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This study aims to describe pre-reproductive movements of Luciobarbus bocagei and Pseudochondrostoma duriense in a regulated canyon-stretch of the Tormes River (Spain), with high environmental variability, semipermeable barriers, and fishway retrofitting actions. The main objectives were to identify peak migration dates and environmental drivers, test ensemble-learning techniques to model fish migration and propose adaptive management measures. To achieve this, fish movements were 5-year monitored in a stepped fishway and Survival Analysis and Random Forest techniques were used for data analysis and modelling. Results showed that migration occurred in May–July, a wider period than the one previously reported in the literature. Movements were triggered by the increase in water temperature and photoperiod, and were strongly affected by the hydraulic river scenario (water levels and discharge) at the semipermeable barriers. Random Forest was able to include the effect of each barrier and predict accurately timing and number of migrants, classifying and ranking the importance of variables. Moreover, developed models allowed to assess fishway retrofitting actions and to predict positive effects in fish number under new, scheduled and variable environmental flow scenarios. Long-term monitoring together with ensemble-learning methods can allow the definition of cost-effective adaptive management strategies to ensure endemic fish conservation.

espanolEl grado de alteracion antropogenica en los ecosistemas fluviales es elevado, debido, fundamentalmente, a la creacion de barreras transversales (presas, azudes, estaciones de aforo, canos de drenaje, etc.), que alteran su conectividad longitudinal, causando, entre otros impactos, la fragmentacion del habitat. Esta problematica afecta a los peces migradores, pues puede suponer un bloqueo o retraso en sus movimientos, reduciendo sus habitats, amenazando la viabilidad y abundancia de sus poblaciones, llegando incluso a su extincion. La capacidad de nado de los peces (velocidad de nado, tiempo de fatiga y distancia recorrida) es uno de los factores que determina su supervivencia. Un sistema que permite establecerla en condiciones naturales son los canales de nado voluntario. En el canal de nado de Vadocondes (Burgos) se ha determinado la capacidad de nado y como influyen en ella factores ambientales y biometricos, de dos de las principales especies de ciprinidos potamodromos de la Peninsula Iberica (barbo iberico –Luciobarbus bocagei– y boga del Duero –Pseudochondrostoma duriense–, con el objetivo de poder desarrollar unas directrices tecnicas y cientificas adecuadas, que garanticen un diseno optimo y eficaz de los pasos para peces, que repercuta positivamente en la conservacion de sus poblaciones. EnglishThe anthropogenic alteration in fluvial ecosystems is high, mainly due to the creation of transverse barriers (dams, weirs, gauging stations, drainage pipes, etc.), which modify their longitudinal connectivity, causing, among other impacts, the fragmentation of the habitat. This problem affects the migrating fish, because it can suppose a blockade or delay in their movements, reducing their habitats, threatening the viability and abundance of their populations, even reaching their extinction. The swimming capacity of the fish (swimming speed, fatigue time and distance traveled) is one of the factors that determine their survival. A system that allows to establish it, in natural conditions, are the voluntary open channel flume. In the flume of Vadocondes (Burgos) the swimming capacity has been determined, and how environmental and biometric factors influence in the swimming capacity, of two of the main species of cyprinids potamodromous of the Iberian Peninsula (Iberian barbel –Luciobarbus bocagei– and northern straight-mouth nase –Pseudochondrostoma duriense–, with the aim of being able to develop adequate technical and scientific guidelines, which guarantee an optimal and effective design of the fishways, which has a positive impact on the conservation of their populations.

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.

Knowing the relationship between size, morphological traits and swimming performance of fish is essential to understand the swimming capacity to successfully surpass these obstacles and the selective pressure that barriers in rivers and streams could exert on fish. Northern straight-mouth nase, an endemic potamodromous cyprinid fish species from the Northwest of the Iberian Peninsula, was selected to carry out volitionally swimming performance experiments in an open channel against three different flow velocities, using telemetry and video cameras. The use of thin-plate spline, on 10 landmarks, evidenced unknown patterns linked to velocity barriers. At lower flow velocity, size is the main factor explaining the swimming performance; thus, large individuals swim up more efficiently. In contrast, at high flow velocities, shape becomes the essential explanatory variable; thereby, streamlined body shapes with a higher relative position of the tail and a narrower caudal peduncle are more efficient. The obtained results show the existence of a relationship between fish morphology and swimming performance, with potential consequences due to selection pressures associated with velocity barriers and their implications on behavioural and dispersal processes. To sum up, velocity barriers could exert a selection pressure on nase populations, so the fishway design and removal should be (re)considered.

