Species tree analyses and speciation-based species delimitation support new species in the relict catfish family Diplomystidae and provide insights on recent glacial history in Patagonia.

Diplomystidae is an early-diverged family of freshwater catfish endemic to southern South America. We have recently collected five juvenile specimens belonging to this family from the Bueno River Basin, a basin which the only previous record was a single juvenile specimen collected in 1996. This finding confirms the distribution of the family further South in northern Patagonia, but poses new questions about the origin of this population in an area with a strong glacial history. We used phylogenetic analyses to evaluate three different hypotheses that could explain the origin of this population in the basin. First, the population could have originated in Atlantic basins (East of the Andes) and dispersed to the Bueno Basin after the Last Glacial Maximum (LGM) via river reversals, as it has been proposed for other population of Diplomystes as well as for other freshwater species from Patagonia. Second, the population could have originated in the geographically close Valdivia Basin (West of the Andes) and dispersed south to its current location in the Bueno Basin. Third, regardless of its geographic origin (West or East of the Andes), the Bueno Basin population could have a longer history in the basin, surviving in situ through the LGM. In addition, we conducted species delimitation analyses using a recently developed method that uses a protracted model of speciation. Our goal was to test the species status of the Bueno Basin population along with another controversial population in Central Chile (Biobío Basin), which appeared highly divergent in previous studies with mtDNA. The phylogenetic analyses showed that the population from the Bueno Basin is more related to Atlantic than to Pacific lineages, although with a deep divergence that predated the LGM, supporting in situ survival rather than postglacial dispersal. In addition, these analyses also showed that the species D. nahuelbutaensis is polyphyletic, supporting the need for a taxonomic reevaluation. The species delimitation analyses supported two new species which are described using molecular diagnostic characters: Diplomystes arratiae sp. nov. from the Biobío, Carampangue, and Laraquete basins, maintaining D. nahuelbutaensis valid only for the Imperial Basin, and Diplomystes habitae sp. nov. from the Bueno Basin. This study greatly increases the number of species within both the family Diplomystidae and Patagonia, and contributes substantially to the knowledge of the evolution of southern South American freshwater biodiversity during its glacial history. Given the important contribution to the phylogenetic diversity of the family, we recommend a high conservation priority for both new species. Finally, this study highlights an exemplary scenario where species descriptions based only on DNA data are particularly valuable, bringing additional elements to the ongoing debate on DNA-based taxonomy.

Functional response of fish assemblage to multiple stressors in a highly regulated Mediterranean river system.

Mediterranean rivers are characterised by strong environmental constrains and species-poor, highly endemic fish fauna. In Europe, these systems are exposed to multiple stressors due to extensive human activities. Studies on the effects of some stressors on riverine fish are available but complex responses of fish assemblages to interplay of flow alteration with physical habitat changes and invasive species have not been evaluated up to date. This study analysed the response of functional diversity of fish assemblages to multiple stressors in the Segura River system in the southern Spain. Fish assemblages were sampled in 16 sites in two consecutive periods (2009-2010 and 2013-2015). Subsequently, we assessed the responses of functional specialisation, originality and entropy (based on nine functional traits and abundances) as well as species richness and abundance to interplay of flow regime alteration and ecological status, fragmentation as well as non-native species abundance across spatial and temporal scales. The governing role of flow regime in structuring fish assemblage was superimposed on physical habitat changes, water quality deterioration and fragmentation as well as the presence of non-native fish species. We found an increase of species richness and abundance but decrease of functional specialisation and originality in river reaches with high level of base flow and more stable hydrological conditions. Opposite pattern was observed in reaches with severe reduction of base flow and marked inversion in the seasonal pattern of high and low flows. We postulate that the use of tools that consider the functional identity of the species as method to assess the effects of environmental alterations on fish biodiversity could improve conservation measures for Mediterranean fish fauna. Furthermore, design flows that mimic natural flow regime patterns characteristic for Mediterranean rivers are a promising tool to provide environmental conditions that would favour native fish within the assemblage and benefit their conservation.

