Iberian Freshwater Research Articles

Why and when do freshwater fish migrate? Observations of migration patterns of the native fishes from the Iberian Peninsula (SW Europe)

We reviewed information on Iberian freshwater fish to characterize their migratory status and identify migration reasons and periods. Most species migrate (87.3 %; 62 species from 15 families). A large number are potamodromous species (45 species from 6 families) but diadromous species (17 species from 11 families) also exist, including anadromous (9 species) and cata¬dromous (8 species). The spawning period is a primary driver of fish migration but feeding and refuge-associated migrations also take place. Sexual maturity is the most important cue triggering fish migration, and other important factors include water temperature, river flow, currents, salinity and photoperiod. Spawning and migrating periods are in general prolonged and vary among years, as a response to the environmental variability of Mediterranean river systems, which are the most frequent in the Iberian Peninsula. Migratory movements of the various native species of each site cover almost the whole or the whole year. Therefore, to allow fish migration, Iberian freshwaters should always be connected, or their fish passes should be permanently, or practically always, in operation.

The influence of databases enrichment using local macroinvertebrate genetic references for metabarcoding based biodiversity studies in river monitoring

Water resources play a vital role in sustaining ecosystems, human life, food security, and biodiversity. However, these resources currently face serious threats caused primarily by human activities. Because of that, European Union have mandated that all its member states monitor the ecological status of their water bodies. One of the most commonly used bioindicator groups for routine river biomonitoring is benthic macroinvertebrates, which rely mainly on morphological identification at the family level and the use of diverse indices to estimate the ecological status of the river ecosystem. The morphological approach has certain limitations, and metabarcoding is now considered an alternative technique for biodiversity studies and biomonitoring of surface waters. However, the efficacy of this approach depends on the comparison of sequencing data with genetic databases, which are often incomplete and geographically biased. In this study, we collected 92 macroinvertebrate samples from rivers in the northern Iberian Peninsula to identify all possible species and generate new DNA sequences for the COI-5P region. The objectives were to perform region-specific enrichment of reference databases for the geographical area of northern Spain and to assess the impact, before and after this strategy, on the taxonomic assignments used for ecological status inference in metabarcoding samples. The results showed that 21% of morphospecies lacked reference sequences in the BOLD or GenBank databases, indicating that a significant number of macroinvertebrate species in the northwestern Iberian Peninsula lack reference sequences in major genetic databases. Enriching the BOLD databases with new sequences led to the detection of an average of 17.25 more BINs (species-assimilable gene cluster) and 1.54 more IBMWP taxa (river ecological status indicator macroinvertebrate taxa) per sample, causing changes in the ecological status determination for 16% of the samples, which clearly shows that improving the databases with local species sequences could enhance the detection and identification of new IBMWP taxa, potentially impacting ecological status inference from molecular data. Additionally, a historical analysis of Iberian macroinvertebrate addition in the BOLD database using public BOLD records revealed an asymptotic trend that can be partially mitigated by region-specific enrichment. These results indicate that a local biota sequencing effort can considerably improve metabarcoding results rather than waiting for a gradual improvement of genetic databases. In conclusion, we advocate for a creation an Iberian freshwater macroinvertebrate checklist and then the initiation of a coordinated effort for sequencing any unsequenced species from the region.

A multi-taxa assessment of aquatic non-indigenous species introduced into Iberian freshwater and transitional waters

