Population trajectories for the Antarctic bivalve Laternula elliptica: identifying demographic bottlenecks in differing environmental futures

The world’s oceans are changing, and dramatic shifts have been documented in the Southern Ocean. The consequences of these shifts to coastal benthic organisms are difficult to predict at present, as ocean warming may increase primary production and food resources for benthic consumers, whilst OA may have negative impacts that differentially affect various species and life stages. A model was developed to investigate how different scenarios of change may influence population size of the Antarctic bivalve Laternula elliptica. The model describes potential implications of both pH and temperature change on survivorship and reproductive output of a population of this bivalve species in McMurdo Sound, Ross Sea. Implications of increases and decreases in mortality rate across different life stages of the population (early, mid and late) were assessed. Additionally, effects on energetic resource partitioning and dictating reproductive potential (RP) were also investigated. Significant declines in RP, due to increased basal metabolic demand, were associated with even relatively small changes in temperature and pH, resulting in populations declining to 25 % of the starting equilibrium density within 60 years. As L. elliptica is a pivotal species to the functionality of the Antarctic coastal benthic ecosystem, wide spread repercussions are expected if populations are impacted as the model predicts. Although further model development is required to explore the ecosystem implications of the population decline described in this paper, this work allows a better understanding of the consequences of change as soon as data on the direction and magnitude of the changes affecting Antarctic seas become available.     

Metabolic demand,oxygen supply,and critical temperatures in the Antarctic bivalve Laternula elliptica

Oxygen consumption (Mo(2)), heartbeat rate and form, and circulating hemolymph oxygen content were measured in relation to temperature in the large Antarctic infaunal bivalve Laternula elliptica. After elevations in temperature from 0 degrees to 3 degrees, 6 degrees, and then 9 degrees C, Mo(2) and heartbeat rate rose to new levels, whereas maximum circulating hemolymph oxygen content fell. At 0 degrees C, Mo(2) was 19.6 micromol O(2) h(-1) for a standard animal of 2-g tissue ash-free dry mass, which equates to a 8.95-g tissue dry-mass or 58.4-g tissue wet-mass animal. Elevation of metabolism following temperature change had acute Q(10) values between 4.1 and 5, whereas acclimated figures declined from 3.4 (between 0 degrees and 3 degrees C) to 2.2 (3 degrees -6 degrees C) and 1.9 (6 degrees -9 degrees C). Heartbeat rate showed no acclimation following temperature elevations, with Q(10) values of 3.9, 3.2, and 4.3, respectively. Circulating hemolymph oxygen content declined from 0 degrees to 3 degrees and 6 degrees C but stayed at a constant Po(2) (73-78 mmHg) and constant proportion ( approximately 50%) of the oxygen content of the ambient water. At 9 degrees C, Mo(2) and heartbeat rate both peaked at values 3.3 times those measured at 0 degrees C, which may indicate aerobic scope in this species. After these peaks, both measures declined rapidly over the ensuing 5 d to the lowest measured in the study, and the bivalves began to die. Hemolymph oxygen content fell dramatically at 9 degrees C to values between 2% and 12% of ambient water O(2) content and had a maximum Po(2) of around 20 mmHg. These data indicate an experimental upper lethal temperature of 9 degrees C and a critical temperature, where a long-term switch to anaerobic metabolism probably occurs, of around 6 degrees C for L. elliptica. Concurrent measures of mitochondrial function in the same species had indicated strong thermal sensitivity in proton leakage costs, and our data support the hypothesis that as temperature rises, mitochondrial maintenance costs rapidly outstrip oxygen supply mechanisms in cold stenothermal marine species.     

Cadmium bioaccumulation and detoxification in the gill and digestive gland of the Antarctic bivalve Laternula elliptica

Exposure to a sublethal concentration of cadmium (Cd; 50 microg L(-1)) resulted in significantly increased Cd concentrations in the gill and digestive gland of the Antarctic bivalve Laternula elliptica. Continuous accumulation of Cd in the two organs during the 14-day exposure period was associated with sequestration of Cd to both the soluble cytosolic and insoluble particulate cell fractions. However, the contribution of each cell fraction to Cd sequestration differed between the two organs; in the gill, a larger portion of Cd was associated with the insoluble fraction, while in the digestive gland, both the soluble and insoluble fractions sequestered similar amounts of Cd. Metal-binding components in the insoluble cell fraction were not identified in this study. On the other hand, a metallothionein-like protein (MTLP) was the major Cd-detoxifying component in the soluble cell fraction of the gill and digestive gland. The amount of MTLP increased linearly with exposure time and the amount of Cd accumulated in the tissue, which suggests a potential utility of MTLP as a biomarker for exposure to Cd and possibly other metals.     

