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Background Historically, Hawai’i has had few massive coral bleaching events, until two consecutive heatwaves elevated seawater temperatures in 2014 and 2015. Consequent mortality and thermal stress were observed in the reef systems of Kane’ohe Bay (O’ahu). Two of the dominant species exhibited a phenotypic dichotomy of either bleaching resistance or susceptibility (Montipora capitata and Porites compressa), while a third species (Pocillopora acuta) was broadly susceptible to bleaching. In order to survey temporal shifts in the coral microbiomes during bleaching and recovery in all three of these coral species, 50 colonies were tagged and periodically monitored. Metabarcoding of three genetic markers (16S rRNA gene ITS1 and ITS2) followed by compositional approaches for community structure analysis, differential abundance and correlations for longitudinal data were used to temporally track and compare Bacteria/Archaea, Fungi and Symbiodiniaceae dynamics of the tagged colonies. Results Bleaching susceptible P. compressa corals recovered faster than P. acuta and Montipora capitata. Prokaryotic and algal microbiomes were majorly shaped by host species, and had no apparent pattern of temporal acclimatization in the overall community structure. Symbiodiniaceae signatures were identified at the colony scale, and were at times related to bleaching susceptibility, revealing higher intraspecific resolution than previously reported. Bacterial compositions were practically constant between bleaching phenotypes, and more diverse in P. acuta and M. capitata. P. compressa’s prokaryotic community was dominated by a single symbiotic bacterium. Fungal associates remained unexplored due to untargeted DNA co-amplification. Compositional approaches (via microbial balances) allowed the identification of fine-scale differences in the abundance of a consortium of microbes, driving changes by bleaching susceptibility and time across all hosts. Conclusions The three major coral reef founders in Kane’ohe Bay revealed different physiological and microbiome responses after 2014–2015 heatwaves. It is difficult to forecast though, a more successful strategy towards future scenarios of global warming. Differentially abundant microbial taxa across time and/or bleaching susceptibility were broadly shared between all hosts, suggesting that locally, the same microbes may modulate stress responses in sympatric coral species. Our study highlights the potential of investigating microbial balances to identify fine-scale microbiome changes, serving as local diagnostic tools of coral reef fitness.

DNA barcoding is a versatile approach that has revolutionized taxonomy and other akin topics in biology and ecology, due to its simplicity and relatively costless procedures. The method consists in the production of one or a few amplicons from informative genetic regions via Sanger sequencing. These markers are selected because they tend to evolve at a similar pace as speciation, allowing to discriminate organismal species. The applicability of this technique is here portrayed for the taxonomical identification of marine sponges (phylum: Porifera) as an exemplification.

34 pages, 4 figures.-- Distributed under Creative Commons CC-BY 4.0, Host-microbe interactions play crucial roles in marine ecosystems. However, we still have very little understanding of the mechanisms that govern these relationships, the evolutionary processes that shape them, and their ecological consequences. The holobiont concept is a renewed paradigm in biology that can help to describe and understand these complex systems. It posits that a host and its associated microbiota with which it interacts, form a holobiont, and have to be studied together as a coherent biological and functional unit to understand its biology, ecology, and evolution. Here we discuss critical concepts and opportunities in marine holobiont research and identify key challenges in the field. We highlight the potential economic, sociological, and environmental impacts of the holobiont concept in marine biological, evolutionary, and environmental sciences. Given the connectivity and the unexplored biodiversity specific to marine ecosystems, a deeper understanding of such complex systems requires further technological and conceptual advances, e.g., the development of controlled experimental model systems for holobionts from all major lineages and the modeling of (info)chemical-mediated interactions between organisms. Here we propose that one significant challenge is to bridge cross-disciplinary research on tractable model systems in order to address key ecological and evolutionary questions. This first step is crucial to decipher the main drivers of the dynamics and evolution of holobionts and to account for the holobiont concept in applied areas, such as the conservation, management, and exploitation of marine ecosystems and resources, where practical solutions to predict and mitigate the impact of human activities are more important than ever, This paper is based on the results of a foresight workshop funded by the EuroMarine network, Sorbonne University, and the UMRs 8227 and 7144 of the Roscoff Biological Station. Ezequiel M. Marzinelli was funded by an Australian Research Council Discovery Project (DP180104041), and José Pintado Valverde was funded by the Galician Innovation Agency (IN607A 2017/4). The work of Simon M. Dittami ad Catherine Leblanc was funded by the ANR project IDEALG (ANR-10-TBR-04). Claire M.M. Gachon, Catherine Leblanc, and SimonMDittami received funding from the European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement number 624575 (ALFF). The work of Fabrice Not was funded by the ANR project IMPEKAB (ANR-15-CE02-001). Ulisse Cardini was funded by the Research Council of Lithuania project INBALANCE (09.3.3-LMT-K-712-01-0069). Johan Decelle was supported by the CNRS and the ATIP-Avenir program, the LabEx GRAL (ANR-10-LABX-49-01) and Pôle Dittami et CBS from the University of Grenoble Alpes. Paco Cardenas received support from the European Union's Horizon 2020 research and innovation program through the SponGES project (grant agreement No. 679849). Elena Kazamia was funded by a Marie Curie Individual Fellowship (Horizon 2020, IRONCOMM). Aschwin H Engelen was supported by Portuguese national funds from FCT - Foundation for Science and Technology through projects UID/Multi/04326/2019 and UIDB/04326/2020

Polar marine biota have adapted to thrive under one of the ocean’s most inhospitable scenarios, where extremes of temperature, light photoperiod and ice disturbance, along with ecological interactions, have selected species with a unique suite of secondary metabolites. Organisms of Arctic and Antarctic oceans are prolific sources of natural products, exhibiting wide structural diversity and remarkable bioactivities for human applications. Chemical skeletons belonging to terpene families are the most commonly found compounds, whereas cytotoxic antimicrobial properties, the capacity to prevent infections, are the most widely reported activities from these environments. This review firstly summarizes the regulations on access and benefit sharing requirements for research in polar environments. Then it provides an overview of the natural product arsenal from Antarctic and Arctic marine organisms that displays promising uses for fighting human disease. Microbes, such as bacteria and fungi, and macroorganisms, such as sponges, macroalgae, ascidians, corals, bryozoans, echinoderms and mollusks, are the main focus of this review. The biological origin, the structure of terpenes and terpenoids, derivatives and their biotechnological potential are described. This survey aims to highlight the chemical diversity of marine polar life and the versatility of this group of biomolecules, in an effort to encourage further research in drug discovery.