Aquatic ecosystems are particularly vulnerable to the introduction of non-indigenous species (NIS), leading to multi-faceted ecological, economic and health impacts worldwide. The Iberian Peninsula comprises an exceptionally biodiverse Mediterranean region with a high number of threatened and endemic aquatic species, most of them strongly impacted by biological invasions. Following a structured approach that combines a systematic review of available information and expert opinion, we provide a comprehensive and updated multi-taxa inventory of aquatic NIS (fungi, macroalgae, vascular plants, invertebrates and vertebrates) in Iberian inland waters. Moreover, we assess overall patterns in the establishment status, introduction pathways, native range and temporal introduction trends of listed NIS. In addition, we discuss the legal coverage provided by both national (Spanish and Portuguese) and European NIS regulations. We inventoried 326 aquatic NIS in Iberian inland waters, including 215 established, 96 with uncertain establishment status and 15 cryptogenic taxa. Invertebrates (54.6%) and vertebrates (24.5%) were the groups with the highest number of NIS, with Arthropoda, Mollusca, and Chordata being the most represented phyla. Recorded NIS originated from diverse geographic regions, with North and South America being the most frequent. Vertebrates and vascular plants were mostly introduced through intentional pathways (i.e. release and escape), whereas invertebrates and macroalgae arrived mostly through unintentional ways (i.e. contaminant or stowaway). Most of the recorded NIS were introduced in Iberian inland waters over the second half of the 20th century, with a high number of NIS introductions being reported in the 2000s. While only 8% of the recorded NIS appear in the European Union list of Invasive Alien Species of Union concern, around 25% are listed in the Spanish and Portuguese NIS regulations. This study provides the most updated checklist of Iberian aquatic NIS, meeting the requirements set by the EU regulation and providing a baseline for the evaluation of its application. We point out the need for coordinated transnational strategies to properly tackle aquatic invasions across borders of the EU members.

Environmental DNA metabarcoding of water samples as a tool for monitoring Iberian freshwater fish composition.

Environmental DNA (eDNA) metabarcoding has been increasingly used to monitor the community assemblages of a wide variety of organisms. Here, we test the efficacy of eDNA metabarcoding to assess the composition of Iberian freshwater fishes, one of the most endangered groups of vertebrates in Spain. For this purpose, we sampled 12 sampling sites throughout one of Spain's largest basins, the Duero, which is home to approximately 70% of the genera and 30% of the primary freshwater fish in Spain. We sampled these sampling sites in the summer by using electrofishing, a traditional sampling method, and eDNA metabarcoding of river water samples using the mitochondrial 12S rRNA gene (12S) as a marker. We also resampled four of these sampling sites in autumn by eDNA. We compared the results obtained through eDNA metabarcoding with those of electrofishing surveys (ones conducted for the present study and past ones) and assessed the suitability of 12S as an eDNA metabarcoding marker for this group of freshwater fishes. We found that the 12S fragment, analysed for 25 Iberian species, showed sufficient taxonomic resolution to be useful for eDNA approaches, and even showed population-level differences in the studied populations across the tissue samples for Achondrostoma arcasii. In most cases, a greater number of species was detected through eDNA metabarcoding than through electrofishing. Based on our results, eDNA metabarcoding is a powerful tool to study the freshwater fish composition in the Iberian Peninsula and to unmask cryptic diversity. However, we highlight the need to generate a local genetic database for 12S gene for such studies and to interpret the results with caution when studying only mitochondrial DNA. Finally, our survey shows that the high detection sensitivity of eDNA metabarcoding and the non-invasiveness of this method allows it to act as a detection system for species of low abundance, such as early invasive species or species in population decline, two key aspects of conservation management of Spanish freshwater fishes.

Ecological role of the Eurasian otter, Lutra lutra (Mustelidae, Carnivora) as a seed dispersal species for riparian vegetation in Iberian fresh waters

Endozoochory is a mutualistic interaction between plants and animals. Such a relationship has rarely been examined in the Eurasian otter Lutra lutra. This study aimed to assess the use and electivity of fruit by this carnivore, along with the viability of ingested seeds. Otter spraints and fruit were collected from the River Bullaque (Guadiana River basin, central Spain) in June and September 2018-2019. A high occurrence of fruit (> 40%) was found in spraints during September. Otters ingested fruit in June from only one plant species: Iberian bushweed Flueggea tinctoria (Phyllanthaceae); whereas seeds ingested in September belonged to four plant species: apple mint Mentha suaveolens (Lamiaceae), common hawthorn Crataegus monogyna, dog rose Rosa canina and elmleaf blackberry Rubus ulmifolius (three Rosaceae species). According to the environmental availability, otters displayed avoidance for mint and rose, neutral selection for hawthorn and preference for blackberry. Germination was unsuccessful for mint seeds, whereas germination was the highest for blackberry (37%; 49% for blackbird Turdus merula, a well-known frugivorous species). Otters appear to display an ecological role as seed dispersers for riparian vegetation in Iberian fresh waters. These findings provide insights into this endozoochorous co-evolution between plants and Carnivora.

