More than one million barriers fragment Europe's rivers.

Rivers support some of Earth's richest biodiversity1 and provide essential ecosystem services to society2, but they are often fragmented by barriers to free flow3. In Europe, attempts to quantify river connectivity have been hampered by the absence of a harmonized barrier database. Here we show that there are at least 1.2 million instream barriers in 36 European countries (with a mean density of 0.74 barriers per kilometre), 68 per cent of which are structures less than two metres in height that are often overlooked. Standardized walkover surveys along 2,715 kilometres of stream length for 147 rivers indicate that existing records underestimate barrier numbers by about 61 per cent. The highest barrier densities occur in the heavily modified rivers of central Europe and the lowest barrier densities occur in the most remote, sparsely populated alpine areas. Across Europe, the main predictors of barrier density are agricultural pressure, density of river-road crossings, extent of surface water and elevation. Relatively unfragmented rivers are still found in the Balkans, the Baltic states and parts of Scandinavia and southern Europe, but these require urgent protection from proposed dam developments. Our findings could inform the implementation of the EU Biodiversity Strategy, which aims to reconnect 25,000 kilometres of Europe's rivers by 2030, but achieving this will require a paradigm shift in river restoration that recognizes the widespread impacts caused by small barriers.

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.

Elucidating Biodiversity Shifts in Ballast Water Tanks during a Cross-Latitudinal Transfer: Complementary Insights from Molecular Analyses.

In this study, the evolution of ballast water (BW) assemblages across different trophic levels was characterized over a 21 day cross-latitudinal vessel transit using a combination of molecular methods. Triplicate BW samples were collected every second day and size-fractionated (<2.7, 10, >50 µm). Measurements of adenosine triphosphate (ATP) and metabarcoding of environmental nucleic acid (DNA and RNA) analyses, complemented by microscopy and flow cytometry, were performed on each sample. Measured ATP concentrations exhibited high variance between replicates and a strong negative trend in the large (≥50 µm) fraction over the voyage. In concert with microscopy, the metabarcoding data indicated a die-off of larger metazoans during the first week of study and gradual reductions in dinoflagellates and ochrophytes. The ATP and metabarcoding data signaled persistent or increased cellular activity of heterotrophic bacteria and protists in the BW, which was supported by flow cytometry. The metabarcoding showed the presence of active bacteria in all size fractions, suggesting that the sequential filtration approach does not ensure taxonomical differentiation, which has implications for BW quality assessment. Although our data show that ATP and metabarcoding have potential for indicative BW screening for BW compliance monitoring, further research and technological development is needed to improve representativeness of sampling and deliver the unequivocal response criteria required by the international Ballast Water Management Convention.

Northern areas as refugia for temperate species under current climate warming: Atlantic salmon (Salmo salar L.) as a model in northern Europe.

In this work, patterns of geographical genetic diversity in Atlantic salmon Salmo salar were studied across the whole Atlantic Arc; whether these patterns (and thus genetic population structure) were affected by water temperatures was also evaluated. Salmo salar populations were characterized using microsatellite loci and then analysed with reference to ocean surface temperature data from across the region. Analysis showed the presence of a latitudinal cline of genetic variability (higher in northern areas) and water temperatures (sea surface temperatures) determining genetic population structure (the latter in combination with genetic drift in southern populations). Under the current global change scenario, northern areas of Europe would constitute refugia for diversity in the future. This is effectively the inverse of what appears to have happened in glacial refugia during the last glacial maximum. From this perspective, the still abundant and large northern populations S. salar should be considered as precious as the small almost relict southern ones and given appropriate protection. Careful management of the species, coordinated across countries and latitudes, is needed in order to avoid its extinction in Europe.

Authentication of commercial candy ingredients using DNA PCR-cloning methodology.

BACKGROUND: Commercial candies are consumed by all population age sectors worldwide. Methods for quality control and composition authentication are therefore needed for best compliance with consumers' preferences. In this study applications of DNA-based methodology for candy quality control have been tested. Eighteen samples of commercial candies (marshmallows, gumdrops, jelly, sherbet, gelatin-based desserts) produced by five countries were analyzed to identify the component species by polymerase chain reaction, cloning and sequencing of 16S rRNA and ribulose -1,5-diphosphate carboxylase oxygenase genes, and the species determined from BLAST comparison with universal databases and phylogenetic analysis. RESULTS: Positive DNA extraction and amplification of the target genes were obtained for 94% of candies assayed, even those containing as little as <0.0005 ng µL(-1) DNA concentration. The results demonstrated that the species detected from DNA were compatible with the information provided on candy labels only in a few products. DNA traces of undeclared species, including fish, were found in most samples, and two products were labeled as vegetarian but contained porcine DNA. CONCLUSION: Based on the inaccuracy found on the labels of sweets we recommend the use of DNA tests for quality control of these popular sweets. DNA tests have been useful in this field but next-generation sequencing methods could be more effective.

