Unidirectional trans-Atlantic gene flow and a mixed spawning area shape the genetic connectivity of Atlantic bluefin tuna.

The commercially important Atlantic bluefin tuna (Thunnus thynnus), a large migratory fish, has experienced notable recovery aided by accurate resource assessment and effective fisheries management efforts. Traditionally, this species has been perceived as consisting of eastern and western populations, spawning respectively in the Mediterranean Sea and the Gulf of Mexico, with mixing occurring throughout the Atlantic. However, recent studies have challenged this assumption by revealing weak genetic differentiation and identifying a previously unknown spawning ground in the Slope Sea used by Atlantic bluefin tuna of uncertain origin. To further understand the current and past population structure and connectivity of Atlantic bluefin tuna, we have assembled a unique dataset including thousands of genome-wide single-nucleotide polymorphisms (SNPs) from 500 larvae, young of the year and spawning adult samples covering the three spawning grounds and including individuals of other Thunnus species. Our analyses support two weakly differentiated but demographically connected ancestral populations that interbreed in the Slope Sea. Moreover, we also identified signatures of introgression from albacore (Thunnus alalunga) into the Atlantic bluefin tuna genome, exhibiting varied frequencies across spawning areas, indicating strong gene flow from the Mediterranean Sea towards the Slope Sea. We hypothesize that the observed genetic differentiation may be attributed to increased gene flow caused by a recent intensification of westward migration by the eastern population, which could have implications for the genetic diversity and conservation of western populations. Future conservation efforts should consider these findings to address potential genetic homogenization in the species.

Blue-Green Algae as Stimulating and Attractive Feeding Substrates for a Mediterranean Commercial Sea Urchin Species, Paracentrotus lividus.

Sea urchins rely on chemical senses to localize suitable food resources, therefore representing model species for chemosensory studies. In the present study, we investigated the chemical sensitivity of the Mediterranean sea urchin Paracentrotus lividus to the blue-green alga Aphanizomenon flos-aquae, namely "Klamath", and to a few amino acids chosen from the biochemical composition of the same algae. To this end, we used the "urchinogram" method, which estimates the movement rate of the sea urchins in response to chemicals. Our results showed that Klamath represents a strong chemical stimulus for P. lividus as it elicits an overall movement of spines, pedicellariae, and tube feet coupled, in some cases, to a coordinated locomotion of the animals. Sea urchins also displayed a sensitivity, even if to a lesser extent, to leucine, threonine, arginine, and proline, thus implying that the amino acids contained in Klamath may account, at least in part, for the stimulating effects exerted by the whole algae. Additionally, our results show that Klamath, as well as spirulina, another blue-green alga with high nutritional value, is very attractive for this sea urchin species. These findings gain further importance considering the potential profit of echinoderms for commercial consumers and their growing role in aquaculture. Klamath and spirulina combine high nutritional profiles with attractive and stimulating abilities and may be considered potential valuable feed supplements in sea urchin aquaculture.

Isotopic life-history signatures are retained in modern and ancient Atlantic bluefin tuna vertebrae.

Isotopic, tagging and diet studies of modern-day teleosts lacked the ability to contextualise life-history and trophic dynamics with a historical perspective, when exploitation rates were lower and climatic conditions differed. Isotopic analysis of vertebrae, the most plentiful hard-part in archaeological and museum collections, can potentially fill this data-gap. Chemical signatures of habitat and diet use during growth are retained by vertebrae during bone formation. Nonetheless, to fulfil their potential to reveal life-history and trophic dynamics, we need a better understanding of the time frame recorded by vertebrae, currently lacking due to a poor understanding of fish bone remodelling. To address this issue, the authors serially-sectioned four vertebral centra of the highly migratory Atlantic bluefin tuna (Thunnus thynnus; BFT) captured off Sardinia (Italy) and analysed their isotopic composition. They show how carbon (δ13 C), nitrogen (δ15 N) and sulphur (δ34 S) isotope values can vary significantly across BFT vertebrae growth-axes, revealing patterning in dietary life histories. Further, they find that similar patterns are revealed through incremental isotopic analysis of inner and outer vertebrae centra samples from 13 archaeological BFT vertebrae dating between the 9th and13th centuries CE. The results indicate that multi-year foraging signatures are retained in vertebrae and allow for the study of life histories in both modern and paleo-environments. These novel methods can be extended across teleost taxa owing to their potential to inform management and conservation on how teleost trophic dynamics change over time and what their long-term environmental, ecological and anthropological drivers are.