Multiple-stressors effects on Iberian freshwaters: A review of current knowledge and future research priorities

Freshwater ecosystems are exposed to an increasing number of stressors, challenging their biomonitoring and management. Despite recent advances in multiple-stressor research, regional-scale assessments in areas with high freshwater biodiversity and increasing anthropogenic pressure are urgently needed. We reviewed 61 studies focused on freshwater individuals, populations and communities from the Iberian Peninsula to (i) quantify the frequency of experimental approaches used (manipulative, observational), aquatic systems, biological organization levels, and types of organisms and modelled responses, (ii) identify key individual stressors and the frequency of significant positive (increase in response magnitude) and negative (decrease) effects and (iii) determine types of interacting stressors and the frequency of their combined effects. Our dataset comprised 409 unique responses to 13 types of individual stressors, 34 stressor pairs and 12 high-order interactions (3- and 4-way). We found a higher prevalence of manipulative (85 %) respect to observational studies, and a greater focus on lotic systems (59 %) and heterotrophic organisms (58 %). The most studied stressors were nutrient (Nutr), thermal (Therm), hydrologic (Hydr), ultraviolet radiation (UVR), toxic (Toxic) and salinity (Sal) stress and land-use pressure. Individual stressors showed a higher proportion of negative (34 %) than positive effects (26 %). Nutr × UVR, Toxic × Toxic, Therm × Toxic, Hydr × Toxic, Sal × Therm, and Nutr × Therm were the most studied stressor pairs. Non-interactive (50 %) and interactive responses (50 %) were balanced. Antagonistic effects (18 %) were slightly more common than synergisms (15 %), reversal or opposing (13 %) and high-order interactions (4 %). Such proportions varied within experimental approaches, biological organization levels and organism types. Our findings are helpful to manage certain stressor combinations in Iberian freshwaters. Further efforts in Iberian multiple-stressor research should be directed to (i) intensify the study of lentic systems, (ii) explore more observational data, (iii) autotrophic organisms and (vi) biodiversity-ecosystem functioning responses, and (v) cover a wider range of stressors and (vi) more complex interacting stressor scenarios.

Quaternary geomorphological and climatic changes associated with the diversification of Iberian freshwater fishes: The case of the genus Cobitis (Cypriniformes, Cobitidae).

We studied the population genetic structure of Cobitis vettonica, an endangered freshwater fish species endemic to the Iberian Peninsula, in order to propose a biogeographic model of the responses of species to the multiple changes that occurred in the Iberian hydrological system during the Quaternary period. We also deciphered the relationship of C. vettonica with its sister species C. paludica, particularly in sympatric areas, and provide genetic information for conservation purposes. To achieve this goal, we analyzed both mitochondrial and nuclear data (the cytochrome b and the nuclear recombination activating 1 genes) and a battery of single‐nucleotide polymorphisms (SNPs) of 248 individuals of C. vettonica or C. paludica from 38 localities, including some sympatric ones, covering the entire distribution area of C. vettonica. We highlight the important role played by the hydrogeomorphological processes and climatic changes that occurred in the Iberian Peninsula during the Quaternary on both the population structure of C. vettonica and its relationship with its sister species C. paludica. Our results support the genetic introgression of populations at the eastern limit of the distribution of C. vettonica. Furthermore, we postulate genetic introgression in sympatric areas. Finally, we propose the establishment or expansion of four Operational Conservation Units (OCUs) for C. vettonica, and highlight the threat faced by its populations due to the low level of genetic diversity detected for some of its populations and genetic introgression with C. paludica, which could eventually displace C. vettonica, resulting in a loss of diversity in this species.

