Multi-omics for studying and understanding polar life.

Polar ecosystems are experiencing amongst the most rapid rates of regional warming on Earth. Here, we discuss 'omics' approaches to investigate polar biodiversity, including the current state of the art, future perspectives and recommendations. We propose a community road map to generate and more fully exploit multi-omics data from polar organisms. These data are needed for the comprehensive evaluation of polar biodiversity and to reveal how life evolved and adapted to permanently cold environments with extreme seasonality. We argue that concerted action is required to mitigate the impact of warming on polar ecosystems via conservation efforts, to sustainably manage these unique habitats and their ecosystem services, and for the sustainable bioprospecting of novel genes and compounds for societal gain.

Antimicrobial resistance in Antarctica: is it still a pristine environment?

Although the rapid spread of antimicrobial resistance (AMR), particularly in relation to clinical settings, is causing concern in many regions of the globe, remote, extreme environments, such as Antarctica, are thought to be relatively free from the negative impact of human activities. In fact, Antarctica is often perceived as the last pristine continent on Earth. Such remote regions, which are assumed to have very low levels of AMR due to limited human activity, represent potential model environments to understand the mechanisms and interactions underpinning the early stages of evolution, de novo development, acquisition and transmission of AMR. Antarctica, with its defined zones of human colonisation (centred around scientific research stations) and large populations of migratory birds and animals, also has great potential with regard to mapping and understanding the spread of early-stage zoonotic interactions. However, to date, studies of AMR in Antarctica are limited. Here, we survey the current literature focussing on the following: i) Dissection of human-introduced AMR versus naturally occurring AMR, based on the premise that multiple drug resistance and resistance to synthetic antibiotics not yet found in nature are the results of human contamination ii) The potential role of endemic wildlife in AMR spread There is clear evidence for greater concentrations of AMR around research stations, and although data show reverse zoonosis of the characteristic human gut bacteria to endemic wildlife, AMR within birds and seals appears to be very low, albeit on limited samplings. Furthermore, areas where there is little, to no, human activity still appear to be free from anthropogenically introduced AMR. However, a comprehensive assessment of AMR levels in Antarctica is virtually impossible on current data due to the wide variation in reporting standards and methodologies used and poor geographical coverage. Thus, future studies should engage directly with policymakers to promote the implementation of continent-wide AMR reporting standards. The development of such standards alongside a centralised reporting system would provide baseline data to feedback directly into wastewater treatment policies for the Antarctic Treaty Area to help preserve this relatively pristine environment. Video Abstract.

Metabarcoding the Antarctic Peninsula biodiversity using a multi-gene approach.

Marine sediment communities are major contributors to biogeochemical cycling and benthic ecosystem functioning, but they are poorly described, particularly in remote regions such as Antarctica. We analysed patterns and drivers of diversity in metazoan and prokaryotic benthic communities of the Antarctic Peninsula with metabarcoding approaches. Our results show that the combined use of mitochondrial Cox1, and 16S and 18S rRNA gene regions recovered more phyla, from metazoan to non-metazoan groups, and allowed correlation of possible interactions between kingdoms. This higher level of detection revealed dominance by the arthropods and not nematodes in the Antarctic benthos and further eukaryotic diversity was dominated by benthic protists: the world's largest reservoir of marine diversity. The bacterial family Woeseiaceae was described for the first time in Antarctic sediments. Almost 50% of bacteria and 70% metazoan taxa were unique to each sampled site (high alpha diversity) and harboured unique features for local adaptation (niche-driven). The main abiotic drivers measured, shaping community structure were sediment organic matter, water content and mud. Biotic factors included the nematodes and the highly abundant bacterial fraction, placing protists as a possible bridge for between kingdom interactions. Meiofauna are proposed as sentinels for identifying anthropogenic-induced changes in Antarctic marine sediments.

Temperatures leading to heat escape responses in Antarctic marine ectotherms match acute thermal limits.

