An ancient, Antarctic-specific species complex: large divergences between multiple Antarctic lineages of the tardigrade genus Mesobiotus.

Antarctica has been isolated and progressively glaciated for over 30 million years, with only approximately 0.3 % of its area currently ice-free and capable of supporting terrestrial ecosystems. As a result, invertebrate populations have become isolated and fragmented, in some cases leading to speciation. Terrestrial invertebrate species currently found in Antarctica often show multi-million year, and even Gondwanan, heritage, with little evidence of recent colonisation. Mesobiotus is a globally distributed tardigrade genus. It has commonly been divided into two "groups", referred to as harmsworthi and furciger, with both groups currently considered cosmopolitan, with global reports including from both the Arctic and the Antarctic. However, some authors considered that Meb. furciger, as originally described, may represent an Antarctic-specific lineage. Using collections of tardigrades from across the Antarctic continent and publicly available sequences obtained from online databases, we use mitochondrial and nuclear ribosomal sequence data to clarify the relationships of Antarctic Mesobiotus species. Our analyses show that all Antarctic members belong to a single lineage, evolving separately from non-Antarctic representatives. Within this Antarctic lineage there are further deep divisions among geographic regions of the continent, consistent with the presence of a species complex. Based on our data confirming the deep divisions between this Antarctic lineage, which includes representatives of both groups, we recommend that the use of furciger and harmsworthi group terminology is now abandoned, as it leads to systematic and biogeographical confusion.

Effects of elevational range shift on the morphology and physiology of a carabid beetle invading the sub-Antarctic Kerguelen Islands.

Climatic changes can induce geographic expansion and altitudinal shifts in the distribution of invasive species by offering more thermally suitable habitats. At the remote sub-Antarctic Kerguelen Islands, the predatory insect Merizodus soledadinus (Coleoptera: Carabidae), introduced in 1913, rapidly invaded coastal habitats. More recent colonisation of higher elevation habitats by this species could be underlain by their increased thermal suitability as the area has warmed. This study compared the effect of elevational range shift on the morphology and physiology of adult M. soledadinus sampled along two altitudinal transects (from the foreshore to 250 m a.s.l.) and a horizontal lowland transect orthogonal to the seashore (400 m length). Although high inter-individual and inter-transect variations in the traits examined were present, we observed that body mass of males and females tended to decrease with elevation, and that triglyceride contents decreased with distance from the shore. Moreover, protein contents of females as well as those of 26 metabolites were influenced significantly by distance to the foreshore. These results suggest that future climate change at the Kerguelen Islands will further assist the colonisation of lowland inland and higher altitude habitats by this aggressively invasive predator, by making previously sub-optimal habitats progressively more suitable.

Parasites in soil samples from Signy island, Antarctica.

Studies on parasite populations in Antarctic soils are scarce and thus little is known about the threat of these parasites towards either the natural fauna or human visitors. However, human presence in Antarctica, mainly through research and tourism, keeps increasing over time, potentially exposing visitors to zoonotic infections from Antarctic wildlife and environment. Most available literature to date has focused on faecal samples from Antarctic vertebrates. Therefore, this study addressed the possible presence of parasites in Antarctic soil that may be infectious to humans. Soil samples were obtained from five locations on Signy Island (South Orkney Islands, maritime Antarctic), namely North Point and Gourlay Peninsula (penguin rookeries), Pumphouse (relic coal-powered pump house), Jane Col (barren high altitude fellfield) and Berntsen Point (low altitude vegetated fellfield close to current research station). Approximately 10% of the soil samples (14/135) from 3 out of the 5 study sites had parasites which included Diphyllobotridae spp. eggs, Cryptosporidium sp., an apicomplexan protozoa (gregarine), Toxoplasma gondii, helminths (a cestode, Tetrabothrius sp., and a nematode larva) and mites. The presence of parasites in the 3 sites are most likely due to the presence of animal and human activities as two of these sites are penguin rookeries (North Point and Gourlay Peninsula) while the third site (Pumphouse Lake) has human activity. While some of the parasite species found in the soil samples appear to be distinctive, there were also parasites such as Cryptosporidium and Toxoplasma gondii that have a global distribution and are potentially pathogenic.

