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Soil vulnerability and environmental land use conflicts were quantified and cross tabulated in the Sordo River basin (Trás-os-Montes Province, NE Portugal) to provide insights about the risk of soil degradation within that basin. Soil vulnerability was assessed by algorithms of multi criteria analysis and weighted linear combination, whereas environmental land use conflicts were described by an original approach. Three vulnerability scenarios were considered, enhancing the importance of topographic slope, soil class and use and occupation of soils, respectively, as vulnerability factors. Regardless the scenario, approximately two-thirds to the basin was defined as areas with high vulnerability. Fortunately, only 10.9% of the basin is in environmental conflict, where actual land uses deviate from natural uses determined by soil characteristics. The cross tabulation of high vulnerability areas with conflict areas, which defines the areas at risk of soil degradation, reveal that risk areas account for approximately 3.2-8.4% of the basin, depending on the scenario, being concentrated in steep slope hillsides suited for forest or mixed forest and pasture occupations but actually used for irrigated crop lands, vineyards and olive yards. Some conservation practices are implemented in the vineyard and olive yard areas, namely through the construction of terraces. These structures minimize soil erosion but are potentially unstable beyond given intervals of terrain slope. On the other hand, the maintenance of terraces is costly and for that reason eventually unaffordable for many farmers, and they also entail a mischaracterization of the historical and architectural heritage of the Douro Wine Region (UNESCO World Heritage). In this context, it was suggested the approval of legislation as regards the design, construction and maintenance of terraces, while in a broader perspective it was recommended the appreciation of a European Directive for soil protection and its subsequent transposition to the national legislation. [ABSTRACT FROM AUTHOR]

The quality of groundwater was evaluated in a rural watershed of northern Portugal (River Sordo basin) where environmental land use conflicts have developed in the course of a progressive invasion of forest and pasture lands by agriculture, especially by vineyards. The selected groundwater quality parameters were the concentrations of sodium, calcium, bicarbonate, chloride and nitrates, derived from natural and anthropogenic sources. The environmental land use conflicts were revealed by the coupling of land use and land capability raster maps. The land capability evaluation allocated 70.3% of the basin to the practicing of agriculture, 20% to livestock pasturing and 9.7% to a mosaic of land uses including agriculture, livestock pasturing and forestry. The assessment of land use conflicts allocated 93.9% of the basin to no conflict areas. Minor conflict areas (4.1%) were found concentrated in the western region of the watershed. They correspond to an invasion of farmlands towards sectors of the catchment capable for the practicing of livestock pasturing. Moderate (1.6%) and major (0.4%) conflict areas were found limited to the eastern region, matching steep hillsides capable for the practicing of livestock pasturing or forestry but presently occupied with vineyards. The spatial distributions of ion concentrations were generally justified by common geochemical processes. The dominance of high concentration levels in moderate and major conflict areas was justified within the framework of nutrient dynamics in vineyard environment. Nitrate in groundwater was likewise produced via the nitrification of N-fertilizers. Apparently, this process promoted the weathering of plagioclase by the nitric acid reaction, in concurrence with the weathering by the carbonic acid reaction. The impact of nitrification was found more important in moderate and major conflict areas, relative to no conflict areas. [ABSTRACT FROM AUTHOR]

As bioindicators, freshwater bivalves are crucial for the assessment of the contamination impact on different levels of biological integration. Escherichia coli is used as a bioindicator of water fecal contamination, representing a critical global concern, especially with the rise of multidrug-resistant (MDR) strains. Phylogenetic diversity, pathotypic characterization, and antibiotic resistance profiles of E. coli isolated from freshwater bivalves ( Anodonta anatina ) were assessed. Samples were collected from the Tua River in Northern Portugal, from two different sites, Chelas and Barcel, representing different degrees of contamination. Antimicrobial susceptibility testing was performed by the disk diffusion method, and characterizations of the phylogenetic groups and pathotypes were assessed by PCR-multiplex and real-time PCR-multiplex, respectively. Results showed that 60% of isolates were characterized as MDR, including resistance to carbapenems, considered the last resort against multidrug-resistant bacteria. Within this study, it was also possible to verify the antimicrobial resistance (AMR) profile differences between the two sampling sites, with bivalve isolates from the Chelas site showing a higher percentage of antibiotic resistance. Among the E. coli isolates, the highest prevalence (55%) was observed in group B1, followed by group D or E (15%), group A (10%), and group E or Clade I (10%). None of the isolates were classified as diarrheagenic E. coli (DEC). This work highlights the potential transmission of antimicrobial-resistant bacteria through bivalves in the food chain. The 'One Health' approach is crucial for combating antimicrobial resistance, namely in edible freshwater species, emphasizing active surveillance to protect human, animal, and environmental health against the spread of antibiotic-resistant bacteria in aquatic environments.