Deception Island is an active volcano located at the southern end of the South Shetland Archipelago, in the Antarctic Ocean. After the last eruption in 1970, benthic recolonization took place within the bay, with echinoderms being the dominant epifauna (e.g., the ophiuroid Ophionotus victoriae, the echinoid Sterechinus neumayeri and the sea star Odontaster validus), together with dense infaunal communities (mostly composed by oligochaetes, polychaetes, and bivalves). Here, we aim to describe the actual status of the marine benthic ecosystems inhabiting the shallow subtidal areas of this volcanic island. Benthic species were qualitatively scored as presence versus absence, considering the different sampling effort between localities done over the years. A total of 139 species of macroorganisms, belonging to 16 phyla were found, including fauna and flora, increasing the species richness values previously reported in all sites surveyed within the volcano caldera. Moreover, a dramatic increase in biodiversity was found towards the entrance of the bay. We suggest, however, that recolonization from external waters may not be the only reason for this pattern. In fact, sediment flux rates and substrate instability are common disturbances within the bay, probably being among the major factors determining benthic community assemblages. These processes probably favour deposit feeding communities at the innermost locations of the bay. This study provides a remarkably increased and updated species inventory from previous reports, altogether with a description of the main communities inhabiting the bay and the abiotic factors regulating this, mainly the bottom type.

Bryozoans are among the most abundant and diverse members of the Antarctic benthos, however the role of bioactive metabolites in ecological interactions has been scarcely studied. To extend our knowledge about the chemical ecology of Antarctic bryozoans, crude ether extracts (EE) and butanol extracts (BE) obtained from two Antarctic common species (Cornucopina pectogemma and Nematoflustra flagellata), were tested for antibacterial and repellent activities. The extracts were screened for quorum quenching and antibacterial activities against four Antarctic bacterial strains (Bacillus aquimaris, Micrococcus sp., Oceanobacillus sp. and Paracoccus sp.). The Antarctic amphipod Cheirimedon femoratus and the sea star Odontaster validus were selected as sympatric predators to perform anti-predatory and substrate preference assays. No quorum quenching activity was detected in any of the extracts, while all EE exhibited growth inhibition towards at least one bacterium strain. Although the species were not repellent against the sea star, they caused repellence to the amphipods in both extracts, suggesting that defence activities against predation derive from both lipophilic and hydrophilic metabolites. In the substrate preference assays, one EE and one BE deriving from different specimens of the species C. pectogemma were active. This study reveals intraspecific variability of chemical defences and supports the fact that chemically mediated interactions are common in Antarctic bryozoans as means of protection against fouling and predation.

The Ha Long Bay (Vietnam) is a shallow area located in the northern part of the Tonkin Gulf, in the South China Sea. It includes more than 3000 islands of variable sizes. A continuous action of karstic processes, initiating around 280 million of years ago, shaped the formation of extraordinary caves and shallow salt-water lakes. These impressive structures recall the marine lakes found in Indonesia and Palau, creating one of the most important UNESCO’s World Heritage Sites. The protected area covers a surface of 434 km2 and comprises 775 islands, forming a triangle with the Dau Go Island (Driftwood Island) to the west, the Ba Ham Lake (Three Shelter Lake) to the south, and the Cong Tay Island to the east. During the Pleistocene, multiple sea level rise episodes that span for 10,000 – 15,000 years contributed to the geographic isolation of marine taxa. Along the coasts of Ha Long Bay islands, several coral species formed very shallow reefs that host peculiar sponge assemblages adapted to low depth and murky waters. In the marine lakes instead, corals can be quite rare while sponges are common. Their diversity though, is only known for a few lakes. Extreme variations in environmental conditions occur yearly in the lakes, due to heavy monsoon rains that cause stratification of the water column and drastic alterations in the communities. The aim of this study is to review the sponge diversity inside marine lakes and along the coastal areas of this peculiar region, comparing data collected more than fifteen years ago with recent data. During three expeditions between 2003 and 2004 we recorded 63 demosponges, of which 46 were living in and outside marine lakes, 17 were characteristic outside the lakes, and 23 were exclusive inside the lakes. After 15 years, a survey in August 2018 was performed to combine morphological data with DNA barcoding, to reveal and enhance the value of the biodiversity of the area. The diversity of sponges displayed drastic recession, showing the decline of more than half of the species typically present. In lieu, the exploration of a new lake (with very low salinity and high-water temperature) unveiled a seemingly new sponge species able to endure these conditions. Our findings suggest the urgency to develop measures of protection for these extraordinary but endangered environments, where biodiversity is still poorly explored. Indeed, these characteristic systems subjected to such intense geophysical isolating mechanisms represent optimal natural laboratories for the study of evolution and speciation of marine biota developing from segregated peripheral populations.