Cross-disciplinarity in the advance of Antarctic ecosystem research.

The biodiversity, ecosystem services and climate variability of the Antarctic continent and the Southern Ocean are major components of the whole Earth system. Antarctic ecosystems are driven more strongly by the physical environment than many other marine and terrestrial ecosystems. As a consequence, to understand ecological functioning, cross-disciplinary studies are especially important in Antarctic research. The conceptual study presented here is based on a workshop initiated by the Research Programme Antarctic Thresholds - Ecosystem Resilience and Adaptation of the Scientific Committee on Antarctic Research, which focussed on challenges in identifying and applying cross-disciplinary approaches in the Antarctic. Novel ideas and first steps in their implementation were clustered into eight themes. These ranged from scale problems, through risk maps, and organism/ecosystem responses to multiple environmental changes and evolutionary processes. Scaling models and data across different spatial and temporal scales were identified as an overarching challenge. Approaches to bridge gaps in Antarctic research programmes included multi-disciplinary monitoring, linking biomolecular findings and simulated physical environments, as well as integrative ecological modelling. The results of advanced cross-disciplinary approaches can contribute significantly to our knowledge of Antarctic and global ecosystem functioning, the consequences of climate change, and to global assessments that ultimately benefit humankind.

Parasites in soil samples from Signy island, Antarctica

@#Studies on parasite populations in Antarctic soils are scarce and thus little is known about the threat of these parasites towards either the natural fauna or human visitors. However, human presence in Antarctica, mainly through research and tourism, keeps increasing over time, potentially exposing visitors to zoonotic infections from Antarctic wildlife and environment. Most available literature to date has focused on faecal samples from Antarctic vertebrates. Therefore, this study addressed the possible presence of parasites in Antarctic soil that may be infectious to humans. Soil samples were obtained from five locations on Signy Island (South Orkney Islands, maritime Antarctic), namely North Point and Gourlay Peninsula (penguin rookeries), Pumphouse (relic coal-powered pump house), Jane Col (barren high altitude fellfield) and Berntsen Point (low altitude vegetated fellfield close to current research station). Approximately 10% of the soil samples (14/135) from 3 out of the 5 study sites had parasites which included Diphyllobotridae spp. eggs, Cryptosporidium sp., an apicomplexan protozoa (gregarine), Toxoplasma gondii, helminths (a cestode, Tetrabothrius sp., and a nematode larva) and mites. The presence of parasites in the 3 sites are most likely due to the presence of animal and human activities as two of these sites are penguin rookeries (North Point and Gourlay Peninsula) while the third site (Pumphouse Lake) has human activity. While some of the parasite species found in the soil samples appear to be distinctive, there were also parasites such as Cryptosporidium and Toxoplasma gondii that have a global distribution and are potentially pathogenic.

Global biogeographic patterns in bipolar moss species.

A bipolar disjunction is an extreme, yet common, biogeographic pattern in non-vascular plants, yet its underlying mechanisms (vicariance or long-distance dispersal), origin and timing remain poorly understood. Here, combining a large-scale population dataset and multiple dating analyses, we examine the biogeography of four bipolar Polytrichales mosses, common to the Holarctic (temperate and polar Northern Hemisphere regions) and the Antarctic region (Antarctic, sub-Antarctic, southern South America) and other Southern Hemisphere (SH) regions. Our data reveal contrasting patterns, for three species were of Holarctic origin, with subsequent dispersal to the SH, while one, currently a particularly common species in the Holarctic (Polytrichum juniperinum), diversified in the Antarctic region and from here colonized both the Holarctic and other SH regions. Our findings suggest long-distance dispersal as the driver of bipolar disjunctions. We find such inter-hemispheric dispersals are rare, occurring on multi-million-year timescales. High-altitude tropical populations did not act as trans-equatorial 'stepping-stones', but rather were derived from later dispersal events. All arrivals to the Antarctic region occurred well before the Last Glacial Maximum and previous glaciations, suggesting that, despite the harsh climate during these past glacial maxima, plants have had a much longer presence in this southern region than previously thought.

