Testing the physiological capacity of the mussel Mytilus chilensis to establish into the Southern Ocean.

The Southern Ocean and the Antarctic Circumpolar Current create environmental conditions that serve as an efficient barrier to prevent the colonization of non-native species (NNS) in the marine ecosystems of Antarctica. However, warming of the Southern Ocean and the increasing number of transport opportunities are reducing the physiological and physical barriers, increasing the chances of NNS arriving. The aim of this study was to determine the limits of survival of the juvenile mussels, M. chilensis, under current Antarctic conditions and those projected under climate change. These assessments were used to define the mussels potential for establishment in the Antarctic region. Experimental mussels were exposed to four treatments: -1.5 °C (Antarctic winter), 2 °C (Antarctic summer), 4 °C (Antarctic projected) and 8 °C (control) for 80 days and a combination of physiological and transcriptomics approaches were used to investigate mussel response. The molecular responses of mussels were congruent with the physiological results, revealing tolerance to Antarctic winter temperatures. However, a higher number of regulated differentially expressed gene (DEGs) were reported in mussels exposed to Antarctic winter temperatures (-1.5 °C). This tolerance was associated with the activation of the biological processes associated with apoptosis (up regulated) and both cell division and cilium assembly (down regulated). The reduced feeding rate and the negative scope for growth, for a large part of the exposure period at -1.5 °C, suggests that Antarctic winter temperatures represents an environmental barrier to M. chilensis from the Magellanic region settling in the Antarctic. Although M. chilensis are not robust to current Antarctica thermal conditions, future warming scenarios are likely to weaken these physiological barriers. These results strongly suggest that the West Antarctic Peninsula could become part of Mytilus distributional range, especially with dispersal aided by increasing maritime transport activity across the Southern Ocean.

On the horns of a dilemma: Evaluation of synthetic and natural textile microfibre effects on the physiology of the pacific oyster Crassostrea gigas.

Fast fashion and our daily use of fibrous materials cause a massive release of microfibres (MF) into the oceans. Although MF pollution is commonly linked to plastics, the vast majority of collected MF are made from natural materials (e.g. cellulose). We investigated the effects of 96-h exposure to natural (wool, cotton, organic cotton) and synthetic (acrylic, nylon, polyester) textile MF and their associated chemical additives on the capacity of Pacific oysters Crassostrea gigas to ingest MF and the effects of MF and their leachates on key molecular and cellular endpoints. Digestive and glycolytic enzyme activities and immune and detoxification responses were determined at cellular (haemocyte viability, ROS production, ABC pump activity) and molecular (Ikb1, Ikb2, caspase 1 and EcSOD expression) levels, considering environmentally relevant (10 MF L-1) and worst-case scenarios (10 000 MF L-1). Ingestion of natural MF perturbed oyster digestive and immune functions, but synthetic MF had few effects, supposedly related with fibers weaving rather than the material itself. No concentration effects were found, suggesting that an environmental dose of MF is sufficient to trigger these responses. Leachate exposure had minimal effects on oyster physiology. These results suggest that the manufacture of the fibres and their characteristics could be the major factors of MF toxicity and stress the need to consider both natural and synthetic particles and their leachates to thoroughly evaluate the impact of anthropogenic debris. Environmental Implication. Microfibres (MF) are omnipresent in the world oceans with around 2 million tons released every year, resulting in their ingestion by a wide array of marine organisms. In the ocean, a domination of natural MF- representing more than 80% of collected fibres-over synthetic ones was observed. Despite MF pervasiveness, research on their impact on marine organisms, is still in its infancy. The current study aims to investigate the effects of environmental concentrations of both synthetic and natural textile MF and their associated leachates on a model filter feeder.

Acclimation of the Antarctic sea urchin Sterechinus neumayeri to warmer temperatures involves a modulation of cellular machinery.

