics for L. h. antarctica and provide key insights into the transcriptomic signals of pteropod maturation and growth during this key seasonal transition.

Environmental controls on pteropod biogeography along the Western Antarctic Peninsula

AbstractPteropods are abundant zooplankton in the Western Antarctic Peninsula (WAP) and important grazers of phytoplankton and prey for higher trophic levels. We analyzed long‐term (1993–2017) trends in summer (January–February) abundance of WAP pteropods in relation to environmental controls (sea ice, sea surface temperature, climate indices, phytoplankton biomass and productivity, and carbonate chemistry) and interspecies dynamics using general linear models. There was no overall directional trend in abundance of thecosomes, Limacina helicina antarctica and Clio pyramidata, throughout the entire WAP, although L. antarctica abundance increased in the slope region and C. pyramidata abundance increased in the South. High L. antarctica abundance was strongly tied to a negative Multivariate El Niño Southern Oscillation Index the previous year. C. pyramidata abundance was best explained by early sea ice retreat 1‐yr prior. Abundance of the gymnosome species, Clione antarctica and Spongiobranchaea australis, increased over the time series, particularly in the slope region. Gymnosome abundance was positively influenced by abundance of their prey, L. antarctica, during the same season, and late sea ice advance 2‐yr prior. These trends indicate a shorter ice season promotes longer periods of open water in spring/summer favoring all pteropod species. Weak relationships were found between pteropod abundance and carbonate chemistry, and no long‐term trend in carbonate parameters was detected. These factors indicate ocean acidification is not presently influencing WAP pteropod abundance. Pteropods are responsive to the considerable environmental variability on both temporal and spatial scales—key for predicting future effects of climate change on regional carbon cycling and plankton trophic interactions.

Southern Ocean pteropods at risk from ocean warming and acidification

Early life stages of marine calcifiers are particularly vulnerable to climate change. In the Southern Ocean aragonite undersaturation events and areas of rapid warming already occur and are predicted to increase in extent. Here, we present the first study to successfully hatch the polar pteropod Limacina helicina antarctica and observe the potential impact of exposure to increased temperature and aragonite undersaturation resulting from ocean acidification (OA) on the early life stage survival and shell morphology. High larval mortality (up to 39%) was observed in individuals exposed to perturbed conditions. Warming and OA induced extensive shell malformation and dissolution, respectively, increasing shell fragility. Furthermore, shell growth decreased, with variation between treatments and exposure time. Our results demonstrate that short-term exposure through passing through hotspots of OA and warming poses a serious threat to pteropod recruitment and long-term population viability.

Additive effects of pCO2 and temperature on respiration rates of the Antarctic pteropod Limacina helicina antarctica.

The Antarctic pteropod, Limacina helicina antarctica, is a dominant member of the zooplankton in the Ross Sea and supports the vast diversity of marine megafauna that designates this region as an internationally protected area. Here, we observed the response of respiration rate to abiotic stressors associated with global change-environmentally relevant temperature treatments (-0.8°C, 4°C) and pH treatments reflecting current-day and future modeled extremes (8.2, 7.95 and 7.7 pH at -0.8°C; 8.11, 7.95 and 7.7 pH at 4°C). Sampling repeatedly over a 14-day period in laboratory experiments and using microplate respirometry techniques, we found that the metabolic rate of juvenile pteropods increased in response to low-pH exposure (pH 7.7) at -0.8°C, a near-ambient temperature. Similarly, metabolic rate increased when pteropods were exposed simultaneously to multiple stressors: lowered pH conditions (pH 7.7) and a high temperature (4°C). Overall, the results showed that pCO2 and temperature interact additively to affect metabolic rates in pteropods. Furthermore, we found that L. h. antarctica can tolerate acute exposure to temperatures far beyond its maximal habitat temperature. Overall, L. h. antarctica appears to be susceptible to pH and temperature stress, two abiotic stressors which are expected to be especially deleterious for ectothermic marine metazoans in polar seas.