Freshwater mussels (Bivalvia, Unionida) play essential roles in the well-functioning of ecosystems, even providing essential services to humans. However, these bivalves face numerous threats (e.g. habitat loss and fragmentation, pollution, introduction of invasive species, and climate change) which have already led to the extinction of many populations. This underscores the need to fully characterize the biology of these species, particularly those, such as Potomida acarnanica , that are still poorly studied. This study presents the first mitogenome of P. acarnanica (Kobelt, 1879), an endemic species of Greece with a distribution limited to only two river basins. The mitochondrial genome of a P. acarnanica specimen, collected at Pamisos River (Peloponnese, Greece), was sequenced by Illumina high-throughput sequencing. This mitogenome (16,101 bp) is characterized by 13 protein-coding genes, 22 transfer RNA and 2 ribosomal RNA genes. The size of this mitogenome is within the range of another Potomida mitogenome already published for the species Potomida littoralis . In the phylogenetic inference, P. acarnanica was recovered as monophyletic with P. littoralis mitogenome in the Lamprotulini tribe, as expected. This genomic resource will assist in genetically characterizing the species, potentially benefiting future evolutionary studies and conservation efforts., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Freshwater bivalves are widely used as accumulation indicators and monitoring tools for assessing contaminant effects on different levels of biological integration. This pilot study aimed to explore the phylogenetic diversity of Escherichia coli isolated from freshwater mussels ( Margaritifera margaritifera and Potomida littoralis ) and characterize their phenotypes and antibiotic resistance profiles. Samples were collected in the Rabaçal and Tua Rivers, in the Douro basin, Portugal-two sites representing different levels of anthropogenic contamination. Antimicrobial susceptibility testing was performed via the disk diffusion method with 21 antibiotics. Results showed that 31% of isolates were multidrug-resistant (MDR). Thus, freshwater mussels provide an effective and time-integrated approach for identifying/quantifying fecal indicators, including MDR bacteria. PCR-based assays were designed for assessing phylogenetic E. coli groups. Among the E. coli isolates, the highest prevalence (44%) was observed in group D or E, followed by group E or Clade I (25%), group A (19%), and group B1 (13%). E. coli isolated from M. margaritifera predominantly exhibited a higher prevalence of phylogroups D or E, whereas E. coli from P. littoralis showed associations with phylogroups E or clade I, B1, A, and D or E. Our results provide new insights into the phylogenetic diversity of E. coli in freshwater bivalves. Additionally, the findings highlight the possible linkage of phylogroups with the host species, the geographical location in the water stream, and human activity. Using E. coli as a bioindicator isolated from freshwater mussels helps us grasp how human activities affect the environment. This study has important implications for those interested in safeguarding water resources, especially in tackling antibiotic resistance in aquatic ecosystems.

Climate change is becoming the leading driver of biodiversity loss. The Mediterranean region, particularly southwestern Europe, is already confronting the consequences of ongoing global warming. Unprecedented biodiversity declines have been recorded, particularly within freshwater ecosystems. Freshwater mussels contribute to essential ecosystem services but are among the most threatened faunal groups on Earth. Their poor conservation status is related to the dependence on fish hosts to complete the life cycle, which also makes them particularly vulnerable to climate change. Species Distribution Models (SDMs) are commonly used to predict species distributions, but often disregard the potential effect of biotic interactions. This study investigated the potential impact of future climate on the distribution of freshwater mussel species while considering their obligatory interaction with fish hosts. Specifically, ensemble models were used to forecast the current and future distribution of six mussel species in the Iberian Peninsula, including environmental conditions and the distribution of fish hosts as predictors. We found that climate change is expected to severely impact the future distribution of Iberian mussels. Species with narrow ranges, namely Margaritifera margaritifera and Unio tumidiformis, were predicted to have their suitable habitats nearly lost and could potentially be facing regional and global extinctions, respectively. Anodonta anatina, Potomida littoralis, and particularly Unio delphinus and Unio mancus, are expected to suffer distributional losses but may gain new suitable habitats. A shift in their distribution to new suitable areas is only possible if fish hosts are able to disperse while carrying larvae. We also found that including the distribution of fish hosts in the mussels' models avoided the underprediction of habitat loss under climate change. This study warns of the imminent loss of mussel species and populations and the urgent need of management actions to reverse current trends and mitigate irreversible damage to species and ecosystems in Mediterranean regions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Europe has a long history of human pressure on freshwater ecosystems. As pressure continues to grow and new threats emerge, there is an urgent need for conservation of freshwater biodiversity and its ecosystem services. However, whilst some taxonomic groups, mainly vertebrates, have received a disproportionate amount of attention and funds, other groups remain largely off the public and scientific radar. Freshwater mussels (Bivalvia, Unionida) are an alarming example of this conservation bias and here we point out six conceptual areas that need immediate and long-term attention: knowledge, threats, socioeconomics, conservation, governance and education. The proposed roadmap aims to advance research, policy and education by identifying the most pressing priorities for the short- and long-term conservation of freshwater mussels across Europe., (© 2022 Society for Conservation Biology.)

