Marine Invertebrate Species Research Articles

Retention of prions in the polychaete Hediste diversicolor and black soldier fly, Hermetia illucens, larvae after short-term experimental immersion and feeding with brain homogenate from scrapie infected sheep

Finding alternative protein and lipid sources for aquafeeds is crucial for the sustainable growth of fed aquaculture. Upcycling industrial side streams and byproducts using extractive species can reduce waste and help reduce the sector's dependence on fish meal and fish oils. Polychaete worms (Hediste diversicolor) and black soldier fly (Hermetia illucens) larvae (BSFL) are promising candidates for converting waste materials into valuable protein and lipid sources. However, further research and evaluations are needed to ensure the safety and regulatory compliance of these alternative feed sources, especially regarding prions spreading potential in the unlikely case that prions would be introduced in the value chain via feedstocks. In the present investigation, BSFL and juvenile polychaetes that had received a massive dose of scrapie prions through immersion and oral inoculation were found to harbour detectable prions using an ultrasensitive amplification method known as PMCA. This observation suggests that both H. diversicolor and BSFL have the potential to serve as mechanical vectors for prions diseases. However, it is important to note that insects, lacking the prion protein gene, are incapable of propagating prions. Therefore, the quantity of prions present in the larvae will inevitably be lower than the amount of prions they encountered. This is the first study to report on the fate of prions through ingestion by these marine and terrestrial invertebrate species.

Acetylcholinesterase and dopamine inhibition suppress the filtration rate, burrowing behaviours, and immunological responses induced by bisphenol A in the hemocytes and gills of date mussels, Lithophaga lithophaga

Bisphenol A (BPA), a common industrial chemical with estrogenic activity, has recently gained attention due to its well-documented negative effects on humans and other organisms in the environment. The potential immunotoxicity and neurotoxicity of BPA remain poorly understood in marine invertebrate species. Therefore, the impacts of exposure to BPA on a series of behaviours, immune responses, oxidative stress, neural biomarkers, histology, and the ultrastructure of gills were investigated in the date mussel, Lithophaga lithophaga. After 28 days of exposure to 0.25, 1, 2, and 5 µg/L BPA, hemolymphs from controls and exposed date mussels were collected, and the effects of BPA on immunological parameters were evaluated. Moreover, oxidative stress and neurochemical levels were measured in the gills of L. lithophaga. BPA reduced filtration rates and burrowing behaviour, whereas a 2 µg/L BPA resulted in an insignificant increase after 24 h. The exposure of date mussels to BPA significantly increased total hemocyte counts, a significant reduction in the diameter and phagocytosis of hemocytes, as well as gill lysozyme level. BPA increased lipid peroxidation levels and SOD activity in gills exposed to 2 and 5 µg/L BPA, but decreased GSH levels and SOD activity in 0.25 and 1 µg/L BPA-treated date mussels. Dose-dependent dynamics were observed in the inhibition of acetylcholinesterase activity and dopamine levels. Histological and scanning electron microscope examination revealed cilia erosion, necrosis, inflammation, and hyperplasia formation in the gills. Overall, our findings suggest a relationship between BPA exposure and changes in the measured immune parameters, oxidative stress, and neurochemical disturbances, which may be factored into the mechanisms underlying BPA toxicity in marine molluscs, providing a scientific foundation for marine BPA risk assessment and indicating immunosuppression in BPA-exposed date mussels.

Molecular taxonomy of fish larvae in the Northwestern Arabian gulf: A baseline study from Kuwait's first marine protected area

