Antarctic Limpet Nacella Concinna Research Articles

Evaluating the effects of ocean warming and freshening on the physiological energetics and transcriptomic response of the Antarctic limpet Nacella concinna

In the Southern Ocean, warming and freshening are expected to be prominent signals of climate change and the reduced ability of Antarctic marine organisms to cope with changing environmental conditions could challenge their future survival. The Antarctic limpet Nacella concinna is a macroinvertebrate of rocky ecosystems, which occurs in high densities in the shallow subtidal zone. Subtidal individuals were exposed to a combination of temperatures (1, 4, 8, 11, 14 °C) and salinities (20 and 30 psu) for a 60-day period. A drastic increment in mortality was observed with seawater warming, showing that N. concinna is highly stenothermal, with limited ability to survive at temperatures warmer than 4 °C, although there was some degree of acclimation at 4 °C and ambient salinity (30 psu). This study confirmed the stenohaline characteristic of this species, with mortality reaching 50% and lower scope for growth at low salinity (20 psu) even at the control temperature (1 °C). At the sub-cellular level, limpets' low tolerance to out-of range salinity is illustrated by the activation of cell remodelling processes whereas the down-regulation of chaperones proteins and plasma membrane ATPase suggest that under the combination of warming and freshening N. concinna experiences a severe level of stress and devote much of its energy to somatic maintenance and survival. The drastic effect observed can be explained by its subtidal origin, an environment with more stable conditions. The surviving individuals at 1 °C and lowered salinity (20 psu) were either more tolerant or showing signs of acclimation after 60 days, but the combination of warming and freshening have a greater combined stress. Projections of climate change for end of the century for this part of the Antarctic can, therefore, result in a significant diminution of the subtidal population of N. concinna, affecting ecological interactions and diversity of the food web.

Can DEB models infer metabolic differences between intertidal and subtidal morphotypes of the Antarctic limpet Nacella concinna (Strebel, 1908)?

Studying the influence of changing environmental conditions on Antarctic marine benthic invertebrates is strongly constrained by limited access to the region, which poses difficulties to performing long-term experimental studies. Ecological modelling has been increasingly used as a potential alternative to assess the impact of such changes on species distribution or physiological performance.Among ecological models, the Dynamic Energy Budget (DEB) approach represents each individual through four energetic compartments (i.e. reserve, structure, maturation and reproduction) from which energy is allocated in contrasting proportions according to different life stages and to two forcing environmental factors (food resources and temperature).In this study, the example of an abundant coastal limpet, Nacella concinna (Strebel 1908), was studied. The species is known to have intertidal and subtidal morphotypes, genetically similar but physiologically and morphologically contrasting.The objectives of this paper are (1) to evaluate the potential of the DEB approach, and assess whether a DEB model can be separately built for the intertidal and subtidal morphotypes, based on a field experiment and data from literature and (2) to analyse whether models are contrasting enough to reflect the known physiological and morphological differences between the morphotypes.We found only minor differences in temperature-corrected parameter values between both populations, meaning that the observed differences can be only explained by differences in environmental conditions (i.e. DEB considered variables, food resources and temperature, but also other variables not considered by DEB). Despite the known morphological difference between the populations, the difference in shape coefficients was small.This study shows that even with the amount of data so far available in the literature, DEB models can already be applied to some Southern Ocean case studies, but, more data are required to accurately model the physiological and morphological differences between individuals.

Phenotypic selection by kelp gulls against pear-shaped shells of the Antarctic limpet Nacella concinna

Abstract The Antarctic limpet (Nacella concinna) presents two ecotypes related to different water depths: littoral individuals have a robust and taller shell while sublittoral individuals are thinner and flatter. Among the environmental factors possibly causing this divergence, avian predation upon littoral individuals has been mooted but has received little research attention. The kelp gull (Larus dominicanus) is the principal consumer of littoral limpets in the study area. We used shells from littoral and sublittoral zones, and from gull middens to examine – through linear morphometrics and elliptic Fourier analysis – whether selective predation exists, and to evaluate the mode of selection that could be operating on different traits. We found that limpet individuals with the apex displaced towards the anterior side were more likely to be predated. However, a remarkable result was that gulls select dorsal pear-shaped rather than elliptical shells and that there is a directional selection against pear-shaped limpets. However, in contrast to previous studies, we did not find that the proportions of elliptical or pear-shaped individuals differed between the littoral and sublittoral zones. This discrepancy could be related to variations in predatory intensity in different locations. These results suggest that where gulls have a strong influence, their effect could be a key factor in dorsal differentiation in limpet shells.