Transcriptomic response of the Antarctic pteropod Limacina helicina antarctica to ocean acidification

BackgroundOcean acidification (OA), a change in ocean chemistry due to the absorption of atmospheric CO2 into surface oceans, challenges biogenic calcification in many marine organisms. Ocean acidification is expected to rapidly progress in polar seas, with regions of the Southern Ocean expected to experience severe OA within decades. Biologically, the consequences of OA challenge calcification processes and impose an energetic cost.ResultsIn order to better characterize the response of a polar calcifier to conditions of OA, we assessed differential gene expression in the Antarctic pteropod, Limacina helicina antarctica. Experimental levels of pCO2 were chosen to create both contemporary pH conditions, and to mimic future pH expected in OA scenarios. Significant changes in the transcriptome were observed when juvenile L. h. antarctica were acclimated for 21 days to low-pH (7.71), mid-pH (7.9) or high-pH (8.13) conditions. Differential gene expression analysis of individuals maintained in the low-pH treatment identified down-regulation of genes involved in cytoskeletal structure, lipid transport, and metabolism. High pH exposure led to increased expression and enrichment for genes involved in shell formation, calcium ion binding, and DNA binding. Significant differential gene expression was observed in four major cellular and physiological processes: shell formation, the cellular stress response, metabolism, and neural function. Across these functional groups, exposure to conditions that mimic ocean acidification led to rapid suppression of gene expression.ConclusionsResults of this study demonstrated that the transcriptome of the juvenile pteropod, L. h. antarctica, was dynamic and changed in response to different levels of pCO2. In a global change context, exposure of L. h. antarctica to the low pH, high pCO2 OA conditions resulted in a suppression of transcripts for genes involved in key physiological processes: calcification, metabolism, and the cellular stress response. The transcriptomic response at both acute and longer-term acclimation time frames indicated that contemporary L. h. antarctica may not have the physiological plasticity necessary for adaptation to OA conditions expected in future decades. Lastly, the differential gene expression results further support the role of shelled pteropods such as L. h. antarctica as sentinel organisms for the impacts of ocean acidification.

Portable tomographic PIV measurements of swimming shelled Antarctic pteropods

A portable tomographic particle image velocimetry (tomographic PIV) system is described. The system was successfully deployed in Antarctica to study shelled Antarctic pteropods (Limacina helicina antarctica)—a delicate organism with an unusual propulsion mechanism. The experimental setup consists of a free-standing frame assembled with optical rails, thus avoiding the need for heavy and bulky equipment (e.g. an optical table). The cameras, lasers, optics, and tanks are all rigidly supported within the frame assembly. The results indicate that the pteropods flap their parapodia (or “wings”) downward during both power and recovery strokes, which is facilitated by the pitching of their shell. Shell pitching significantly alters the flapping trajectory, allowing the pteropod to move vertically and/or horizontally. The pronation and supination of the parapodia, together with the figure-eight motion during flapping, suggest similarities with insect flight. The volumetric velocity field surrounding the freely swimming pteropod reveals the generation of an attached vortex ring connecting the leading-edge vortex to the trailing-edge vortex during power stroke and a presence of a leading-edge vortex during recovery stroke. These vortex structures play a major role in accelerating the organism vertically and indicate that forces generated on the parapodia during flapping constitute both lift and drag. After completing each stroke, two vortex rings are shed into the wake of the pteropod. The complex combination of body kinematics (parapodia flapping, shell pitch, sawtooth trajectory), flow structures, and resulting force balance may be significantly altered by thinning of the pteropod shell, thus making pteropods an indicator of the detrimental effects of ocean acidification.

Pteropod eggs released at high pCO2 lack resilience to ocean acidification.

The effects of ocean acidification (OA) on the early recruitment of pteropods in the Scotia Sea, was investigated considering the process of spawning, quality of the spawned eggs and their capacity to develop. Maternal OA stress was induced on female pteropods (Limacina helicina antarctica) through exposure to present day pCO2 conditions and two potential future OA states (750 μatm and 1200 μatm). The eggs spawned from these females, both before and during their exposure to OA, were incubated themselves in this same range of conditions (embryonic OA stress). Maternal OA stress resulted in eggs with lower carbon content, while embryonic OA stress retarded development. The combination of maternal and embryonic OA stress reduced the percentage of eggs successfully reaching organogenesis by 80%. We propose that OA stress not only affects the somatic tissue of pteropods but also the functioning of their gonads. Corresponding in-situ sampling found that post-larval L. helicina antarctica concentrated around 600 m depth, which is deeper than previously assumed. A deeper distribution makes their exposure to waters undersaturated for aragonite more likely in the near future given that these waters are predicted to shoal from depth over the coming decades.