Sea urchin harvest inside marine protected areas: an opportunity to investigate the effects of exploitation where trophic upgrading is achieved.

Background: Marine protected areas (MPAs) usually have both positive effects of protection for the fisheries' target species and indirect negative effects for sea urchins. Moreover, often in MPAs sea urchin human harvest is restricted, but allowed. This study is aimed at estimating the effect of human harvest of the sea urchin Paracentrotus lividus within MPAs, where fish exploitation is restricted and its density is already controlled by a higher natural predation risk. The prediction we formulated was that the lowest densities of commercial sea urchins would be found where human harvest is allowed and where the harvest is restricted, compared to where the harvest is forbidden. Methods: At this aim, a collaborative database gained across five MPAs in Sardinia (Western Mediterranean, Italy) and areas outside was gathered collecting sea urchin abundance and size data in a total of 106 sites at different degrees of sea urchin exploitation: no, restricted and unrestricted harvest sites (NH, RH and UH, respectively). Furthermore, as estimates made in past monitoring efforts (since 2005) were available for 75 of the sampled sites, for each of the different levels of exploitation, the rate of variation in the total sea urchin density was also estimated. Results: Results have highlighted that the lowest sea urchin total and commercial density was found in RH sites, likely for the cumulative effects of human harvest and natural predation. The overall rate of change in sea urchin density over time indicates that only NH conditions promoted the increase of sea urchin abundance and that current local management of the MPAs has driven towards an important regression of populations, by allowing the harvest. Overall, results suggest that complex mechanisms, including synergistic effects between natural biotic interactions and human pressures, may occur on sea urchin populations and the assessment of MPA effects on P. lividus populations would be crucial to guide management decisions on regulating harvest permits. Overall, the need to ban sea urchin harvest in the MPAs to avoid extreme reductions is encouraged, as inside the MPAs sea urchin populations are likely under natural predation pressures for the trophic upgrading.

Effect of High-Pressure Processing on Fresh Sea Urchin Gonads in Terms of Shelf Life, Chemical Composition, and Microbiological Properties.

Paracentrotus lividus is a widespread sea urchin species appreciated worldwide for the taste of its fresh gonads. High-pressure processing (HPP) can provide a thermal equivalent to pasteurization, maintaining the organoleptic properties of the raw gonads. This study evaluated HPP technology's effect at 350 MPa and 500 MPa on microbial inactivation and biochemical characteristics of P. lividus gonads. HPP at 350 MPa resulted in a higher decrease in protein and free amino acids associated with a loss of olfactory, color, and gustatory traits and a visual alteration of the texture. On the other hand, gonad samples stored for 40 days after treatments at 500 MPa showed a good organoleptic profile similar to fresh gonads. Furthermore, only 500 MPa effectively reduced mesophilic bacteria contamination among the two HPP treatments carried out. Total lipids increased during storage; however, the SAFA/PUFA rate was homogeneous during HPP trials ranging from 2.61-3.91 g/100 g. Total protein decreased more than 40% after HPP at 350 MPa, whereas, after 500 MPa, it remained stable for 20 days. The amount of free amino acid constantly decreased during storage after HPP at 350 MPa and remained constant at 500 MPa. HPP can effectively remove the bacterial flora and inactivate enzymes, maintaining the properties of the fresh sea urchin gonads.

Influence of Different Light Sources on the Biochemical Composition of Arthrospira spp. Grown in Model Systems.