The Distribution of the Antarctic Limpet Nacella concinna (Nacellidae) on Underwater Landscapes of the Meek Channel, Argentine Islands, Graham Land

Objective(s). To clarify the patterns of Nacella concinna (Nacellidae) distribution in the water area of the Argentine Islands Archipelago (Meek Channel). Methods. Morphometric, statistical, cartographic methods as well as geoinformation technologies were used. Results. Analyzed the patterns of mollusc distribution on different transects, taking into account their morphometric characteristics — the length of the shell, the mollusс weight, the population density. It is determined that the distribution of Nacella concinna subpopulations does not have any visible strict regularity. At different transects with an insignificant exception the morphometric characteristics of the mollusсs are distributed both in classical and non-classical models. At the MK1 transect is shown the classical model of the distribution of morphometric characteristics, namely with an increase in the mollusс population density, the morphometric characteristics (shell length and weight of the mollusс) tend to decrease. At the MK2 transect is shown non-classical distribution model. The morphometric characteristics (shell length, mollusс weight) increase with an increase in the population density of the molluscs. At the MK3 transect is shown the classical distribution model, but a characteristic feature is an increase in the population density up to 5 m depth and a gradual decrease in the population density from 5 to 20 m. Geoinformation models of the Nacella concinna distribution by depth in the water area of the Ukrainian Antarctic Akademik Vernadsky station were constructed. Conclusions. Maps of the distribution of the Nacella concinna population were created for this region for the first time. The dimensional classes of the Nacella concinna subpopulation from selected water areas of the Argentine Islands were described.

Gonadal histology and gametogenesis of the Antarctic limpet Nacella concinna (Patellogastropoda, Nacellidae) collected at Potter Cove, 25 de Mayo (King George) Island, during austral summer

The limpet Nacella concinna is one of the most widely distributed gastropods along the Antarctic Peninsula. This species has been a useful tool in ecological and physiological studies for understanding Antarctic trophic interactions. Although the reproduction of limpets has been thoroughly studied, very little is known about gametogenesis in the genus Nacella. The purpose of this study is to describe the gonadal morphology and gametogenesis in N. concinna observed by light microscopy and to determine its chromosome complement. All the limpets were sexually mature at the time of sampling. Oocytes were separated by trabeculae with abundant glycogen reserves. There was a predominance of late vitellogenic oocytes. Many previtellogenic oocytes were pyriform in shape. Oogonia were clustered near the ovary wall or the trabeculae. Only two meiotic stages were found: metaphase I and anaphase I, both of which showed a regular arrangement of chromosomes. The oviduct contained mature oocytes surrounded by sperm. Testicular tubules were filled with spermatozoa. The elongated head of the spermatozoon resembles that of other Nacellidae members. One of the bivalents is heteropycnotic. The haploid complement is n = 4 and the sex determination system is XO/XX. Although N. concinna has been commonly characterized as a broadcast-spawner, its unique spawning behavior, the presence of oocytes at anaphase I and spermatozoa within the oviduct suggests internal fertilization.