Global warming is threatening marine Antarctic fauna, which has evolved in isolation in a cold environment for millions of years. Facing increasing temperatures, marine Antarctic invertebrates can either tolerate or develop adaptations to these changes. On a short timescale, their survival and resistance to warming will be driven by the efficiency of their phenotypic plasticity through their capacity for acclimation. The current study aims at evaluating the capacity for acclimation of the Antarctic sea urchin Sterechinus neumayeri to predicted ocean warming scenarios (+2, RCP 2.6 and + 4 °C, RCP 8.5, IPCC et al., 2019) and deciphering the subcellular mechanisms underlying their acclimation. A combination of transcriptomics, physiological (e.g. growth rate, gonad growth, ingestion rate and oxygen consumption), and behavioral-based approaches were used on individuals incubated at 1, 3 and, 5 °C for 22 weeks. Mortality was low at warmer temperatures (20%) and oxygen consumption and ingestion rate seemed to reach a stable state around 16 weeks suggesting that S. neumayeri might be able to acclimate to warmer temperatures (until 5 °C). Transcriptomic analyses highlighted adjustments of the cellular machinery with the activation of replication, recombination, and repair processes as well as cell cycle and division and repression of transcriptional and signal transduction mechanisms and defense processes. These results suggest that acclimation to warmer scenarios might require more than 22 weeks for the Antarctic Sea urchins S. neumayeri but that projections of climate change for the end of the century may not strongly affect the population of S. neumayeri of this part of the Antarctic.

Evaluation of Sub-Antarctic and Antarctic sea urchins' thermal reaction norm through righting behavior and comparison with in situ measurements.

Sea urchin's survival may depend on their capacity to recover proper orientation rapidly and effectively after inversion, enabling escape from predator and preventing desiccation. This righting behavior has been used as a repeatable and reliable indicator to assess echinoderms performance across environmental conditions, including thermal sensitivity and thermal stress. The current study aims at evaluating and comparing the thermal reaction norm for righting behavior (time for righting (TFR) and capacity to self-right) of three common sea urchins from high latitude, the Patagonian sea urchins Loxechinus albus and Pseudechinus magellanicus, and the Antarctic sea urchin Sterechinus neumayeri. In addition, to infer the ecological implications of our experiments, we compared laboratory-based and in situ TFR of these three species. We observed that populations of the Patagonian sea urchins L. albus and P. magellanicus presented similar trend of righting behavior, overly accelerating with increasing temperature (from 0 to 22°C). Little variations and high inter-individual variability were observed below 6°C in the Antarctic sea urchin TFR, and righting success strongly decreased between 7 and 11°C. For the three species, TFR was lower in in situ experiments compared to the laboratory. Overall, our results suggest that the populations of Patagonian sea urchin exhibit a wide thermal tolerance and, based on S. neumayeri's TFR, aligning with the narrow thermal tolerance of Antarctic benthos. Finally, the differences between laboratory and in situ experiments highlights the importance of considering the complexity of marine environments for future predictions.

Evolutionary constraints on physiology confound range shift predictions of two nacellid limpets.

Physiological comparisons are fundamental to quantitative assessments of the capacity of species to persist within their current distribution and to predict their rates of redistribution in response to climate change. Yet, the degree to which physiological traits are conserved through evolutionary history may fundamentally constrain the capacity for species to adapt and shift their geographic range. Taxa that straddle major climate transitions provide the opportunity to test the mechanisms underlying evolutionary constraints and how such constraints may influence range shift predictions. Here we focus on two abundant and shallow water nacellid limpets which have representative species on either side of the Polar front. We test the thermal thresholds of the Southern Patagonian limpet, Nacella deaurata and show that its optimal temperatures for growth (4 °C), activity (-1.2 to -0.2 °C) and survival (1 to 8 °C) are mismatched to its currently experienced annual sea surface temperature range (5.9 to 10 °C). Comparisons with the congeneric Antarctic limpet, N. concinna, reveal an evolutionary constraint on N. deaurata physiology, with overlapping thermal capacities, suggesting that a cold climate legacy has been maintained through the evolution of these species. These physiological assessments predict that the South American range of N. deaurata will likely decline with continued warming. It is, however, one of the first species with demonstrated physiological capacity to successfully colonize the cold Southern Ocean. With the expected increase in opportunities for transport within high southern latitudes, N. deaurata has the potential to establish and drive ecological change within the shallow Southern Ocean.