Downstream of small-scale hydropower plants (SHPs), the intensity, frequency and persistence of hydropeaking events often cause an intolerable stress on fish of all life stages. Rapid increases in flow velocity result in fish avoiding unstable habitats and seeking refuge to reduce energy expenditure. To understand fish movements and the habitat use of native Iberian cyprinids in a high-gradient peaking river, 77 individuals were PIT tagged downstream of Bragado SHP in the North of Portugal. Tagged fish species included Pseudochondrostoma duriense and Squalius carolitertii. Fish positions were recorded manually on two di erent occasions: during hydropeaking events (HP) and without hydropeaking events (NHP). From the 77 tagged fish, we were able to record habitat use for 33 individuals (20 P. duriense and 13 S. carolitertii) in a total of 125 relocations. Fish species were distributed along the river reach with high density in the upstream area in the vicinity of the SHP tailrace, in particular during HP. Fish locations were associated with velocity for P. duriense and S. carolitertii. The latter tended to use faster flowing waters than P. duriense. Our findings on the habitat use in peaking rivers are a valuable tool to help in the selection and design of mitigation measures

River fragmentation by large hydropower plants (LHP) has been recognized as a major threat for potamodromous fish. Fishways have thus been built to partially restore connectivity, with fish lifts representing the most cost-effective type at high head obstacles. This study assessed the effectiveness with which a fish lift in a LHP on the River Lima (Touvedo, Portugal), allows potamodromous fish&mdash, Iberian barbel (Luciobarbus bocagei), Northern straight-mouth nase (Pseudochondrostoma duriense) and brown trout (Salmo trutta fario)- to migrate upstream. Most fish (79.5%) used the lift between summer and early-fall. Water temperature was the most significant predictor of both cyprinids&rsquo, movements, whereas mean daily flow was more important for trout. Movements differed according to peak-flow magnitude: nase (67.8%) made broader use of the lift in the absence of turbined flow, whereas a relevant proportion of barbel (44.8%) and trout (44.2%) passed when the powerhouse was operating at half (50 m3s&minus, 1) and full-load (100 m3s&minus, 1), respectively. Size-selectivity found for barbel and trout could reflect electrofishing bias towards smaller sizes. The comparison of daily abundance patterns in the river with fish lift records allowed the assessment of the lift&rsquo, s efficacy, although biological requirements of target species must be considered. Results are discussed in the context of management strategies, with recommendations for future studies.

A repulsive effect, that some induced primary stimuli, like sound and light, is known to be provoked in fish behavior. In the present study, two strobe light frequencies, 350 flashes/minute and 600 flashes/minute, were tested in laboratorial conditions, using three native freshwater fish species of northern Portugal: Brown trout (Salmo trutta), Northern straight-mouth nase (Pseudochondrostoma duriense) and Iberian barbel (Luciobarbus bocagei). The results showed a differential repulsive behavior of the fish species to light stimulus, and particularly to a frequency of 600 flashes/minute. S. trutta presented the most repulsive behavior, whereas the L. bocagei showed less repulsion to the light stimulus. No relevant differences were found between pre-test and post-assessments, confirming a rapid recovery of natural fish behavior after the deterrent effect. The results highlighted the potential of behavioral barriers, particularly in salmonid streams, based on strobe light stimulus. The project n 13737: Original Solutions—ENI and CITAB-UTAD was funded project: ANI/QREN/FEDER. European Investment Funds by FEDER/COMPETE/POCI—Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145- FEDER-006958 and National Funds by FCT— Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013. Acknowledgments: The present study was technically supported by Instituto da Conservação da Natureza e Florestas (ICNF—Departamento Norte), namely the facilities at the fish farm “Posto Aquicola de Castrelos”, for the execution of the experimental design. info:eu-repo/semantics/publishedVersion

The Vilariça River (located in the northeast of Portugal) is inserted in an agricultural basin and it was chosen to replace the spawning grounds for fish, that was lost due to the construction of dams in the Sabor River. Thus, it is essential to study the effect of agricultural practices on water quality and in the health status of fish. The barbel (Luciobarbus bocagei) and Douro nase (Pseudochondrostoma duriense) were the selected species and the work was developed in two seasons (Summer 2016 and Winter 2017). For that, the histopathological changes of fish gill were used as biomarkers, through a semi-quantitative approach that considers the injuries severity. And the water quality assessment criteria followed the methodologies proposed for classifying the status of surface water bodies from Portugal. The current study showed severe histopathological changes in both species and both seasons, and the water was classified as polluted and extremely polluted in Summer and Winter respectively. The pollution in Summer was due to high temperatures, low dissolved oxygen and major concentration of As and Mn, and in Winter is due to the high concentration of Total Suspended Solids, nitrites and Cd. The increase of values of physico-chemical parameters on the water was caused by the less streamflow and excessive agricultural fertilization in Summer which arrive the river by irrigation, and by the erosion of soil particles with heavy metals associated in Winter. Also, the canonical analysis showed that physico-chemical parameters concentrations in Summer justify the major prevalence of aneurism in barbel and exudate in nase and Winter the major prevalence of hypertrophy in barbel. In conclusion, the study showed that the gill injuries of barbel and Douro nase was correlated with the water quality and it is influenced by seasonal agricultural practices and the flow regime.