A Century after! Rediscovery of the ancient catfish Diplomystes Bleeker 1858 (Siluriformes: Diplomystidae) in coastal river basins of Chile and its implications for conservation

The ancient catfish family Diplomystidae, with seven species endemic to rivers of southern South America, represents one of the oldest branches of the diverse order Siluriformes. With most species endangered, new reports of these species become extremely valuable for conservation. Currently, it is assumed that Diplomystes species inhabit only Andean (large) basins, and that they are extinct from coastal (small) basins from which their presence have not been recorded since 1919. Here, we document new records of the family Diplomystidae in the Laraquete and Carampangue basins, two coastal basins from the Nahuelbuta Coast Range, Chile, with no previous reports. This finding represents the rediscovery of the genus in coastal basins in more than a Century. Based on analysis of mitochondrial DNA sequences, the collected specimens were found to be closely related to Diplomystes nahuelbutaensis from the Andean Biobío Basin, but sufficiently differentiated to suggest that coastal basin populations are a different management unit. These populations are important because, contrary to previous thoughts, they prove these catfish can survive in small river networks, providing unique opportunities for research and conservation. The conservation category of Critically Endangered (CE) is recommended for the populations from the Laraquete and Carampangue basins.(AU)

La familia de bagres Diplomystidae, con siete especies endémicas de ríos del sur de Sudamérica, es uno de los linajes mas antiguos del diverso orden Siluriformes. Al estar la mayoría de las especies amenazadas, nuevos registros de éstas son extremadamente valiosos para su conservación. Actualmente, se ha asumido que los Diplomystidos se distribuyen solo en cuencas Andinas (más grandes), y que sus especies estarían extintas en cuencas de menor tamaño como las costeras, sin registros desde 1919. En este trabajo documentamos la familia Diplomistidae en las cuencas de Carampangue y Laraquete, dos cuencas costeras de la Cordillera de Nahuelbuta, Chile, lo que representa el primer registro de esta familia en estas cuencas costeras. Además, este hallazgo representa el re-descubrimiento de la familia en cuencas costeras después de un siglo. Sobre la base de análisis de ADN mitocondrial, los especímenes colectados se relacionaron más cercanamente con poblaciones de la especie Diplomystes nahuelbutaensis presente en la cuenca del Biobío. Sin embargo, existen diferencias genéticas suficientes entre las poblaciones costeras y las del Biobío para justificar su separación como unidad de manejo distinta. Estas poblaciones costeras son importantes porque demuestran que los Diplomístidos pueden sobrevivir en cuencas de pequeño tamaño, ofreciendo oportunidades únicas para su investigación y conservación. Se recomienda la categoría de conservación En Peligro Critico de Extinción (CR) para las poblaciones de las cuencas Laraquete y Carampangue.(AU)

Assessing the impact of waste water effluents on native fish species from a semi-arid region, NE Spain.

Mediterranean rivers are strongly affected by pollution and water scarcity. Over the summer period, urban and industrial effluents arrive into the rivers with little dilution. In order to assess the water quality, two native fish species, Barbus meridionalis and Squalius laietanus, were collected from six sites along the Ripoll River (Spain). PAH metabolites, alkylphenols (nonylphenol and octylphenol) and the musk galaxolide levels were determined in bile. 7-Ethoxyresorufin-O-deethylase (EROD) and 7-benzyloxy-4-trifluoromethyl-coumarin O-debenzyloxylase (BFCOD) were measured as metabolic biomarkers, and the activity of CYP19 aromatase was determined in the ovaries of B. meridionalis as a biomarker of endocrine disruption. The analysis of bile indicated that fish from the lower course of the river were highly exposed to different pollutants. Accordingly, a significant induction of EROD (9 to 10-fold) and BFCOD (3 to 5-fold) activities were detected in both fish species together with an increased aromatase activity in females of B. meridionalis from the most polluted sites. Considering that sewage treatment plant (STP) effluents are essential for maintaining environmental flows in small Mediterranean rivers, this study highlights the need to improve the efficiency of STPs to protect fish health.

Functional diversity measures revealed impacts of non-native species and habitat degradation on species-poor freshwater fish assemblages.