Unveiling the unknown phylogenetic position of the scallop Austrochlamys natans and its implications for marine stewardship in the Magallanes Province.

Two species of scallop, Austrochlamys natans ("Ostión del Sur") and Zygochlamys patagonica ("Ostión patagonico") are presently exploited in the southern part of the Magallanes Province (MP). The lack of clarity in taxonomic identification and ecological aspects is generating both erroneous extraction statistics and an unperceived harvesting pressure on A. natans and Z. patagonica. We aim to discriminate these Magallanes scallops accurately, improve our understanding of their complex natural history and discuss possible implications for their management and conservation status, given the current fisheries statistics. To achieve these goals, we present a complete review of the historical identification of the Magallanes scallop and a multi-locus molecular phylogeny which allowed us to recover the phylogenetic position of A. natans. We sampled 54 individuals from five localities across the southern Pacific coast of the MP. We calculated the depth of the byssal notch (BND) and shell height (VH) ratio from morphological characters and conducted phylogenetic reconstructions with mitochondrial (12S and 16S) and nuclear markers (28S) using Bayesian and maximum likelihood analyses. Both morphology and molecular phylogeny identified two distinct entities, Z. patagonica and a distinct, highly divergent lineage that corresponds to A. natans. Our study provides integrative evidence to alert the current fishery management and the need for further conservation studies.

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

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

Species vulnerability under climate change: Study of two sea urchins at their distribution margin.

In order to develop powerful predictions on the impact of climate change on marine organisms, it is critical to understand how abiotic drivers such as temperature can directly and indirectly affect marine organisms. Here, we evaluated and compared the physiological vulnerability of the leading-edge populations of two species of sea urchins Loxechinus albus and Pseudechinus magellanicus in response to predicted ocean warming and food limitation. After exposing sea urchins to a 60-day experimental period to contrasting temperature (1 °C, 7 °C and 14 °C corresponding respectively to the actual average summer temperature in Antarctica, the control treatment temperature and the predicted future temperature in the Strait of Magellan) and diet levels (ad libitum or food limitation), sea urchin stress tolerance was assessed. Sea urchins' physiology was measured at the organismal and sub-cellular level by studying the organisms energy balance (behavior, growth, gonad index, ingestion rate, O2 uptake, energy reserves) and the expression of genes associated with aerobic metabolism. Our results showed that at their distribution edge, and despite their distinct geographical repartition, both species might be resilient to ocean warming. However, the combination of ocean warming and food limitation reduced the stress tolerance of sea urchins. In a warming ocean, another strategy could be to migrate toward the pole to a cooler environment but incubation at 1 °C resulted in a diminution of both species' aerobic scope. Overall, if these engineer species are unable to acclimate to food limitation under future climate, population fitness could be affected with ecological and economic consequences.

First mussel settlement observed in Antarctica reveals the potential for future invasions.

Global biodiversity is both declining and being redistributed in response to multiple drivers characterizing the Anthropocene, including synergies between biological invasions and climate change. The Antarctic marine benthos may constitute the last biogeographic realm where barriers (oceanographic currents, climatic gradients) have not yet been broken. Here we report the successful settlement of a cohort of Mytilus cf. platensis in a shallow subtidal habitat of the South Shetland Islands in 2019, which demonstrates the ability of this species to complete its early life stages in this extreme environment. Genetic analyses and shipping records show that this observation is consistent with the dominant vectors and pathways linking southern Patagonia with the Antarctic Peninsula and demonstrates the potential for impending invasions of Antarctic ecosystems.

Combined effects of warming and freshening on the physiological energetics of the edible whelk Trophon geversianus.