Life in the intertidal: Cellular responses, methylation and epigenetics

Abstract Phenotypic plasticity is essential for the persistence of organisms under changing environmental conditions but the control of the relevant cellular mechanisms including which genes are involved and the regulation of those genes remains unclear. One way to address this issue is to evaluate links between gene expression, methylation and phenotype using transplantation and common garden experiments within genetically homogeneous populations. This approach was taken using the Antarctic limpet Nacella concinna. In this species, two distinct phenotypes are associated with the intertidal and subtidal zones. The in situ gene expression and methylation profiles of intertidal and subtidal cohorts were directly compared before and after reciprocal transplantation as well as after a common garden acclimation to aquarium conditions for 9 months. Expression profiles showed significant modulation of cellular metabolism to habitat zone with the intertidal profile characterised by transcription modules for antioxidant production, DNA repair and the cytoskeleton reflecting the need to cope with continually fluctuating and stressful conditions including wave action, UV irradiation and desiccation. Transplantation had an effect on gene expression. The subtidal animals transplanted to the intertidal zone modified their gene expression patterns towards that of an intertidal profile. In contrast, many of the antioxidant genes were still differentially expressed in the intertidal animals several weeks after transplantation into the relatively benign subtidal zone. Furthermore, a core of genes involved in antioxidation was still preferentially expressed in intertidal animals at the end of the common garden experiment. Thus, acclimation in an aquarium tank for 9 months did not completely erase the intertidal gene expression profile. Significant methylation differences were measured between intertidal and subtidal animals from the wild and after transplantation, which were reduced on common garden acclimation. This suggests that epigenetic factors play an important role in physiological flexibility associated with environmental niche. A plain language summary is available for this article.

Time course of lead induced proteomic changes in gill of the Antarctic limpet Nacella Concinna (Gastropoda: Patellidae)

The effect of increasing levels of metals from anthropogenic sources on Antarctic invertebrates is poorly understood. Here we exposed limpets (Nacella concinna) to 0, 0.12 and 0.25μgL−1 lead for 12, 24, 48 and 168h. We subsequently quantified the changes in protein abundance from gill, using 2D gel electrophoresis and mass spectrometry. We identified several antioxidant proteins, including the metal binding Mn-superoxide dismutase and ferritin, increasing abundances early on. Chaperones involved in the redox-dependent maturation of proteins in the endoplasmic reticulum (ER) showed higher abundance with lead at 48h. Lead also increased the abundance of Zn-binding carbonic anhydrase at 12h, suggesting a challenge to acid-base balance. Metabolic proteins increased abundance at 168h, suggesting a greater ATP demand during prolonged exposure. Changes in abundance of the small G-protein cdc42, a signaling protein modifying cytoskeleton, increased early and subsequently reversed during prolonged exposure, possibly leading to the modification of thick filament structure and function. We hypothesize that the replacement of metals initially affected antioxidant proteins and increased the production of reactive oxygen species. This disrupted the redox-sensitive maturation of proteins in the ER and caused increased ATP demand later on, accompanied by changes in cytoskeleton. SignificanceProteomic analysis of gill tissue in Antarctic limpets exposed to different concentrations of lead (Pb) over a 168h time period showed that proteomic changes vary with time. These changes included an increase in the demand of scavenging reactive oxygen species, acid-base balance and a challenge to protein homeostasis in the endoplasmic reticulum early on and subsequently an increase in energy metabolism, cellular signaling, and cytoskeletal modifications. Based on this time course, we hypothesize that the main mode of action of lead is a replacement of metal-cofactors of key enzymes involved in the scavenging of reactive oxygen species and the regulation of acid-base balance.

Fe, oxidative and nitrosative metabolism in the Antarctic limpet Nacella concinna

The hypothesis of this work was that oxidative and nitrosative metabolism in the digestive gland (DG) of two limpet populations (intertidal and subtidal) of the Antarctic species Nacella concinna show different behavior when they were exposed to either intermittent (intertidal) or constant (subtidal) natural Fe. Total Fe content and labile Fe pool were higher in the DG of the subtidal compared to the intertidal population. However, no significant differences between populations were seen on the Fe atoms content of the isolated ferritin. Ascorbyl radical content was 2.0±0.4 and 6.5±0.8pmol/mg FW in the DG of the intertidal and subtidal animals, respectively. Lipid damage, assessed as content of thiobarbituric reactive substances, was different between the tissues of intertidal and subtidal samples, 491±102 and 1242±367pmol/mg FW, respectively. Catalase and superoxide dismutase activities showed no differences between the limpets. Nitric oxide (NO) content was 25±3 and 22±2pmol/mg FW in DG from intertidal and subtidal animals, respectively. NO synthase-like (NOS-like) activity was evaluated supplementing the samples with the enzyme co-factors, and the inhibitory effect of Nω-nitro-L-arginine methyl ester hydrochloride was tested. NO generation rate was 3.4±0.3 and 4.7±0.6pmol/minmg FW in DG from the intertidal and subtidal population, respectively. These results showed that the oxidative condition of the limpet population constantly covered by the Fe enriched water is more affected than the intertidal population. However, the nitrosative metabolism seems to be independent of the environmental high Fe content since similar NO steady state concentration and NOS-like activity were measured in both populations.