Stable isotopes prove shell growth bands in the Antarctic bivalve Laternula elliptica to be formed annually

Growth marks in calcareous shells or skeleton parts of benthic invertebrates are often assumed to be formed annually and hence are used for ageing purposes. For some Antarctic species this assumption has been validated by tagging experiments or direct monitoring of growth throughout the year. Here we use the record of stable isotope ratios (δ¹⁸O and δ¹³C) deposited in shell carbonate of Laternula elliptica from King George Island to demonstrate that shell growth bands are formed annually in this species. Received: 16 July 1995 / Accepted: 27 October 1996     

Hyperoxia alleviates thermal stress in the Antarctic bivalve, Laternula elliptica: evidence for oxygen limited thermal tolerance

Understanding thermal limits and the ability of species to cope with changing temperatures is crucial for a cause and effect understanding of climate effects on organisms and ecosystems. Data available for marine species from various phyla and climates led to the hypothesis that a mismatch between oxygen demand and limited capacity of oxygen supply to tissues is the first mechanism to restrict survival at thermal extremes. Here we show that doubling the oxygen content of the ambient seawater from 160 mmHg partial pressure to 350 mmHg raised the upper temperature limits of the Antarctic marine bivalve Laternula elliptica by about 2.5°C. It reduced the accumulation of the anaerobic end product succinate or of total CO₂ as a sign of respiratory distress. These findings provide further evidence that oxygen supply does limit thermal tolerance in marine animals. As water temperatures rise animals will face a double problem of progressively reduced oxygen solubility in the water and enhanced costs reflected in increased metabolic rates.     

Physiological plasticity, long term resistance or acclimation to temperature, in the Antarctic bivalve, Laternula elliptica

To further investigate the previously reported limited acclimation capacities of Antarctic marine stenotherms, the Antarctic mud clam, Laternula elliptica (King and Broderip, 1830-1831), was incubated at 3.0°C for 89days. The thermal windows of a suite of biochemical and physiological metrics that characterise tissue aerobic status, were then measured in response to acute temperature elevation (2-2.5°C increase per week). To test if acclimation had occurred at the higher temperature, results were compared with published data, from the preceding year, for L. elliptica which had been incubated at ambient temperature (0.0°C) and then subjected to the same acute temperature treatments. Incubation to 3.0°C led to a temperature induced increase of tissue aerobic status (reduced intracellular cCO(2) with increased O(2) consumption, PLA (phospho-L-arginine) and ATP). At the highest acute temperature (7.5°C) the increase in anaerobic pathways (summed acetate/succinate and propionate) was less after 3.0°C than 0.0°C incubation. No other metric shifted its reaction norm in response to acute temperature elevation and so whole animal acclimation had not occurred, even after 3months at 3.0°C. Combined with the constant mortality throughout the 3.0°C incubation period, these data suggest that the recorded physiological changes were either the early stages of acclimation or, more likely, time limited resistance mechanisms.     

Iron and nitrosative metabolism in the Antarctic mollusc Laternula elliptica

The objective of this work was to study Fe distribution, and oxidative and nitrosative metabolism in Laternula elliptica for physiological analysis and interspecific comparisons. Lipid peroxidation, superoxide dismutase and catalase activity and total Fe content were estimated in the digestive glands (DG) of L. elliptica. The labile Fe pool (LIP) represents the amount of cellular Fe responsible for catalyzing radical-dependent reactions. LIP assessed by the calcein assay, represents 3.5% of the total Fe in L. elliptica. Experimental isolation of ferritin (Ft) was performed. Subunit analyses of the protein by SDS-polyacrilamide gel electrophoresis indicated that the protein was composed of 20.6kDa protein subunits, consistent with the horse spleen Ft and the molecular weight markers, however, a higher molecular mass subunit could appear. The identity of the protein was confirmed by Western blot analysis. The nitrate+nitrite content was 73±7pmol/mg fresh mass (FW). The nitric oxide (NO) content in DG homogenates, assessed by electronic paramagnetic resonance (EPR) spin trapping measurements using the NO trap sodium-N-methyl-D-glucamine dithiocarbamate-Fe at room temperature, was 30±2pmol/mg FW. Nitric oxide synthase-like activity (1.50±0.09pmol/mg FW min) was assessed by measuring NO production by EPR in the presence of L-arginine (L-A) and NADPH. This activity was significantly inhibited by L-A analogs such as Nω-nitro-L-arginine methyl ester hydrochloride (-77%) and Nω-nitro-L-arginine (-62%), or by the lack of added L-A (-55%). The data presented here documented the physiological presence of labile Fe, Ft and highly reactive nitrogen species, and are the first evidence that support the hypothesis that NO being generated in L. elliptica might contribute to restrict oxidative damage by a close link with Fe metabolism.     