The interacting effects of climate change pressures and human use of natural resources are increasingly affecting marine biodiversity. Variations in key abiotic factors such as temperature and salinity may therefore negatively influence marine organisms that are already threatened by intensive fisheries. Herein, we tested the hypothesis that future ocean warming and freshening will affect the fitness and survival of the overexploited snail Trophon geversianus in Southern Patagonia. To test this hypothesis, we investigated the effect of a 50 day incubation period of five temperatures (1, 5, 9, 12 and 15 °C) and two salinities (25 and 30 psu), (which correspond to current and projected conditions for Antarctic and Sub-Antarctic regions), on the physiological energetics (ingestion rate, absorption efficiency, oxygen uptake and scope for growth (SFG)) of the edible whelk T. geversianus. Our results showed no significant effects for salinity or the combination of temperature and salinity on T. geversianus bioenergetics. On the contrary, incubation at low temperatures (1 and 5 °C) was shown to affect the ingestion rate, absorption efficiency, oxygen uptake and SFG for T. geversianus, whereas for specimens incubated at 12 and 15 °C, physiological rates remained similar to control. Our data suggests that T. geversianus might be robust to warming and future variations of salinity, but longer term experiments are needed to ensure that no reduction of performance will occur after an extended incubation time from an increase in temperature.

Transcriptome mining of apoptotic mechanisms in response to density and functional diets in Oncorhynchus mykiss and role in homeostatic regulation.

Husbandry conditions often expose fish to several stressors, compromising organism's global homeostasis, which has consequences on aquaculture production. In order to depict intestinal homeostatic responses to deleterious conditions and the potential beneficial effects of functional diets in aquaculture fish, the effects of chronic suboptimal density condition on the neuro-immune endocrine system of rainbow trout was investigated through a transcriptomic survey. After 30 days under high stocking density, 67% of genes involved in homeostasis were found down-regulated whereas when fed with a diet supplemented with the probiotic Saccharomyces cerevisiae, <1% of these genes were found down-regulated. Genes involved in apoptotic mechanisms were found to be particularly responsive to both density and diets. At high density, several genes involved in the intrinsic and extrinsic apoptotic pathway (Fadd, Fas, Bcl-2 and Bax) as well as caspases (Casp8 and Casp3) were found down-regulated (Fold change <-7). However, fish fed with a diet supplemented with the probiotic yeast had an opposite response, indicating mitigation of stress effects. Overall, these results indicate the activation of homeostatic machinery and particularly genes involved in the apoptotic pathway as a result of probiotic feeding, counteracting apoptotic molecular repression derived from chronic stress.

Global host molecular perturbations upon in situ loss of bacterial endosymbionts in the deep-sea mussel Bathymodiolus azoricus assessed using proteomics and transcriptomics.

BACKGROUND: Colonization of deep-sea hydrothermal vents by most invertebrates was made efficient through their adaptation to a symbiotic lifestyle with chemosynthetic bacteria, the primary producers in these ecosystems. Anatomical adaptations such as the establishment of specialized cells or organs have been evidenced in numerous deep-sea invertebrates. However, very few studies detailed global inter-dependencies between host and symbionts in these ecosystems. In this study, we proposed to describe, using a proteo-transcriptomic approach, the effects of symbionts loss on the deep-sea mussel Bathymodiolus azoricus' molecular biology. We induced an in situ depletion of symbionts and compared the proteo-transcriptome of the gills of mussels in three conditions: symbiotic mussels (natural population), symbiont-depleted mussels and aposymbiotic mussels. RESULTS: Global proteomic and transcriptomic results evidenced a global disruption of host machinery in aposymbiotic organisms. We observed that the total number of proteins identified decreased from 1118 in symbiotic mussels to 790 in partially depleted mussels and 761 in aposymbiotic mussels. Using microarrays we identified 4300 transcripts differentially expressed between symbiont-depleted and symbiotic mussels. Among these transcripts, 799 were found differentially expressed in aposymbiotic mussels and almost twice as many in symbiont-depleted mussels as compared to symbiotic mussels. Regarding apoptotic and immune system processes - known to be largely involved in symbiotic interactions - an overall up-regulation of associated proteins and transcripts was observed in symbiont-depleted mussels. CONCLUSION: Overall, our study showed a global impairment of host machinery and an activation of both the immune and apoptotic system following symbiont-depletion. One of the main assumptions is the involvement of symbiotic bacteria in the inhibition and regulation of immune and apoptotic systems. As such, symbiotic bacteria may increase their lifespan in gill cells while managing the defense of the holobiont against putative pathogens.