Moss survival through in situ cryptobiosis after six centuries of glacier burial.

Cryptobiosis is a reversible ametabolic state of life characterized by the ceasing of all metabolic processes, allowing survival of periods of intense adverse conditions. Here we show that 1) entire moss individuals, dated by 14C, survived through cryptobiosis during six centuries of cold-based glacier burial in Antarctica, 2) after re-exposure due to glacier retreat, instead of dying (due to high rates of respiration supporting repair processes), at least some of these mosses were able to return to a metabolically active state and remain alive. Moss survival was assessed through growth experiments and, for the first time, through vitality measurements. Future investigations on the genetic pathways involved in cryptobiosis and the subsequent recovery mechanisms will provide key information on their applicability to other systematic groups, with implications for fields as divergent as medicine, biodiversity conservation, agriculture and space exploration.

Are the Antarctic dipteran, Eretmoptera murphyi, and Arctic collembolan, Megaphorura arctica, vulnerable to rising temperatures?

Polar terrestrial invertebrates are suggested as being vulnerable to temperature change relative to lower latitude species, and hence possibly also to climate warming. Previous studies have shown Antarctic and Arctic Collembola and Acari to possess good heat tolerance and survive temperature exposures above 30 °C. To test this feature further, the heat tolerance and physiological plasticity of heat stress were explored in the Arctic collembolan, Megaphorura arctica, from Svalbard and the Antarctic midge, Eretmoptera murphyi, from Signy Island. The data obtained demonstrate considerable heat tolerance in both species, with upper lethal temperatures ≥35 °C (1 h exposures), and tolerance of exposure to 10 and 15 °C exceeding 56 days. This tolerance is far beyond that required in their current environment. Average microhabitat temperatures in August 2011 ranged between 5.1 and 8.1 °C, and rarely rose above 10 °C, in Ny-Ålesund, Svalbard. Summer soil microhabitat temperatures on Signy Island have previously been shown to range between 0 and 10 °C. There was also evidence to suggest that E. murphyi can recover from high-temperature exposure and that M. arctica is capable of rapid heat hardening. M. arctica and E. murphyi therefore have the physiological capacity to tolerate current environmental conditions, as well as future warming. If the features they express are characteristically more general, such polar terrestrial invertebrates will likely fare well under climate warming scenarios.

The effect of acclimation temperature on thermal activity thresholds in polar terrestrial invertebrates.

In the Maritime Antarctic and High Arctic, soil microhabitat temperatures throughout the year typically range between -10 and +5 °C. However, on occasion, they can exceed 20 °C, and these instances are likely to increase and intensify as a result of climate warming. Remaining active under both cool and warm conditions is therefore important for polar terrestrial invertebrates if they are to forage, reproduce and maximise their fitness. In the current study, lower and upper thermal activity thresholds were investigated in the polar Collembola, Megaphorura arctica and Cryptopygus antarcticus, and the mite, Alaskozetes antarcticus. Specifically, the effect of acclimation on these traits was explored. Sub-zero activity was exhibited in all three species, at temperatures as low as -4.6 °C in A. antarcticus. At high temperatures, all three species had capacity for activity above 30 °C and were most active at 25 °C. This indicates a comparable spread of temperatures across which activity can occur to that seen in temperate and tropical species, but with the activity window shifted towards lower temperatures. In all three species following one month acclimation at -2 °C, chill coma (=the temperature at which movement and activity cease) and the critical thermal minimum (=low temperature at which coordination is no longer shown) occurred at lower temperatures than for individuals maintained at +4 °C (except for the CTmin of M. arctica). Individuals acclimated at +9 °C conversely showed little change in their chill coma or CTmin. A similar trend was demonstrated for the heat coma and critical thermal maximum (CTmax) of all species. Following one month at -2 °C, the heat coma and CTmax were reduced as compared with +4 °C reared individuals, whereas the heat coma and CTmax of individuals acclimated at +9 °C showed little adjustment. The data obtained suggest these invertebrates are able to take maximum advantage of the short growing season and have some capacity, in spite of limited plasticity at high temperatures, to cope with climate change.

