Environmental DNA for the surveillance of biosecurity threats in Mediterranean lagoons.

Invasive species that outcompete endemic ones and toxic harmful algae that cause algal blooms threaten marine resources like fisheries, aquaculture, and even tourism. Environmental DNA (eDNA) metabarcoding can help as a method for early alert. In this study, we have analyzed communities inhabiting six lagoons within the Gulf of Lion (northwest Mediterranean Sea) with spatial protection as RAMSAR and Natura 2000 sites. Employing the COI gene as the only metabarcode, we found 15 genera that have caused recognized algal bloom outbreaks in the studied lagoons since 2000. In addition, seven alien invasive species that can pose risks to the rich marine resources of the zone and lagoons were also found. The results found from eDNA are consistent with events of toxic algae blooms before and after the sampling moment and with reported occurrences of the invasive species in nearby Mediterranean areas. Multivariate multiple analysis showed the importance of anthropic pressure in the abundance of these nuisance species. Mitigation actions and routine eDNA metabarcoding in zones of special interest like these fragile French Mediterranean lagoons are recommended for early alert of nuisance species in order to plan timely management actions.

Procambarus virginalis Lyko, 2017: A new threat to Iberian inland waters.

An eco-monitoring programme to assess faunal biodiversity in the main rivers of the northern Iberian Peninsula (Spain) reveals the first occurrence of the marbled crayfish Procambarus virginalis (Decapoda: Cambaridae) in Iberian inland waters. Iberian specimens have been identified by combining morphological and genetic traits. We discuss the most plausible pathways and introduction vectors, its potential invasiveness and subsequent impacts on host localities. Our preliminary results raise concern about the potential threat of P. virginalis to native fauna and ecosystem dynamics, as P. virginalis was found in an area of great cultural and ecological importance with relevant populations of endangered species. Due to the invasive history of the marbled crayfish, eradication of these individuals is urgent. This study confirms the importance of early warning systems for exotic species, keeping the population, forest guards and field technicians informed about potential invasive species to execute a rapid and effective response.

New dockside eDNA based protocol to detect the seaweed Asparagopsis armata evaluated by stakeholders.

Early detection of invasive species is crucial to deal effectively with biological invasions in ports, which are hotspots of species introductions. In this study, a simplified end-time PCR methodology conducted on eDNA from water samples was developed for rapid detection of the invasive seaweed Asparagopsis armata (four hours from water collection to result visualization). It was tested dockside in four international Spanish ports in presence of stakeholders, whose feedback was obtained to explore the real applicability of this biotechnology. Although biological invasions were not a main concern for them, results indicate a unanimous approval of the methodology by the stakeholders, having detected the presence of A. armata in three of the ports. Stakeholders suggested further developments for easier application of the tool and multiple species detection, to be adopted for the control of invasive species in ports.

Plankton assessment across the distribution of West African hake and tuna based on eDNA metabarcoding.

The richness of plankton communities determines the fish productivity in the ocean, including important resources that rely on extractive fisheries, such as hakes (genus Merluccius) and tunas (genus Thunnus). Their preys forage on zooplankton, and the latter feed on phytoplankton. Inventories of plankton communities for scientific advice to sustainable fishing are essential in this moment of climate change. Plankton is generally inventoried using conventional methodologies based on large water volumes and visual morphological analyses of samples. In this study, we have employed metabarcoding on environmental DNA (eDNA) samples extracted from small water volumes for plankton inventory from twelve distant sampling stations in the East Atlantic Ocean. Zones rich in hake and tuna prey were detected from eDNA, and multivariate multiple regression analysis was able to predict those zones from diatom-based indices and planktonic diversity based on functional groups. Salinity was negatively correlated with the proportion of diatoms in phytoplankton, highlighting expected impacts of current global change on marine plankton communities. The results emphasise the importance of the plankton richness for fish productivity and support the utility of environmental DNA as a tool to monitor plankton composition changes.

High microplastics concentration in liver is negatively associated with condition factor in the Benguela hake Merluccius polli.

Microplastics (MPs) affect both marine and terrestrial biota worldwide for their harmful effects, which range from physical cell damage to physiological deterioration. In this research, microplastics were quantified from gills, liver and muscle of demersal Benguela hakes Merluccius polli (n = 94), caught by commercial trawling from northwest African waters. Plastic polymers were identified using Fourier Transformed-infraRed spectroscopy (FT-iR). Fulton's k condition factor and the degree of DNA degradation in liver were measured. None of the individuals were free of MPs, whose concentration ranged from 0.18 particles/g in muscle to 0.6 in liver. Four hazardous polymers were identified: 2-ethoxyethylmethacrylate, polyester, polyethylene terephthalate, and poly-acrylics. MP concentration in liver was correlated negatively with the condition factor, suggesting physiological damage. Positive association of MP concentration and liver DNA degradation was explained from cell breakage during trawl hauls during decompression, suggesting an additional way of MPs harm in organisms inhabiting at great depth. This is the first report of potential MPs-driven damage in this species; more studies are recommended to understand the impact of MP pollution on demersal species.