Sulaibikhat Bay, a shallow embayment dominated by tides and extensive mud flats along the northern coast of Kuwait Bay, serves as a crucial nursery ground for various commercially important species of finfish and shellfish in the Northwestern Arabian Gulf (NWAG). As the location of Kuwait's inaugural marine protected area (MPA), Sulaibikhat Bay plays a vital role in fostering numerous fish and marine invertebrate species of commercial importance in the Arabian/Persian Gulf. Accurate identification of larval specimens from this habitat is essential for understanding their spawning dynamics and recruitment process. However, limitations in taxonomic identification hinder scientific understanding of fish larval recruitment. To address this, our baseline study employed an integrated taxonomic approach, combining conventional morphology with molecular techniques. Fish larvae collected from the MPA and surrounding waters in Spring (2022) were identified up to the genus and species levels using partial gene sequencing of 16 S rRNA and mitochondrial cytochrome oxidase (COI) genes. Morphological analysis indicated the prevalence of Sparidae, Sillaginidae, and Dorosomatidae families. Molecular analysis confirmed the presence of these families and identified two species, Crenidens crenidens (Sparidae) and Nematalosa nasus (Dorosomatidae), along with two genera, Diplodus sp. (Sparidae) and Sillago sp. (Sillaginidae), which exhibited maximum similarity to available COI sequences of Diplodus sargus, and Sillago maculata, respectively. This represents the first baseline molecular study on fish larvae from NWAG, laying the groundwork for future in-depth analyses with more larval specimens and spatiotemporal samplings from the entire exclusive economic zone (EEZ) of Kuwait in the Arabian/Persian Gulf.

Living in a dynamic environment: The effects of multi-ways temperature variation on embryo and newborn juveniles of a shallow-water octopus (Amphioctopus fangsiao)

Shallow waters are characterized by fluctuating environmental conditions, modulating marine life cycles and biological phenomena. Multiple variations in water temperature could affect eggs and embryos during spawning events of many marine invertebrate species, yet most of the findings on embryonic development in invertebrates come from experiments based on the constant temperature. In this study, to examine the effects of temperature variation on octopus embryos, Amphioctopus fangsiao, a common shallow-water octopus along the coast of China, was exposed to the constant temperature (18 °C, in situ temperature of the seawater in Lianyungang), ramping temperatures (from 18 to 24 °C), diel oscillating temperatures (18 °C and 20 °C for 12 h each day), and acute increasing temperatures (the temperature increased sharply from 18 °C to 24 °C at embryonic development stage XIX) for 47 days (from embryogenesis to settlement). The results demonstrated that the temperature variations accelerated the development time of A. fangsiao embryos. Temperature fluctuations could cause embryonic oxidative damage and disorder of glycolipid metabolism, thereby affecting the growth performance of embryos and the survival rate of hatchings. Through transcriptome sequencing, the mechanistic adaption of the embryo to environmental temperature variations was revealed. The pathways involved in the TCA cycle, DNA replication and repair, protein synthesis, cell signaling, and nervous system damage repair were significantly enriched, indicating that the embryo could improve heat tolerance to thermal stress by regulating gene expression. Moreover, acute warming temperatures posed the most detrimental effects on A. fangsiao embryos, which could cause embryos to hatch prematurely from the vegetal pole, further reducing the survival of hatchings. Meanwhile, the diel oscillating temperature was observed to affect the normal morphology of the embryo, resulting in embryo deformities. Thus, the constant temperature is critical for balanced growth and defense status in octopuses by maintaining metabolism homeostasis. For the first time, this study evaluates the effects of multiple temperature fluctuations on embryos of A. fangsiao, providing new insights into the physiological changes and molecular responses of cephalopod embryos following dynamic temperature stress.

Dealing with biases introduced by lipids in stable carbon and nitrogen isotope analyses: a solution based on 28 marine invertebrate, fish, and mammal species

Dealing with biases introduced by lipids in stable carbon and nitrogen isotope analyses: a solution based on 28 marine invertebrate, fish, and mammal species

Comparative study of domoic acid accumulation, isomer content and associated digestive subcellular processes in five marine invertebrate species