Multi-decadal survival of an Antarctic nematode, Plectus murrayi, in a -20°C stored moss sample.

It is not clear for how long Antarctic soil nematodes might tolerate freezing. Samples of the Antarctic moss, Bryum argenteum, were collected on 1 October 1983 at Langhovde, Soya coast, eastern Antarctica and were stored at -20°C. After 25.5 years of storage, living nematodes were recovered from the samples and were identified as Plectus murrayi by morphological examination and nucleotide sequencing of ribosomal RNA loci. The nematodes can grow and reproduce in a water agar plate with bacteria (mainly Pseudomonas sp.) cultured from the moss extract. They showed freezing tolerance at -20°C and -80°C and their survival rate after exposure to -20°C, but not -80°C, was increased if they were initially frozen slowly at a high sub-zero temperature. They also showed some ability to tolerate desiccation stress.

Pre-adapted to the maritime Antarctic?--rapid cold hardening of the midge, Eretmoptera murphyi.

During the 1960s, the midge, Eretmoptera murphyi, was transferred from sub-Antarctic South Georgia (55°S 37°W) where it is endemic to a single location on maritime Antarctic Signy Island (60°S 45°W). Its distribution has since expanded considerably, suggesting that it is pre-adapted to the more severe conditions further south. To test one aspect of the level of its pre-adaptation, the rapid cold hardening (RCH) response in this species was investigated. When juvenile (L1-L2) and mature (L3-L4) larvae of E. murphyi were directly exposed to progressively lower temperatures for 8h, they exhibited Discriminating Temperatures (DTemp, temperature at which there is 10-20% survival of exposed individuals) of -11.5 and -12.5°C, respectively. The mean SCP was above -7.5°C in both larval groups, confirming the finding of previous studies that this species is freeze-tolerant. Following gradual cooling (0.2°Cmin(-1)), survival was significantly greater at the DTemp in both larval groups. The response was strong, lowering the lower lethal temperature (LLT) by up to 6.5°C and maintaining survival above 80% for at least 22h at the DTemp. RCH was also exhibited during the cooling phase of an ecologically relevant thermoperiodic cycle (+4°C to -3°C). Mechanistically, the response did not affect freezing, with no alteration in the supercooling point (SCP) found following gradual cooling, and was not induced while the organism was in a frozen state. These results are discussed in light of E. murphyi's pre-adaptation to conditions on Signy Island and its potential to colonize regions further south in the maritime Antarctic.

Mite dispersal among the Southern Ocean Islands and Antarctica before the last glacial maximum.

It has long been maintained that the majority of terrestrial Antarctic species are relatively recent, post last glacial maximum, arrivals with perhaps a few microbial or protozoan taxa being substantially older. Recent studies have questioned this 'recolonization hypothesis', though the range of taxa examined has been limited. Here, we present the first large-scale study for mites, one of two dominant terrestrial arthropod groups in the region. Specifically, we provide a broad-scale molecular phylogeny of a biologically significant group of ameronothroid mites from across the maritime and sub-Antarctic regions. Applying different dating approaches, we show that divergences among the ameronothroid mite genera Podacarus, Alaskozetes and Halozetes significantly predate the Pleistocene and provide evidence of independent dispersals across the Antarctic Polar Front. Our data add to a growing body of evidence demonstrating that many taxa have survived glaciation of the Antarctic continent and the sub-Antarctic islands. Moreover, they also provide evidence of a relatively uncommon trend of dispersals from islands to continental mainlands. Within the ameronothroid mites, two distinct clades with specific habitat preferences (marine intertidal versus terrestrial/supralittoral) exist, supporting a model of within-habitat speciation rather than colonization from marine refugia to terrestrial habitats. The present results provide additional impetus for a search for terrestrial refugia in an area previously thought to have lacked ice-free ground during glacial maxima.

Gene expression associated with changes in cold tolerance levels of the Antarctic springtail, Cryptopygus antarcticus.