Coding/non-coding cross-talk in intestinal epithelium transcriptome gives insights on how fish respond to stocking density.

Fish respond to increasing stocking density as a stressor, adjusting physiological functions to increase energy supply for coping with deleterious effects and adapting. These responses are complex and systemic, and the molecular mechanisms involved remain undetermined. One of the most sensitive organs to environmental and homeostatic disruptions is the intestine, and since it plays several vital functions, understanding the molecular underpinnings of this organ under deleterious conditions is imperative for health improvement in aquaculture systems. This study aimed to understand how different stocking densities of rainbow trout (Oncorhynchus mykiss) modulates the coding and non-coding RNAs profiling, that in turn, play key roles to maintain the fish homeostasis. For this, the intestine tissue of juvenile trout stocked for 30 days either at low (LD: 3 kg m-3) or high density (HD: 40 kg m-3) were sampled to isolate total RNA and then construct cDNA libraries for an illumina sequencing platform. Differential gene expression analysis revealed a generalized downregulation of transcripts, including coding and long non-coding RNAs (lncRNAs). Notably, significant differences in transcripts involved in metabolic pathways, as well as immune and epithelium integrity and stability related pathways were found. A high number of downregulated transcripts enriched these pathways, and a strong correlation was observed between the most differentially expressed transcripts and the highly expressed lncRNAs. This study suggests a cross-talk between coding and non-coding RNAs in the intestine of fish exposed to suboptimal conditions, providing new insights into the regulatory role of the lncRNAs on fish response to stressors.

Single and repetitive microplastics exposures induce immune system modulation and homeostasis alteration in the edible mussel Mytilus galloprovincialis.

Seashore invertebrates such as mussels are exposed to multiple bouts of pollution related to human activities. Plastic debris originating from land-based activities are a concerning issue as they may be fragmented in smaller pieces (microplastics, < 5 mm diameter) which have an excellent potential for uptake by a large variety of animals. Here, we set out to explore the whole transcriptome profiling of Mytilus galloprovincialis associated with temporal variability of microplastics concentrations. Mussels were submitted to (i) a single 18 days-exposure to a concentration of microplastics found during pollution events (4.6 E+5 polyethylene microbeads L-1), (ii) a recovery period to investigate the reversibility of microplastics effects and (iii) a repeated exposure to microplastics to evidence acclimation to microplastics pollution events. Overall, 18 days-exposure to microplastics was mostly associated with disruption of mussel global homeostasis resulting in the production of stress and immune-related proteins and as a consequence, a diminution of energy allocated to growth. During the recovery period, a contrasting response was observed with the activation of apoptotic processes and the up-regulation of immune-receptors and stress-related proteins (glutathione peroxidase, hsp70) in mussels previously exposed to microplastics. These divergent responses, suggest that the establishment of compensatory mechanism as an attempt to recover, is not sufficient to counteract physiological stress induced by the first exposure. Finally, the differences observed in gene expression between single and repeated exposures to microplastics suggest, under the experimental conditions tested, that mussels may be able to establish a stress-memory upon microplastics exposure.

Transcriptome mining of immune-related genes in the muricid snail Concholepas concholepas.