Information about biotic interactions (e.g. competition, predation, parasitism, diseases, mutualism, allelopathy) is fundamental to better understand species distribution and abundance, ecosystem functioning, and ultimately guide conservation efforts. However, conservation planning often overlooks these important interactions. Here, we aim to demonstrate a new framework to include biotic interactions into Marxan. For that, we use freshwater mussels and fish interaction (as mussels rely on fishes to complete their life cycle) in the Douro River basin (Iberian Peninsula) as a case study. While doing that, we also test the importance of including biotic interactions into conservation planning exercises, by running spatial prioritisation analysis considering either: 1) only the target species (freshwater mussels); 2) freshwater mussels and their obligatory hosts (freshwater fishes); 3) freshwater mussels, fishes and their interactions. With this framework we found that biotic interactions tend to be underrepresented when the data on both freshwater mussels and fishes is not simultaneously included in the spatial prioritisation. Overall, the priority areas selected across all scenarios are mostly located in the western part of the Douro River basin, where most freshwater mussels and fishes still occur. Given the low overlap of priority areas identified here and the current Natura 2000 network, our approach may be useful for establishing (or enlarging) protected areas, especially in light of the EU Biodiversity Strategy for 2030. Also, this work may provide guidance for future habitat restoration and management of main threats to freshwater biodiversity., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Freshwater mussels are among the most endangered groups of fauna anywhere in world. The indiscriminate use of antibiotics has led to the emergence of resistant strains. These antibiotic-resistant bacteria play a key role in increasing the risk allied with the use of surface water and in spread of resistance genes. Two endangered freshwater mussel species, Margaritifera margaritifera and Potomida littoralis, were sampled at 4 sampling sites along a 50 km stretch of River Tua. Water samples were taken at same sites. Of the total of 135 isolates, 64.44% (39.26% from water and 25.19% from mussels) were coliform bacteria. Site T1, with the lowest concentration of coliform bacteria, and site T2 were the only ones where M. margaritifera was found. No E. coli isolates were found in this species and the pattern between water and mussels was similar. P. littoralis, which was present at T3/T4 sites, is the one that faces the highest concentration of bacterial toxins, which are found in treated wastewater effluents and around population centers. Sites T3/T4 have the isolates (water and mussels) with the highest resistance pattern, mainly to β-lactams. Water and P. littoralis isolates (T3/T4) showed resistance to penicillins and their combination with clavulanic acid, and to cephalosporins, precisely to a fourth generation of cephalosporin antibiotics. The analysis provides important information on the risk to water systems, as well as the need to investigate possible management measures. It is suggested that future studies on the health status of freshwater bivalves should incorporate measures to indicate bacteriological water quality., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The Paiva River is considered one of the least polluted rivers in Europe and its watershed has a high conservation value. However, the Paiva River basin suffers pressures related with recurrent disturbances in land use, such as forest fires, agricultural activities, urbanization and pressures that affect the natural hydromorphological conditions and the continuity of watercourses. Blue and Green Infrastructures (BGINs) emerge to improve biodiversity, sustainability and the supply of ecosystem services while improving socioeconomic aspects. Thus, this article aims to identify priority areas in the basin, for intervention with these infrastructures. For that, a spatial multicriteria decision analysis (MDCA) was carried out according to several data related to the Paiva River Basin. As local politicians and responsible entities for the natural resources management are the main experts on the problems and their possible solutions at the local level, they were involved in this decision-making model. Therefore, these specialized stakeholders did the weighting assignment according to the most or least importance of the same for the work. The map of priority locations to implement BGINs was obtained in the sequel. To the top 5 priority areas, stakeholders attributed the best solutions based on nature. The most recommended BGINs were recovery/maintenance of riparian vegetation and conservation and reforestation of the native forest, both presented in four of the five areas, and introduction of fuel management strips presented in three of the five areas. Thus, we concluded that it is extremely important to include the communities and the competent entities of nature and environment management in scientific projects related to conservation, forming a synergy that makes it possible to combine scientific knowledge with local experience acquired in the field. This project uses a very flexible methodology of local data and can be a great example to be implemented in other hydrographic basins anywhere in the world., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Theidentification of key biodiversity areas (KBA) was initiated by the International Union for Conservation of Nature in 2004 to overcome taxonomic biases in the selection of important areas for conservation, including freshwater ecosystems. Since then, several KBAs have been identified mainly based on the presence of trigger species (i.e., species that trigger either the vulnerability and or the irreplaceability criterion and thus identify a site as a KBA). However, to our knowledge, many of these KBAs have not been validated. Therefore, classical surveys of the taxa used to identify freshwater KBAs (fishes, molluscs, odonates, and aquatic plants) were conducted in Douro (Iberian Peninsula) and Sebou (Morocco) River basins in the Mediterranean Biodiversity Hotspot. Environmental DNA analyses were undertaken in the Moroccan KBAs. There was a mismatch between the supposed and actual presence of trigger species. None of the trigger species were found in 43% and 50% of all KBAs surveyed in the Douro and Sebou basins, respectively. Shortcomings of freshwater KBA identification relate to flawed or lack of distribution data for trigger species. This situation results from a misleading initial identification of KBAs based on poor (or even inaccurate) ecological information or due to increased human disturbance between initial KBA identification and the present. To improve identification of future freshwater KBAs, we suggest selecting trigger species with a more conservative approach; use of local expert knowledge and digital data (to assess habitat quality, species distribution, and potential threats); consideration of the subcatchment when delineating KBAs boundaries; thoughtful consideration of terrestrial special areas for conservation limits; and periodic field validation., (© 2021 The Authors. Conservation Biology published by Wiley Periodicals LLC on behalf of Society for Conservation Biology.)

Anthropogenic freshwater habitats may provide undervalued prospects for long-term conservation as part of species conservation planning. This fundamental, but overlooked, issue requires attention considering the pace that humans have been altering natural freshwater ecosystems and the accelerated levels of biodiversity decline in recent decades. We compiled 709 records of freshwater mussels (Bivalvia, Unionida) inhabiting a broad variety of anthropogenic habitat types (from small ponds to large reservoirs and canals) and reviewed their importance as refuges for this faunal group. Most records came from Europe and North America, with a clear dominance of canals and reservoirs. The dataset covered 228 species, including 34 threatened species on the IUCN Red List. We discuss the conservation importance and provide guidance on how these anthropogenic habitats could be managed to provide optimal conservation value to freshwater mussels. This review also shows that some of these habitats may function as ecological traps owing to conflicting management practices or because they act as a sink for some populations. Therefore, anthropogenic habitats should not be seen as a panacea to resolve conservation problems. More information is necessary to better understand the trade-offs between human use and the conservation of freshwater mussels (and other biota) within anthropogenic habitats, given the low number of quantitative studies and the strong biogeographic knowledge bias that persists., (© 2021 John Wiley & Sons Ltd.)