The biology and ecology of the Antarctic limpet Nacella concinna

Physiological studies suggest that Antarctic marine organisms are adversely affected by rising global temperatures and ocean acidification and have poor prospects for survival. However, according to ecological studies, their vulnerability might be less severe than initially thought. Thus, a realistic forecast of species survival and Antarctic biodiversity should be based on studies from a variety of species under consideration of ecological factors. The limpet Nacella concinna is often found in the rocky intertidal and sublittoral zones of the Antarctic Peninsula and adjacent subantarctic islands. This review summarizes most of the available information on the biology of this limpet, one of the most conspicuous invertebrates of the intertidal zone. There is some evidence that adult N. concinna are physiologically flexible and can acclimate to 3 °C. However, the requirements of the larval stage are poorly known, thus precluding realistic predictions of how elevated temperatures will affect N. concinna populations. Data on physiological performance (righting ability, tenacity and radula rasping rate) under different temperatures could provide a useful baseline for further field investigations on the effects of warming. The species could be used as model organism for investigating the biological effects of ongoing global warming on slow-growing Antarctic ectotherms. Nacella concinna might also be a better biomonitor for polycyclic aromatic hydrocarbons than other Antarctic mollusks.

Out of Antarctica: quaternary colonization of sub-Antarctic Marion Island by the limpet genus Nacella (Patellogastropoda: Nacellidae)

The distribution of the Southern Ocean nearshore marine benthic fauna is the consequence of major geologic, oceanographic, and climatic changes during the last 50 Ma. As a result, a main biogeographic principle in the Southern Ocean is the clear distinction of the Antarctic biota. The Antarctic Polar Front (APF) represents an important barrier between Antarctica and other sub-Antarctic provinces. However, the high degree of genetic affinity between populations of the Antarctic limpet Nacella concinna and its sub-Antarctic relative Nacella delesserti from Marion Island stands against this tenet. Here, we performed new phylogenetic reconstructions in Nacella with special emphasis on the relationship between N. concinna and N. delesserti. Similarly, we performed population-based analyses in N. concinna and N. delesserti to further understand the genetic legacy of the Quaternary glacial cycles. Phylogenetic reconstructions recognized N. concinna and N. delesserti as two closely but distinct monophyletic entities and therefore as valid evolutionary units. The cladogenetic process separating them occurred ~0.35 Ma and is consistent with the origin of Marion Island (~0.45 Ma). Exceptional long-distance dispersal between provinces located inside and outside the APF, rather than revealing the permeability of the Antarctic Polar Front, seems to be related to latitudinal shift in the position of the APF during coldest periods of the Quaternary. Diversity indices, neutrality tests, haplotype networks, and demographic inference analysis showed that the demography of both species exhibits a clear signal of postglacial expansion.

Extinction and recolonization of maritime Antarctica in the limpet Nacella concinna (Strebel, 1908) during the last glacial cycle: toward a model of Quaternary biogeography in shallow Antarctic invertebrates

Quaternary glaciations in Antarctica drastically modified geographical ranges and population sizes of marine benthic invertebrates and thus affected the amount and distribution of intraspecific genetic variation. Here, we present new genetic information in the Antarctic limpet Nacella concinna, a dominant Antarctic benthic species along shallow ice-free rocky ecosystems. We examined the patterns of genetic diversity and structure in this broadcast spawner along maritime Antarctica and from the peri-Antarctic island of South Georgia. Genetic analyses showed that N.concinna represents a single panmictic unit in maritime Antarctic. Low levels of genetic diversity characterized this population; its median-joining haplotype network revealed a typical star-like topology with a short genealogy and a dominant haplotype broadly distributed. As previously reported with nuclear markers, we detected significant genetic differentiation between South Georgia Island and maritime Antarctica populations. Higher levels of genetic diversity, a more expanded genealogy and the presence of more private haplotypes support the hypothesis of glacial persistence in this peri-Antarctic island. Bayesian Skyline plot and mismatch distribution analyses recognized an older demographic history in South Georgia. Approximate Bayesian computations did not support the persistence of N.concinna along maritime Antarctica during the last glacial period, but indicated the resilience of the species in peri-Antarctic refugia (South Georgia Island). We proposed a model of Quaternary Biogeography for Antarctic marine benthic invertebrates with shallow and narrow bathymetric ranges including (i) extinction of maritime Antarctic populations during glacial periods; (ii) persistence of populations in peri-Antarctic refugia; and (iii) recolonization of maritime Antarctica following the deglaciation process.