Despite the deleterious effects of the phycotoxin domoic acid (DA) on human health, and the permanent threat of blooms of the toxic Pseudo-nitzschia sp. over commercially important fishery-resources, knowledge regarding the physiological mechanisms behind the profound differences in accumulation and depuration of this toxin in contaminated invertebrates remain very scarce. In this work, a comparative analysis of accumulation, isomer content, and subcellular localization of DA in different invertebrate species was performed. Samples of scallops Pecten maximus and Aequipecten opercularis, clams Donax trunculus, slippersnails Crepidula fornicata, and seasquirts Asterocarpa sp. were collected after blooms of the same concentration of toxic Pseudo-nitzschia australis. Differences (P < 0.05) in DA accumulation were found, wherein P. maximus showed up to 20-fold more DA in the digestive gland than the other species. Similar profiles of DA isomers were found between P. maximus and A. opercularis, whereas C. fornicata was the species with the highest biotransformation rate (∼10 %) and D. trunculus the lowest (∼4 %). DA localization by immunohistochemical analysis revealed differences (P < 0.05) between species: in P. maximus, DA was detected mainly within autophagosome-like vesicles in the cytoplasm of digestive cells, while in A. opercularis and C. fornicata significant DA immunoreactivity was found in post-autophagy residual bodies. A slight DA staining was found free within the cytoplasm of the digestive cells of D. trunculus and Asterocarpa sp. The Principal Component Analysis revealed similarities between pectinids, and a clear distinction of the rest of the species based on their capabilities to accumulate, biotransform, and distribute the toxin within their tissues. These findings contribute to improve the understanding of the inter-specific differences concerning the contamination-decontamination kinetics and the fate of DA in invertebrate species.

Genomic DNA extraction optimization and validation for genome sequencing using the marine gastropod Kellet’s whelk

Next-generation sequencing technologies, such as Nanopore MinION, Illumina Hiseq and Novaseq, and PacBio Sequel II, hold immense potential for advancing genomic research on non-model organisms, including the vast majority of marine species. However, application of these technologies to marine invertebrate species is often impeded by challenges in extracting and purifying their genomic DNA due to high polysaccharide content and other secondary metabolites. In this study, we help resolve this issue by developing and testing DNA extraction protocols for Kellet’s whelk (Kelletia kelletii), a subtidal gastropod with ecological and commercial importance, by comparing four DNA extraction methods commonly used in marine invertebrate studies. In our comparison of extraction methods, the Salting Out protocol was the least expensive, produced the highest DNA yields, produced consistent high DNA quality, and had low toxicity. We validated the protocol using an independent set of tissue samples, then applied it to extract high-molecular-weight (HMW) DNA from over three thousand Kellet’s whelk tissue samples. The protocol demonstrated scalability and, with added clean-up, suitability for RAD-seq, GT-seq, as well as whole genome sequencing using both long read (ONT MinION) and short read (Illumina NovaSeq) sequencing platforms. Our findings offer a robust and versatile DNA extraction and clean-up protocol for supporting genomic research on non-model marine organisms, to help mediate the under-representation of invertebrates in genomic studies.

Long-term evolution of the structure of the St. Lawrence (Canada) marine ecosystem in the context of climate change and anthropogenic activities: An isotopic perceptive.

Documenting long-term changes in the trophic structure of food webs and how species respond to these changes is essential to forecast their vulnerability and resilience to environmental stressors. Over the past decades, the St. Lawrence marine ecosystem (Canada) has experienced major changes in its physical, chemical, and biological conditions from overfishing, acoustic and chemical pollution, climate change, and the increased abundance of some top predators. These changes have likely affected the trophodynamics of the ecosystem, and are suspected to have deleterious effects on endangered species of mammals and other components of the ecosystem, such as blue whales (Balaenoptera musculus), fin whales (B. physalus), and beluga (Delphinapterus leucas). This study examined the trophic structure of the St. Lawrence marine ecosystem, including the isotopic niche of various species, over two periods of contrasting pressures from anthropogenic and climatic stressors (1995-2003 vs. 2019-2021). Stable isotope ratios were measured in 1240 samples of 21 species of marine invertebrates, fishes, and mammals sampled during both periods. A significant change in the isotopic value and niche position between periods is observed in most of the sampled species. While the direction of change and effect size were not uniform among species, these changes confirmed that substantial modifications in community structure have occurred over time. Niche overlap decreased considerably among some of the pelagic and demersal fishes, and among whale species during the most recent period. Combined with a concomitant reduction in niche breadth in several species, these observations suggested that resource sharing was limited among these species. This study highlighted some degree of dietary plasticity in several species, and a long-term change in the trophic structure of the St. Lawrence marine ecosystem, with likely effects on diet composition and energetics of several populations, including endangered species.