Freshwater mussels are undergoing rapid global declines due to habitat loss and fragmentation, among other factors, but little is known about the effects of small hydropower plants. Here we assessed the impact of small hydropower plants on the abundance and size structure of the imperilled pearl mussel Margaritifera margaritifera. For this, we sampled 66 sites in three Portuguese rivers (Mente, Rabaçal and Tuela) located upstream and downstream of dams and within the reservoirs. Pearl mussels were significantly more abundant upstream than downstream of dams (97.4% more) or within reservoirs (98.5% more). In addition, juveniles were mostly found upstream of dams. The most significant environmental alterations that explained the observed patterns were related to changes in sediment characteristics (accumulation of fine sediments and organic matter in reservoirs) and water chemistry, most notably suspended solids (highest values in reservoirs) and dissolved oxygen (lowest values in reservoirs). Overall, results show that small hydropower plants can deeply affect pearl mussel populations: specimens almost disappeared from the areas within the reservoirs and sites located downstream only retained adults without signs of recent recruitment. Future management measures devoted to the conservation of pearl mussels should take into account the results reported here to avoid the construction of new dams in pearl mussel rivers; improve management of the river flow in downstream areas; and consider the decommissioning of dams in pearl mussel rivers with a particular attention devoted to the re-naturalization of river sections under the influence of the reservoir and downstream areas., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interests in this work., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Using a new fossil-calibrated mitogenome-based approach, we identified macroevolutionary shifts in mitochondrial gene order among the freshwater mussels (Unionoidea). We show that the early Mesozoic divergence of the two Unionoidea clades, Margaritiferidae and Unionidae, was accompanied by a synchronous split in the gene arrangement in the female mitogenome (i.e., gene orders MF1 and UF1). Our results suggest that this macroevolutionary jump was completed within a relatively short time interval (95% HPD 201-226 Ma) that coincided with the Triassic-Jurassic mass extinction. Both gene orders have persisted within these clades for ~200 Ma. The monophyly of the so-called "problematic" Gonideinae taxa was supported by all the inferred phylogenies in this study using, for the first time, the M- and F-type mitogenomes either singly or combined. Within Gonideinae, two additional splits in the gene order (UF1 to UF2, UF2 to UF3) occurred in the Mesozoic and have persisted for ~150 and ~100 Ma, respectively. Finally, the mitogenomic results suggest ancient connections between freshwater basins of East Asia and Europe near the Cretaceous-Paleogene boundary, probably via a continuous paleo-river system or along the Tethys coastal line, which are well supported by at least three independent but almost synchronous divergence events.

This study focused on the use of sound playbacks as acoustic deterrents to direct native potamodromous migratory species away from all kind of traps. The effects of two acoustic treatments, a repeated sine sweep up to 2 kHz (sweep-up stimulus) and an intermittent 140 Hz tone, were tested in three fish species native to Iberia: Salmo trutta, Pseudochondrostoma duriense and Luciobarbus bocagei. In contrast with S. trutta, the endemic cyprinids P. duriense and L. bocagei exhibited a strong repulse reaction to the frequency sweep-up sound. The 140 Hz stimulus did not seem to alter significantly the behaviour of any of the studied species. These results highlight the potential of acoustic stimuli as fish behavioural barriers and their application to in situ conservation measures of native Iberian fish populations, to protect them from hydropower dams. In addition, this study shows that acoustic deterrents can be used selectively on target species., (© 2018 The Fisheries Society of the British Isles.)

The freshwater pearl mussel Margaritifera margaritifera is a highly threatened species in Europe. Several mechanisms may be responsible for the decline in distribution and abundance of European pearl mussel populations, but almost no quantitative data exists about the possible negative impacts of invasive alien species (IAS). In this study, we clearly demonstrate that the invasive signal crayfish Pacifastacus leniusculus predates pearl mussels, using a laboratorial experiment followed by in situ validation in four rivers in the North of Portugal (Mente, Rabaçal, Tuela and Baceiro Rivers; Douro Basin). In the laboratory, the crayfish had a clear preference for small-sized pearl mussels but no differences in predation were found in mesocosms with and without sediment. In addition, we clearly demonstrated that the signal crayfish predates pearl mussels in natural conditions and detected a significant density dependent effect (i.e., sites with more crayfish presented higher number of pearl mussel shells with marks of predation). Given the recent introduction of the signal crayfish and the potential negative impacts on pearl mussel populations we also investigated its autoecology (distribution, abundance, size structure and sex-ratio) in the four studied rivers. Significant differences in average abundance and size of the crayfish were detected between sites and the sex-ratio was highly skewed to females. In view of the widespread distribution of signal crayfish (and other invasive crayfish species) in Europe, future management actions devoted to the conservation of pearl mussels should take in consideration the possible negative effects of these predators, especially on juveniles., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Gill histopathology is a valuable tool to evaluate ecosystems health, because the gill is a key site of waterborne pollutant uptake and the first target organ to their toxicity. Consequently, this makes it important in biomonitoring programs. This study aims to evaluate gill histopathological differences in Douro basin native fish species and determine possible associations with water quality and the ecological status classifications. Two native fish species (Pseudochondrostoma duriense and Luciobarbus bocagei) were sampled in four points of the Douro basin: two reference points, Ameixiosa (Paiva River) and Covelas (Bestança River), both classified with an excellent ecological status; and two disturbed points, Castro Daire (Paiva River) and Alvações do Corgo (Corgo River), categorized with a good and a moderate ecological status, respectively. Gill histopathological differences were qualitative and quantitatively analyzed. The histological analysis showed that, in all sampling locations, both species presented some degree of gill differences, such as epithelial lifting, lamellar fusion, and/or necrosis. The histopathological differences evaluation emphasized some variances in the responses between the two species. In nase, the filament and lamellar epithelium proliferation were the histopathological differences that better reflected the river ecological status classification, proving their usefulness in biomonitoring programs.