Arthrospira platensis and Arthrospira maxima are prokaryotic microalgae commercially marketed as spirulina. The pigments extracted from these algae are widely used for cosmetic and nutraceutical applications. This work aimed to evaluate the influence of three light-emitting lamps (white, orange and blue) on the growth and biomass composition of two strains of A. platensis (M2 and M2M) and one of A. maxima. The obtained data show strain- and light-dependent responses of the microalgae. In addition, white and orange lights led to a similar overall effect by increasing the levels of chlorophyll a and carotenoids. However, exposure to orange light resulted in the highest dry weight (5973.3 mg L-1 in M2M), whereas white light stimulated an increase in the carbohydrate fraction (up to 42.36 g 100 g-1 in A. maxima). Conversely, blue light led to a constant increase in the concentration of phycocyanin (14 g 100 g-1 in A. maxima) and a higher content of proteins in all strains. These results provide important environmental information for modulating the growth of different spirulina strains, which can be used to address the synthesis of biochemical compounds of strategic importance for the development of new nutraceutical foods.

Regional patterns of δ 13C and δ 15N for European common cuttlefish (Sepia officinalis) throughout the Northeast Atlantic Ocean and Mediterranean Sea.

Stable isotope compositions of carbon and nitrogen (expressed as δ 13C and δ 15N) from the European common cuttlefish (Sepia officinalis) were measured in order to evaluate the utility of using these natural tracers throughout the Northeast Atlantic Ocean and Mediterranean Sea (NEAO-MS). Mantle tissue was obtained from S. officinalis collected from 11 sampling locations spanning a wide geographical coverage in the NEAO-MS. Significant differences of both δ 13C and δ 15N values were found among S. officinalis samples relative to sampling location. δ 13C values did not show any discernable spatial trends; however, a distinct pattern of lower δ 15N values in the Mediterranean Sea relative to the NEAO existed. Mean δ 15N values of S. officinalis in the Mediterranean Sea averaged 2.5‰ lower than conspecifics collected in the NEAO and showed a decreasing eastward trend within the Mediterranean Sea with the lowest values in the most eastern sampling locations. Results suggest δ 15N may serve as a useful natural tracer for studies on the population structure of S. officinalis as well as other marine organisms throughout the NEAO-MS.

Flash flood simulation and valve behavior of Mytilus galloprovincialis measured with Hall sensors.

Mussels close their shell as a protective strategy and the quantification of this behavioral marker may represent an alarm signal when they are exposed to environmental stressors. In the present study, we investigated the ability of the Mediterranean mussel Mytilus galloprovincialis to recover and then the resilience or inertia of valve activity after a pulsing exposition to diverse levels of salinity (5, 10, 20, and 35 PSU as reference value). The trial simulated an event of drastic and sudden reduction of seawater salinity thus mimicking an event of flash flood from intense rain. Valve gaping and movements were measured in continuous cycle for 10 days using a customized magneto-electric device which uses Hall sensors. Results showed that under normal conditions of salinity (35 PSU), the general pattern of valve movements was a continuously open state with sporadic spikes indicating a closing motion. At salinity of 5, PSU mussels reacted by closing their valves, leading to a 77% mortality on the 4th day. At salinity of 10, PSU animals were observed with closed valves for the entire duration of the exposure and no mortality occurred, they showed a significant reduction in the valve activity once the reference value of salinity was re-established. In contrast, salinity of 20 PSU did not trigger a significant behavioral response. Interestingly, there no define rhythms of valve movements were recorded during salinity challenges.

Influence of the Technological Process on the Biochemical Composition of Fresh Roe and Bottarga from Liza ramada and Mugil cephalus.

Bottarga is a high-priced delicacy with high nutritional value, and, in Italy, bottarga from mullets has been recognized to be a traditional food product. The flathead grey mullet Mugil cephalus and the thinlip grey mullet Liza ramada are the main cultured grey mullets in the Mediterranean Sea. In this study, fresh roe and bottarga from these two species were investigated to evaluate the influence of the technological process and the species on their biochemical composition and health advantages. The 1 h/200 g salting-out step did not increase the levels of NaCl in the bottarga, although it highly decreased the levels of some heavy metals like Cu and Al. Processing of fresh roe in bottarga led to an essential modification of the lipid fraction, following a general series of monousatturated fatty acid (MUFA)> poliunsutturated fatti acid (PUFA) > saturated fatty acid (SAFA) and an increase in both ω3 and ω6 in Liza ramada. Moreover, bottarga showed higher levels of squalene and cholesterol and an increased Essential Amino Acid/Total Amino Acid ratio (EAA/TAA) in both species. In addition to the nutritional benefits for the consumer, the process proposed in this study may represent a reliable tool for local producers to obtain a final bottarga with both a reproducible biochemical composition and organoleptic characteristics.