Comparative transcriptome analysis reveals key genes and pathways related to gonad development in the sea cucumber Apostichopus japonicus

The sea cucumber Apostichopus japonicus is an economically important marine species in China, and understanding the mechanisms underlying its gonad development is crucial for successful reproduction and breeding. In this study, we performed transcriptome comparisons and analyses of A. japonicus gonadal and non-gonadal tissues to identify genes and molecular pathways associated with gonadal development. We also supplemented the annotation of the A. japonicus genome. Collectively, results revealed a total of 941 ovary-specific genes and 2499 testis-specific genes through different expression analysis and WGCNA analysis. The most enriched pathways in ovary and testis were “DNA replication” and “purine metabolism”, respectively. Additionally, we identified key candidate gene modules that control gonad development and germ cell maturation, with CDT1 and DYNC2LI1 serving as hub genes. Our findings provide important insights into the gonadal development system of A. japonicus and offer valuable references for further research on reproductive biology in this marine invertebrate species.

Fertilization Mode Covaries with Body Size.

AbstractThe evolution of internal fertilization has occurred repeatedly and independently across the tree of life. As it has evolved, internal fertilization has reshaped sexual selection and the covariances among sexual traits, such as testes size, and gamete traits. But it is unclear whether fertilization mode also shows evolutionary associations with traits other than primary sex traits. Theory predicts that fertilization mode and body size should covary, but formal tests with phylogenetic control are lacking. We used a phylogenetically controlled approach to test the covariance between fertilization mode and adult body size (while accounting for latitude, offspring size, and offspring developmental mode) among 1,232 species of marine invertebrates from three phyla. Within all phyla, external fertilizers are consistently larger than internal fertilizers: the consequences of fertilization mode extend to traits that are only indirectly related to reproduction. We suspect that other traits may also coevolve with fertilization mode in ways that remain unexplored.

Marine heatwave temperatures enhance larval performance but are meditated by paternal thermal history and inter-individual differences in the purple sea urchin, Strongylocentrotus purpuratus.

Marine heatwave (MHW) events, characterized by periods of anomalous temperatures, are an increasingly prevalent threat to coastal marine ecosystems. Given the seasonal phenology of MHWs, the full extent of their biological consequences may depend on how these thermal stress events align with an organism's reproductive cycle. In organisms with more complex life cycles (e.g., many marine invertebrate species) the alignment of adult and larval environments may be an important factor determining offspring success, setting the stage for MHW events to influence reproduction and development in situ. Here, the influence of MHW-like temperatures on the early development of the California purple sea urchin, Strongylocentrotus purpuratus, were explored within the context of paternal thermal history. Based on temperature data collected during MHW events seen in Southern California from 2014-2020, adult urchins were acclimated to either MHW or non-MHW temperatures for 28days before their sperm was used to produce embryos that were subsequently raised under varying thermal conditions. Once offspring reached an early larval stage, the impact of paternal and offspring environments were assessed on two aspects of offspring performance: larval size and thermal tolerance. Exposure to elevated temperatures during early development resulted in larger, more thermally tolerant larvae, with further influences of paternal identity and thermal history, respectively. The alignment of paternal and offspring exposure to MHW temperatures had additional positive benefits on larval thermal tolerance, but this tolerance significantly decreased when their thermal experience mismatched. As the highest recorded temperatures within past MHW events have occurred during the gametogenesis of many kelp forest benthic marine invertebrate species, such as the purple sea urchin, such parental mediated impacts may represent important drivers of future recruitment and population composition for these species.

Toxicity and mutagenicity studies of 6PPD-quinone in a marine invertebrate species and bacteria.