Thermal tolerance windows are key indicators of the range of temperatures tolerated by animals and therefore, a measure of resilience to climate change. In the ocean, where ectotherms are immersed, body temperatures are tightly coupled to environmental temperature and species have few options for thermoregulation. However, mobile species do have the ability to orientate towards optimal temperatures and move away from sub-optimal or dangerous temperatures. Escape responses are one such locomotory behavior, which typically manifests as a series of violent flicking movements that move individuals out of dangerous environments. We tested 11 species of Antarctic marine ectotherms, from one of the most stable shallow water marine environments, with an annual temperature range of -2°C to +2°C, that are vulnerable to small degrees of warming. Three species, the clam Laternula elliptica, the sea cucumber Cucumaria georgiana, and the brittlestar Ophionotus victoriae, showed no, or virtually no, escape response to temperature. Escape responses from a further eight species had a median response temperature of 11.2 (interquartile range, 10°C-15.7°C), which is well above current environmental temperatures but close to the range for acute lethal limits of Antarctic marine ectotherms (CTmax range, 17.2°C-26.6°C). This highlights that both acute tolerance limits and escape responses, fall outside current environmental temperatures, but also those predicted for 100s of years in the Southern Ocean. In a warmer Southern Ocean Antarctic fauna may not have the capacity to use temperature to select optimal thermal conditions, which leaves adaptation as a primary mechanism for their persistence.

Sources of elevated heavy metal concentrations in sediments and benthic marine invertebrates of the western Antarctic Peninsula.

Antarctica is one of the least anthropogenically-impacted areas of the world. Metal sources to the marine environment include localised activities of research stations and glacial meltwater containing metals of lithogenic origin. In this study, concentrations of nine metals (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn) were examined in three species of benthic invertebrates collected from four locations near Rothera Research Station on the western Antarctic Peninsula: Laternula elliptica (mudclam, filter feeder), Nacella concinna (limpet, grazer) and Odontaster validus (seastar, predator and scavenger). In addition, metals were evaluated in sediments at the same locations. Metal concentrations in different body tissues of invertebrates were equivalent to values recorded in industrialized non-polar sites and were attributed to natural sources including sediment input resulting from glacial erosion of local granodioritic rocks. Anthropogenic activities at Rothera Research Station appeared to have some impact on metal concentrations in the sampled invertebrates, with concentrations of several metals higher in L. elliptica near the runway and aircraft activities, but this was not a trend that was detected in the other species. Sediment analysis from two sites near the station showed lower metal concentrations than the control site 5 km distant and was attributed to differences in bedrock metal content. Differences in metal concentrations between organisms were attributed to feeding mechanisms and habitat, as well as depuration routes. L. elliptica kidneys showed significantly higher concentrations of eight metals, with some an order of magnitude greater than other organs, and the internal structure of O. validus had significantly higher Ni. This study supports previous assessments of N. concinna and L. elliptica as good biomonitors of metal concentrations and suggests O. validus as an additional biomonitor for use in future Antarctic metal monitoring programs.

Persistent Organic Pollutants in two species of migratory birds from Rothera Point, Adelaide Island, Antarctica.

Carcasses of South Polar Skuas (Catharacta maccormicki) and Kelp gulls (Larus dominicanus) were opportunistically collected around of Rothera Research station (67°35'8″S and 68°7'59″W) during the 2016/2017 austral summer. Samples of their tissues (muscle, liver and subcutaneous fat) were analysed for Persistent Organic Pollutants (POPs). Organochlorine pesticides (OCPs) showed the highest concentrations, notably for pp'-DDE and HCB. The Polychlorinated biphenyls (PCBs)-profiles demonstrated a clear dominance of hexa- and hepta-CBs, while concentrations of polybrominated diphenyl ethers (PBDEs) remained low. The concentrations of some POPs (e.g. HCB) were lower than in past studies on similar species, however others were within the previous range (PCBs) or even higher than previous reported values (DDE). Although no major interspecific differences in the absolute concentrations of POPs were detected, their profiles varied, being likely related to feeding and migration patterns of each species. The current study provides important baseline data for future monitoring of POPs in Antarctica.