Multiple Non-Species-Specific Pathogens Possibly Triggered the Mass Mortality in Pinna nobilis.

The fan mussel, Pinna nobilis, represents the largest bivalve endemic to the Mediterranean Sea. Since 2016, dramatic mass mortality of this species has been observed in several areas. The first surveys suggested that Haplosporidium pinnae (currently considered species-specific) was the main etiological agent, but recent studies have indicated that a multifactorial disease may be responsible for this phenomenon. In this study, we performed molecular diagnostic analyses on P. nobilis, P. rudis, and bivalve heterologous host species from the island of Sardinia to shed further light on the pathogens involved in the mass mortality. The results support the occurrence of a multifactorial disease and that Mycobacterium spp. and H. pinnae are not necessarily associated with the illness. Indeed, our analyses revealed that H. pinnae is not species-specific for P. nobilis, as it was present in other bivalves at least three years before the mass mortality began, and species of Mycobacterium were also found in healthy individuals of P. nobilis and P. rudis. We also detected the species Rhodococcus erythropolis, representing the first report in fan mussels of a bacterium other than Mycobacterium spp. and Vibrio spp. These results depict a complicated scenario, further demonstrating how the P. nobilis mass mortality event is far from being fully understood.

Integrated environmental evaluation of heavy metals and metalloids bioaccumulation in invertebrates and seaweeds from different marine coastal areas of sardinia, mediterranean sea.

In this work, three gastropods Patella vulgata, Osilinus turbinata, and Tahis clavigera, one echinoderm Parancetrotus lividus, one coelenterate Anemonia sulcata, and two seaweed Padina pavonica, and Cystoseira mediterranea were collected from three different marine areas of Sardinia in the Mediterranean sea and studied for heavy metals and metalloid content and accumulation trends. Inductively coupled plasma optical emission spectrometry (ICP-OES) was used for the determination of Al, AS, B, Ba, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Te, Ti, V and Zn in the selected samples. The results showed that gastropods were capable of accumulating Al, Ba, Cu, Fe, Sr, and Zn; seaweeds can better concentrate Al, Fe, and Zn than all other species. At the same time, echinoderms and coelenterate had limited ability to store specific metals, showing a much more homogeneous distribution. PCA analysis allowed us to discriminate among the sites and the species. Cala Zafferano was the area with the higher values of accumulation of all metals in all species as expected, considering its proximity to industrial sites. The results of the analysis showed clearly that heavy metal and metalloid accumulation was different for each species studied. Therefore, for a correct environmental assessment of a given area, a comprehensive approach is strongly recommended by exploiting the different properties of both accumulation and concentration of the metals by different aquatic species.

Tracking a mass mortality outbreak of pen shell Pinna nobilis populations: A collaborative effort of scientists and citizens.

A mass mortality event is devastating the populations of the endemic bivalve Pinna nobilis in the Mediterranean Sea from early autumn 2016. A newly described Haplosporidian endoparasite (Haplosporidium pinnae) is the most probable cause of this ecological catastrophe placing one of the largest bivalves of the world on the brink of extinction. As a pivotal step towards Pinna nobilis conservation, this contribution combines scientists and citizens' data to address the fast- and vast-dispersion and prevalence outbreaks of the pathogen. Therefore, the potential role of currents on parasite expansion was addressed by means of drift simulations of virtual particles in a high-resolution regional currents model. A generalized additive model was implemented to test if environmental factors could modulate the infection of Pinna nobilis populations. The results strongly suggest that the parasite has probably dispersed regionally by surface currents, and that the disease expression seems to be closely related to temperatures above 13.5 °C and to a salinity range between 36.5-39.7 psu. The most likely spread of the disease along the Mediterranean basin associated with scattered survival spots and very few survivors (potentially resistant individuals), point to a challenging scenario for conservation of the emblematic Pinna nobilis, which will require fast and strategic management measures and should make use of the essential role citizen science projects can play.