Host-microbe interactions play crucial roles in marine ecosystems, but we still have very little understanding of the mechanisms that govern these relationships, the evolutionary processes that shape them, and their ecological consequences. The holobiont concept is a renewed paradigm in biology that can help describe and understand these complex systems. It posits that a host and its associated microbiota, living together in a long-lasting relationship, form the holobiont, and have to be studied together, as a coherent biological and functional unit, in order to understand the biology, ecology and evolution of the organisms. Here we discuss critical concepts and opportunities in marine holobiont research and identify key challenges in the field. We highlight the potential economic, sociological, and environmental impacts of the holobiont concept in marine biological, evolutionary, and environmental sciences with comparisons to terrestrial science whenever appropriate. A deeper understanding of such complex systems, however, will require further technological and conceptual advances. The most significant challenge will be to bridge functional research on simple and tractable model systems and global approaches. This will require scientists to work together as an (inter)active community in order to address, for instance, ecological and evolutionary questions and the roles of holobionts in biogeochemical cycles., PeerJ Preprints, Computer Science Preprints., 2019

Cahong in Ha Long Bay (Vietnam) is a small lake with a reduced, invisible connection with the open sea. The water column conditions locally experience notable fluctuations across the year, mostly driven by biannual monsoon seasons. Salinity, temperature, and pH often reach extreme values, unsustainable for the majority of the marine fauna. Therefore, the biodiversity of the benthic macrofauna in this peculiar habitat is remarkably low. In particular, a single sponge species new to science was found solely populating this characteristic brackish lake during our last survey in August 2018. Spongilla manconiae sp. nov. is a new Porifera species described here. It belongs to an exclusively freshwater taxon and seems to have acquired adaptive traits to tolerate extreme peaks of temperature and salinity. The mitochondrial Cytochrome C Oxidase subunit 1 (COI) and the nuclear Internal Transcribed Spacers 1 and 2 (ITSs) gene markers were used for barcoding tagging and phylogenetic analyses. The new species revealed large genetic distances and separate clustering in the tree topology, with respect to other reference spongillid sequences from various geographic areas. The study provides evidence for an urgency to protect these unique marine lake systems because they represent rare, fluctuant, fragile habitats that may speed up speciation processes.

The Gram-negative Pantoea eucrina D2 was isolated from the marine sponge Chondrosia reniformis. Sponges were collected in a shallow volcanic vents system in Ischia island (South Italy), influenced by CO2 emissions and lowered pH. The chemical diversity of the secondary metabolites produced by this strain, under different culture conditions, was explored by a combined approach including molecular networking, pure compound isolation and NMR spectroscopy. The metabolome of Pantoea cf. eucrina D2 yielded a very complex molecular network, allowing the annotation of several metabolites, among them two biosurfactant clusters: lipoamino acids and surfactins. The production of each class of metabolites was highly dependent on the culture conditions, in particular, the production of unusual surfactins derivatives was reported for the first time from this genus, interestingly the production of these metabolites only arises by utilizing inorganic nitrogen as a sole nitrogen source. Major components of the extract obtained under standard medium culture conditions were isolated and identified as N-lipoamino acids by a combination of 1D and 2D NMR spectroscopy and HRESI-MS analysis. Assessment of the antimicrobial activity of the pure compounds towards some human pathogens, indicated a moderate activity of leucine containing N-lipoamino acids towards Staphylococcus aureus, Staphylococcus epidermidis and a clinical isolate of the emerging food pathogen Listeria monocytogenes.

This review is an overview on marine bioactive peptides with promising activities for the development of alternative drugs to fight human pathologies. In particular, we focus on potentially prolific producers of peptides in microorganisms, including sponge-associated bacteria and marine photoautotrophs such as microalgae and cyanobacteria. Microorganisms are still poorly explored for drug discovery, even if they are highly metabolically plastic and potentially amenable to culturing. This offers the possibility of obtaining a continuous source of bioactive compounds to satisfy the challenging demands of pharmaceutical industries. This review targets peptides because of the variety of potent biological activities demonstrated by these molecules, including antiviral, antimicrobial, antifungal, antioxidant, anticoagulant, antihypertensive, anticancer, antidiabetic, antiobesity, and calcium-binding bioactivities. Several of these peptides have already gained recognition as effective drug agents in recent years. We also focus on cutting-edge omic approaches for the discovery of novel compounds for pharmacological applications. With rapid depletion of natural resources, omic technologies may be the solution to efficiently produce a vast variety of novel peptides with unique pharmacological potential.

“Opisthobranch” mollusks, comprising the commonly known sea slugs and sea hares, present a wide variety of bioactive compounds protecting them against potential predators and competitors, and enhancing their ecological performance. Sea slug defenses include chemicals obtained directly from their prey, transformed dietary metabolites, or even de novo biosynthesized bioactive compounds. In this chapter, we describe what is known about the chemical ecology of marine slugs (“Opisthobranchs”) from a biological perspective and the molecules involved, focusing on the origins, bioactivities, and ecological roles in combination with other defensive strategies. Biogeographical and evolutionary aspects are also considered here. Often, chemical defenses are associated with warning (aposematic) colorations, allowing species to survive in exposed habitats, where predators learn to associate bright colorations with their bad taste. Many pigments possess bioactive properties themselves (e.g., alkaloids) while being part of photosynthetic systems, or may act as sunscreens protecting from UV light. Chemical defense allocation is particularly common in sea slugs, with extensive literature reporting bioactive products stored in exposed, vulnerable areas, such as the mantle, foot, gills, and rhinophores; within mucus or ink secretions; in specialized glands, like mantle dermal formations (MDFs); and also occasionally in eggs, embryos, and larval stages.