Condition and size of the non-native pikeperch Sander lucioperca (Linnaeus, 1758) in Portuguese river basins.

We studied life‐history traits focusing on the growth and condition of the pikeperch Sander lucioperca to evaluate its phenotypic plasticity when introduced to new environments. Pikeperch is a non‐native fish introduced to Iberian freshwater fauna in 1998 that quickly spread to other river basins through human‐mediated activities, occupying now a wide variety of habitats along mainland Portugal. Condition (K and SMI), fork length at age, and length–weight relationships were studied for Portuguese populations. Pikeperch fork length for ages 1, 2, 3, and 4 was different between several populations. We applied generalized linear models (GLM) to study the influence of habitat type, latitude, altitude, time after first detection, and fish prey richness on pikeperch populations size at age 4 and condition. We observed higher condition values on populations from lower altitudes at lentic systems more recently introduced. But higher fork length at age 4 was found in populations from higher altitudes, on older populations with higher prey richness. Habitat type, time since first detection, and fish fauna composition are discussed as the main environmental factors explaining the observed phenotypic plasticity with concerns on predatory impact on native fauna.

Historical biogeography of the Iberian Peninsula: multilocus phylogeny and ancestral area reconstruction for the freshwater fish genus Squalius (Actinopterygii, Leuciscidae)

The Iberian Peninsula is one of the most ancient geological areas in Europe. Its complex orography is a consequence of several geological episodes related with two orogenic episodes that occurred from Late Devonian (Variscan orogeny) to the Cenozoic (Alpine orogeny). Unraveling the evolutionary history of strict freshwater fauna, such as leuciscid fishes, may contribute to our understanding of the paleogeographic history of a region and its configuration along time. Within the Iberian freshwater fish fauna, the genus Squalius is one of the oldest and most diversified leuciscids. Here, we used phylogenetic, phylogeographic, and ancestral area reconstruction approaches to infer relationships among Iberian drainages and the biogeographic history of these Squalius species, particularly of two of the most widely distributed species, S. carolitertii and S. pyrenaicus. Our results confirmed, in agreement with previous studies, an underestimation of the real diversity of Squalius in the Iberian Peninsula and identified divergent evolutionary lineages, highlighting the reticulate evolutionary history of the Iberian Squalius species. The ancestral reconstruction analyses indicated that several vicariance and dispersal processes occurred among different Squalius lineages, linked to the paleogeographic configuration of the Iberian Peninsula during the Cenozoic period.

First steps towards a barcoding database for the Iberian cladocerans

Due to similarities in morphological features together with strong dispersal abilities, it was thought that some groups of zooplankton (e.g. rotifers, copepods, and cladocerans) have cosmopolitan distributions. In the particular case of cladocerans, recent molecular studies using DNA barcode regions have indicated a different picture, including the existence of multiple regional endemic species and geographical phylogroups; even at very small geographical scales. This has demostrated that cladocera species are less widely distributed than assumed. Morphological identifications of these animals require expertise and high taxonomic specialization. Even so, species identifications are hampered by the small size of the organisms (especially from the littoral zone) and by the sampling cost for obtaining rare species and both parthenogenetic and gamogenetic specimens. The use of molecular techniques can provide new tools to identify cryptic diversity, and by being added to taxonomical approaches, provide more precise data of biodiversity. However, the accuracy of species assignment in metabarcoding relies on the availability of a DNA reference library, which is challenging in areas with high endemicity rates such as the Iberian Peninsula. A preliminary compilation of the available molecular data for the cytochrome (COI) in public repositories (Barcode of Life Data Systems and NCBI GenBank) shows that the available sequences only cover ~60% of the Iberian freshwater cladocerans. The family Daphniidae is very well represented, while the family Chydoridae, which contained most of the Iberian endemism, is underrepresented. We have identified the gaps, and are now focusing on collecting the target organisms to fill the missing taxa of the Iberian library. A compendium of the sampling points, species recovered so far, pitfalls, and future strategy is presented here. The effective completion of a DNA database for cladocerans will have applications not only in biomonitoring programs but to develop DNA-based methods in paleolimnology.