Isotopic shift for defining habitat exploitation by the Antarctic limpet Nacella concinna from rocky coastal habitats (Marian Cove, King George Island)

δ 13C and δ 15N of the Antarctic limpet Nacella concinna tissues and their potential food sources were used to determine their dietary origins and their movements between diverse habitats of intertidal and subtidal rocky shores and tide pools of Marian Cove, King George Island, Antarctica in the austral summer. δ 13C and δ 15N of the organic matter sources of epilithic microalgae, macroalgae and suspended particulate organic matter (SPOM) were readily distinguishable to discern their relative contribution to the limpet diets, with the most depleted values being found in SPOM and the most enriched in macroalgae. The limpets exhibited a spatial trend in distribution due to their seasonal migration, with smaller individuals in the subtidal zone compared with larger ones on the intertidal sites. The limpet isotopes had relatively broad ranges of δ 13C and δ 15N (−26.6 to −12.8‰ and 2.6–7.1‰, respectively), suggesting a dietary shift between habitats as well as size classes. The stable isotope ratios for each habitat seem likely to reflect the differing availabilities of the three potential food sources. Isotope mixing model results indicate a spatial shift in dietary mixture between habitats as well as limpet size classes. Epilithic microalgae and phytoplankton made great contributions to the diet of the subtidal limpets. Together with epilithic microalgae, macroalgae were significant contributors to the intertidal limpets where macroalgae were abundant. A higher contribution of macroalgae to the limpet diets was found in the tide pools. In contrast, while phytoplankton was an important food source for the limpet spat, a great dietary dependence on epilithic microalgae was found in the small-size limpets from the lower intertidal zone. Our results suggest that limpet grazing can determine microalgal and/or macroalgal abundance and coverage on the Antarctic rocky-shore ecosystem, and trophic structure of benthic food web can change along environmental gradients even at spatial scales of tens or hundreds of metres in the Antarctic.

Effects of brooding and broadcasting reproductive modes on the population genetic structure of two Antarctic gastropod molluscs

Life-history characteristics exert a profound influence upon the population structure of many marine organisms. However, relatively few genetic studies have compared direct with indirect-developing species in the same ecosystem or geographical region, and none to our knowledge within an Antarctic setting. To address this issue, we have collected novel amplified fragment length polymorphism (AFLP) data from the direct-developing top shell Margarella antarctica to form a comparison with previously published data for the broadcast-spawning Antarctic limpet Nacella concinna. We scored 270 loci in 240 M. antarctica individuals sampled from five populations spanning the full length of the Antarctic Peninsula. Profound differences were identified in the strength and pattern of population structure between the two species, consistent with gene flow being highly restricted in M. antarctica relative to N. concinna.

Strong Population Genetic Structure in a Broadcast-Spawning Antarctic Marine Invertebrate

Although studies of population genetic structure are commonplace, a strong bias exists toward species from low latitudes and with relatively poor dispersal capabilities. Consequently, we used 280 amplified fragment length polymorphism bands to explore patterns of genetic differentiation among 8 populations of a high latitude broadcast-spawning marine mollusc, the Antarctic limpet Nacella concinna. Over 300 individuals were sampled along a latitudinal gradient spanning the Antarctic Peninsula from Adelaide Island to King George Island (67°-62°S), then to Signy Island (60°S) and South Georgia (54°S). Populations from the Antarctic Peninsula exhibited little genetic structure but were themselves strongly differentiated from both Signy and South Georgia. This finding was analytically highly robust and implies the presence of significant oceanographic barriers to gene flow in a species long regarded as a classic example of a widely dispersing broadcast spawner.