Trait-based ecology has been developed for decades to infer ecosystem responses to stressors based on the functional structure of communities, yet its value in species-poor systems is largely unknown. Here, we used an extensive dataset in a Spanish region highly prone to non-native fish invasions (15 catchments, N=389 sites) to assess for the first time how species-poor communities respond to large-scale environmental gradients using a taxonomic and functional trait-based approach in riverine fish. We examined total species richness and three functional trait-based indices available when many sites have ≤3 species (specialization, FSpe; originality, FOri and entropy, FEnt). We assessed the responses of these taxonomic and functional indices along gradients of altitude, water pollution, physical habitat degradation and non-native fish biomass. Whilst species richness was relatively sensitive to spatial effects, functional diversity indices were responsive across natural and anthropogenic gradients. All four diversity measures declined with altitude but this decline was modulated by physical habitat degradation (richness, FSpe and FEnt) and the non-native:total fish biomass ratio (FSpe and FOri) in ways that varied between indices. Furthermore, FSpe and FOri were significantly correlated with Total Nitrogen. Non-native fish were a major component of the taxonomic and functional structure of fish communities, raising concerns about potential misdiagnosis between invaded and environmentally-degraded river reaches. Such misdiagnosis was evident in a regional fish index widely used in official monitoring programs. We recommend the application of FSpe and FOri to extensive datasets from monitoring programs in order to generate valuable cross-system information about the impacts of non-native species and habitat degradation, even in species-poor systems. Scoring non-native species apart from habitat degradation in the indices used to determine ecosystem health is essential to develop better management strategies.

Movement patterns and home range in Diplomystes camposensis (Siluriformes: Diplomystidae), an endemic and threatened species from Chile

We document movement patterns and home range of Diplomystes camposensis, an endemic and threatened freshwater catfish from Chile. We tracked the movements of seven individuals of different body size (13.5 to 19 cm SL) using portable radio telemetry equipment to investigate movement patterns in relation to day/night activity and habitat use in the San Pedro River (Valdivia Basin). Tracked movements and model-based analyses revealed that D. camposensis has a large home range and high mobility. The average home range was 0.068163 ± 0.033313 km2, and the average area of higher activity was 0.005646 ± 0.011386 km2. The mean linear home range was 387.4 m. The results also showed that movements were longer during the night, supporting nocturnal habits. Movements tended to be in an upstream direction for some individuals, although these differences were not significant when data was pooled. Large home range and movements suggest that the species may require large river areas to meet ecological demands, an aspect that could be severely affected by fragmentation. These results, along with previously published genetic data, suggest that the conservation of D. camposensis would be seriously threatened by hydromorphological alterations (e.g. lack of connectivity), such as those resulting from dam building.(AU)

En este trabajo documentamos patrones de movimiento y estimación de ámbito de hogar de Diplomystes camposensis, un siluriforme endémico y amenazado del Sur de Chile. Por medio de radio telemetría, se monitorearon 7 individuos con un rango de tamaño entre 13.5 y 19 cm de longitud estándar, para evaluar patrones de movimiento con respecto al uso de hábitat y tiempo de actividad (dia/noche) en la zona del Río San Pedro, Cuenca del Río Valdivia. Los resultados muestran que D. camposensis tiene un ámbito de hogar grande y una alta movilidad. El ámbito de hogar fue de 0.068163 ± 0.033313 km2 con un área promedio de mayor actividad de 0.005646 ± 0.011386 km2. El ámbito de hogar lineal medio fue de 387.4 m. Los resultados también mostraron que la especie presenta una mayor actividad por la noche y una tendencia hacia un mayor flujo de movimiento en dirección aguas arriba, aunque esto último no fue significativo. Un ámbito de hogar grande y su alta movilidad sugieren que la especie podría requerir de amplias zonas del río para satisfacer sus demandas ecológicas. Al igual que estudos previos con datos genéticos, estos resultados sugieren que la especie D. camposensis se vería perjudicada por alteraciones en la hidromorfología del cauce (e.g. falta de conectividad) tales como aquellas que resulten de la construcción de represas.(AU)

Trends in biomarkers, biotic indices, and fish population size revealed contrasting long-term effects of recycled water on the ecological status of a Mediterranean river.