Solar radiation represents a key abiotic factor in the evolution of life in the oceans. In general, marine, biota—particularly in euphotic and dysphotic zones—depends directly or indirectly on light, but ultraviolet radiation (UV-R) can damage vital molecular machineries. UV-R induces the formation of reactive oxygen species (ROS) and impairs intracellular structures and enzymatic reactions. It can also affect organismal physiologies and eventually alter trophic chains at the ecosystem level. In Antarctica, physical drivers, such as sunlight, sea-ice, seasonality and low temperature are particularly influencing as compared to other regions. The springtime ozone depletion over the Southern Ocean makes organisms be more vulnerable to UV-R. Nonetheless, Antarctic species seem to possess analogous UV photoprotection and repair mechanisms as those found in organisms from other latitudes. The lack of data on species-specific responses towards increased UV-B still limits the understanding about the ecological impact and the tolerance levels related to ozone depletion in this region. The photobiology of Antarctic biota is largely unknown, in spite of representing a highly promising reservoir in the discovery of novel cosmeceutical products. This review compiles the most relevant information on photoprotection and UV-repair processes described in organisms from the Southern Ocean, in the context of this unique marine polar environment.

Antarctic benthic communities are largely structured by predation, which leads to the development of mechanisms of repellence. Among those mechanisms, chemical defences are quite extensive, yet poorly understood. To increase knowledge about the role of chemical defences in the Southern Ocean ecosystems, we assessed the incidence of feeding repellents in sessile and vagile invertebrates from nine phyla: Porifera, Cnidaria, Nemertea, Annelida, Mollusca, Bryozoa, Echinodermata, Hemichordata, and Tunicata (Ascidiacea). Samples were collected at depths of 120–789 m in the eastern Weddell Sea and Bouvet Island, and at depths ranging 0–100 m in the South Shetland Islands. When possible, specimens were dissected to study anatomical allocation of repellents. The common, eurybathic sea star Odontaster validus was chosen to perform feeding repellence bioassays, using diethyl ether (lipophilic) and butanol (hydrophilic) extracts from these samples. Among the 75 species tested, 52 % were studied for the first time for anti-predatory properties. Results provide further evidence of the prevalence of defensive metabolites in Antarctic organisms, with 47 % of the species exhibiting significant repellence within their lipophilic extracts. They also suggest a wider use of nonpolar defensive chemicals. Sessile taxa displayed highest repellence activities, with ascidians, cnidarians, and sponges being the most chemically protected. Overall, the present study indicates that natural products by mediating trophic interactions between prey and their potential predators play an important role in structuring Antarctic benthic ecosystems., SI

This review is an overview on marine bioactive peptides with promising activities for the development of alternative drugs to fight human pathologies. In particular, we focus on potentially prolific producers of peptides in microorganisms, including sponge-associated bacteria and marine photoautotrophs such as microalgae and cyanobacteria. Microorganisms are still poorly explored for drug discovery, even if they are highly metabolically plastic and potentially amenable to culturing. This offers the possibility of obtaining a continuous source of bioactive compounds to satisfy the challenging demands of pharmaceutical industries. This review targets peptides because of the variety of potent biological activities demonstrated by these molecules, including antiviral, antimicrobial, antifungal, antioxidant, anticoagulant, antihypertensive, anticancer, antidiabetic, antiobesity, and calcium-binding bioactivities. Several of these peptides have already gained recognition as effective drug agents in recent years. We also focus on cutting-edge omic approaches for the discovery of novel compounds for pharmacological applications. With rapid depletion of natural resources, omic technologies may be the solution to efficiently produce a vast variety of novel peptides with unique pharmacological potential.

This review is an overview on marine bioactive peptides with promising activities for the development of alternative drugs to fight human pathologies. In particular, we focus on potentially prolific producers of peptides in microorganisms, including sponge-associated bacteria and marine photoautotrophs such as microalgae and cyanobacteria. Microorganisms are still poorly explored for drug discovery, even if they are highly metabolically plastic and potentially amenable to culturing. This offers the possibility of obtaining a continuous source of bioactive compounds to satisfy the challenging demands of pharmaceutical industries. This review targets peptides because of the variety of potent biological activities demonstrated by these molecules, including antiviral, antimicrobial, antifungal, antioxidant, anticoagulant, antihypertensive, anticancer, antidiabetic, antiobesity, and calcium-binding bioactivities. Several of these ...

Taking into account the broad biological activities found in the meridianin indole alkaloids isolated to date, we have re-examined the organic extracts of an Antarctic collection of the tunicates Aplidium meridianum and A. falklandicum (Chordata: Ascidiacea) by HPLC in conjunction with a high-resolution mass spectrometer (HPLC-MS). A new set of analogs of meridianins A–G has been detected, and their structures are proposed on the basis of the molecular formulae identified by LC-HRMS analysis using a C18 column with a gradient of water/acetonitrile and an LTQ-FT-MS Orbitrap detector. Remarkably, dimers derived from meridianin A and from meridianin B or E were also detected. Our findings provide further evidence of the broad variability within the meridianin-like derivatives of this highly bioactive alkaloid family. Copyright © 2015 John Wiley & Sons, Ltd.

Many bioactive products from benthic invertebrates mediating ecological interactions have proved to reduce predation, but their mechanisms of action, and their molecular identities, are usually unknown. It was suggested, yet scarcely investigated, that nutritional quality interferes with defensive metabolites. This means that antifeedants would be less effective when combined with energetically rich prey, and that higher amounts of defensive compounds would be needed for predator avoidance. We evaluated the effects of five types of repellents obtained from Antarctic invertebrates, in combination with diets of different energetic values. The compounds came from soft corals, ascidians and hexactinellid sponges; they included wax esters, alkaloids, a meroterpenoid, a steroid, and the recently described organic acid, glassponsine. Feeding repellency was tested through preference assays by preparing diets (alginate pearls) combining different energetic content and inorganic material. Experimental diets contained various concentrations of each repellent product, and were offered along with control compound-free pearls, to the Antarctic omnivore amphipod Cheirimedon femoratus. Meridianin alkaloids were the most active repellents, and wax esters were the least active when combined with foods of distinct energetic content. Our data show that levels of repellency vary for each compound, and that they perform differently when mixed with distinct assay foods. The natural products that interacted the most with energetic content were those occurring in nature at higher concentrations. The bioactivity of the remaining metabolites tested was found to depend on a threshold concentration, enough to elicit feeding repellence, independently from nutritional quality.