The population of the Chilean endemic marine gastropod Concholepas concholepas locally called "loco" has dramatically decreased in the past 50 years as a result of intense activity of local fisheries and high environmental variability observed along the Chilean coast, including episodes of hypoxia, changes in sea surface temperature, ocean acidification and diseases. In this study, we set out to explore the molecular basis of C. concholepas to cope with biotic stressors such as exposure to the pathogenic bacterium Vibrio anguillarum. Here, 454pyrosequencing was conducted and 61 transcripts related to the immune response in this muricid species were identified. Among these, the expression of six genes (CcNFκß, CcIκß, CcLITAF, CcTLR, CcCas8 and CcCath) involved in the regulation of inflammatory, apoptotic and immune processes upon stimuli, were evaluated during the first 33 h post challenge (hpc). The results showed that CcTLR, CcCas8 and CcCath have an initial response at 4 hpc, evidencing an up-regulation from 4 to 24 hpc. Notably, the response of CcNFKB occurred 2 h later with a statistically significant up-regulation at 6 hpc and 10 hpc. Furthermore, the challenge with V. anguillarum induced a statistically significant down-regulation of CcIKB between 2 and 10 hpc as well as a down-regulation of CcLITAF between 2 and 4 hpc followed in both cases by an up-regulation between 24 and 33 hpc. This work describes the first transcriptomic effort to characterize the immune response of C. concholepas and constitutes a valuable transcriptomic resource for future efforts to develop sustainable aquaculture and conservations tools for this endemic marine snail species.

Ocean acidification and pathogen exposure modulate the immune response of the edible mussel Mytilus chilensis.

Ocean acidification (OA) is one of the main consequences of increasing atmospheric carbon dioxide (CO2), impacting key biological processes of marine organisms such as development, growth and immune response. However, there are scarce studies on the influence of OA on marine invertebrates' ability to cope with pathogens. This study evaluated the single and combined effects of OA and bacterial infection on the transcription expression of genes related to antioxidant system, antimicrobial peptides and pattern recognition receptors in the edible mussel Mytilus chilensis. Individuals of M. chilensis were exposed during 60 days at two concentrations of pCO2 (550 and 1200 µatm) representing respectively current and future scenario of OA and were then injected with the pathogenic bacterium Vibrio anguillarum. Results evidenced an immunomodulation following the OA exposure with an up-regulation of C-type Lectin and Mytilin B and a down-regulation of Myticin A and PGRP. This immunomodulation pattern is partially counteracted after challenge with V. anguillarum with a down-regulation of the C-type lectin and Mytilin B and the up-regulation of Myticin A. In turn, these results evidence that pCO2-driven OA scenarios might triggers specific immune-related genes at early stages of infection, promoting the transcription of antimicrobial peptides and patterns recognition receptors. This study provides new evidence of how the immune response of bivalves is modulated by higher CO2 conditions in the ocean, as well one factor for the resilience of marine population upon global change scenarios.

Functional Diets Modulate lncRNA-Coding RNAs and Gene Interactions in the Intestine of Rainbow Trout Oncorhynchus mykiss.

The advent of functional genomics has sparked the interest in inferring the function of non-coding regions from the transcriptome in non-model species. However, numerous biological processes remain understudied from this perspective, including intestinal immunity in farmed fish. The aim of this study was to infer long non-coding RNA (lncRNAs) expression profiles in rainbow trout (Oncorhynchus mykiss) fed for 30 days with functional diets based on pre- and probiotics. For this, whole transcriptome sequencing was conducted through Illumina technology, and lncRNAs were mined to evaluate transcriptional activity in conjunction with known protein sequences. To detect differentially expressed transcripts, 880 novels and 9067 previously described O. mykiss lncRNAs were used. Expression levels and genome co-localization correlations with coding genes were also analyzed. Significant differences in gene expression were primarily found in the probiotic diet, which had a twofold downregulation of lncRNAs compared to other treatments. Notable differences by diet were also evidenced between the coding genes of distinct metabolic processes. In contrast, genome co-localization of lncRNAs with coding genes was similar for all diets. This study contributes novel knowledge regarding lncRNAs in fish, suggesting key roles in salmons fed with in-feed additives with the capacity to modulate the intestinal homeostasis and host health.

Identification and gene expression of multiple peptidoglycan recognition proteins (PGRPs) in the deep-sea mussel Bathymodiolus azoricus, involvement in symbiosis?