Recycled water is important for maintaining river flow in semi-arid regions. However, it has ecological risk, as suggested by comparison of habitat and white and red blood cell count in two wild fish species (Barbus meridionalis and Squalius laietanus) before and after an input of recycled water in Ripoll River (NE Spain) in 2009. Due to the lack of normal ranges for blood variables in wild fish, we surveyed seasonally the same river reaches in 2013 to test if blood alterations from 2009 compromised the viability of the fish populations. By examining other indicators of river health in baseline and polluted sites (fish abundance, mass-length relationships, and community indices in fish, diatoms and invertebrates), we tested for the superior utility of blood tests in biomonitoring. The comparison of water quality and scores of diatoms and invertebrate indices between polluted and reference sites showed that polluted sites improved from 2009 to 2013. The abundance of B. meridionalis also increased in polluted sites, but that of S. laietanus declined in 2013 compared to 2009. These results contrast with results of blood analyses in 2009, which suggested that B. meridionalis was more seriously affected by pollution than S. laietanus. The fish index did not reveal the risk of recycled water to fish health, whereas fish mass-length relationships suggested that S. laietanus individuals in 2013 had a better body condition in polluted than in reference sites. Given that the two fish species had opposite results in reference sites, and that the physical habitat was more suitable for B. meridionalis in polluted sites in 2013 than was for S. laietanus, trends in population size are not only explained by pollution. The role of phenology is suggested by peaks in blood disorders during the breeding season. However, more long-term studies combining indicators of river health at the individual and community scales are needed to fully assess the ecological risk of recycled water in this river. These studies will also help to develop blood tests as reliable health indicators in wild fish populations.

Ecological impact and recovery of a Mediterranean river after receiving the effluent from a textile dyeing industry.

The textile industry is one of the largest sectors globally, representing up to 20% of industrial water pollution. However, there is limited insight into how fluvial ecosystems respond and recover from this impact. From summer 2012 to spring 2013, we examined water quality and ecological status upstream and 1.5km downstream the input of a textile industry wastewater treatment plant (WWTP) in Ripoll River, NE Spain. The ecological status was determined via diversity measures and 10 biotic indices based on diatoms, macrophytes, macroinvertebrates and fish. Our results showed that the WWTP severely deteriorated water quality and biological communities at the discharge site, but that they improved at 1.5km downstream. Severity also varied across taxa and seasons, being fish the most affected taxa and spring the season with the best ecological status. The strong correlation amongst water quality variables and many biotic indices across taxa indicated that this is a chronic pollution event affecting multiple trophic levels. Thus, this study suggests that there is an urgent need to invest in wastewater treatment in this industry to preserve the ecological integrity of Ripoll River and especially its fish fauna. Likewise, it illustrates the diagnostic power of biotic indices based on diatoms, macroinvertebrates and fish, as driven by the European Water Framework Directive.

Ecological relevance of biomarkers in monitoring studies of macro-invertebrates and fish in Mediterranean rivers.

Mediterranean rivers are probably one of the most singular and endangered ecosystems worldwide due to the presence of many endemic species and a long history of anthropogenic impacts. Besides a conservation value per se, biodiversity is related to the services that ecosystems provide to society and the ability of these to cope with stressors, including climate change. Using macro-invertebrates and fish as sentinel organisms, this overview presents a synthesis of the state of the art in the application of biomarkers (stress and enzymatic responses, endocrine disruptors, trophic tracers, energy and bile metabolites, genotoxic indicators, histopathological and behavioural alterations, and genetic and cutting edge omic markers) to determine the causes and effects of anthropogenic stressors on the biodiversity of European Mediterranean rivers. We also discuss how a careful selection of sentinel species according to their ecological traits and the food-web structure of Mediterranean rivers could increase the ecological relevance of biomarker responses. Further, we provide suggestions to better harmonise ecological realism with experimental design in biomarker studies, including statistical analyses, which may also deliver a more comprehensible message to managers and policy makers. By keeping on the safe side the health status of populations of multiple-species in a community, we advocate to increase the resilience of fluvial ecosystems to face present and forecasted stressors. In conclusion, this review provides evidence that multi-biomarker approaches detect early signs of impairment in populations, and supports their incorporation in the standardised procedures of the Water Frame Work Directive to better appraise the status of European water bodies.