Shape differentiation and characterization in the two morphotypes of the Antarctic limpet Nacella concinna using Elliptic Fourier analysis of shells

The Antarctic limpet Nacella concinna (Strebel 1908) presents two ecotypes (littoral and sublittoral) differing in morphological and behavioral characteristics and more recently discovered, in physiological traits and genetic population structure. Previous works, based on traditional morphometry, used only three measurements (length, width and height of the shell) and their relationships. However, this methodology could not describe in detail the shape of the morphotypes. In the present work, Elliptic Fourier analysis (EFA) of shells was used to study the shape of the two morphotypes in six localities along the Antarctic Peninsula. The use of EFA confirmed the morphometric differences. The littoral morphotype has higher and globose shells with the apex displaced to the anterior part; the sublittoral individuals are more flat and pointed, and have the apex very well defined. Low allometric effect was detected in SL individuals, whereas L specimens did not show an allometric relationship within the examined size range. Differences in shell shape among localities were recorded. EFA analysis reflected the overall shell shape and allowed to characterize the main differences in shell shape between ecotypes that were difficult to quantify using the standard morphometric approach.

No evidence for genetic differentiation between Antarctic limpet Nacella concinna morphotypes

The extent to which genetic divergence can occur in the absence of physical barriers to gene flow is currently one of the most controversial topics in evolutionary biology, with implications for our understanding of speciation, phenotypic plasticity and adaptive potential. This is illustrated by a recent study reporting a surprising pattern of genetic differentiation between intertidal and subtidal morphotypes of the broadcast-spawning Antarctic limpet Nacella concinna. To explore this further, we collected almost 400 Antarctic limpets from four depths (intertidal, 6, 15 and 25 m) at Adelaide island, Antarctica, and conducted a combined morphometric and genetic analysis using 168 polymorphic amplified fragment length polymorphism (AFLP) loci. Morphological analysis revealed not only pronounced differences between the two morphotypes, but also a continuous cline in shell shape from the intertidal zone down to 25 m depth, suggesting that the distinction between the morphotypes may be artificial. Moreover, genetic analysis using both Fst and a Bayesian analogue found no evidence for differentiation either between the two morphotypes or by depth, and a Bayesian cluster analysis did not detect any cryptic genetic structure. Our findings lend support to the notion that limpets can be phenotypically highly plastic, although further studies are required to determine unequivocally whether there is any genetic basis to the observed variation in shell morphology.

Differences in heavy metal concentrations and in the response of the antioxidant system to hypoxia and air exposure in the Antarctic limpet Nacella concinna

During the austral spring and summer months, the Antarctic limpet Nacella concinna colonizes intertidal environments in the Western Antarctica Peninsula region. The species is divided into a permanently sub-littoral and a seasonally intertidal, migratory subpopulation. We investigate the physiological differentiation between the two limpet groups to identify cellular and molecular changes that accompany adaptation of stenothermal Antarctic invertebrates to life under more stressful intertidal habitat conditions. A major difference between the two groups is the significantly higher concentrations of heavy metals (Fe, Al, Zn) from ingested sediments in sub-littoral limpet digestive glands (DG), associated with higher rates of reactive oxygen species (ROS) formation in this organ. ROS formation is accompanied by significantly higher SOD activity in sub-littoral limpet DG. These high SOD activities are, however, not conserved during either air exposure or hypoxic stress exposure of the sub-littoral limpets, when ROS production is slowed due to the absence of oxygen. The intertidal animals maintain higher levels of SOD and also conserve catalase activity at higher levels during hypoxia or air exposure compared to sub-littoral individuals under the same exposure conditions. More oxidized redox potential in gills and foot muscle and higher antioxidant enzyme activities in gills indicate that intertidal limpets maintain more oxygenated tissues during air exposure, in keeping with shell-lifting for oxygen up-take by the gills of intertidal limpets which migrate up the shore in the spring and down in the autumn. An increase of the redox ratio (GSSG/GSH) and accumulation of the lipid oxidation derived malonedialdehyde in intertidal limpet foot muscle during 12h of exposure to air shows that indeed this tissue becomes more oxidized before the limpets eventually contract their shells tightly to minimize water loss and eventually become anaerobic. Intertidal limpets obviously avoid early onset of anaerobic energy production seen in their sub-littoral congeners when exposed to air and are still able to maintain tissue redox ratio balance when exposed to air.