Freshwater bivalves have suffered major global declines, being the introduction of invasive alien species (IAS) an important, but not well studied, mechanism of threat. This study assessed the predator-prey relationship between two non-native crayfish species (Procambarus clarkii and Pacifastacus leniusculus) and three native (Anodonta anatina, Potomida littoralis and Unio delphinus) and one non-native (Corbicula fluminea) freshwater bivalve species through experiments in laboratory and validation under natural conditions (Sabor River basin, Portugal). All native bivalve species were preyed both in laboratory and in the field; however, both crayfish species were unable to prey C. fluminea. Predation was dependent on crayfish and bivalve species but was not affected neither by crayfish nor bivalve sizes. In the laboratory, the most preyed species by both crayfishes was A. anatina. On average, this species was preyed at least 12% more than other species, when crayfishes had a choice. Similar results were found in the field. We also found signs of competition between both crayfishes, being P. clarkii more dominant and aggressive as this species, on average, manipulated the bivalves 63.6% more times and 24:33 min longer than P. leniusculus, and initiated 55.8% more agnostic bouts. Our results support the idea that P. clarkii and P. leniusculus can affect native freshwater bivalves, but clear interspecific differences were detected. Both crayfishes may have direct and indirect impacts on bivalve populations by increasing mortality or by reducing their fitness. In addition, since both crayfishes do not prey C. fluminea, they offer this IAS another advantage over native bivalves. Given the widespread distribution of both P. clarkii and P. leniusculus and the threatened status of many freshwater bivalves, the dynamics and impacts of this relationship should be taken in account in the implementation of management measures devoted to the conservation of native freshwater bivalves., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Two Unionida (freshwater mussel) families are present in the Northern Hemisphere; the Margaritiferidae, representing the most threatened of unionid families, and the Unionidae, which include several genera of unresolved taxonomic placement. The recent reassignment of the poorly studied Lamprotula rochechouartii from the Unionidae to the Margaritiferidae motivated a new search for other potential species of margaritiferids from members of Gibbosula and Lamprotula. Based on molecular and morphological analyses conducted on newly collected specimens from Vietnam, we here assign Gibbosula crassa to the Margaritiferidae. Additionally, we reanalyzed all diagnostic characteristics of the Margaritiferidae and examined museum specimens of Lamprotula and Gibbosula. As a result, two additional species are also moved to the Margaritiferidae, i.e. Gibbosula confragosa and Gibbosula polysticta. We performed a robust five marker phylogeny with all available margaritiferid species and discuss the taxonomy within the family. The present phylogeny reveals the division of Margaritiferidae into four ancient clades with distinct morphological, biogeographical and ecological characteristics that justify the division of the Margaritiferidae into two subfamilies (Gibbosulinae and Margaritiferinae) and four genera (Gibbosula, Cumberlandia, Margaritifera, and Pseudunio). The systematics of the Margaritiferidae family is re-defined as well as their distribution, potential origin and main biogeographic patterns., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The impacts of river damming on aquatic fauna are assessed and then integrated across hierarchical scales, portrayed as nested circles. The outer circles characterize the studied site for habitat disturbance and hydrologic regime during the construction and filling phases. The inner circles characterize the construction and filling phase zones for water quality parameters and aquatic fauna. The procedure for integrating the impacts comprises two consecutive stages: in the task stage, inner circles are characterized through field and laboratory work involving sampling and analyses of water aliquots and aquatic fauna; in the impact assessment stage, circle data are processed in ecological and statistical algorithms, which allow identification of changes in abundance and composition of aquatic fauna communities and their integration with changes in water quality parameters across the construction and filling phase zones. The integrative assessment of river damming impacts on aquatic fauna was carried out in the Sabor River dam (Portugal). This dam created two reservoirs: primary and secondary. Changes in water quality caused by dam construction and stream water impoundment were significant, marked by increases in temperature and electric conductivity downstream, accumulation of phosphorus and nitrogen in the reservoirs triggering the growth of algae and the increase of chlorophyll a, and drop of transparency. These changes were aggravated in the secondary reservoir. The consequences of water deterioration for aquatic fauna were severe, marked by abrupt declines of native fish species and invasion of exotic species even upwards the reservoirs. The ecological status determined from ecological quality ratios of benthic macroinvertebrate assemblages were also affected, changing from good-fair in the unaffected watercourses to fair-poor in the lakes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The natural variation of environmental factors in freshwater basins determines their biodiversity. Among them, calcium is a key physiological compound for freshwater invertebrates. It is required for shell formation, muscle contraction, it mediates gene expression and allows counteracting acidosis during stress periods, among other functions. Although the distribution of different freshwater species has been suggested to be linked with the environmental calcium concentration, as yet, no research studies have confirmed this. Identifying whether environmental calcium concentrations might determine the invasion success of alien species would be critical in developing and implementing effective management strategies to control them. Here, a multidisciplinary approach integrating field surveys, analytical chemistry techniques, molecular biology analyses and a lab-scale experiment was taken to decipher whether the environmental calcium concentration might hamper the establishment of Corbicula fluminea in northwestern Iberian rivers. A Principal Component Analysis on water chemistry variables from 13 water bodies identified environmental calcium concentration, among others, as one key factor that best characterized the distribution area of C. fluminea. The calcium content in animals' bodies from two representative rivers was dependent on the environmental calcium concentration of freshwater basins; the lower the concentration, the lower the body's content. The expression of stress- and calcium homeostasis-related genes was higher in C. fluminea from low calcium concentration environments than in those from calcium-rich freshwater basins. Finally, under experimental conditions, lower water calcium concentrations decreased C. fluminea growth rates. The present data suggest, for the first time, that environmental calcium concentration may act as a determinant factor on the invasion success of C. fluminea in freshwater environments. Our results provide new clues for the identification of basins with increased risk of potential invasion by C. fluminea based on environmental calcium levels., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Freshwater mussels of the order Unionida are key elements of freshwater habitats and are responsible for important ecological functions and services. Unfortunately, these bivalves are among the most threatened freshwater taxa in the world. However, conservation planning and management are hindered by taxonomic problems and a lack of detailed ecological data. This highlights the urgent need for advances in the areas of systematics and evolutionary relationships within the Unionida. This study presents the most comprehensive phylogeny to date of the larger Unionida family, i.e., the Unionidae. The phylogeny is based on a combined dataset of 1032bp (COI+28S) of 70 species in 46 genera, with 7 of this genera being sequenced for the first time. The resulting phylogeny divided the Unionidae into 6 supported subfamilies and 18 tribes, three of which are here named for the first time (i.e., Chamberlainiini nomen novum, Cristariini nomen novum and Lanceolariini nomen novum). Molecular analyses were complemented by investigations of selected morphological, anatomical and behavioral characters used in traditional phylogenetic studies. No single morphological, anatomical or behavioral character was diagnostic at the subfamily level and few were useful at the tribe level. However, within subfamilies, many tribes can be recognized based on a subset of these characters. The geographical distribution of each of the subfamilies and tribes is also presented. The present study provides important advances in the systematics of these extraordinary taxa with implications for future ecological and conservation studies., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Many methods in freshwater biomonitoring tend to be restricted to a few levels of biological organization, limiting the potential spectrum of measurable of cause-effect responses to different anthropogenic impacts. We combined distinct organisational levels, covering biological biomarkers (histopathological and biochemical reactions in liver and fish gills), community based bioindicators (fish guilds, invertebrate metrics/traits and chironomid pupal exuviae) and ecosystem functional indicators (decomposition rates) to assess ecological status at designated Water Framework Directive monitoring sites, covering a gradient of human impact across several rivers in northern Portugal. We used Random Forest to rank the variables that contributed more significantly to successfully predict the different classes of ecological status and also to provide specific cut levels to discriminate each WFD class based on reference condition. A total of 59 Biological Quality Elements and functional indicators were determined using this procedure and subsequently applied to develop the integrated Multiple Ecological Level Index (MELI Index), a potentially powerful bioassessment tool., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Freshwater mussels of the family Unionidae exhibit a particular form of mitochondria inheritance called double uniparental inheritance (DUI), in which the mitochondria are inherited by both male and female parents. The (M)ale and (F)emale mitogenomes are highly divergent within species. In the present study, we determine and describe the complete M and F mitogenomes of the Endangered freshwater mussel Potomida littoralis (Cuvier, 1798). The complete M and F mitogenomes sequences are 16 451 bp and 15 787 bp in length, respectively. Both F and M have the same gene content: 13 protein-coding genes (PCGs), 22 transfer RNA (trn) and 2 ribosomal RNA (rrn) genes. Bayesian analyses based on the concatenated nucleotide sequences of 12 PCGs and 2 rrn genes of both genomes, including mitogenome sequences available from related species, were performed. Male and Female lineages are monophyletic within the family, but reveal distinct phylogenetic relationships.