Towards a plastic-less planet. Gender and individual responsibility predict the effect of imagery nudges about marine (micro)plastic pollution on R-behavior intentions.

Emerging microplastics (MP) pollution is one of the biggest threats for the oceans today. Consumers could reduce MP pollution adopting R-behaviors such as reducing consumption of plastic, refusing products with MP, replacing them for green products, and recycling. Here we tested the efficiency of online nudges (images and short messages) for promoting MP-conscious behavior in Spain (n = 671). The perceived level of environmental responsibility and the willingness to adopt R-behaviors were measured. Messages about seafood with MP and plastic-polluted marine environment were more efficient than images of animals killed by plastics. Feeling responsible for MP pollution predicted R-behavior intention. Women would adopt more R-behaviors than men, while men were more sensitive than females to the proposed nudges. Raising the sense of environmental responsibility would be priority in education campaigns. For different cultural sensitivities to animal suffering, evoking environmental health instead of threats to wildlife would be generally recommended.

Microplastics across biomes in diadromous species. Insights from the critically endangered Anguilla anguilla.

Microplastic pollution affects freshwater and marine biota worldwide, microplastics occurring even inside the organisms. With highly variable effects, from physical damage to toxicity of plastic compounds, microplastics are a potential threat to the biodiversity, community composition and organisms' health. This emerging pollutant could overstress diadromous species, which are exposed to both sea and river water in their life cycle. Here we have quantified microplastics in young European eel Anguilla anguilla, a critically endangered catadromous fish, entering three rivers in southwestern Bay of Biscay. River water, sediments and seawater were also analysed for microplastics. The microplastic type was identified using Fournier-Transform Infrared spectroscopy and then searched for their hazard potential at the European Chemical Agency site. Both riverine and sea microplastic pollution were predictors of eels' microplastic profile (types of microplastics by shape and colour): A. anguilla juveniles entering European rivers already carry some marine microplastics and acquire more from river water. Potentially hazardous plastic materials were found from eels, some of them dangerous for aquatic life following the European Chemical Agency. This confirms microplastics as a potential threat for the species. Between-rivers differences for microplastics profiles persistent over years highlight the convenience of analysing and preventing microplastics at a local spatial scale, to save diadromous species from this stressor. Since the origin of microplastics present in glass eels seems to be dual (continental + seawater), new policies should be promoted to limit the entry of microplastics in sea and river waters.

Microplastics in seafood: Relative input of Mytilus galloprovincialis and table salt in mussel dishes.

Due to current marine pollution, microplastics ingestion through seafood is an increasing risk for consumers. In this study, microplastics from mussels (Mytilus galloprovincialis) and table salt employed in popular dishes in Bay of Biscay (Spain) were quantified and analysed by Fourier-Transformed Infrared spectroscopy. Microplastics varied in mussels (mean 0.55-3.20 items/g) depending on the environmental pollution of the collection point (seawater, 0.002-0.015 items/mL; sand, 0.06-0.38 items/g). Microplastics content in table salt (0.1-0.38 items/gr) was much lower than in mussels. Chemical substances found from microplastics in mussels and salt are catalogued as hazardous for human health. Significant correlation between microplastics in sand and mussels was found, suggesting that consumers' risk of microplastics ingestion depends on the harvesting area. Routine microplastics analysis in mussels and disclosure of microplastics content on seafood labels are recommended for conscious, informed consumption.

Heavy metals in fish nearby electronic waste may threaten consumer's health. Examples from Accra, Ghana.

Electronic waste sites are rich in heavy metals contained in electronic and electric equipment waste and pose a risk of pollution if metals enter in the environment nearby. The Korle lagoon, located in the center of Accra, is receiving waste effluents from industries, households and the adjacent e-waste burning site Agbogbloshie which is the biggest in the country. Thus, the risk of heavy metal contamination of the water body and subsequent uptake in the aquatic food chain is particularly relevant. Small-scale fishing, not entering the commercial chain, occurs in the lagoon despite its consideration of biologically dead. We assessed if the exposure to heavy metals through these fish consumption is posing higher health risks than fish sold on Ghanaian markets. Using ICP-MS technology, we quantified concentrations of As, Cd, Co, Cr, Cu, Hg, Ni, Pb and Zn in fish caught from the Korle Lagoon (Trachinotus ovatus, Mugil curema and Mugil cephalus) and compared them to fish from the Tema Newtown fishing market (Scomber colias, Pseudotolithus senegallus). Cobalt and lead concentrations, typical e-waste metals, were higher in fish from the Korle lagoon, even though they were of lower trophic level. Calculated risk indices revealed risk of elevated arsenic and mercury exposure, particularly through T. ovatus from the Korle lagoon, if consumed daily as it is common in the region. This study suggests the need of monitoring programs of Ghanaian catch, with a special focus in environmental risk areas like Korle lagoon to ensure human food safety.