The role of bioactive metabolites in ecological interactions involving Antarctic bryozoans has been scarcely studied. Bryozoans are one of the most abundant and diverse members of the Antarctic benthos and are preyed upon by diverse kinds of predators. They seem to be casual food items of the common Antarctic sea urchin Sterechinus neumayeri and the ubiquitous omnivorous amphipod Cheirimedon femoratus. In this study, the cytotoxic activity against embryos and sperm of the sea urchin S. neumayeri and the substrate preferences of the amphipod C. femoratus were assessed using organic extracts from Antarctic bryozoans, in order to determine the presence of chemical defenses. New adapted protocols were designed using a solidifying gel for simulating bryozoans’ surface. We analyzed 32 organic extracts from 16 samples that belonged to 13 different bryozoan species. No cytotoxicity was detected against embryos of S. neumayeri, while 12 of the 13 bryozoan species were cytotoxic to sperm at natural concentrations. In the substrate preference assays, the amphipod C. femoratus was repelled by ten species. The variable bioactivities found in both types of organic partitions of extracts indicated the presence of both lipophilic and hydrophilic defenses. Inter- and intraspecific variability of chemical defenses was detected also, suggesting environmental-induced responses, symbiotic production, and/or genetic variability. Possible alternative defensive mechanisms are also discussed for species with low or no chemical defense. Our results clearly support the fact that chemically mediated bioactivity in Antarctic bryozoans is common, and there is a likely ecological role of cytotoxic and repellent compounds for their protection.

Alcyonacean soft corals lack physical or skeletal defenses and their nematocyst system is weak, leading to the conclusion that soft corals mainly rely on chemistry for protection from predators and microbes. Defensive chemicals of primary and secondary metabolic origin are exuded in the mucus surface layer, explaining the general lack of heavy fouling and predation in corals. In Antarctic ecosystems, where generalist predation is intense and mainly driven by invertebrate consumers, the genus Alcyonium is represented by eight species. Our goal was to investigate the understudied chemical ecology of Antarctic Alcyonium soft corals. We obtained six samples belonging to five species: A. antarcticum, A. grandis, A. haddoni, A. paucilobulatum, and A. roseum, and assessed the lipid-soluble fractions for the presence of defensive agents in these specimens. Ethyl ether extracts were tested in feeding bioassays with the sea star Odontaster validus and the amphipod Cheirimedon femoratus as putative sympatric predators. Repellent activities were observed towards both consumers in all but one of the samples assessed. Moreover, three of the extracts caused inhibition to a sympatric marine bacterium. The ether extracts afforded characteristic illudalane sesquiterpenoids in two of the samples, as well as particular wax esters subfractions in all the colonies analyzed. Both kinds of metabolites displayed significant deterrent activities demonstrating their likely defensive role. These results suggest that lipophilic chemicals are a first line protection strategy in Antarctic Alcyonium soft corals against predation and bacterial fouling.

Warming oceans menace reef ecosystems by disrupting symbiosis between cnidarians and Symbiodinium zooxanthellae, thus triggering bleach episodes. Temperature fluctuations promote adjustments in physiological variables and symbiont composition, which can cause stress responses, but can also yield adaptation if fitter host–symbiont homeostasis are achieved. To understand such processes manipulative studies are required, but many reef-building cnidarians pose limitations to experimental prospects. We exposed Exaiptasia anemones to Gradual Thermal Stress (GTS) and Heat Shock (HS) exposures and monitored chlorophyll and symbiont dynamics to test the phenotypic plasticity of these photosynthetic holobionts. GTS enhanced chlorophyll concentrations and decreased Symbiodinium proliferation. A recovery period after GTS returned chlorophyll to lower concentrations and symbiont divisions to higher rates. HS triggered a stress response characterized by intense symbiont declines through degradation and expulsion, algal compensatory proliferation, and chlorophyll accumulation. Anemones pre-exposed to GTS displayed more acute signs of symbiont paucity after HS, demonstrating that recurrent stress does not always induce bleaching-resistance. Our study is the first documenting Symbiodinium C and D, along with the predominant Clade B1 in Exaiptasia anemones. C subclades found in outdoor specimens faded under laboratory exposures. Clade D emerged after HS treatments, and especially after GTS pre-exposure. This highlights the thermotolerance of D subclades found in E. pallida and shows that bleaching-recovery can involve shifts of background symbiont phylotypes. This study enlightens the capability of Exaiptasia anemones to acclimate to gradually increased temperatures, and explores into how thermal history influences in subsequent stress tolerance in symbiotic cnidarians.

Antarctic and sub-Antarctic benthic invertebrates are subjected to intense predation by mobile macroinvertebrates. Accordingly, chemical protection as well as other defensive mechanisms are expected to be common in organisms inhabiting these ecosystems. In order to evaluate anti-predation activities and allocation of chemical defenses within the anatomy of marine benthic Antarctic and sub-Antarctic invertebrates, 55 species were tested for feeding repellence against the sea star Odontaster validus, a common eurybathic sympatric predator. The invertebrates tested were collected from the deep waters of two poorly surveyed areas in terms of chemical ecology studies: the eastern Weddell Sea (Antarctica) and the vicinities of Bouvet Island (sub-Antarctica). Experiments were conducted at the Spanish Antarctic Base in Deception Island. In the feeding deterrence experiments, shrimp pieces were treated with crude lipophilic fractions obtained from each species, and were offered to the sea stars. A total of 29 species (53 %) from 7 different phyla (Porifera, Cnidaria, Chordata, Bryozoa, Echinodermata, Mollusca, and Annelida) showed feeding repellence against O. validus, and are therefore chemically protected against this keystone predator. Furthermore, 25 species were dissected into parts to investigate the possible allocation of defensive compounds. Some of the results obtained from these analyses support the prediction that the most exposed/vulnerable tissues concentrate chemical defenses to avoid predation against the sea stars. In summary, the results obtained in our survey support the hypothesis that deep-water Antarctic and sub-Antarctic benthic invertebrates are well protected chemically against sympatric predators, similarly to what has been reported in previous studies investigating shallow-water Antarctic species.