In the Uberaba River basin (state of Minas Gerais, Brazil), pastures for livestock production have invaded areas of native vegetation (Cerrado biome), while already existing pastures were invaded by crop agriculture, with an expansion of sugar cane plantations in the most recent years. In some areas of the basin, these land use changes were classified as environmental land use conflicts because the new uses were not conforming to land capability, i.e. the soil's natural use. Where the areas in conflict became dense, some soil properties have changed significantly, namely the organic matter content and the exchangeable potassium concentration, which have decreased drastically (5kg/m(3) per 10% increase in the conflict area) threatening the fertility of soil. Besides, these changes may have triggered a cascade of other environmental damages, specifically the increase of soil erosion and the degradation of water quality with negative impacts on aquatic biodiversity, related to a disruption of soil organic matter structural functions. Because half the Uberaba catchment has been considered is a state of accentuated environmental degradation, not only caused by environmental land use conflicts, conservation measures have been proposed and requested for immediate implementation across the watershed., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The Potomida genus (Bivalvia, Unionida) has a Circum-Mediterranean distribution and like other freshwater mussel species, its populations have suffered dramatic declines. Although this genus is currently considered as monotypic, it has a long history of taxonomic revisions and presently many aspects of its systematics and evolutionary history are unclear. We sampled a total of 323 individuals from 39 different sites across the Potomida genus distribution, and sequenced two mitochondrial (16S rDNA and Cytochrome c Oxidase Subunit I) and one nuclear (28S rDNA) genes to clarify its phylogeny and phylogeographic history. Our results show that the genus includes two well-supported clades, one comprising solely the western Mediterranean species Potomida littoralis, and the other including two eastern Mediterranean species, the Greek endemic P. acarnanica and the Anatolian and Middle Eastern P. semirugata. We suggest that Potomida started radiating during the upper Miocene, and that both vicariance and dispersal events shaped the diversification and distribution of the genus along the Mediterranean region. P. littoralis is further divided in two mitochondrial lineages, one restricted to Europe and the other occurring mostly in North Africa. Moreover, some European basins present both lineages in sympatry. The conservation status of the three recognized species should be reevaluated, particularly P. acarnanica, since it is restricted to two Greek river basins presenting a high risk of extinction. Overall, our results clarify some important gaps in knowledge concerning the phylogeny, phylogeography and evolution of the Potomida genus in the Mediterranean region with important taxonomical, ecological and conservational implications., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Margaritifera marocana is one of the rarest freshwater mussel species (listed as critically endangered), and is endemic to Morocco. Despite its constrained distribution and low abundance, to date there are no quantitative studies addressing the conservation status of this species. Surveys were conducted in 36 sites along the Oum Er Rbia river basin (Rivers Derna, Laabid and Oum Er Rbia) to assess the distribution, abundance, population structure and genetic diversity of M. marocana. Just one specimen was found on River Oum Er Rbia and none on River Derna; however, a high abundance was found in the lower section of River Laabid (e.g., site Laabid 6 reached a mean density of 11.0 ± 6.8 ind.m(-2)). Contrary to earlier information, which reported an overall population size fewer than 250 individuals in a restricted area and no juvenile presence, this study showed that a much higher abundance exists in River Laabid alone. In addition, the species is present in more than 50 km of this river and is still recruiting since small specimens were found. Regarding genetic diversity, six of nine loci previously used in Margaritifera margaritifera were polymorphic and suitable in M. marocana. The spatial range contraction of this species is likely to be very recent, since no strong signature was detected by the molecular diversity indices. Information gathered in this study can be used as a reference to the present conservation status of M. marocana, and guide future research and management initiatives to better conserve it. We conclude discussing the potential major threats for the future survival of M. marocana and suggest some management measures (and research needs) that should be urgently applied., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Unionoid freshwater mussels have a unique life cycle with a form of parental care where the larvae are developed and kept inside the gills until release, followed by an obligate parasitic stage on fish. The size and location of the marsupium have been used as important phylogenetic characters in unionoids and in Anodontini its location was described exclusively on the outer demibranchs. Two recent surveys in a lake in the North of Portugal revealed large anodontine mussels morphological identical to Anodonta anatina but with glochidia in both demibranchs and with an unusual large size. In order to establish the identity of these mussels, a barcoding approach was used and an anatomical description of the gills and glochidia was performed. These mussels were identified as A. anatina and presented an inner demibranch pair with tripartite tubes. The glochidial sizes were much higher than previously reported for the species reaching maximum (length × height) values of 566 × 552 μm. This species reveals a high ecological plasticity being able to change its size and anatomy to increase its fertility as well as infestation performance. J. Exp. Zool. 325A:52-56, 2016. © 2015 Wiley Periodicals, Inc., (© 2015 Wiley Periodicals, Inc.)