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. The project nº 13737: Original Solutions - ENI and CITAB-UTAD was funded project: ANI/QREN/FEDER. The Science and Technology Foundation, Portugal funded M.C.P.A. (strategic projects UID/MAR/04292/2013 granted to MARE) and P.J.F. (UID/BIA/00329/2013 granted to cE3c). The scientific plan was achieved under European Investment Funds by FEDER/COMPETE/POCI– Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UID/AGR/04033. The present study was technically supported by Instituto da Conservação da Natureza e Florestas (ICNF – Delegação Norte), namely the facilities at the fish farm “Estação Aquicola de Castrelos”, info:eu-repo/semantics/publishedVersion

This paper presents sprinting data from Iberian barbel (Luciobarbus bocagei) and northern straight-mouth nase (Pseudochondrostoma duriense), volitionally swimming against high velocity flows (1.5, 2.5 and 3 m s−1) in an open channel flume. Swimming endurance and speed greatly exceeded previously published observations with both species attaining swim speeds >20 body lengths s−1. Flow velocity was the primary variable limiting the distance both species were able to traverse. Barbel swam greater distances than nase at higher flow velocities, with longer individuals attaining greater distances than smaller ones. The results challenge established fish passage guidelines, suggesting that in some cases these species are capable of passing much higher velocities than was previously believed.

A simple, non-lethal method for assigning conservation status is proposed based on three conservation classes (favourable, unfavourable/inadequate and unfavourable/bad) to fish species based on their natural population variability. To demonstrate this approach, conservation classes were allocated to three native fish species (brown trout Salmo trutta L., northern Iberian chub Squalius carolitertii (Doadrio), and northern straight-mouth nase, Pseudochondrostoma duriense (Coelho) based on basic population variables (density and biomass) using a 9-year dataset (2004–2012) from 15 different sampling sites in Spain. The classification of the population variables into conservation classes was made using previously described nonparametric methods and percentiles categorisation, and the current conservation status of the studied species was established. Temporal trend analyses showed that S. carolitertii experienced a progressive significant decrease during the study period, whereas both S. trutta and P. duriense exhibited weak increases in density. The study exemplifies the need to quantify the natural variability of fish resources to reveal their current conservation status and to determine whether short-term and long-term management actions are needed to rehabilitate native fish assemblages. The proposed approach could easily be employed by fisheries managers to monitor the conservational status of other native species.

Approximately 6% of the freshwater living northern straight-mouth nase Pseudochondrostoma duriense in two Spanish rivers had attached post-metamorphic sea lamprey Petromyzon marinus. Minimum prey size was 16·1 cm fork length and 56·3 g mass. The condition factor of attacked P. duriense was 16% lower than that of unattacked conspecifics.

Knowledge of food resource partitioning between sympatric fish species is critical for understanding the fish communities functioning. Four sympatric fish species (Salmo trutta, Pseudochondrostoma duriense, Squalius carolitertii and Barbus bocagei) were captured in August 2010 in the River Tormes (Avila, Central Spain) in order to study food resource partitioning between fish species such us as a possible mechanism enabling their coexistence, using a fuzzy principal component analysis (FPCA). Diet comparison among species shows that detritus are present in P. duriense, B. bocagei and S. carolitertii, although in different occurrence, showing that these species present typically omnivorous feeding habits. Regarding prevalent food, in all species, benthic prey constituted the most important prey in abundance terms, and terrestrial invertebrates were only consumed by S. trutta, S. carolitertii and P. duriense. High overlap values (Schoener's index from 0.8 up to 1) were found, but it may not indicate competition, since species can adopt different strategies to overcome competence. Thus, prey traits analyses suggest that differences in macrohabitat use, drift behaviour of prey and prey size are important adaptive features that may reduce the inter-specific competition in the fish community and permit the food partitioning that allows coexistence.