Expansion of Non-Native Brown Trout in South Europe May Be Inadvertently Driven by Stocking: Molecular and Social Survey in the North Iberian Narcea River.

The biological and anthropogenic (management) factors that may contribute to the expansion of non-native lineages in managed fish have been studied in this work taking brown trout (Salmo trutta) as a model species. The changes of users' opinion about stocking was studied employing social science methodology (surveys). The evolution of hatchery stocks together with the outcome of stocking were analysed with two genetic tools: the LDH-C1* locus (marker of non-native stocks) and six microsatellite loci (for assignment of wild trout to the natural population or putative hatchery stocks). Consulted stakeholders were convinced of the correctness of releasing only native stocks, although in practice the hatcheries managed by them contained important proportions of non-native gene carriers. Our results suggest that allochthonous individuals perform better and grow faster in hatchery conditions than the native ones. We also find a dilution of the impact of this kind of suplementation in wild conditions. The use of only native individuals as hatchery breeders tested for the presence of non-native alleles previously to the artificial crosses must be a priority. Surveys can help steer policy making toward decisions that will be followed by the public, but they should not be used to justify science.

Evaluation of large-scale genetic structure in complex demographic and historical scenarios: the mitochondrial DNA and Y-chromosome pools of the Iberian Atlantic façade.

Genetic structural patterns of human populations are usually a combination of long-term evolutionary forces and short-term social, cultural, and demographic processes. Recently, using mitochondrial DNA and Y-chromosome loci, various studies in northern Spain have found evidence that the geographical distribution of Iron Age tribal peoples might have influenced current patterns of genetic structuring in several autochthonous populations. Using the wealth of data that are currently available from the whole territory of the Iberian Peninsula, we have evaluated its genetic structuring in the spatial scale of the Atlantic façade. Hierarchical tree modeling procedures, combined with a classic analysis of molecular variance (AMOVA), were used to model known sociocultural divisions from the third century BCE to the eighth century CE, contrasting them with uniparental marker data. Our results show that, while mountainous and abrupt areas of the Iberian North bear the signals of long-term isolation in their maternal and paternal gene pools, the makeup of the Atlantic façade as a whole can be related to tribal population groups that predate the Roman conquest of the Peninsula. The maintenance through time of such a structure can be related to the numerous geographic barriers of the Iberian mainland, which have historically conditioned its settlement patterns and the occurrence of genetic drift processes.

Over the sands and far away: interpreting an Iberian mitochondrial lineage with ancient Western African origins.

OBJECTIVES: There is an ongoing effort to characterize the genetic links between Africa and Europe, mostly using lineages and haplotypes that are specific to one continent but had an ancient origin in the other. Mitochondrial DNA has been proven to be a very useful tool for this purpose since a high number of putatively European-specific variants of the African L* lineages have been defined over the years. Due to their geographic locations, Spain and Portugal seem to be ideal places for searching for these lineages. METHODS: Five members of a minor branch of haplogroup L3f were found in recent DNA samplings in the region of Asturias (Northern Spain), which is known for its historical isolation. The frequency of L3f in this population (≈1%) is unexpectedly high in comparison with other related lineages in Europe. Complete mitochondrial DNA sequencing of these L3f lineages, as well phylogenetic and phylogeographic comparative analyses have been performed. RESULTS: The L3f variant found in Asturias seems to constitute an Iberian-specific haplogroup, distantly related to lineages in Northern Africa and with a deep ancestry in Western Africa. Coalescent algorithms estimate the minimum arrival time as 8,000 years ago, and a possible route through the Gibraltar Strait. CONCLUSIONS: Results are concordant with a previously proposed Neolithic connection between Southern Europe and Western Africa, which might be key to the proper understanding of the ancient links between these two continents.

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.

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.

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.

Universal primers for species authentication of animal foodstuff in a single polymerase chain reaction.

BACKGROUND: There are many DNA-based systems for detecting animal species present in food and food products, applicable for food quality control and authentication. However, most (if not all) methods require more than one pair of primers and cannot be applied over a wide taxonomic range, e.g. identifying vertebrates and invertebrates with the same primers and protocols. RESULTS: A pair of primers is described here that allows in a single polymerase chain reaction the identification of animal species in food and processed (precooked, canned or smoked) food products over a wide taxonomic range. CONCLUSION: These primers permit the identification of most animal taxa employed in human nutrition, from invertebrates such as molluscs to higher vertebrates, distinguishing between species of the same genus. The short fragment amplified within the 16S rDNA exhibits phylogenetic value and could be considered universal based on the wide taxonomic range assayed. The primers are easy to use and accessible for laboratories with a modest budget, as well as being valuable for consumer information and to reveal food fraud.

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.

Individual-specific transgenerational marking of fish populations based on a barium dual-isotope procedure.