A transcriptome resource for the Antarctic pteropod Limacina helicina antarctica

The pteropod Limacina helicina antarctica is a dominant member of the zooplankton assemblage in the Antarctic marine ecosystem, and is part of a relatively simple food web in nearshore marine Antarctic waters. As a shelled pteropod, Limacina has been suggested as a candidate sentinel organism for the impacts of ocean acidification, due to the potential for shell dissolution in undersaturated waters. In this study, our goal was to develop a transcriptomic resource for Limacina that would support mechanistic studies to explore the physiological response of Limacina to abiotic stressors such as ocean acidification and ocean warming. To this end, RNA sequencing libraries were prepared from Limacina that had been exposed to a range of pH levels and an elevated temperature to maximize the diversity of expressed genes. RNA sequencing (RNA-seq) was conducted on an Illumina NextSeq500 which produced 339,000,000 150bp paired-end reads. The de novo transcriptome was produced using Trinity and annotation of the assembled transcriptome resulted in the identification of 81,229 transcripts in 137 KEGG pathways. This RNA-seq effort resulted in a transcriptome for the Antarctic pteropod, Limacina helicina antarctica, that is a major resource for an international marine science research community studying these pelagic molluscs in a global change context.

Microstructural shell strength of the Subantarctic pteropod Limacina helicina antarctica

Anthropogenic inputs of CO2 are changing ocean chemistry and will likely affect calcifying marine organisms, particularly aragonite producers such as pteropods. This work seeks to set a benchmark analysis of pteropod shell properties and variability using nanoindentation and electron microscopy to measure the structural and mechanical properties of Subantarctic pteropod shells (Limacina helicina antarctica) collected in 1998 and 2007. The 1998 shells were collected by a sediment trap deployed at 2000 m, 47°S, 142°E, and the 2007 shells were collected using nets from mixed-layer waters in the region (44°–54°S, 140°–155°E). Transmission electron microscopy revealed that the shells are composed of a polycrystalline structure, and no obvious porosity was visible. The hardness and modulus of the shells were measured using shell cross-section nanoindentation, across various regions of the shell from the inner to outer whorl. No change in mechanical properties was found with respect to the region of the shell cross-section probed. There was no statistically significant difference in the mean modulus or hardness of the shells between the 1998 and 2007 data sets. No major changes in the mechanical properties of these pteropod shells were detected between the 1998 and 2007 data sets, and we discuss the possible biases in the sampling techniques in complicating our analysis. However, quantifying the mechanical properties and microstructure of calcified may still provide insights into the responses of calcification to environmental changes, such as ocean acidification.

Dissolution dominating calcification process in polar pteropods close to the point of aragonite undersaturation.

Thecosome pteropods are abundant upper-ocean zooplankton that build aragonite shells. Ocean acidification results in the lowering of aragonite saturation levels in the surface layers, and several incubation studies have shown that rates of calcification in these organisms decrease as a result. This study provides a weight-specific net calcification rate function for thecosome pteropods that includes both rates of dissolution and calcification over a range of plausible future aragonite saturation states (Ωar). We measured gross dissolution in the pteropod Limacina helicina antarctica in the Scotia Sea (Southern Ocean) by incubating living specimens across a range of aragonite saturation states for a maximum of 14 days. Specimens started dissolving almost immediately upon exposure to undersaturated conditions (Ωar∼0.8), losing 1.4% of shell mass per day. The observed rate of gross dissolution was different from that predicted by rate law kinetics of aragonite dissolution, in being higher at Ωar levels slightly above 1 and lower at Ωar levels of between 1 and 0.8. This indicates that shell mass is affected by even transitional levels of saturation, but there is, nevertheless, some partial means of protection for shells when in undersaturated conditions. A function for gross dissolution against Ωar derived from the present observations was compared to a function for gross calcification derived by a different study, and showed that dissolution became the dominating process even at Ωar levels close to 1, with net shell growth ceasing at an Ωar of 1.03. Gross dissolution increasingly dominated net change in shell mass as saturation levels decreased below 1. As well as influencing their viability, such dissolution of pteropod shells in the surface layers will result in slower sinking velocities and decreased carbon and carbonate fluxes to the deep ocean.