Can acclimation of thermal tolerance, in adults and across generations, act as a buffer against climate change in tropical marine ectotherms?

Thermal acclimation capacity was investigated in adults of three tropical marine invertebrates, the subtidal barnacle Striatobalanus amaryllis, the intertidal gastropod Volegalea cochlidium and the intertidal barnacle Amphibalanus amphitrite. To test the relative importance of transgenerational acclimation, the developmental acclimation capacity of A. amphitrite was investigated in F1 and F2 generations reared at a subset of the same incubation temperatures. The increase in CTmax (measured through loss of key behavioural metrics) of F0 adults across the incubation temperature range 25.4-33.4°C was low: 0.00°C (V. cochlidium), 0.05°C (S. amaryllis) and 0.06°C (A. amphitrite) per 1°C increase in incubation temperature (the acclimation response ratio; ARR). Although the effect of generation was not significant, across the incubation temperature range of 29.4-33.4°C, the increase in CTmax in the F1 (0.30°C) and F2 (0.15°C) generations of A. amphitrite was greater than in the F0 (0.10°C). These correspond to ARR's of 0.03°C (F0), 0.08°C (F1) and 0.04°C (F2), respectively. The variability in CTmax between individuals in each treatment was maintained across generations, despite the high mortality of progeny. Further research is required to investigate the potential for transgenerational acclimation to provide an extra buffer for tropical marine species facing climate warming.

Age-related thermal response: the cellular resilience of juveniles.

Understanding species' responses to environmental challenges is key to predicting future biodiversity. However, there is currently little data on how developmental stages affect responses and also whether universal gene biomarkers to environmental stress can be identified both within and between species. Using the Antarctic clam, Laternula elliptica, as a model species, we examined both the tissue-specific and age-related (juvenile versus mature adult) gene expression response to acute non-lethal warming (12 h at 3 °C). In general, there was a relatively muted response to this sub-lethal thermal challenge when the expression profiles of treated animals, of either age, were compared with those of 0 °C controls, with none of the "classical" stress response genes up-regulated. The expression profiles were very variable between the tissues of all animals, irrespective of age with no single transcript emerging as a universal biomarker of thermal stress. However, when the expression profiles of treated animals of the different age groups were directly compared, a very different pattern emerged. The profiles of the younger animals showed significant up-regulation of chaperone and antioxidant transcripts when compared with those of the older animals. Thus, the younger animals showed evidence of a more robust cellular response to warming. These data substantiate previous physiological analyses showing a more resilient juvenile population.

One hundred questions of importance to the conservation of global biological diversity.

We identified 100 scientific questions that, if answered, would have the greatest impact on conservation practice and policy. Representatives from 21 international organizations, regional sections and working groups of the Society for Conservation Biology, and 12 academics, from all continents except Antarctica, compiled 2291 questions of relevance to conservation of biological diversity worldwide. The questions were gathered from 761 individuals through workshops, email requests, and discussions. Voting by email to short-list questions, followed by a 2-day workshop, was used to derive the final list of 100 questions. Most of the final questions were derived through a process of modification and combination as the workshop progressed. The questions are divided into 12 sections: ecosystem functions and services, climate change, technological change, protected areas, ecosystem management and restoration, terrestrial ecosystems, marine ecosystems, freshwater ecosystems, species management, organizational systems and processes, societal context and change, and impacts of conservation interventions. We anticipate that these questions will help identify new directions for researchers and assist funders in directing funds.

Heart rate and ventilation in Antarctic fishes are largely determined by ecotype.