Molecular cloning and thermal stress-induced expression of a pi-class glutathione S-transferase (GST) in the Antarctic bivalve Laternula elliptica

Glutathione S-transferases (GSTs) are multifunctional phase II detoxification enzymes that catalyze the attachment of electrophilic substrates to glutathione. The pi-class GST cDNA (leGSTp) was cloned from the cold-adapted Antarctic bivalve Laternula elliptica. We used degenerated primers designed based on highly conserved regions of known mollusk GSTs to amplify the corresponding L. elliptica mRNA. Full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full sequence of the GST cDNA was 1189 bp in length, with a 5' untranslated region (UTR) of 74 bp, a 3' UTR of 485 bp, and an open reading frame of 630 bp encoding 209 amino acid residues with an estimated molecular mass of 23.9 kDa and an estimated isoelectric point of 8.3. Quantitative RT-PCR confirmed basal expression of leGSTp, which was up-regulated upon heat treatment (10 degrees C for different time periods) by a factor of 2.3 (at 24 h) and 2.7 (at 48 h) in the digestive gland and gill tissues, respectively. The recombinant leGSTp expressed in Escherichia coli was purified by affinity chromatography and characterized. The purified leGSTp exhibited high activity towards the substrates ethacrynic acid (ECA) and 1-chloro-2,4-dinitrobenzene (CDNB). The recombinant leGSTp had a maximum activity at approximately pH 8.0, and its optimum temperature was 35 degrees C.     

Cloning, expression and characterization of metallothionein from the Antarctic clam Laternula elliptica

The genes for two apparent subtypes of metallothionein (MT) isoform were isolated from the Antarctic clam Laternula elliptica. Determination of the nucleotide sequence showed that the gene consists of 222 bp that code a 73-amino acid protein. The comparison between MT cDNA sequences of L. elliptica and other bivalves showed strong homologies on positions of cysteine residues, which are important for their metal binding abilities. The gene for the MT was inserted into a pET vector and overexpressed as a carboxyl terminal extension of glutathionein-S-transferase (GST) in Escherichia coli. After the GST fusion proteins had been purified by glutathione-Sepharose affinity chromatography column and digested with enterokinase, the MT was purified with gel filtration and analyzed for its biochemical properties. Recombinant MTs were reconstituted with Cd, Cu, and Zn, and kinetic studies of the reactions with electrophilic disulphide, DTNB, were investigated to explore their metal binding ability. It is revealed that the Cd-MT and Zn-MT react with DTNB biphasically, and that Zn-MT reacts with DTNB more rapidly, and with a significantly greater pseudo-first-order rate constant. Cu-MT reacts monophasically and releases metal slowly from MT.     

Production of reactive oxygen species by isolated mitochondria of the Antarctic bivalve Laternula elliptica (King and Broderip) under heat stress

Formation of reactive oxygen species (ROS) in mitochondrial isolates from gill tissues of the Antarctic polar bivalve Laternula elliptica was measured fluorimetrically under in vitro conditions. When compared to the rates measured at habitat temperature (1 degrees C), significantly elevated ROS formation was found under temperature stress of 7 degrees C and higher. ROS formation correlated significantly with oxygen consumption in individual mitochondrial preparations over the entire range of experimental temperatures (1-12 degrees C). ROS generation per mg of mitochondrial protein was significantly higher in state 3 at maximal respiration and coupling to energy conservation, than in state 4+, where ATPase-activity is inhibited by oligomycin and only proton leakage is driving the residual oxygen consumption. The percent conversion of oxygen to the membrane permeant hydrogen peroxide amounted to 3.7% (state 3) and 6.5% (state 4+) at habitat temperature (1 degrees C), and to 7% (state 3) and 7.6% (state 4+) under experimental warming to 7 degrees C. This is high compared to 1-3% oxygen to ROS conversion in mammalian mitochondrial isolates and speaks for a comparatively low control of toxic oxygen formation in mitochondria of the polar bivalve. However, low metabolic rates at cold Antarctic temperatures keep absolute rates of mitochondrial ROS production low and control oxidative stress at habitat temperatures. Mitochondrial coupling started to fall beyond 3 degrees C, closely to pejus temperature (4 degrees C) of the bivalve. Accordingly, the proportion of state 4 respiration increased from below 30% at 1 degrees C to over 50% of total oxygen consumption at 7 degrees C, entailing reduced ADP/O ratios under experimental warming. Progressive mitochondrial uncoupling and formation of hazardous ROS contribute to bias mitochondrial functioning under temperature stress in vitro. Deduced from a pejus temperature, heat stress commences already at 5 degrees C, and is linked to progressive loss of phosphorylation efficiency, increased mitochondrial oxygen demand and elevated oxidative stress above pejus temperatures.     