Stones in the road prevents effective implementation of eDNA-based freshwater quality monitoring in the Iberian Peninsula

The Water Framework Directive (WFD) monitoring requires the use of stressor-specific Multimetric Indices, intercalibrated and validated at river basin level, such as the IBMWP, which is one of the most widely used indices for the biomonitoring of rivers in the Iberian Peninsula. This qualitative index is based on the identification mainly at family level of 125 groups of macroinvertebrates. Currently, molecular biomonitoring systems (biomonitoring 2.0) is widely considered as a good alternative to conduct water quality assessments. The problem comes with the requirement of having complete DNA sequences databases for the targeted species within the indicator groups. In order to know the percentage of sequence coverage of the different taxonomic groups, is needed to know the number of freshwater species in each group. The fact that currently it is only necessary to identify them morphologically at family level to calculate the index means that there are no exhaustive lists of lower taxonomic ranges. In this study, we have studied the available taxonomic databases at species level for river macroinvertebrates in the Iberian Peninsula. For this purpose, species geographically referred to the Iberian Peninsula were extracted from three sources: two European (Freshwaterecology.info database and Weigand et al. (2019)) and one Iberian database/list (Múrria et al. 2020). The resultant species list was compared with GBIF (Global Biodiversity Information Facility) using its tool “Species Matching” to find synonyms and fuzzy names. The similarities and differences between databases were studied (considering the synonyms but not the fuzzy names) and a new list of 3586 species was constructed. Our analysis revealed that from 125 taxonomic groups covered by the IBMWP, there are no species collected for 11 of them (9%) and for another 34 groups (27%) there are less than 5 species collected in the databases. Then, using the BAGS software, all sequences of the mitochondrial gene COX1 were mined from BOLD database. From the 3586 species in the list, only 1900 (53%) have at least one sequence in BOLD. Therefore, conducting biomonitoring 2.0 in Iberian freshwater ecosystems is far from effective today. We need to overcome two big obstacles to reach the goal of using molecular biomonitoring in the Iberian Peninsula: a complete taxonomic list of quality indicators species by groups, and to sequence DNA barcodes from all relevant species.

Is it safe to remove a dam at the risk of a sprawl by exotic fish species?

The longitudinal dimension of river connectivity has been significantly disrupted by barriers to compensate for water demand in the long periods of water scarcity in the Iberian Peninsula. The scale of this modification is widespread in the Portuguese part of Douro River network where, side to side with agriculture water demand, there is a constant increase in hydropower production. Thus, native species in Iberian freshwater systems, performing reproductive migrations along the rivers, are strongly affected by the amplification of fragmentation caused by the tremendous density of transversal obstacles in this river basin.We aimed to prioritize dam removal in the Portuguese part of Douro River, mainly considering obsolete barriers (small dams, weirs) based on a spatial multicriteria decision analysis (GIS-MCDA) based on a prioritization procedure. A diversity of parameters were used to prioritize (rank) the dam's suitability for removal, considering the losses of connectivity and fish biodiversity, habitat degradation, negative effects on water quality and ecological conditions, and socio-economic factors. Different weights were assigned to the different attributes in each criterion according to their importance. The analysis also included a significant constraint: the potential spreading of exotic invasive fish species if connection was reestablished through dam removal.This procedure started with the georeferencing of 1201 transversal obstacles that were further characterized for their relative permeability to fish migration. In conclusion the model used allowed to identify 158 priority barriers, as well as the 5 most fragmented tributaries, which means the most impacted by river regulation. In 8 cases the barriers were big dams (> 15 m), whereas in the remaining 150 were weirs. From a final rank of 20 most impacting structures, the MCDA results also identified two cases where potential removal could trigger the additional impact to native fish species related to the sprawl of alien populations.