The present study focuses on the development and evaluation of an individual-specific transgenerational marking procedure using two enriched barium isotopes, (135)Ba and (137)Ba, mixed at a given and selectable molar ratio. The method is based on the deconvolution of the isotope patterns found in the sample into four molar contribution factors: natural xenon (Xe nat), natural barium (Ba nat), Ba135, and Ba137. The ratio of molar contributions between Ba137 and Ba135 is constant and independent of the contribution of natural barium in the sample. This procedure was tested in brown trout ( Salmo trutta ) kept in captivity. Trout were injected with three different Ba137/Ba135 isotopic signatures ca. 7 months and 7 days before spawning to compare the efficiency of the marking procedure at long and short term, respectively. The barium isotopic profiles were measured in the offspring by means of inductively coupled plasma mass spectrometry. Each of the three different isotopic signatures was unequivocally identified in the offspring in both whole eggs and larvae. For 9 month old offspring, the characteristic barium isotope signatures could also be detected in the otoliths even in the presence of a high and variable amount of barium of natural isotope abundance. In conclusion, it can be stated that the proposed dual-isotope marking is inheritable and can be detected after both long-term and short-term marking. Furthermore, the dual-isotope marking can be made individual-specific, so that it allows identification of offspring from a single individual or a group of individuals within a given fish group.

Mitochondrial diversity patterns and the Magdalenian resettlement of Europe: new insights from the edge of the Franco-Cantabrian refuge.

Phylogeography of the mitochondrial lineages commonly found in Western Europe can be interpreted in the light of a postglacial resettlement of the continent. The center of this proposal lies in the Franco-Cantabrian glacial refuge, located in the northern Iberian Peninsula and Southwestern France. Recently, this interpretation has been confronted by the unexpected patterns of diversity found in some European haplogroups. To shed new lights on this issue, research on Iberian populations is crucial if events behind the actual genetics of the European continent are to be untangled. In this regard, the region of Asturias has not been extensively studied, despite its convoluted history with prolonged periods of isolation. As mitochondrial DNA is a kind of data that has been commonly used in human population genetics, we conducted a thorough regional study in which we collected buccal swabs from 429 individuals with confirmed Asturian ancestry. The joint analysis of these sequences with a large continent-wide database and previously published diversity patterns allowed us to discuss a new explanation for the population dynamics inside the Franco-Cantabrian area, based on range expansion theory. This approximation to previously contradictory findings has made them compatible with most proposals about the postglacial resettlement of Western Europe.

Evolution in the block: common elements of 5S rDNA organization and evolutionary patterns in distant fish genera.

The 5S rDNA is organized in the genome as tandemly repeated copies of a structural unit composed of a coding sequence plus a nontranscribed spacer (NTS). The coding region is highly conserved in the evolution, whereas the NTS vary in both length and sequence. It has been proposed that 5S rRNA genes are members of a gene family that have arisen through concerted evolution. In this study, we describe the molecular organization and evolution of the 5S rDNA in the genera Lepidorhombus and Scophthalmus (Scophthalmidae) and compared it with already known 5S rDNA of the very different genera Merluccius (Merluccidae) and Salmo (Salmoninae), to identify common structural elements or patterns for understanding 5S rDNA evolution in fish. High intra- and interspecific diversity within the 5S rDNA family in all the genera can be explained by a combination of duplications, deletions, and transposition events. Sequence blocks with high similarity in all the 5S rDNA members across species were identified for the four studied genera, with evidences of intense gene conversion within noncoding regions. We propose a model to explain the evolution of the 5S rDNA, in which the evolutionary units are blocks of nucleotides rather than the entire sequences or single nucleotides. This model implies a "two-speed" evolution: slow within blocks (homogenized by recombination) and fast within the gene family (diversified by duplications and deletions).

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.

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.

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.

Greater willingness to reduce microplastics consumption in Mexico than in Spain supports the importance of legislation on the use of plastics.

Introduction: Microplastics (MP) threaten all organisms worldwide. MP are produced directly as microbeads in cosmetics and hygiene products, or indirectly from breakage of larger plastics. The control of MP requires consumers' engagement to refuse products containing microbeads. Methods: We conducted a survey on 572 university students from Mexico and Spain, two countries where microbeads are not banned yet. More strict laws for plastic control areenforced in Mexico than in Spain. Results: Controlling for age and education, despite knowing less about MP, Mexicans checked for microbeads on product labels more frequently than Spaniards, and desired to reduce MP consumption more. A stronger correlation between individual awareness and willingness of MP control was found for Mexican than for Spanish students. Discussion: Perhaps more strict legislation against plastics creates an environment favorable to MP control. Unclear statement of microbeads on labels was the main reason for not checking microbead contents; environmental education and a stricter control of plastics and MP were identified as necessary policy changes in the two countries. Corporation engagement on clearer product labeling is also suggested.