Environmental DNA sampling protocols for the surveillance of marine non-indigenous species in Irish coastal waters.

Understanding the spread and distribution of Non-Indigenous Species (NIS) is key when implementing legislation to maintain good ecosystem health. Environmental DNA (eDNA) has shown great potential to detect aquatic organisms in a rapid and cost-effective way, however their applicability to new environments must be validated prior to their implementation. Here, we tested different field sampling methods in combination with eDNA metabarcoding to develop a tool to detect NIS. Large and small volumes of seawater were filtered, in addition to the collection of sediment and horizontal tow net samples at 12 locations across four distinct geographic areas in Ireland. The biggest dissimilarity in the species recovered was found between sediment and town net samples. Tow nets showed to be the most efficient. A total of 357 taxa were identified, including 16 NIS. Fine mesh tow nets were identified as the most cost-efficient for large-scale monitoring and surveillance of NIS.

Virgin Polystyrene Microparticles Exposure Leads to Changes in Gills DNA and Physical Condition in the Mediterranean Mussel Mytilus Galloprovincialis.

The ever-growing concentration of microplastics in the marine environment is leading to a plethora of questions regarding marine organisms' present and future health status. In this article, the Mediterranean mussel (Mytilus galloprovincialis), a commercial species distributed worldwide, has been exposed to 21 daily doses of polystyrene microparticles (10 µm) at four different concentrations that are environmentally realistic (control: no microplastics, C1: 0.02 mg/L, C2: 0.2 mg/L, and C3: 2 mg/L). The physical status through the condition index, and damages in DNA integrity in gills, through DNA fragmentation, were determined. Results showed a minor effect on DNA integrity but a worse physical status at higher doses. Results could be interpreted as a decrease in mussel feeding activity/filtration rates when exposed to high microplastic concentrations, thus reducing the direct exposure to microplastics in gills. These effects could be happening currently and/or may happen in the near future, threatening populations inhabiting microplastics-polluted environments.

DNA Analysis Detects Different Mislabeling Trend by Country in European Cod Fillets.

Atlantic cod, Gadus morhua, is a highly appreciated fish in European seafood markets and is one of the most substituted fish species in the world. Fraud have been detected in European markets in the last decade, finding different substitute species sold as G. morhua or Atlantic cod on the label. In this study, we analyzed 252 samples of fresh and frozen cod fillets sold in Germany, the Netherlands, and France using DNA barcoding. Different trends were found in different countries: while the level of mislabeling found in Germany and the Netherlands remained at zero in the last years, a significant increase was found in the French markets comparing the current results with previous studies on fillets in France. On the one hand, this mislabeling proves the need to encourage European efforts to control seafood authenticity; on the other, zero mislabeling in two countries shows the success of current European regulations.

Fraud in highly appreciated fish detected from DNA in Europe may undermine the Development Goal of sustainable fishing in Africa.

Despite high effort for food traceability to ensure safe and sustainable consumption, mislabeling persists on seafood markets. Determining what drives deliberate fraud is necessary to improve food authenticity and sustainability. In this study, the relationship between consumer's appreciation and fraudulent mislabeling was assessed through a combination of a survey on consumer's preferences (N = 1608) and molecular tools applied to fish samples commercialized by European companies. We analyzed 401 samples of fish highly consumed in Europe and worldwide (i.e. tuna, hake, anchovy, and blue whiting) through PCR-amplification and sequencing of a suite of DNA markers. Results revealed low mislabeling rate (1.9%), with a higher mislabeling risk in non-recognizable products and significant mediation of fish price between consumer´s appreciation and mislabeling risk of a species. Furthermore, the use of endangered species (e.g. Thunnus thynnus), tuna juveniles for anchovy, and still not regulated Merluccius polli hake as substitutes, points towards illegal, unreported and/or unregulated fishing from African waters. These findings reveal a worrying intentional fraud that hampers the goal of sustainable seafood production and consumption, and suggest to prioritize control efforts on highly appreciated species.

Poorer diversity but tougher species in old ballast water: Biosecurity challenges explored from visual and molecular techniques.