N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD-quinone), an oxidation product of the tire additive, 6PPD, has been associated with high mortality of salmonids (0.1 μg/L). The objective of this study was to determine the acute toxicity using neonates and mutagenicity (micronuclei in hemolymph of exposed adults) of 6PPD-quinone in the marine amphipod Parhyale hawaiensis. Also, we studied its mutagenicity in the Salmonella/microsome assay using five strains of Salmonella with and without metabolic system (rat liver S9, 5%). 6PPD-quinone did not present acute toxicity to P. hawaiensis from 31.25 to 500 μg/L. Micronuclei frequency increased after 96 h-exposure to 6PPD-quinone (250 and 500 μg/L) when compared to the negative control. 6PPD-quinone also showed a weak mutagenic effect for TA100 only in the presence of S9. We conclude that 6PPD-quinone is mutagenic to P. hawaiensis and weakly mutagenic to bacteria. Our work provides information for future risk assessment of the presence of 6PPD-quinone in the aquatic environment.

Interactive effects of microplastics and benzo[a]pyrene on two species of marine invertebrates

This study aimed to evaluate B[a]P and low-density polyethylene microplastics (MPs) toxicty, alone and in mixture (0.03 to 30 μg L−1 of B[a]P; and 5, 50 and 500 mg L−1 for MPs). Five mg L−1 of MPs is considerably higher than commonly reported environmental concentrations, although it has been reported for marine environments. Individual (sea urchin embryo-larval development and mortality of mysids) and sub-individual responses (LPO and DNA damage in mysids) were assessed. The toxicity increased as the B[a]P concentration increased, and microplastics alone did not cause toxicity. B[a]P toxicity was not modified by the lowest concentration of MPs (5 mg L−1), but at higher MPs concentrations (50 and 500 mg L−1), the effects of B[a]P on sea urchin development and in biomarkers in mysids were diminished. Microplastics interacted with B[a]P in seawater, reducing its toxicity, probably due to adsorption of B[a]P to the surface of microplastics.

Intraspecific allometric shift in an endemic and endangered marine mollusc: A new minimally intrusive tool to capture shell shape in the field

AbstractPatella ferruginea (Gastropoda, Patellidae) is an endemic and endangered marine invertebrate species from the Western Mediterranean that presents two ecomorphotypes. Currently, there is no objective and nonmanipulative method to determine these ecomorphotypes in the field. The present study uses the geometric morphometrics approach to investigate variation in the shell‐shape allometric trajectories of both ecomorphotypes. In addition, we propose a new manipulation‐free tool to assign ecomorphotype in the field. A total of 190 shells, 123 from the lamarcki (20.0 to 86.7 mm) and 67 from the rouxi (25.3 to 95.9 mm) ecomorphotypes, were analyzed using three different 2D landmark and semilandmark configurations capturing the lateral, internal, and dorsal shell views. We provide compelling graphical and analytical evidence that these ecomorphotypes differ in allometric growth patterns. However, the findings of the current study do not support that lamarcki and rouxi correspond to different ontogenetic states of P. ferruginea. We hypothesize that the difference in the allometric trajectories between the rouxi and the lamarcki ecomorphotypes could be due to environmental constraints, but it might also be due to currently undetected genetic factors. Since the allometry‐free shape lateral view presented the most significant reliability in assigning ecomorphotypes, we evaluate an easy and inexpensive tool to assign individuals in the field to each ecomorphotype using a contour gauge and a simple landmark configuration (only three landmarks). Cross‐validated classification indicated that ecomorphotype assignations for lateral contour present total reliability calculated at 93.15%. Therefore, we encourage it to be implemented in future research to avoid sacrificing or manipulating specimens of this endangered species.

Comparative Genomic Insights into Bacterial Induction of Larval Settlement and Metamorphosis in the Upside-Down Jellyfish Cassiopea.