Trophic interactions are important factors structuring animal communities. We assessed the trophic relations of four fish species that live in sympatry in the River Ladra (NW Spain), and cluster analysis differentiated two feeding strategies: (1) species with omnivorous feeding habits, feeding mainly on detritus and plant material but with aquatic macroinvertebrates as an important complement (Achondrostoma arcasii and Pseudochondrostoma duriense) and (2) species feeding mainly (Salmo trutta) or exclusively (Gasterosteus gymnurus) on aquatic macroinvertebrates. Concerning ingested macroinvertebrates, the trophic overlap was quantified using Schoener’s index and the results obtained revealed a high diet overlap among the species (from 81.3 up to 99.2%). In order to get a deeper insight into mechanisms of fish species coexistence, we used ten biological and ecological traits of macroinvertebrate prey to discriminate feeding preferences. As a result, despite the high similarity among the diets, our analyses suggest that differences in diel activity patterns and drift behaviour of preys, as well as differences in the prey size, are important adaptive features that may reduce the inter-specific competition in the fish community and permit the partitioning of food that allows coexistence.

Pseudochondrostoma duriense and Achondrostoma oligolepis are two Iberian endemic cyprinid fish species that occur in sympatry over most of their distribution range and that are suspected to hybridize in nature. Here, we employed a combination of mitochondrial and microsatellite markers to explore the extent of introgressive hybridization between these fishes. Two natural hybrid zones were identified in different river basins. Introgression was bi-directional and both hybrid zones consisted mostly of parental genotypes/phenotypes (i.e. bimodal hybrid zones). Yet, they appeared to differ in the extent and direction of introgression, which supports the view that they constitute independent outcomes of different hybridization processes probably influenced by environmental features. Several discordances were found between mtDNA and microsatellite results, suggesting that this hybridization process has complex consequences and illustrating the importance of using independent markers to define accurately the hybrid status of individuals in the presence of high levels of backcrossing.

Pseudochondrostoma duriense and Pseudochondrostoma polylepis hybridize extensively with Achondrostoma oligolepis in natural populations. In this first survey, karyotypes were comparatively analyzed by C-, AgNOR- and CMA(3)-banding procedures in pure (non-introgressed) fish specimens. Leuciscinae pattern was evidenced in the three species: metacentrics and submetacentrics dominance, a big subtelo/acrocentric (marker) chromosome pair and a 2n = 50; small macro-structural differences were observed. Heterochromatin was centromere-associated. Exceptions were found at sm1 and st/a1 long arms and at m1, sm3 and sm6 short arms. The st/a1 band was telomeric in the straight-mouth nases and sub-terminal in A. oligolepis. Multiple NORs of heterochromatic nature were found in sm pairs of the three species. Signals were telomeric except for one pair in A. oligolepis. Two to four structural and two functional NORs were found in P. duriense and P. polylepis, and four to six structural and four functional NORs in A. oligolepis. Species-specific markers will prove useful in hybrid zones' cytogenetic characterization and for in-depth studies of genome compatibility-related issues in future studies.

Background Iberian Leuciscinae are greatly diverse comprising taxa of hybrid origin. With highly conservative karyotypes, Iberian Chondrostoma s.l. have recently demonstrated sub-chromosomal differentiation and rapid genome restructuring in natural hybrids, which was confirmed by ribosomal DNA (rDNA) transposition and/or multiplication. To understand the role of repetitive DNAs in the differentiation of their genomes, a genetic and molecular cytogenetic survey was conducted in Achondrostoma oligolepis, Anaecypris hispanica, Iberochondrostoma lemmingii, I. lusitanicum, Pseudochondrostoma duriense, P. polylepis, Squalius pyrenaicus and hybrids between A. oligolepis x (P. duriense/P. polylepis), representing ‘alburnine’, chondrostomine and Squalius lineages. Results Partial Rex3 sequences evidenced high sequence homology among Leuciscinae (≥98 %) and different fish families (80–95 %) proposing a relatively recent activity of these elements in the species inspected. Low nucleotide substitution rates (