Differences in the physiological response of inter- and subtidal Antarctic limpets Nacella concinna to aerial exposure

Antarctic intertidal zones are extremely harsh environments, and the Antarctic limpet Nacella concinna is one of the most conspicuous components of the macrofauna, colonizing such areas at King George Island, South Shetlands. The limpet subpopulation encompasses an intertidal and a subtidal ecomorph; these differ with respect to shell height and inner volume. We studied mor- phological, behavioural and physiological traits that distinguish both N. concinna subpopulations. We found a higher volume to circumference ratio of intertidal versus subtidal limpets to go along with a drastically reduced loss of body water during air exposure in intertidal specimens. Smaller inter- tidal limpets were less susceptible to desiccation than larger individuals. Further, the intertidal snails avoided accumulation of anaerobic metabolites (succinate, lactate and propionate), whereas subtidal limpets switched to anaerobic fermentation when exposed to air for >12 h. Only acetate accumulated in both types of snails after 12 h. Mean pO2 in shell water was higher in intertidal (2.6 kPa) than sub- tidal individuals (1.3 kPa) when submersed under water, and only the intertidal snails appeared able to respire air on emersion. These differences document strong physiological diversification between shore levels and shell morphs in the Antarctic limpet subpopulation at King George Island.

Genetic differentiation between morphotypes in the Antarctic limpet Nacella concinna as revealed by inter-simple sequence repeat markers

The limpet Nacella concinna (Strebel 1908) was the focus of numerous studies dealing with Antarctic benthos. One of the main characteristics of the species is the presence of two distinguishable morphotypes, one inhabiting the intertidal (during summer) and the other inhabiting the subtidal. For a long time these forms were considered as an expression of phenotypic plasticity, since previous studies did not found genetic differences between them. In the present work, we performed both a morphometric and a genetic differentiation analysis (using ISSR-PCR markers) of these two forms in three stations sampled at Potter Cove, South Shetland Islands. The results confirmed the morphological differences between intertidal and subtidal forms reported in other Antarctic localities. The genetic differences detected indicate that the two forms can be considered as genetically distinct populations maintaining low levels of gene flow. The degree of reproductive isolation of the ecotypes is discussed, as well as the possible origin of the divergence. The genetic differentiation observed can also have behavioral and physiological correlates, pointing out the importance of taking into account the potential differences in the response of both populations to different conditions in future studies in this species.

Does starvation influence the antioxidant status of the digestive gland of Nacella concinna in experimental conditions?

In a previous study we analysed the effect of diesel seawater contamination in the digestive gland of the Antarctic limpet Nacella concinna. We observed that antioxidant enzyme activities decreased after one-week starvation prior to the experiment, and this was considered in the analysis of the obtained results. To know whether the digestive gland oxidant–antioxidant status may be altered by starvation and experimental conditions, we evaluated the food deprivation effect in limpets from the nearshore shallow waters of Potter Cove, Antarctica. Organisms were acclimated to laboratory conditions and were divided in fed and starved groups, and maintained in these conditions during one month. Every week 20 limpets were sampled from each group. Digestive glands were dissected and kept frozen until they were processed. Superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) activities, as well as lipid peroxidation (LPO) measured as thiobarbituric reactive substances (TBARS), protein oxidation (PO) and reduced glutathione (GSH) were measured. For both groups of limpets, SOD increased its activity in the first week of the exposure period, with a maximum in the second week. CAT activity increased significantly in the second week, only for the starved group. Similarly, GST activity also increased for starved group in the second week; but maintained this tendency for both groups until the fourth week. In fed and starved limpets, TBARS values increased significantly, during the first week and then returned to normal values. The PO levels in the starved group increased only during the first week. The GSH content, for the fed group, increased significantly after the third week. The obtained results indicate that biochemical or physiological studies conducted with N. concinna should consider the effects of food deprivation and time spent under experimental conditions.