Bacteria are important mediators of the larval transition from pelagic to benthic environments for marine organisms. Bacteria can therefore dictate species distribution and success of an individual. Despite the importance of marine bacteria to animal ecology, the identity of inductive microbes for many invertebrates are unknown. Here, we report the first successful isolation of bacteria from natural substrates capable of inducing settlement and metamorphosis of the planula larvae stage of a true jellyfish, the upside-down jellyfish Cassiopea xamachana. Inductive bacteria belonged to multiple phyla, with various capacity to induce settlement and metamorphosis. The most inductive isolates belonged to the genus Pseudoalteromonas, a marine bacterium known to induce the pelago-benthic transition in other marine invertebrates. In sequencing the genome of the isolated Pseudoalteromonas and a semiinductive Vibrio, we found biosynthetic pathways previously implicated in larval settlement were absent in Cassiopea inducing taxa. We instead identified other candidate biosynthetic gene clusters involved in larval metamorphosis. These findings could provide hints to the ecological success of C. xamachana compared to sympatric congeneric species within mangrove environments and provide avenues to investigate the evolution of animal-microbe interactions. IMPORTANCE The pelagic to benthic transition for the larvae of many marine invertebrate species are thought to be triggered by microbial cues. The microbial species and exact cue that initiates this transition remains unknown for many animals. Here, we identify two bacterial species, a Pseudoalteromonas and a Vibrio, isolated from natural substrate that induce settlement and metamorphosis of the upside-down jellyfish Cassiopea xamachana. Genomic sequencing revealed both isolates lacked genes known to induce the life history transition in other marine invertebrates. Instead, we identified other gene clusters that may be important for jellyfish settlement and metamorphosis. This study is the first step to identifying the bacterial cue for C. xamachana, an ecologically important species to coastal ecosystems and an emerging model system. Understanding the bacterial cues provides insight into marine invertebrate ecology and evolution of animal-microbe interactions.

Marine litter colonization: Methodological challenges and recommendations

Marine litter colonization by marine invertebrate species is a major global concern resulting in the dispersal of potentially invasive species has been widely reported. However, there are still several methodological challenges and uncertainties in this field of research. In this review, literature related to field studies on marine litter colonization was compiled and analyzed. A general overview of the current knowledge is presented. Major challenges and knowledge gaps were also identified, specifically concerning: 1) uncertainties in species identification, 2) lack of standardized sampling methodologies, 3) inconsistencies with the data reported, and 4) insufficient chemical-analytical approaches to understand this phenomenon. Aiming to serve as a guide for future studies, several recommendations are provided for each point, particularly considering the inaccessibility to advanced techniques and laboratories.

Cone snail species off the Brazilian coast and their venoms: a review and update.

The genus Conus includes over 900 species of marine invertebrates known as cone snails, whose venoms are among the most powerful described so far. This potency is mainly due to the concerted action of hundreds of small bioactive peptides named conopeptides, which target different ion channels and membrane receptors and thus interfere with crucial physiological processes. By swiftly harpooning and injecting their prey and predators with such deadly cocktails, the slow-moving cone snails guarantee their survival in the harsh, competitive marine environment. Each cone snail species produces a unique venom, as the mature sequences of conopeptides from the venoms of different species share very little identity. This biochemical diversity, added to the numerous species and conopeptides contained in their venoms, results in an immense biotechnological and therapeutic potential, still largely unexplored. That is especially true regarding the bioprospection of the venoms of cone snail species found off the Brazilian coast - a region widely known for its biodiversity. Of the 31 species described in this region so far, only four - Conus cancellatus, Conus regius, Conus villepinii, and Conus ermineus - have had their venoms partially characterized, and, although many bioactive molecules have been identified, only a few have been actually isolated and studied. In addition to providing an overview on all the cone snail species found off the Brazilian coast to date, this review compiles the information on the structural and pharmacological features of conopeptides and other molecules identified in the venoms of the four aforementioned species, paving the way for future studies.