Table S1. Species names and dissected body-regions of the samples used to obtain extracts for feeding assays. WW: sample wet weight; DW: sample dry weight; EE: dry weight of the diethyl ether extract; NEE: percentage of the natural tissue concentration of EE in the sample calculated by dividing EE by DW; na: not available; body-regions — BAS: basal, API: apical, EXT: external, INT: internal, POL: polyparium, AX: axis body-parts; morphotypes — BW numbers after species names indicate conspecific samples from different collection sites

Ascidians have developed multiple defensive strategies mostly related to physical, nutritional or chemical properties of the tunic. One of such is chemical defense based on secondary metabolites. We analyzed a series of colonial Antarctic ascidians from deep-water collections belonging to the genera Aplidium and Synoicum to evaluate the incidence of organic deterrents and their variability. The ether fractions from 15 samples including specimens of the species A. falklandicum, A. fuegiense, A. meridianum, A. millari and S. adareanum were subjected to feeding assays towards two relevant sympatric predators: the starfish Odontaster validus, and the amphipod Cheirimedon femoratus. All samples revealed repellency. Nonetheless, some colonies concentrated defensive chemicals in internal body-regions rather than in the tunic. Four ascidian-derived meroterpenoids, rossinones B and the three derivatives 2,3-epoxy-rossinone B, 3-epi-rossinone B, 5,6-epoxy-rossinone B, and the indole alkaloids meridianins A–G, along with other minoritary meridianin compounds were isolated from several samples. Some purified metabolites were tested in feeding assays exhibiting potent unpalatabilities, thus revealing their role in predation avoidance. Ascidian extracts and purified compound-fractions were further assessed in antibacterial tests against a marine Antarctic bacterium. Only the meridianins showed inhibition activity, demonstrating a multifunctional defensive role. According to their occurrence in nature and within our colonial specimens, the possible origin of both types of metabolites is discussed.

The chemical analysis of the ascidian Aplidium fuegiense resulted in the isolation of three novel meroterpenoids 2–4, structurally related to the main co-occurring known rossinone B (1). The structures of the new compounds were determined by interpretation of spectroscopic data. Compounds 1–4 were found to be selectively localized in the viscera of the ascidian.

Predation and competition are important factors structuring Antarctic benthic communities and are expected to promote the production of chemical defenses. Tunicates are subject to little predation, and this is often attributed to chemical compounds, although their defensive activity has been poorly demonstrated against sympatric predators. In fact, these animals, particularly the genus Aplidium, are rich sources of bioactive metabolites. In this study, we report the natural products, distribution and ecological activity of two Aplidium ascidian species from the Weddell Sea (Antarctica). In our investigation, organic extracts obtained from external and internal tissues of specimens of A. falklandicum demonstrated to contain deterrent agents that caused repellency against the Antarctic omnivorous predator, the sea star Odontaster validus. Chemical analysis performed with Antarctic colonial ascidians Aplidium meridianum and Aplidium falklandicum allowed the purification of a group of known bioactive indole alkaloids, meridianins A-G. These isolated compounds proved to be responsible for the deterrent activity. Funding was provided by the Ministry of Science and Education of Spain through the ECOQUIM Projects (REN2003-00545, REN2002-12006E ANT and CGL2004-03356/ANT).

Chemical investigation of the lipophilic extract of the Antarctic soft coral Alcyonium grandis led us to the finding of nine unreported sesquiterpenoids, 2-10. These molecules are members of the illudalane class and in particular belong to the group of alcyopterosins, illudalanes isolated from marine organisms. The structures of 2-10 were determined by interpretation of spectroscopic data. Repellency experiments conducted using the omnivorous Antarctic sea star Odontaster validus revealed a strong activity in the lipophilic extract of A. grandis against predation.

Out of the many bioactive compounds described from the oceans, only a small fraction have been studied for their ecological significance. Similarly, most chemically mediated interactions are not well understood, because the molecules involved remain unrevealed. In Antarctica, this gap in knowledge is even more acute in comparison to tropical or temperate regions, even though polar organisms are also prolific producers of chemical defenses, and pharmacologically relevant products are being reported from the Southern Ocean. The extreme and unique marine environments surrounding Antarctica along with the numerous unusual interactions taking place in benthic communities are expected to select for novel functional secondary metabolites. There is an urgent need to comprehend the evolutionary role of marine derived substances in general, and particularly at the Poles, since molecules of keystone significance are vital in species survival, and therefore, in structuring the communities. Here we provide a mini-review on the identified marine natural products proven to have an ecological function in Antarctic ecosystems. This report recapitulates some of the bibliography from original Antarctic reviews, and updates the new literature in the field from 2009 to the present.

Taking into account the broad biological activities found in the meridianin indole alkaloids isolated to date, we have re-examined the organic extracts of an Antarctic collection of the tunicates Aplidium meridianum and A. falklandicum (Chordata: Ascidiacea) by HPLC in conjunction with a high-resolution mass spectrometer (HPLC-MS). A new set of analogs of meridianins A-G has been detected, and their structures are proposed on the basis of the molecular formulae identified by LC-HRMS analysis using a C18 column with a gradient of water/acetonitrile and an LTQ-FT-MS Orbitrap detector. Remarkably, dimers derived from meridianin A and from meridianin B or E were also detected. Our findings provide further evidence of the broad variability within the meridianin-like derivatives of this highly bioactive alkaloid family.