The relationship between the deep-sea mussel Bathymodiolus azoricus and its thiotrophic (SOX) and methanotrophic (MOX) symbionts has been ecologically and functionally well studied. Endosymbiosis is common in deep-sea hydrothermal vent fauna, yet little is known about the molecular mechanisms underlying the regulation of interactions between host and symbionts. In this study we focused on a group of pattern recognition receptors (PRR), called PGRPs that are able to recognize the peptidoglycan of bacterial cell wall. We first characterised the different PGRPs isoforms in B. azoricus gills and identified five paralogs. Among them two displayed a signal peptide. Then, specific probes designed for each paralog were used to perform real-time PCR quantification in gills of individuals showing various bacterial content as a result of in situ experimental procedures. Overall we found a decrease of PGRPs expression when symbionts amount decreases, suggesting an implication of PGRPs in the regulation of symbionts in B. azoricus gills. We therefore hypothesize that secreted proteins could act as cooperation signals to induce colonisation of symbiotic tissue while non-secreted proteins may regulate the density of endosymbionts within the gill tissue.

Long non-coding RNAs are associated with spatiotemporal gene expression profiles in the marine gastropod Tegula atra.

Increasing evidence suggests that long non-coding RNAs (lncRNAs) play diverse roles in cellular processes, including in the regulation of embryogenesis and growth. However, little is known about the role of lncRNAs in marine invertebrates inhabiting changing environments. Therefore, the aim of this study was to present the first characterization of lncRNAs in an intertidal marine gastropod. Specifically, Tegula atra individuals were sampled in four sites of the central-northern Chilean coastline (28-31°) during summer and winter. A pipeline was constructed, and 3524 putative lncRNAs were identified from transcriptome databases specific to T. atra. These lncRNAs exhibited characteristics common to known lncRNAs, including a length shorter than coding sequences, low GC-content, and low sequence conservation. Expression analyses revealed that lncRNAs varied more in the summer. Furthermore, a majority of the differentially expressed lncRNAs were found in the southernmost population, the seasonal temperatures of which varied the greatest among all groups. Additionally, co-expression analysis found some lncRNAs strongly correlated with coding genes involved in the environmental stress response, such as heat shock proteins and metalloproteins. In contrast, other lncRNA expressions were strongly uncorrelated with genes involved in lipid/carbohydrates metabolism and cell-cell communication. This study provides the first large-scale characterization of lncRNAs in a marine gastropod, with results suggesting a putative role of lncRNAs in thermal tolerance, as well as an association with molecular mechanisms involved in the local adaptations of marine invertebrate populations.

Uncovering the Complex Transcriptome Response of Mytilus chilensis against Saxitoxin: Implications of Harmful Algal Blooms on Mussel Populations.

Saxitoxin (STX), a principal phycotoxin contributing to paralytic shellfish poisoning, is largely produced by marine microalgae of the genus Alexandrium. This toxin affects a wide range of species, inducing massive deaths in fish and other marine species. However, marine bivalves can resist and accumulate paralytic shellfish poisons. Despite numerous studies on the impact of STX in marine bivalves, knowledge regarding STX recognition at molecular level by benthic species remains scarce. Therefore, the aim of this study was to identify novel genes that interact with STX in the Chilean mussel Mytilus chilensis. For this, RNA-seq and RT-qPCR approaches were used to evaluate the transcriptomic response of M. chilensis to a purified STX as well as in vivo Alexandrium catenella exposure. Approximately 800 million reads were assembled, generating 138,883 contigs that were blasted against the UniProt Mollusca database. Pattern Recognition Receptors (PRRs) involved in mussel immunity, such as Toll-like receptors, tumor necrosis factor receptors, and scavenger-like receptors were found to be strongly upregulated at 8 and 16 h post-STX injection. These results suggest an involvement of PRRs in the response to STX, as well as identifying potential, novel STX-interacting receptors in this Chilean mussel. This study is the first transcriptomic overview of the STX-response in the edible species M. chilensis. However, the most significant contribution of this work is the identification of immune receptors and pathways potentially involved in the recognition and defense against STX's toxicity and its impact of harmful algae blooms on wild and cultivated mussel populations.