Key factors explaining critical swimming speed in freshwater fish: a review and statistical analysis for Iberian species

Swimming performance is a key feature that mediates fitness and survival in aquatic animals. Dispersal, habitat selection, predator–prey interactions and reproduction are processes that depend on swimming capabilities. Testing the critical swimming speed (Ucrit) of fish is the most straightforward method to assess their prolonged swimming performance. We analysed the contribution of several predictor variables (total body length, experimental water temperature, time step interval between velocity increments, species identity, taxonomic affiliation, native status, body shape and form factor) in explaining the variation of Ucrit, using linear models and random forests. We compiled in total 204 studies testing Ucrit of 35 inland fishes of the Iberian Peninsula, including 17 alien species that are non-native to that region. We found that body length is largely the most important predictor of Ucrit out of the eight tested variables, followed by family, time step interval and species identity. By contrast, form factor, temperature, body shape and native status were less important. Results showed a generally positive relationship between Ucrit and total body length, but regression slopes varied markedly among families and species. By contrast, linear models did not show significant differences between native and alien species. In conclusion, the present study provides a first comprehensive database of Ucrit in Iberian freshwater fish, which can be thus of considerable interest for habitat management and restoration plans. The resulting data represents a sound foundation to assess fish responses to hydrological alteration (e.g. water flow tolerance and dispersal capacities), or to categorize their habitat preferences.

High diversity of fish ectoparasitic monogeneans (Dactylogyrus) in the Iberian Peninsula: a case of adaptive radiation?

The epicontinental fauna of the Iberian Peninsula is strongly influenced by its geographical history. As the possibilities for dispersion of organisms into and from this region were (and still are) limited, the local fauna consists almost exclusively of endemic species. Almost all Iberian freshwater fishes of the families Leuciscidae and Cyprinidae are endemic and on-going research on these taxa continually uncovers new species. Nevertheless, information on their host-specific parasites remains scarce. In this study, we investigate the diversity and phylogenetic relationships in monogeneans of the genus Dactylogyrus (gill ectoparasites specific to cyprinoid fish) in the Iberian Peninsula. Twenty-two species were collected and identified from 19 host species belonging to Cyprinidae and Leuciscidae. A high degree of endemism was observed, with 21 Dactylogyrus species reported from Iberia only and a single species, D. borealis, also reported from other European regions. Phylogenetic analysis split the endemic Iberian Dactylogyrus into two well-supported clades, the first encompassing Dactylogyrus parasitizing endemic Luciobarbus spp. only, and the second including all Dactylogyrus species of endemic leuciscids and four species of endemic cyprinids. Species delimitation analysis suggests a remarkable diversity and existence of a multitude of cryptic Dactylogyrus species parasitizing endemic leuciscids (Squalius spp. and representatives of Chondrostoma s.l.). These results suggest a rapid adaptive radiation of Dactylogyrus in this geographically isolated region, closely associated with their cyprinoid hosts. Moreover, phylogenetic analysis supports that Dactylogyrus parasites colonized the Iberian Peninsula through multiple dispersion events.

Citizen warnings and post checkout molecular confirmations using eDNA as a combined strategy for updating invasive species distributions

Citizen warnings alerting about the presence of invasive species has been claimed as fundamental in the strategies addressed to manage aquatic invasions. A great volume of online information coming from fishermen and ordinary citizens, about detection of new species in their communities is currently available. Unfortunately, it is not always implemented as a useful tool by agencies, and within stakeholders protocols, to prevent and manage biological invasions. In this work, we have used as a case study the wels catfish Silurus glanis Linnaeus, 1758. Online blogs, webpages, videos, magazines, and newspapers, were searched for news about the presence and spread of S. glanis in the Iberian Peninsula until June 2016. This information was compiled with official reports (scientific papers and regional and national governmental reports) to create a map showing the putative current pattern of S. glanis invasion through Iberian freshwater ecosystems. The current situation pointed to Silurus’s presence in six of the seven main river basins of the Iberian territory. Since non-official reports need post-alerts confirmations, we have also set up a molecular pilot study designing genus specific primers to detect this species in environmental DNA (eDNA) from the reported locations. In the pilot study, primers were tested on eDNA samples extracted from experimental aquariums and on real environmental samples taken from different basins in Spain (Ebro, Douro and Tagus). In all these basins S. glanis was detected. Official Silurus reports were confirmed with two molecular markers in five out of the six cases (83%) assayed in this work, and in two out of three non-official reports (66%) coming from fishermen websites and newspaper reports. The proposed combined strategy (citizen alerts and the eDNA detection method) can be a helpful tool in early detection of invasive species allowing fast and effective management actions by stakeholders.