Trace metals in the Antarctic soft-shelled clam Laternula elliptica: implications for metal pollution from Antarctic research stations

The concentrations of Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn were measured in several soft-tissue types of the Antarctic soft-shelled clam, Laternula elliptica, which had been collected from Rothera Research Station, Adelaide Island, Antarctic Peninsula. Concentrations of Mn, Ni, Cr and Pb were significantly higher in the kidney than in any other soft tissue and highest concentrations of Cu were observed in the digestive gland. In general, the total tissue concentrations of heavy metals in L. elliptica were considered to be at baseline levels, except for Cu in organisms near the end of the runway. Copper concentrations were an order of magnitude greater (357 µg/g dry weight) than baseline levels, suggesting anthropogenic contamination or an unidentified natural source. However, there was no indication of anthropogenic metal contamination occurring close to the sewage outfall at Rothera, which is a significant metal source. The results indicate that L. elliptica is a useful long-term biomonitor of heavy metal contamination in Antarctic coastal waters.     

Enhanced sexual reproduction in bryophytes at high latitudes in the maritime Antarctic

Abstract

Contrary to the generally accepted hypothesis that bryophyte fertility decreases with increasing latitude and therefore climatic severity, a detailed study of bryophyte reproductive strategies at sites in the southern maritime Antarctic (68–72°S) has revealed that an unexpectedly high proportion of species is capable of producing sporophytes. Of the regional bryoflora, 43% (19 species; 17 mosses, 2 liverworts) in Marguerite Bay and 47% (17 species; 16 mosses, 1 liverwort) in Alexander Island are known to produce sporophytes, although the number fruiting at comparable latitudes on the colder and more arid Antarctic continent is less (33%). These numbers and proportions are much greater than were previously known at such relatively high southern polar latitudes. Sporophytes of several species are recorded for the first time within the Antarctic biome, while those of two liverworts (Lophozia excisa and Cephaloziella varians) are reported for the first time south of 62°S. High incidence of sporophyte production is attributed to locally favourable microclimatic conditions producing small-scale 'oases'. The large majority of fertile mosses are monoecious short acrocarps growing on rather calcareous soils. Spore production and size data for a number of species are compared with previously published studies of the same or closely related species from the northern maritime Antarctic and sub-Antarctic, but no consistent trends are revealed between species over the latitudinal gradient.     

Prefabrication of shell ornamentation in the bivalve Laternula

The minute, cone-like spikes which cover the valve surfaces of Laternula arc formed by the mantle in advance of the growing shell margin and are subsequently incorporated into the calcarcous shell. This 'prefabrication' process allows the formation of a prismatic spike ultrastructure distinct from other layers of the nacro-prismatic shell and provides ornamentation to the otherwise uncalcified ventral surface. A consideration of a number of morphologic variables, including the mechanism of shell construction, suggests that spikes function to increase the skin friction of the valve surfaces and thereby promote a stable life position. Variations on Laternula's shell construction mechanism arc found throughout the Pandoracea and possibly larger taxonomic groupings.     

The structure of a shellfish specific GST class glutathione S‐transferase from antarctic bivalve Laternula elliptica reveals novel active site architecture

Glutathione‐S‐transferases have been identified in all the living species examined so far, yet little is known about their function in marine organisms. In a previous report, the recently identified GST from Antarctic bivalve Laternula elliptica (LeGST) was classified into the rho class GST, but there are several unique features of LeGST that may justify reclassification, which could represent specific shellfish GSTs. Here, we determined the crystal structure of LeGST, which is a shellfish specific class of GST. The structural analysis showed that the relatively open and wide hydrophobic H‐site of the LeGST allows this GST to accommodate various substrates. These results suggest that the H‐site of LeGST may be the result of adaptation to their environments as sedentary organisms. Proteins 2013. © 2012 Wiley Periodicals, Inc.