The ability of the Antarctic microarthropod Cryptopygus antarcticus (Collembola, Isotomidae) to survive low temperatures has been well studied at the physiological level, with recent investigations indicating the importance of the moulting process in conferring this ability. This study investigated gene expression in groups of C. antarcticus that have distinct differences in their ability to survive low temperatures. A microarray containing c. 5400 C. antarcticus expressed sequence tags was used to investigate gene expression differences between groups of animals with different supercooling points (SCP), and to low temperatures close to their SCP. By demonstrating the involvement of moult-related genes in the differential survival of two groups of C. antarcticus with distinct SCP profiles, the results of this investigation add support to the suggestion that moulting plays a role in conferring cold tolerance in C. antarcticus.

Temporal and spatial metabolic rate variation in the Antarctic springtail Gomphiocephalus hodgsoni.

Spatial and temporal environmental variation in terrestrial Antarctic ecosystems are known to impact species strongly at a local scale, but the ways in which organisms respond (e.g. physiologically, behaviourally) to such variation are poorly understood. Further, very few studies have attempted to assess inter-annual variability of such responses. Building on previous work demonstrating intra-seasonal variation in standard metabolic rate in the springtail Gomphiocephalushodgsoni, we investigated variation in metabolic activity of G. hodgsoni across two austral summer periods at Cape Bird, Ross Island. We also examined the influence of spatial variation by comparing metabolic rates of G. hodgsoni at Cape Bird with those from two other isolated continental locations within Victoria Land (Garwood and Taylor Valleys). We found significant differences between metabolic rates across the 2 years of measurement at Cape Bird. In addition, standard metabolic rates of G. hodgsoni obtained from Garwood and Taylor Valleys were significantly higher than those at Cape Bird where habitats are comparable, but environmental characteristics differ (e.g. microclimatic temperatures are higher). We discuss potential underlying causes of these metabolic rate variation patterns, including those related to differences among individuals (e.g. physiological and genetic differences), locations (e.g. habitat quality and microclimatic regime differences) and populations (e.g. acclimation differences among G. hodgsoni populations in the form of metabolic cold adaptation (MCA)).

Temporal metabolic rate variation in a continental Antarctic springtail.

Terrestrial systems in Antarctica are characterized by substantial spatial and temporal variation. However, few studies have addressed the paucity of data on metabolic responses to the unpredictable Antarctic environment, particularly with regard to terrestrial biota. This study measured metabolic rate variation for individual springtails at a continental Antarctic site using a fiber-optic closed respirometry system incorporating a custom-made respiration chamber. Concurrent measures of (behavioural) activity were made via daily pitfall counts. Metabolic rate of Gomphiocephalus hodgsoni measured at constant temperature varied systematically with progression through the austral summer, and was greatest mid-season. This finding of clear intra-seasonal and temperature-independent variation in mass-specific metabolic rate in G. hodgsoni is one of very few such reports for a terrestrial invertebrate (and the only such study for Antarctica), and parallels physiological studies in the Antarctic marine environment linking metabolic rate elevation with biological function rather than temperature adaptation per se. However, response to temperature at relatively short time-scales is also likely to be an important part of the life history strategy of Antarctic terrestrial invertebrates such as G. hodgsoni, which appears capable of both physiologically and behaviourally 'tuning' in to short-term thermal variability to respond appropriately to the local unpredictable Antarctic habitat.

The significance of the moult cycle to cold tolerance in the Antarctic collembolan Cryptopygus antarcticus.

Research into the ecophysiology of arthropod cold tolerance has largely focussed on those parts of the year and/or the life cycle in which cold stress is most likely to be experienced, resulting in an emphasis on studies of the preparation for and survival in the overwintering state. However, the non-feeding stage of the moult cycle also gives rise to a period of increased cold hardiness in some microarthropods and, as a consequence, a proportion of the field population is cold tolerant even during the summer active period. In the case of the common Antarctic springtail Cryptopygus antarcticus, the proportion of time spent in this non-feeding stage is extended disproportionately relative to the feeding stage as temperature is reduced. As a result, the proportion of the population in a cold tolerant state, with low supercooling points (SCPs), increases at lower temperatures. We found that, at 5 degrees C, about 37% of the population are involved in ecdysis and exhibit low SCPs. At 2 degrees C this figure increased to 50% and, at 0 degrees C, we estimate that 80% of the population will have increased cold hardiness as a result of a prolonged non-feeding, premoult period. Thus, as part of the suite of life history and ecophysiological features that enable this Antarctic springtail to survive in its hostile environment, it appears that it can take advantage of and extend the use of a pre-existing characteristic inherent within the moulting cycle.