Phylogeography and Population Genetic Analyses in the Iberian Toothcarp (Aphanius iberus Valenciennes, 1846) at Different Time Scales.

Secondary freshwater fish species inhabiting fluctuating and extreme environments are susceptible to changes in dispersion, effective population size, and genetic structure. The Iberian toothcarp Aphanius iberus is an endemic cyprinodontid of the Iberian Peninsula restricted to brackish water of salt marshes and coastal lagoons on the eastern Spanish Mediterranean coast. In this study, we analyzed mitochondrial cytochrome b (cyt b) DNA and microsatellite variation to evaluate ways in which the processes of extinction, dispersal, and colonization of A. iberus across its geographic distribution have affected its population genetic structure over time and space. The A. iberus network reconstruction indicated subtle levels of phylogeographic structuring. This, combined with substantial mitochondrial DNA (mtDNA) genetic diversity, suggests that Pleistocene glaciations had a lesser effect on the demographic structure of its populations than was the case for Iberian freshwater species with a similar distribution. Haplotype network, hierarchical analysis of molecular variance, and pairwise ΦST comparisons involving some Levantine samples showed a relatively high degree of mtDNA differentiation, which could be explained by historical isolation of the Villena Lagoon population. Conversely, significant genetic differentiation that follows an isolation-by-distance pattern, and a reduction in Ne though time was detected with microsatellites, suggesting extensive habitat fragmentation on the Mediterranean coast of the Iberian Peninsula over the past hundreds of years. At a smaller geographical scale (Mar Menor Lagoon), habitat fragmentation, probably due to human activity, appears to have resulted in substantially reduced migration and increased genetic drift, as shown by expanded genetic differentiation of populations.

Protein analysis and gene expression indicate differential vulnerability of Iberian fish species under a climate change scenario.

Current knowledge on the biological responses of freshwater fish under projected scenarios of climate change remains limited. Here, we examine differences in the protein configuration of two endemic Iberian freshwater fish species, Squalius carolitertii and the critically endangered S. torgalensis that inhabit in the Atlantic-type northern and in the Mediterranean-type southwestern regions, respectively. We performed protein structure modeling of fourteen genes linked to protein folding, energy metabolism, circadian rhythms and immune responses. Structural differences in proteins between the two species were found for HSC70, FKBP52, HIF1α and GPB1. For S. torgalensis, besides structural differences, we found higher thermostability for two proteins (HSP90 and GBP1), which can be advantageous in a warmer environment. Additionally, we investigated how these species might respond to projected scenarios of 3° climate change warming, acidification (ΔpH = -0.4), and their combined effects. Significant changes in gene expression were observed in response to all treatments, particularly under the combined warming and acidification. While S. carolitertii presented changes in gene expression for multiple proteins related to folding (hsp90aa1, hsc70, fkbp4 and stip1), only one such gene was altered in S. torgalensis (stip1). However, S. torgalensis showed a greater capacity for energy production under both the acidification and combined scenarios by increasing cs gene expression and maintaining ldha gene expression in muscle. Overall, these findings suggest that S. torgalensis is better prepared to cope with projected climate change. Worryingly, under the simulated scenarios, disturbances to circadian rhythm and immune system genes (cry1aa, per1a and gbp1) raise concerns for the persistence of both species, highlighting the need to consider multi-stressor effects when evaluating climate change impacts upon fish. This work also highlights that assessments of the potential of endangered freshwater species to cope with environmental change are crucial to help decision-makers adopt future conservation strategies.