Macrozoobenthos monitoring in Portuguese transitional waters in the scope of the water framework directive using morphology and DNA metabarcoding

Despite the growing use and potential of DNA metabarcoding to improve and expedite macrozoobenthos monitoring, its employment in Water Framework Directive (WFD) monitoring of transitional ecosystems still remains largely unexplored. In the current study, we addressed this research gap by building upon the biomonitoring network program of the Portuguese Environmental Agency (APA). The ecological condition of 20 sites from four major transitional ecosystems in the west coast of Portugal was assessed, namely Minho, Lima, Vouga and Mondego estuaries, by using metabarcoding and the morphology-based approach for characterizing macrozoobenthic communities. A total of 154 marine invertebrate species were detected with both methodologies, distributed by 11 phyla. In the majority of the sites, metabarcoding returned a higher number of species and phyla than the morphology-based approach (up to 2.5 times higher). In parallel, the proportion of species detected concurrently by both methods was low (35 species, 23%). The use of a multi-locus strategy increased recovered diversity through metabarcoding, since 37 species were detected exclusively with COI and 46 with 18S. For about 61% of the species recovered through morphology, metabarcoding failed detection, among which 20% was due to the lack of reference sequences in genetic databases. For the remaining, we did not find any plausible reason for only 10%, which could be due either to inefficient DNA extraction or PCR failure. Although morphological and metabarcoding-derived biotic indices did not match completely, similar responses to the environmental gradient were obtained in morphology and metabarcoding based-datasets. We anticipate that metabarcoding can increase the throughput and quality of the assessments, allowing faster assessments with greater spatial-temporal density and robust identifications of all specimens in a sample including larval stages, juveniles, and cryptic lineages, as well as smaller taxonomic groups that cannot be identified to species level using the traditional approach.

EFFETTI DEGLI INTERFERENTI ENDOCRINI SULLE COMUNITÀ DI INVERTEBRATI MARINI

Organic pollutants with disrupting activity on the endocrine system are known as endocrine disrupting compounds (ECDs) and are broadly diffuse on the Planet. By different pathways, they reach the marine environment that is the ultimate collector of these substances. Several researches pointed out acute and chronic effects of these pollutants on the reproduction and development of fish and marine mammals. Fewer studies have been dedicated to explore the effects of ECDs on marine invertebrates though these represent most of marine biodiversity. Researches are hampered by limited knowledge of endocrinology and physiology of several animal groups, which has been studied only for few phyla such as mollusks, annelids and arthropods. The clearest example of endocrine disruption by a chemical compound in marine environment is the pseudo-hermaphroditism caused in females of prosobranch mollusks by the exposure to tributyltin, an antifouling agent used in the paints for ships. This molecule has an adverse effect also on crustaceans in which it inhibits the signaling pathway of the molting hormone. Several studies evidenced that lot of marine invertebrate species are sensible to the action of estrogenic compounds (xeno-estrogens) such as Bisphenol A and Nonylphenol that are used in the manufacturing of the plastic materials. Nevertheless, molecular mechanisms responsible for the observed effects are still uncertain. In my laboratory, we demonstrated that tunicates, that do not have estrogen receptors, are sensible to the action of xeno-estrogens that act by binding to a different group of receptors, the Estrogen Related Receptors. Since these receptors are present also in mammals, our studies could help to clarify pathogenic mechanisms shared with humans.

Genomic Analyses Implicate the Amazon-Orinoco Plume as the Driver of Cryptic Speciation in a Swimming Crab.

The Amazon-Orinoco plume (AOP) is the world's largest freshwater and sediment discharge into the ocean. Previous studies limited to mtDNA suggest that the swimming crab Callinectes ornatus Ordway, 1863 exists as two distinct genetic clusters separated by the AOP. However, questions concerning migration, diversification time, and species delimitation are unresolved. Densely sampling markers across the genome (SNPs) could elucidate the evolutionary processes within this species. Here, we combined mtDNA data and ddRAD-seq to explore the diversification patterns and processes within the swimming crab C. ornatus. We show great genetic differentiation between groups on the north and south sides of the plume but also signs of hybridization. Demographic modeling indicates the divergence between groups starting around 8 Mya following the AOP's formation. After a period of isolation, we detect two incidences of secondary contact with stronger migration in concordance with the North Brazil Current flow. Our results suggest speciation with gene flow explained by the interplay among the AOP, oceanographic currents, and long larval dispersal. This work represents the first investigation employing ddRAD-seq in a marine invertebrate species with distribution encompassing the north and south Atlantic and sheds light on the role of the AOP in the diversification of a marine species.