Extrinsic neural and humoral influences on heart rate (fH) and ventilation frequency (fV) were examined following varying periods of post-surgical recovery in eight related Antarctic fish species inhabiting an array of inshore niches. Resting fH after recovery from handling was lower than previous reports, and the novel measurement of routine fH in free-swimming Dissostichus mawsoni (6.14 beats min(-1), bpm) is the lowest recorded for any fish. The extent of cardio-depressive cholinergic (vagal) tonus explained the large range of fH among species and varied with behavioural repertoire, being lower in the more active species, apart from Notothenia coriiceps. Adrenergic tonus was low compared with cholinergic tonus, with the exception of Trematomus newnesi. Hence, high cardiac cholinergic tonus may be a genotypic trait of the notothenioids that diverged with ecotype. Power spectral analysis showed that the vagal influence produced comparable spectra among species of similar morphology and ecotype. Removal of autonomic tonus resulted in a remarkably similar intrinsic fH between species. Simultaneous measurements of cardio-respiratory variables and oxygen consumption (M(O(2))) were made in the benthic Trematomus bernacchii and cryopelagic Pagothenia borchgrevinki. The slopes of the relationship between fH and M(O(2)) were similar. Trematomus bernacchii, however, had a higher M(O(2)) for a given fH than P. borchgrevinki, and P. borchgrevinki required a two-fold larger range in fH to reach a similar maximum M(O(2)), suggesting that there is a difference in cardiovascular fitness between the two species. Overall, the data suggest that cardio-respiratory control in Antarctic nototheniids is largely determined by activity levels associated with a given ecotype.

Reduced oxygen at high altitude limits maximum size.

The trend towards large size in marine animals with latitude, and the existence of giant marine species in polar regions have long been recognized, but remained enigmatic until a recent study showed it to be an effect of increased oxygen availability in sea water of a low temperature. The effect was apparent in data from 12 sites worldwide because of variations in water oxygen content controlled by differences in temperature and salinity. Another major physical factor affecting oxygen content in aquatic environments is reduced pressure at high altitude. Suitable data from high-altitude sites are very scarce. However, an exceptionally rich crustacean collection, which remains largely undescribed, was obtained by the British 1937 expedition from Lake Titicaca on the border between Peru and Bolivia in the Andes at an altitude of 3809 m. We show that in Lake Titicaca the maximum length of amphipods is 2-4 times smaller than other low-salinity sites (Caspian Sea and Lake Baikal).

Temperature and Embryonic Development in Relation to Spawning and Field Occurrence of Larvae of Three Antarctic Echinoderms.

The effects of temperature on development and viability were measured at 14 levels between -2{deg}C and +3{deg}C on embryos of two asteroids (Odontaster validus and Odontaster meridionalis) and an echinoid (Sterechinus neumayeri) from Signy Island, Antarctica. Development rates were 2 to 10 times slower than those for temperate or tropical echinoderms, with times to hatching up to 240 h. Development rates for the two asteroids differed by 1.15 x, and rates for both species approximately doubled over the experimental temperature range. In O. validus, embryo viability was independent of temperature, but in O. meridionalis viability declined with increasing temperature. Development rates for S. neumayeri were little affected by temperature above +0.2{deg}C, but declined rapidly at lower temperatures. Conversely, the number of nonviable eggs was low and constant below +1.7{deg}C, but rose rapidly at higher temperatures. A window of optimal temperature, between +0.2{deg}C and +1.7{deg}C, has therefore been proposed for development time and embryo viability in this population of S. neumayeri. Spawning trials and field observations of larvae indicated that the time of gamete release and periods of larval development in S. neumayeri coincided with austral summer sea temperatures in the same window. Embryos of O. meridionalis and O. validus are released in winter, when temperatures are constantly below -1.6{deg}C. Comparison of the different strategies suggests that larval food supply and predation during planktonic phases are not the dominant ecological factors for these species.

Evaluation of CMC and HA solutions for adhesiolysis.