Dietary traits of invasive bleak Alburnus alburnus (Actinopterygii, Cyprinidae) between contrasting habitats in Iberian fresh waters

The bleak Alburnus alburnus (L., 1758) is a limnophilic fish native to most of Europe. This cyprinid species is a successful invader in the Iberian Peninsula, where it threatens the highly endemic fish fauna. However, studies on bleak autoecology are scarce in this region, with few data on foraging strategies. The aim of this work was to compare the dietary traits of bleak populations between contrasting habitats in Iberian fresh waters. For this purpose, bleak were seasonally collected from the small River Gevora and the Sierra Brava Reservoir (Guadiana River Basin, southwestern Spain). As percentages of occurrence and ingested mass, plant material and planktonic Crustacea were the most important food categories in the river and the reservoir, respectively. The intake of pelagic prey was higher in the reservoir, especially during summer. Benthic prey, plant material and detritus were more consumed in the river, especially benthos and detritus during spring, whereas plant intake decreased during this season. Shannon index, as a measure of trophic diversity, was higher in the reservoir, except for spring. Overall results suggest that this wide trophic plasticity will facilitate the bleak invasion process throughout Mediterranean Europe, which poses a serious risk to its highly valuable native fish fauna.

Risk of invasion predicted with support vector machines: A case study on northern pike (Esox Lucius, L.) and bleak (Alburnus alburnus, L.)

The impacts of invasive species are recognised as a major threat to global freshwater biodiversity. The risk of invasion (probability of presence) of two avowed invasive species, the northern pike (Esox Lucius, L.) and bleak (Alburnus alburnus, L.), was evaluated in the upper part of the Cabriel River (eastern Iberian Peninsula). Habitat suitability models for these invasive species were developed with Support Vector Machines (SVMs), which were trained with data collected downstream the Contreras dam (the last barrier impeding the invasion of the upper river segment). Although SVMs gained visibility in habitat suitability modelling, they cannot be considered widespread in ecology. Thus, with this technique, there is certain controversy about the necessity of performing variable selection procedures. In this study, the parameters tuning and the variable selection for the SVMs was simultaneously performed with a genetic algorithm and, contradicting previous studies in freshwater ecology, the variable selection proved necessary to achieve almost perfect accuracy. Further, the development of partial dependence plots allowed unveiling the relationship between the selected input variables and the probability of presence. Results revealed the preference of northern pike for large and wide mesohabitats with vegetated shores and abundant prey whereas bleak preferred deep and slightly fast flow mesohabitats with fine substrate. Both species proved able to colonize the upper part of the Cabriel River but the habitat suitability for bleak indicated a slightly higher risk of invasion. Altogether may threaten the endemic species that actually inhabit that stretch, especially the Júcar nase (Parachondrostoma arrigonis; Steindachner), which is one of the most critically endangered Iberian freshwater fish species.

The role of anglers' perceptions and habits in biological invasions: perspectives from the Iberian Peninsula

Abstract Anglers are a recognized vector for invasive fauna, with both intentional and accidental introductions reported worldwide. The purpose of this study was to investigate the Iberian freshwater angler's habits and perceptions related to biological invasions, using an international and bilingual survey in Spain and Portugal. The results showed that anglers from both countries cover great distances to fish, and that they commonly visit several places and fish during consecutive days, representing a major risk for invasions. Yet, anglers' activity patterns throughout the year are not constant: the summer months involve more intense activity and, hence, entail higher risk. In both countries, the preferred fish species were invasive, and their introductions were reported more often than native species, with motivations being country‐dependent. The use of live bait was not frequent, but sometimes invasive species were used and the release of unused bait in the water was reported very frequently. Most anglers recognize that introductions have environmental impacts and that anglers have an active role in intentional introductions. However, only a minority is aware of the angler's role in accidental transportation of invasive species. These findings on anglers' behaviour patterns and perceptions may be used to model invasion risks and also improve monitoring and awareness programmes led by government agencies. Copyright © 2016 John Wiley & Sons, Ltd.