Xavier, José C. et al., Polar marine ecosystems have global ecological and economic importance because of their unique biodiversity and their major role in climate processes and commercial fisheries, among others. Portugal and Spain have been highly active in a wide range of disciplines in marine biology of the Antarctic and the Arctic. The main aim of this paper is to provide a synopsis of some of the results and initiatives undertaken by Portuguese and Spanish polar teams within the field of marine sciences, particularly on benthic and pelagic biodiversity (species diversity and abundance, including microbial, molecular, physiological and chemical mechanisms in polar organisms), conservation and ecology of top predators (particularly penguins, albatrosses and seals), and pollutants and evolution of marine organisms associated with major issues such as climate change, ocean acidification and UV radiation effects. Both countries have focused their polar research more in the Antarctic than in the Arctic. Portugal and Spain should encourage research groups to continue increasing their collaborations with other countries and develop multi-disciplinary research projects, as well as to maintain highly active memberships within major organizations, such as the Scientific Committee for Antarctic Research (SCAR), the International Arctic Science Council (IASC) and the Association of Polar Early Career Scientists (APECS), and in international research projects. © 2013 Elsevier B.V.

The Antarctic sea star Odontaster validus and the amphipod Cheirimedon femoratus are important predators in benthic communities. Some bryozoans are part of the diet of the asteroid and represent both potential host biosubstrata and prey for this omnivorous lysianassid amphipod. In response to such ecological pressure, bryozoans are expected to develop strategies to deter potential predators, ranging from physical to chemical mechanisms. However, the chemical ecology of Antarctic bryozoans has been scarcely studied. In this study we evaluated the presence of defenses against predation in selected species of Antarctic bryozoans. The sympatric omnivorous consumers O. validus and C. femoratus were selected to perform feeding assays with 16 ether extracts (EE) and 16 butanol extracts (BE) obtained from 16 samples that belonged to 13 different bryozoan species. Most species (9) were active (12 EE and 1 BE) in sea star bioassays. Only 1 BE displayed repellence, indicating that repellents against the sea star are mainly lipophilic. Repellence toward C. femoratus was found in all species in different extracts (10 EE and 12 BE), suggesting that defenses against the amphipod might be both lipophilic and hydrophilic. Interspecific and intraspecific variability of bioactivity was occasionally detected, suggesting possible environmental inductive responses, symbiotic associations, and/or genetic variability. Multivariate analysis revealed similarities among species in relation to bioactivities of EE and/or BE. These findings support the hypothesis that, while in some cases alternative chemical or physical mechanisms may also provide protection, repellent compounds play an important role in Antarctic bryozoans as defenses against predators.

Ascidians have developed multiple defensive strategies mostly related to physical, nutritional or chemical properties of the tunic. One of such is chemical defense based on secondary metabolites. We analyzed a series of colonial Antarctic ascidians from deep-water collections belonging to the genera Aplidium and Synoicum to evaluate the incidence of organic deterrents and their variability. The ether fractions from 15 samples including specimens of the species A. falklandicum, A. fuegiense, A. meridianum, A. millari and S. adareanum were subjected to feeding assays towards two relevant sympatric predators: the starfish Odontaster validus, and the amphipod Cheirimedon femoratus. All samples revealed repellency. Nonetheless, some colonies concentrated defensive chemicals in internal body-regions rather than in the tunic. Four ascidian-derived meroterpenoids, rossinones B and the three derivatives 2,3-epoxy-rossinone B, 3-epi-rossinone B, 5,6-epoxy-rossinone B, and the indole alkaloids meridianins A–G, along with other minoritary meridianin compounds were isolated from several samples. Some purified metabolites were tested in feeding assays exhibiting potent unpalatabilities, thus revealing their role in predation avoidance. Ascidian extracts and purified compound-fractions were further assessed in antibacterial tests against a marine Antarctic bacterium. Only the meridianins showed inhibition activity, demonstrating a multifunctional defensive role. According to their occurrence in nature and within our colonial specimens, the possible origin of both types of metabolites is discussed.

Antarctic marine ecosystems are immersed in an isolated, relatively constant environment where the organisms inhabiting their benthos are mainly sessile suspension feeders. For these reasons, physical and chemical biotic interactions play an essential role in structuring these marine benthic communities (Dayton et al., 1974; Orejas et al., 2000). These interactions may include diverse strategies to avoid predation (e.g. Iken et al., 2002), competition for space or food (e.g. Bowden et al., 2006) and avoiding fouling (e.g. Rittschof, 2001; Peters et al., 2010). For instance, in the marine benthos, one of the most extended effective strategies among sessile soft-bodied organisms is chemical defense, mediated by several bioactive natural products mostly considered secondary metabolites (e.g. Paul et al., 2011). The study of the “chemical network” (chemical ecology interactions) structuring the communities provides information about the ecology and biology of the involved species, the function and the structure of the community and, simultaneously, it may lead to the discovery of new compounds useful to humans for their pharmacological potential (e.g. Avila, 1995; Bhakuni, 1998; Munro et al., 1999; Faulkner, 2000; Lebar et al., 2007; Avila et al., 2008). In the last three decades, the study of marine chemical ecology has experienced great progress, thanks to the new technological advances for collecting and studying marine samples, and the possibility of identification of molecules with smaller amounts of compounds (e.g. Paul et al., 2006, 2011; Blunt et al., 2011).