Millions of tons of water cross the oceans inside ships' ballast tanks every day. Planktonic species hitch-hike with water and some may pose risks to ecosystems and economies if get released and establish outside their native range. We monitored ballast water in different trans-equatorial travels, visually and using molecular techniques, and found significant increases of potential nuisance taxa over travel duration, despite evident diversity depletion. Thus, less diverse but more resistant and potentially more harmful communities persist in ballast water over long voyages. If we consider the enormous volume transported every day, the persistence of resistant species in ballast water would be threating the global marine biodiversity. This should be taken into account when modeling and assessing the bioinvasion risks associated with the ballast water and transfer considered in the future research.

From the ocean to jellies forth and back? Microplastics along the commercial life cycle of red algae.

Red algae are increasingly exploited for direct consumption and for production of gelling agents like agar and carrageenan, widely employed in food and personal care products. In this article we identify knowledge gaps about microplastics in the whole commercial life cycle of gelling red algae, from their marine production to the final wastewater treatment. Recommendations for new research include studies of microplastics deposition on red algae at sea, during the industrial process of production of gelling agents, and indeed about improvements of microplastics retention in wastewater treatment plants.

eDNA metabarcoding of small plankton samples to detect fish larvae and their preys from Atlantic and Pacific waters.

Zooplankton community inventories are the basis of fisheries management for containing fish larvae and their preys; however, the visual identification of early-stage larvae (the "missing biomass") is difficult and laborious. Here, eDNA metabarcoding was employed to detect zooplankton species of interest for fisheries from open and coastal waters. High-Throughput sequencing (HTS) from environmental samples using small water volumes has been proposed to detect species of interest whose DNA is the most abundant. We analyzed 6-L water samples taken from subtropical and tropical waters using Cytochrome oxidase I (COI) gene as metabarcode. In the open ocean, several commercial fish larvae and invertebrate species important in fish diet were found from metabarcodes and confirmed from individual barcoding. Comparing Atlantic, Mediterranean, Red Sea, and Pacific samples we found a lower taxonomic depth of OTU assignments in samples from tropical waters than in those from temperate ones, suggesting large gaps in reference databases for those areas; thus a higher effort of zooplankton barcoding in tropical oceans is highly recommended. This and similar simplified sampling protocols could be applied in early detection of species important for fisheries.

Ship-driven biopollution: How aliens transform the local ecosystem diversity in Pacific islands.

Ships moving species across the oceans mix marine communities throughout latitudes. The introduction of new species may be changing the ecosystems even in remote islands. In tropical Pacific islands where maritime traffic is principally local, eDNA metabarcoding and barcoding revealed 75 introduced species, accounting in average for 28% of the community with a minimum of 13% in the very remote Rangiroa atoll. The majority of non-native species were primary producers -from diatoms to red algae, thus the ecosystem is being transformed from the bottom. Primary producers were more shared among sites than other exotics, confirming ship-mediated dispersal in Pacific marine ecosystems. Limited alien share and an apparent saturation of aliens (similar proportion in ports of very different size) suggests the occurrence of "alien drift" in port communities, or random retention of newly introduced aliens that reminds genetic drift of new mutations in a population.

Maritime ports and beach management as sources of coastal macro-, meso-, and microplastic pollution.

Plastic pollution is a major environmental issue that affects coasts all around the world. Many studies point out the importance of a better management of this type of pollution. In this pioneering study, we have analyzed the distribution of macro-, meso-, and microplastics along the coast of Asturias (southwest Bay of Biscay, Spain). Significant correlation analysis suggests that mesoplastics are produced in situ by fragmentation of macroplastics. Differences between groups of beaches revealed the importance of maritime ports and fishing activities as sources of macroplastics and likely of microplastics as well. Another source of microplastics could be atmospheric deposition, especially for fibers. Multiple regression model allowed to confirm the utility of beach services like cleaning and trash bins to control macroplastics. These results emphasize the need of an integral treatment of marine plastic pollution involving fishers and maritime ports, as well as the importance of providing beach services.

Stress resistance for unraveling potential biopollutants. Insights from ballast water community analysis through DNA.

In marine settings, anthropogenic disturbances and climate change increase the rate of biological invasions. Predicting still undescribed invasive alien species (IAS) is needed for preparing timely management responses. We tested a strategy for discovering new potential IAS using DNA in a trans-equatorial expedition onboard RV Polarstern. During one-month travel, species inside ballast water experienced oxygen depletion, warming, darkness and ammonium stress. Many organisms died but several phytoplankton and zooplankton survivors resisted and were detected through a robust combination of individual sampling, DNA barcoding and metabarcoding, new in ballast water studies. Ammonium was identified as an important influential factor to explain diversity changes in phytoplankton and zooplankton. Some species reproduced until the end of the travel. These species tolerant to travel stress could be targeted as potential IAS and prioritized for designing control measures. Introducing resistance to travel stress in biosecurity risk analysis would be recommended.