Metabolomic fingerprint of cryo-stress in a freeze tolerant insect.

This study employed H-NMR spectroscopy to assay the metabolome of the high Arctic freeze-tolerant dipteran larvae, Heleomyza borealis, after recovery from exposure to a range of sub-zero temperature treatments. Our data demonstrate the resilience of freeze tolerance in individuals of this permanently freeze-tolerant species that were acclimated to summer temperatures (5 degree C): recovery of homeostasis after 48 h was not significantly disturbed by 2h exposures to -3, -12, or -20 degree C. Evidence of homeostatic perturbation to cryo-stress - both in terms of changes in specific metabolite concentrations as well as systemic changes in metabolism determined using multivariate pattern recognition techniques - was expressed almost entirely at a temperature coincident with the significant onset of mortality (-25 degree C) and considerably below the minimum winter temperatures of its over-wintering habitat (c.-12 degree C).

Plasticity and superplasticity in the acclimation potential of the Antarctic mite Halozetes belgicae (Michael).

The plasticity of an organism's phenotype may vary spatially and temporally, and across levels of physiological organisation. Given the adaptive value of plasticity in heterogeneous environments, it might be expected that it will be expressed most in a phenotype's most significant adaptive suites; at high latitudes, one of these is low temperature adaptation. This study examines the phenotypic plasticity of cold acclimation in the Antarctic mite, Halozetes belgicae (Michael). Both plastic and 'superplastic' (extreme plasticity) acclimation responses were found. Plastic responses were evident in responses to laboratory acclimation and field acclimatisation. 'Superplasticity' was found in its ability to rapidly cold harden (RCH) at 0, -5 and -10 degrees C. For example, after just 2 h of acclimation at 0 degrees C, mites acclimated at 10 degrees C shifted their supercooling points (SCPs) by approx. 15 degrees C. In terms of the combined speed of induction and lowering of lethal temperature, this is the most potent RCH response yet reported for a terrestrial arthropod. RCH was also expressed in thermal activity thresholds. Mechanisms responsible for significant differences in recovery from chill torpor are unknown; however, analysis of gut nucleator abundance suggest that the dynamic management of supercooling potential is largely achieved behaviourally, via evacuation. Comparisons with the literature reveal that plasticity in this species varies latitudinally, as well as temporally. The high degree of plasticity identified here is coincident with H. belgicae's occupation of the most exposed spatial niche available to Antarctic terrestrial arthropods.

Influence of temperature on the hygropreference of the Collembolan, Cryptopygus antarcticus, and the mite, Alaskozetes antarcticus from the maritime Antarctic.

The hygropreference of adult Cryptopygus antarcticus and Alaskozetes antarcticus was investigated over 2 h at 5, 10 and 20 degrees C, along humidity gradients (9-98% RH) established by means of different salt solutions. Two chamber arrangements were employed, linear and grid, to determine any influence of thigmotactic behaviour on distribution within the RH gradient. The humidity preference of both species varied with temperature. At 5 and 10 degrees C, C. antarcticus distributed homogeneously showing no clear RH preference. At 20 degrees C, this species preferred the highest humidity (98% RH). A. antarcticus demonstrated a preference for the lowest humidity (9% RH) at 5 degrees C, but at 10 degrees C its distribution differed between the two arena types. At 20 degrees C, A. antarcticus showed no clear humidity preference. Assays to control for experimental asymmetries along the gradient; thigmotactic behaviour; and aggregative behaviour exclude these factors as explanations for the observed results. The mean initial water content of samples did not differ significantly between temperature regimes (C. antarcticus: 68.6, 71.1 and 74.3%; A. antarcticus: 68.1, 70.1 and 68.6% at 5, 10 and 20 degrees C respectively), but the level of water loss increased significantly with temperature. The influence of desiccation tolerance and the ecological significance of the observed humidity preferences are discussed.