Spatial dynamics and mixing of bluefin tuna in the Atlantic Ocean and Mediterranean Sea revealed using next-generation sequencing.

The Atlantic bluefin tuna is a highly migratory species emblematic of the challenges associated with shared fisheries management. In an effort to resolve the species' stock dynamics, a genomewide search for spatially informative single nucleotide polymorphisms (SNPs) was undertaken, by way of sequencing reduced representation libraries. An allele frequency approach to SNP discovery was used, combining the data of 555 larvae and young-of-the-year (LYOY) into pools representing major geographical areas and mapping against a newly assembled genomic reference. From a set of 184,895 candidate loci, 384 were selected for validation using 167 LYOY. A highly discriminatory genotyping panel of 95 SNPs was ultimately developed by selecting loci with the most pronounced differences between western Atlantic and Mediterranean Sea LYOY. The panel was evaluated by genotyping a different set of LYOY (n = 326), and from these, 77.8% and 82.1% were correctly assigned to western Atlantic and Mediterranean Sea origins, respectively. The panel revealed temporally persistent differentiation among LYOY from the western Atlantic and Mediterranean Sea (FST  = 0.008, p = .034). The composition of six mixed feeding aggregations in the Atlantic Ocean and Mediterranean Sea was characterized using genotypes from medium (n = 184) and large (n = 48) adults, applying population assignment and mixture analyses. The results provide evidence of persistent population structuring across broad geographic areas and extensive mixing in the Atlantic Ocean, particularly in the mid-Atlantic Bight and Gulf of St. Lawrence. The genomic reference and genotyping tools presented here constitute novel resources useful for future research and conservation efforts.

Amino-modified polystyrene nanoparticles affect signalling pathways of the sea urchin (Paracentrotus lividus) embryos.

Polystyrene nanoparticles have been shown to pose serious risk to marine organisms including sea urchin embryos based on their surface properties and consequently behaviour in natural sea water. The aim of this study is to investigate the toxicity pathways of amino polystyrene nanoparticles (PS-NH2, 50 nm) in Paracentrotus lividus embryos in terms of development and signalling at both protein and gene levels. Two sub-lethal concentrations of 3 and 4 µg/mL of PS-NH2 were used to expose sea urchin embryos in natural sea water (PS-NH2 as aggregates of 143 ± 5 nm). At 24 and 48 h post-fertilisation (hpf) embryonic development was monitored and variations in the levels of key proteins involved in stress response and development (Hsp70, Hsp60, MnSOD, Phospho-p38 Mapk) as well as the modulation of target genes (Pl-Hsp70, Pl-Hsp60, Pl-Cytochrome b, Pl-p38 Mapk, Pl-Caspase 8, Pl-Univin) were measured. At 48 hpf various striking teratogenic effects were observed such as the occurrence of cells/masses randomly distributed, severe skeletal defects and delayed development. At 24 hpf a significant up-regulation of Pl-Hsp70, Pl-p38 Mapk, Pl-Univin and Pl-Cas8 genes was found, while at 48 hpf only for Pl-Univin was observed. Protein profile showed different patterns as a significant increase of Hsp70 and Hsp60 only after 48 hpf compared to controls. Conversely, P-p38 Mapk protein significantly increased at 24 hpf and decreased at 48 hpf. Our findings highlight that PS-NH2 are able to disrupt sea urchin embryos development by modulating protein and gene profile providing new understandings into the signalling pathways involved.

Correction: Atlantic Bluefin Tuna (Thunnus thynnus) Biometrics and Condition.

[This corrects the article DOI: 10.1371/journal.pone.0141478.].

Atlantic Bluefin Tuna (Thunnus thynnus) Biometrics and Condition.