Aqueous hyaluronic acid (HA) and carboxymethylcellulose (CMC) solutions were tested as tissue-protective coatings during lysis of surgical adhesions by blunt dissection or electrocautery in a rat cecal abrasion model. Phosphate-buffered saline (PBS) was used as a tissue coating solution in 200 female Sprague-Dawley rats prior to controlled cecal abrasion with a surgical gauze-tipped rotary abrader (four 1.5-cm-diameter areas; 70 g weight/60 revolutions/130 rpm). One-week after this initial cecal abrasion, rats were operated on again and adhesions were scored and lysed. The rats were randomly assigned to receive experimental tissue coating solutions either before (prelysis; n = 160) or after (postlysis; n = 40) adhesiolysis. Animals with prelysis coatings were further divided into blunt dissection or electrocautery adhesiolysis groups and were tested with 2 mL cecal coating of PBS, 0.4% HA, 0.5% CMC, or 1.0% CMC tissue coating solutions (n = 20/group). Rats treated postlysis received 2 mL cecal coating plus 2 mL intraperitoneal instillation of PBS, 1.8, 1.9, or 2.0% CMC. One week after adhesiolysis, rats were operated on again for final adhesion scoring. Prelysis tissue coating with 0.5 or 1.0% CMC solution appeared to inhibit adhesion reformation after blunt dissection, whereas 0.4% HA was not effective in this model. Solutions applied before electrocautery dissection or after blunt dissection were ineffective.

Melatonin rhythms in Pony mares and foals.

Melatonin concentrations in intact (N = 3) and sham-operated (N = 3) mares during March were greater (P less than 0 . 05) during the night than during the day, but this pattern was not seen in 3 mares from which the superior cervical ganglia had been removed bilaterally. When 4 Pony mares were exposed to a photoperiod of 10L:14D for 3 weeks and then to continuous darkness (0L:24D) for another 3 weeks, melatonin levels were greater (P less than 0 . 05) at the end of the 0L:24D period than during the earlier period and still displayed rhythmic fluctuations but were no longer co-ordinated with equivalent day/night rhythms or among mares. When melatonin rhythms were monitored in 3 mares and their foals housed in open pens exposed to natural lighting, significant time trends in melatonin concentrations were observed in mares when the foals were aged 1-3, 4-6 and 7-11 weeks, but foals did not display significant times trends in melatonin until they were 7-11 weeks old.

Effects of pinealectomy in Pony mares.

When mares were pinealectomized in summer or in winter there was no difference in the annual ovulation date of these animals and of pineal-intact controls during the first post-operative breeding season but in the second season the ovulation date of the winter pinealectomized mares was significantly delayed, suggesting a long-term but clear pineal involvement in seasonal reproductive patterns. Exposure of pinealectomized mares to a stimulatory photoperiod failed to advance the average date of first annual ovulation and was similar to that of intact mares receiving no stimulatory lighting. The results indicate that pinealectomy blocks the ability to respond to photostimulation. In intact mares plasma melatonin concentrations were significantly increased during darkness (P less than 0 . 003) but pinealectomized mares demonstrated no significant time trends.

Effects of pituitary stalk transection on endocrine function in Pony mares.

The pituitary stalk was transected in 10 Pony mares by a surgical approach that involved dorsal reflection of the brain and micro-dissection from the ventro-lateral aspect of the pituitary. Diabetes insipidus was the most immediate and marked result, requiring extensive electrolyte and antidiuretic therapy for approximately 48 h after operation. Fluid stasis then developed and no further supportive measures were necessary. Endocrine challenge tests with GnRH and TRH before and after stalk transection indicated a loss of responsiveness (GnRH) or suppressed responsiveness (TRH) after the operation. This technique permits isolation of the pituitary from its hypothalmic releasing and/or inhibiting hormones and therefore permits more refined study of the hypothalamic-pituitary axis.