In this study, we assess the impacts of future climate and land-use in the Beça River (northern Portugal) under different scenarios and how this will translate into the conservation status of the endangered pearl mussel Margaritifera margaritifera (Linnaeus, 1758). This species is currently present in several stretches of the Beça River that still hold adequate ecological conditions. However, the species is threatened by projected declines in precipitation for the 21st century, with implication on the river flows and water depths that might decrease below the species requisites. This situation could be especially critical during summer conditions since the ecological flows may not be assured and several river stretches may be converted into stagnant isolated pools. The habitat connectivity will also be affected with reverberating effects on the mobility of Salmo trutta, the host of M. margaritifera, with consequences in the reproduction and recruitment of pearl mussels. In addition, human-related threats mostly associated with the presence of dams and an predicted increases in wildfires in the future. While the presence of dams may decrease even further the connectivity and river flow, with wildfires the major threat will be related to the wash out of burned areas during storms, eventually causing the disappearance of the mussels, especially the juveniles. In view of future climate and land-use change scenarios, conservation strategies are proposed, including the negotiation of ecological flows with the dam promoters, the replanting of riparian vegetation along the water course and the reintroduction of native tree species throughout the catchment., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Soil losses were calculated in a rural watershed where environmental land use conflicts developed in the course of a progressive invasion of forest and pasture/forest lands by agriculture, especially vineyards. The hydrographic basin is located in the Douro region where the famous Port wine is produced (northern Portugal) and the soil losses were estimated by the Universal Soil Loss Equation (USLE) in combination with a Geographic Information System (GIS). Environmental land use conflicts were set up on the basis of land use and land capability maps, coded as follows: 1-agriculture, 2-pasture, 3-pasture/forest, and 4-forest. The difference between the codes of capability and use defines a conflict class, where a negative or nil value means no conflict and a positive i value means class i conflict. The reliability of soil loss estimates was tested by a check of these values against the frequency of stone wall instabilities in vineyard terraces, with good results. Using the USLE, the average soil loss (A) was estimated in A=12.2 t·ha(-1)·yr(-1) and potential erosion risk areas were found to occupy 28.3% of the basin, defined where soil losses are larger than soil loss tolerances. Soil losses in no conflict regions (11.2 t·ha(-1)·yr(-1)) were significantly different from those in class 2 (6.8 t·ha(-1)·yr(-1)) and class 3 regions (21.3 t·ha(-1)·yr(-1)) that in total occupy 2.62 km(2) (14.3% of the basin). When simulating a scenario of no conflict across the entire basin, whereby land use in class 2 conflict regions is set up to permanent pastures and in class 3 conflict regions to pine forests, it was concluded that A=0.95 t·ha(-1)·yr(-1) (class 2) or A=9.8 t·ha(-1)·yr(-1) (class 3), which correspond to drops of 86% and 54% in soil loss relative to the actual values., (Copyright © 2014 Elsevier B.V. All rights reserved.)