The compiled data for this study represents the first Atlantic and Mediterranean-wide effort to pool all available biometric data for Atlantic bluefin tuna (Thunnus thynnus) with the collaboration of many countries and scientific groups. Biometric relationships were based on an extensive sampling (over 140,000 fish sampled), covering most of the fishing areas for this species in the North Atlantic Ocean and Mediterranean Sea. Sensitivity analyses were carried out to evaluate the representativeness of sampling and explore the most adequate procedure to fit the weight-length relationship (WLR). The selected model for the WLRs by stock included standardized data series (common measurement types) weighted by the inverse variability. There was little difference between annual stock-specific round weight-straight fork length relationships, with an overall difference of 6% in weight. The predicted weight by month was estimated as an additional component in the exponent of the weight-length function. The analyses of monthly variations of fish condition by stock, maturity state and geographic area reflect annual cycles of spawning and feeding behavior. We update and improve upon the biometric relationships for bluefin currently used by the International Commission for the Conservation of Atlantic Tunas, by incorporating substantially larger datasets than ever previously compiled, providing complete documentation of sources and employing robust statistical fitting. WLRs and other conversion factors estimated in this study differ from the ones used in previous bluefin stock assessments.

Metazoan gill parasites of the Atlantic bluefin tuna Thunnus thynnus (Linnaeus) (Osteichthyes: Scombridae) from the Mediterranean and their possible use as biological tags.

The gills of 63 specimens of the Atlantic bluefin tuna Thunnus thynnus (Linnaeus) (Osteichthyes: Scombridae) from three localities of the Mediterranean (Sardinian, Tyrrhenian and Levantine Seas) were examined for metazoan parasites. The parasite fauna of T. thynnus from the Sea of Sardinia included 11 species: five didymozoid trematodes, three capsalid and one hexostomid monogeneans, and one caligid and one pseudocycnid copepods. Four didymozoids were found in fish from the Levantine Sea and only one didymozoid was recorded in fish from the Tyrrhenian Sea. Dividing the hosts into four size-groups (small, medium-sized, large and extra large), the pairwise comparison of prevalence and mean abundance of the new and literary data) showed differences according to host size. The differences in the composition of the parasitic faunas and in the prevalence of parasites, observed between the small tunas from the Tyrrhenian Sea and the medium-sized tunas from the Adriatic Sea, Levantine Sea and the North-East (NE) Atlantic Ocean, indicated that these groups form discrete units. The parasite fauna of the large tunas from the Sea of Sardinia is the richest among the bluefin tuna populations of the Mediterranean and the NE Atlantic, due to the presence of species not found elsewhere in bluefin tunas, such as Caligus coryphaenae Steenstrup et Lütken, 1861, Capsala magronum (Ishii, 1936) and C. paucispinosa (Mamaev, 1968). This fact and the prevalence of some parasites of this group (lower than those of medium-sized fish from the NE Atlantic and higher than the small and medium-sized tunas from the Mediterranean) suggest that the large-sized tuna group in the western Mediterranean is formed by Mediterranean resident tunas (poorly infected), and by tunas migrating from the Atlantic Ocean (heavily infected).

GC-ITMS analysis of PAH contamination levels in the marine sea urchin Paracentrotus lividus in Sardinia.

This paper describes the results of a two-year monitoring study examining the pollution of the sea urchin Paracentrotus lividus by polycyclic aromatic hydrocarbons (PAHs) in Sardinia. GC-ITMS analysis of sea urchin gonads showed the presence of 11 and 12 PAHs in the samples of Capo Pecora, and Capitana, respectively. Fluorene, naphthalene and its two degradation products, 1-methyl-naphthalene, and 2-methyl-naphthalene, were detected in all samples analyzed. The ΣPAH residues showed a similar trend over the two-year sampling period. Furthermore, the residues in the first year were slightly higher than in the second year. The information obtained by the multivariate statistical analysis PLS-DA allowed for the determination of samples based on field site and varying habitat types (rocky reef, and Posidonia seabed). The results of this study showed that Posidonia sea urchins are contaminated by high molecular weight PAHs and that Capitana samples are more contaminated due to a higher level of human activity in the area.