Understanding the mechanisms underlying diversification and speciation by introgressive hybridization is currently one of the major challenges in evolutionary biology. Here, the analysis of hybridization between two pairs of Iberian Leuciscinae provided new data on independent hybrid zones involving Achondrostoma oligolepis (AOL) and Pseudochondrostoma duriense (PDU), and confirmed the occurrence of hybrids between AOL and Pseudochondrostoma polylepis (PPO). A multilevel survey combining morphological, genetic and cytogenomic markers on a vast population screening successfully sorted the selected fishes as admixed. Results were similar in both AOL × PDU and AOL × PPO systems. Overall, hybrid morphotypes, cytogenomic data and genetic profiling indicated preferential backcrossing and suggested AOL as a major genomic contributor. Moreover, results implied AOL as more permissive to introgression than PDU or PPO. Although PDU- and PPO-like individuals appeared more resilient to genome modifications, AOL appeared to be more involved and affected by the ongoing hybridization events, as chromosomal translocations were only found in AOL-like individuals. All hybrids analysed evidenced extensive ribosomal DNA (rDNA) polymorphism that was not found in parental species, but usually seen falling within the range of possible parental combinations. Yet, transgressive phenotypes that cannot be explained by normal recombination, including more rDNA clusters than expected or the occurrence of syntenic rDNAs, were also detected. Present results proved rapid genomic evolution providing the genetic novelty for species to persist. In addition, although the ultimate consequences of such apparently extensive and recurrent events remain unknown, modern genome-wide methodologies are of great promise towards answering questions concerning the causes, dynamics and impacts of hybridization.

Knowledge of the distribution of species life stages at multiple spatial scales is fundamental to both a proper assessment of species management and conservation programmes and the ability to predict the consequences of human disturbances for river systems. The habitat requirements of three native cyprinid species—the Iberian barbel Barbus bocagei Steindachner, the Iberian straight-mouth nase Pseudochondrostoma polylepis (Steindachner), and the Northern straight-mouth nase Pseudochondrostoma duriense (Coelho)—were examined at 174 undisturbed or minimally disturbed sites in 8 river catchments across western Iberia, by modelling occurrence and counts of species life stages at two spatial scales—large (regional) and instream (local)—using hurdle models. All the life stages of the barbel showed a negative association with upstream high-gradient river reaches, whereas juvenile P. duriense favoured such areas. Stream width and openness were negatively related with the occurrence of juvenile and small adult barbel, but not with large adults. Juvenile nase, on the other hand, were found to be mainly confined to fast-flowing habitats with high instream cover and coarser substrata. Advanced life stages of the barbel were mainly associated with the “pure” regional and shared components, whereas the purely local attributes accounted for much of the model variation among nases, in particular juveniles, and juvenile barbel. The results of this study are useful for setting or refining management goals, and highlight the need to separately consider life stages when performing conservation-related studies of species distribution.

A new distribution range is proposed for Pseudochondrostoma polylepis and Pseudochondrostoma duriense based on the phylogeny of two nuclear and one mitochondrial gene sequences, with implications on the geographic history of the north-western Iberian drainages.

There is a need to quantify the multivariate quality of a recreational fishery at the site scale to better communicate the relative quality among sites to the public and anglers. Borrowing on the general approach of multimetric indices of biotic integrity (IBIs), we developed fishery quality indices (FQIs) from species quality indices (SQIs) based on measures of fish abundance and size structure for northern and central Portuguese streams. Our FQIs showed regional patterns indicating a range in fishery quality. Higher coldwater FQI scores were mostly found in the northwestern (Minho and Lima), northeastern Douro, and northern Tagus basins. Higher warmwater FQI scores occurred in the eastern Tagus basin. The species that contributed the most to warmwater FQI scores were largemouth bass Micropterus salmoides, pumpkinseed Lepomis gibbosus, the cyprinid Luciobarbus bocagei, chubs Squalius carolitertii and S. pyrenaicus, and nases Pseudochondrostoma duriense and P. polylepis. The chubs, nases, and brown trout Salmo trutta contributed the most to coldwater FQI scores. As expected, our indices were correlated with river size and with disturbance at the catchment, segment, and site scales. Regression models for separate coldwater and warmwater FQIs were stronger than those for the individual SQIs and for an all-site FQI. The correlation was positive between the coldwater FQI and a coldwater IBI but negative between the warmwater FQI and warmwater IBIs. The proposed FQIs offer a quantitative approach for assessing relative fishery quality among sites and for making regional assessments given an appropriate study design. The component SQIs and SQI metrics of the FQIs can be disassociated to determine the population and species characteristics most affected by various environmental variables.

In order to contribute to a better understanding of feeding behaviour of adult twaite shad, Alosa fallax ,i n freshwater, we studied the stomach content composition of 147 A. fallax during their upstream spawning migration in the River Ulla (Galicia, NW Spain). The diet composition included 34 types of prey, aquatic invertebrates being the most important food. The present study is, to the best of our knowledge, the first record of A. fallax feeding on potamodromous Pseudochondrostoma duriense. Additional keywords: Alosa fallax, diet, Iberian Peninsula, piscivorous, Pseudochondrostoma duriense.