An important population of the critically endangered pearl mussel Margaritifera margaritifera (Linnaeus, 1758) was surveyed at the edge of its southern distribution (River Paiva, Portugal). Although an earlier study suggested that this population had a very low number of individuals (<500), a narrow distribution, and was mainly comprised by old specimens our data contradict these findings. Our assessment estimated a population with probably more than 5,000 individuals distributed across 80 km of the river length. From the 32 sites surveyed, 19 contained M. margaritifera with higher abundances verified in the middle and upper parts of the river (a maximum of 78 ind. per 100 m of river stretch was recorded). The pearl mussels showed a clear preference for areas near the banks, in shallow water, sandier and gravel sediments, and a high degree of riparian vegetation cover. The population structure was skewed with a very high percentage of large (and old) animals but 3.7 % of the individuals collected were juveniles (<60 mm in length); therefore, this population can be considered functional. Environmental characterization indicated that this river is still in excellent or good condition although some areas showed deterioration due to discharge of domestic effluents. The main conservation requirements of M. margaritifera in the River Paiva include maintaining the water quality (and if possible stopping the discharge of domestic effluents), increasing riparian vegetation cover, removing several weirs to increase connectivity, and increasing trout density.

Freshwater mussels have decreased dramatically in Iberia over the last decades. These animals are responsible for important ecosystem services such as recycling nutrients and improving water clarity. Under this view a better knowledge on the biological features of these animals is extremely important for future conservation and management actions. In this study the reproductive and gametogenic cycle of Anodonta anatina were studied during 2 years in one population as well as the sex ratio and hermaphroditism in six distinct populations, using standard histology. Gametogenesis was continuous in both sexes and germinal epithelium in early stages of development. Gametes were present throughout the reproductive cycle. Oogenesis and spermatogenesis occurred mainly between January and May. Larvae brooding occurred between September and March and main glochidia discharge occurred over a short period (2-3 weeks) in March. For the sex-ratio and hermaphroditism assessments a variable number of individuals were collected from several populations from lakes and rivers. Previous studies described A. anatina as mainly dioecious with only a few populations presenting occasional hermaphroditism. However, the present study indicates that A. anatina sexual behavior is influenced by environmental conditions, being mainly dioecious in rivers with increased hermaphroditism in standing waters. Although self-fertilization was not confirmed, additional studies with molecular characterization of larvae using fast evolving markers should be used in future studies to enlighten this process. Overall, this study indicates that for more efficient conservation actions and management plans, freshwater mussel reproductive biology should be studied at the population level mainly in the subfamily Anodontinae., (Copyright © 2013 Wiley Periodicals, Inc.)

The increased pollution in ecosystems reinforces the importance of both chemical monitoring and biological monitoring of streams and rivers, as an effective water quality-based approach to assess aquatic ecosystem health. In this study, gudgeon (Gobio gobio) and mullet (Mugil cephalus) liver histopathology (biomarker) and some macroinvertebrate community indexes and metrics (bioindicator) were used to evaluate the effect of the wastewater treatment plant (WWTP) of Febros (Avintes) in Febros River water quality and ecosystem health. Regarding macroinvertebrate communities, the Belgian Biotic Index (BBI) and Iberian Biological Monitoring Working Party (IBMWP) indexes suggested that Febros water was slightly polluted, even though the worst situation was found downstream the WWTP discharge. Concerning community metrics, upstream percent of individuals in five numerically dominant taxa (80%) was slightly more superior than the downstream (78%). The presence of intolerant or sensible individuals, determined by percent of Ephemeroptera, Plecoptera, and Trichoptera individuals and number of Ephemeroptera, Plecoptera, and Trichoptera families metrics, was higher upstream WWTP, reflecting a better water quality. The histopathology shows the presence of hepatic lesions in gudgeon and mullet. The statistical analysis of the lesion gradation showed that only necrosis was significantly higher in gudgeon captured downstream the WWTP, while differences were not observed for mullet. The multivariate analysis of data confirmed the existence of differences in hepatic lesions between gudgeon and mullet and between sampling sites. Regarding macroinvertebrate community, this analysis showed that the organic contamination reflected by the BBI and IBMWP indexes values was a determinant factor in the spatial distribution of macroinvertebrates. This work showed that the study of different biological organization levels can be used for a better assessment of ecosystem ecological integrity and can be used as a tool to reveal anthropogenic activity effects in macroinvertebrate diversity and in fish liver pathology from Febros River.