Hexactinellids (glass sponges) are an understudied class with syncytial organization and poor procariotic associations, thought to lack defensive secondary metabolites. Poriferans, though, are outstanding sources of bioactive compounds; nonetheless, a growing suspicion suggests that many of these chemicals could be symbiont-derived. In Polar latitudes, sponges are readily invaded by diatoms, which could provide natural products. Hexactinellids are typical of deep waters; but in Antarctica, they dominate the upper shelf providing shelter and food supply to many opportunistic mesograzers and macroinvertebrates, which exert strong ecological pressures on them. Aiming to examine the incidence of defensive activities of hexactinellids against consumption, feeding experiments were conducted using their lipophilic fractions. Antarctic hexactinellid and demosponge extracts were tested against the asteroid Odontaster validus and the amphipod Cheirimedon femoratus as putative sympatric, omnivorous consumers. Hexactinellids yielded greater unpalatable activities towards the amphipod, while no apparent allocation of lipophilic defenses was noted. After chemical analyses on the lipophilic fractions from these Antarctic glass sponges, quite similar profiles were revealed, and no peculiar secondary metabolites, comparable to those characterizing other poriferans, were found. Instead, the lipidic compounds 5α(H)-cholestan-3-one and two glycoceramides were isolated for their particular outspread presence in our samples. The isolated compounds were further assessed in asteroid feeding assays, and their occurrence was evaluated for chemotaxonomical purposes in all the Antarctic samples as well as in glass sponges from other latitudes by NMR and MS. Characteristic sphingolipids are proposed as chemical markers in Hexactinellida, with possible contributions to the classification of this unsettled class.

71 páginas, 1 tabla, 3 figuras., Antarctic ecosystems are exposed to unique environmental characteristics resulting in communities structured both by biotic interactions such as predation and competition, as well as abiotic factors such as seasonality and ice-scouring. It is important to understand how ecological factors may trigger chemical mechanisms in marine Antarctic organisms as a response for survival. However, very little is known yet about the evolution of chemical compounds in Antarctic organisms. Investigations in chemical ecology have demonstrated over the last several years that defensive metabolites have evolved in numerous representative Antarctic species. This contradicts earlier theories concerning biogeographic variation in predation and chemical defenses. As reviewed here, a number of interesting natural products have been isolated from Antarctic organisms. However, we believe many more are still to be discovered. Currently, many groups such as microorganisms, planktonic organisms and deepsea fauna remain almost totally unknown regarding their natural products. Furthermore, for many described compounds, ecological roles have yet to be evaluated. In fact, much of the research carried out to date has been conducted in the laboratory, and only in a few cases in an ecologically relevant context. Therefore, there is a need to extend the experiments to the field, as done in tropical and temperate marine ecosystems, or at least, to test the activity of the chemicals in natural conditions and ecologically meaningful interactions. Defense against predators is always one of the main topics when talking about the roles of natural products in species interactions, but many other interesting aspects, such as competition, chemoattraction, fouling avoidance and ultraviolet (UV) protection, also deserve further attention. In our opinion, challenging future developments are to be expected for Antarctic marine chemical ecology in the years to come., This work would not have been possible without the financial support of the Ministry of Science and Education of Spain through different grants along recent years in the general frame of our ECOQUIM projects (ANT97-1590-E, ANT97-0273, REN2002-12006-E ⁄ANT, REN2003-00545 and CGL2004- 03356 ⁄ANT).

Generalist predation constitutes a driving force for the evolution of chemical defences. In the Antarctic benthos, asteroids and omnivore amphipods are keystone opportunistic predators. Sessile organisms are therefore expected to develop defensive mechanisms mainly against such consumers. However, the different habits characterizing each predator may promote variable responses in prey. Feeding-deterrence experiments were performed with the circumpolar asteroid macropredator Odontaster validus to evaluate the presence of defences within the apolar lipophilic fraction of Antarctic invertebrates and macroalgae. A total of 51% of the extracts were repellent, yielding a proportion of 17 defended species out of the 31 assessed. These results are compared with a previous study in which the same fractions were offered to the abundant circum-Antarctic amphipod Cheirimedon femoratus . Overall, less deterrence was reported towards asteroids (51%) than against amphipods (80.8%), principally in sponge and algal extracts. Generalist amphipods, which establish casual host–prey sedentary associations with biosubstrata (preferentially sponges and macroalgae), may exert more localized predation pressure than sea stars on certain sessile prey, which would partly explain these results. The nutritional quality of prey may interact with feeding deterrents, whose production is presumed to be metabolically expensive. Although optimal defence theory posits that chemical defences are managed and distributed as to guarantee protection at the lowest cost, we found that only a few organisms localized feeding deterrents towards most exposed and/or valuable body regions. Lipophilic defensive metabolites are broadly produced in Antarctic communities to deter opportunistic predators, although several species combine different defensive traits. Keywords: Antarctic invertebrates; Antarctic algae; chemical ecology; sea star Odontaster validus ; amphipod Cheirimedon femoratus ; chemical defence. (Published: 7 April 2014) Citation: Polar Research 2014, 33 , 21624, http://dx.doi.org/10.3402/polar.v33.21624

Corrigendum to “Rossinone-related meroterpenes from the Antarctic ascidian Aplidium fuegiense” [Tetrahedron 68 (2012) 3541e3544] Marianna Carbone , Laura N u~ nez-Pons , Miriam Paone , Francesco Castelluccio , Conxita Avila , Margherita Gavagnin a,* Consiglio Nazionale delle Ricerche, Istituto di Chimica Biomolecolare, Via Campi Flegrei 34, 80078-I Pozzuoli, Napoli, Italy Departament de Biologia Animal (Invertebrats), Facultat de Biologia, Universitat de Barcelona, Av. Diagonal 6, 08028-E Barcelona, Catalunya, Spain

The n-butanol extract of an Antarctic hexactinellid sponge, Anoxycalyx (Scolymastra) joubini, was found to contain a taurine-conjugated anthranilic acid, never reported so far either as a natural product or by synthesis. The compound was inactive against human cancer cells in an in vitro growth inhibitory test, and also showed no antibacterial activity.