Invertebrate community responses to pollutants in mediterranean basins : insights from ecotoxicological approaches = Respostes de la comunitat d'invertebrats als contaminants en conques mediterrànies: aproximacions ecotoxicològiques

Els sistemes aquàtics estan subjectes a la pressió de múltiples pertorbacions que afecten l'estructura i el funcionament de les comunitats i ecosistemes. Una de les principals pressions antropogèniques que afecten als ecosistemes aquàtics és la contaminació. La producció química a gran escala ha donat lloc a l'alliberament de substàncies d’origen agrícola, industrial i urbà als sistemes fluvials. Aquesta contaminació és una amenaça complexa i múltiple, degut a la diversitat de compostos presents i a la dinàmica i interacció d'aquests amb els compartiments físics (aigua, sediments) i biològics (organismes). A les conques fluvials mediterrànies, on la variabilitat hidrològica és important, aquesta contaminació és motiu d'especial preocupació especialment en el període d’estiatge, quan disminueix la capacitat de dilució dels rius. A més dels contaminants prioritaris legislats, molts estudis han posat de manifest la presència generalitzada de microcontaminants, també anomenats contaminants emergents. Aquesta tesi proporciona evidència dels efectes dels contaminants emergents i prioritaris en les comunitats d'invertebrats en quatre conques mediterrànies de la Península Ibèrica: l'Ebre, el Llobregat, el Xúquer i el Guadalquivir, emprant mètodes ecològics, biomarcadors, i bioassaigs in vitro i in vivo. Els efectes ecotoxicològics s’han detectat a nivell de comunitat, de població i d’individu, i s’ha vist que tant els contaminants presents a l’aigua com els acumulats en els sediments superficials afecten l’estructura de la comunitat (riquesa específica i abundàncies). També s’ha vist que la contaminació afecta la capacitat de reproducció (nombre d’ous), el desenvolupament, l’alimentació, el creixement i l’activitat d’enzims antioxidatius (catalasa). En concret, els principals responsables de l’alt risc ecotoxicològic identificats han estat els plaguicides, els alquilfenols industrials, i els metalls. Addicionalment, la tesi es completa amb un experiment de laboratori on es testen els efectes directes i indirectes de dos contaminants concrets en el detritívor Gammarus pulex. Aquesta tesi integra aproximacions observacionals i experimentals de múltiples disciplines ambientals (química ambiental, ecotoxicologia i ecologia) per dilucidar els efectes dels diferents contaminants. La combinació dels assajos experimentals amb estudis de les comunitats naturals permet comprendre millor els efectes de les pertorbacions a diferents escales i prevenir la degradació dels ecosistemes aquàtics, així com millorar la seva gestió.Aquatic systems are under pressure from multiple stressors that affect the structure, the functioning, and the health of species, communities and ecosystems. One of the main human pressures affecting the Earth’s ecosystems is chemical pollution. Large-scale chemical production has led to the release of these compounds from agricultural, industrial, and municipal waste, into riverine, transitional, and marine waters. These pollutants can cause harmful effects on aquatic species and communities, in turn causing severe negative impacts on the structure and functioning of ecosystems, and the goods and services they provide. Chemical pollution is not only one of the most important threats to aquatic systems, it is also a complex threat due to the diversity of compounds present in these systems and the dynamics and interaction of these compounds with physical (water, sediment) and biological (amphibian, fish, invertebrate, algae) compartments. Furthermore, aquatic pollution is of special concern in Mediterranean basins, which suffer high flow variability and receive significant contaminant inputs from diffuse and point sources. Future scenarios predict that these basins will be increasingly affected by both the scarcity of water, enhanced by climate change, and high pollution. Apart from well-known pollutants, called priority substances, many studies have revealed the widespread occurrence of low level concentrations of organic micropollutants, also known as emerging contaminants (ECs), and awareness is growing with regard to the importance of mixtures of several contaminants, priority and emerging, as found in the more polluted water bodies of Europe. Different tools can be used in order to link the chemical and ecological quality of aquatic ecosystems: ecological methods (Chapter 3, 4 and 5), biomarkers of field-exposed organisms (Chapter 4 and 6), or in vitro and in vivo bioassays (Chapter 4, 5, 6 and 7). By combining these different tools, this thesis provides evidence of the effects of chemical stressors (emerging and priority pollutants) on invertebrate communities in four threatened Mediterranean rivers of the Iberian Peninsula: the Ebro, the Llobregat, the Júcar and the Guadalquivir. The ecotoxicological effects have been detected at community level, but also at the population and individual levels. It has been found that contaminants in water and also contaminants accumulated in surficial sediments affect community structure (richness and abundance). It has also been seen these contaminants affect the reproductive capacity (number of eggs), the development, the feeding, growth and the activity of antioxidative enzymes (catalase) of diferent species. The general picture obtained shows that priority and emerging pollutants represent a serious toxicity risk for invertebrates in these basins, and that the main specific drivers of this risk are pesticides, alkylphenols, and metals. Additionally, the thesis is completed with a laboratory experiment where the direct and indirect effects of two specific contaminants in the detritivorous Gammarus pulex have been tested. Research based on field studies and laboratory toxicity tests, combining experimental assays with natural community studies, is essential to understand the effects of stressors at different scales and to prevent future degradation of aquatic ecosystems. This thesis integrates observational and experimental approaches of diferent environmental disciplines (environmental chemistry, ecotoxicology and ecology) to elucidate the effects of pollution. The combination of experimental tests to study natural communities can help to understand the effects of disturbances at different scales and prevent the degradation of aquatic ecosystems and improve its management

Axonal plasticity in response to active forces generated through magnetic nano-pulling

Mechanical force is crucial in guiding axon outgrowth before and after synapse formation. This process is referred to as "stretch growth." However, how neurons transduce mechanical input into signaling pathways remains poorly understood. Another open question is how stretch growth is coupled in time with the intercalated addition of new mass along the entire axon. Here, we demonstrate that active mechanical force generated by magnetic nano-pulling induces remodeling of the axonal cytoskeleton. Specifically, the increase in the axonal density of microtubules induced by nano-pulling leads to an accumulation of organelles and signaling vesicles, which, in turn, promotes local translation by increasing the probability of assembly of the "translation factories." Modulation of axonal transport and local translation sustains enhanced axon outgrowth and synapse maturation

Increasing the genetic diagnosis yield in inherited retinal dystrophies: assigning pathogenicity to novel non-canonical splice site variants

Aims: We aimed to validate the pathogenicity of genetic variants identified in inherited retinal dystrophy (IRD) patients, which were located in non-canonical splice sites (NCSS). Methods: After next generation sequencing (NGS) analysis (target gene panels or whole exome sequencing (WES)), NCSS variants were prioritized according to in silico predictions. In vivo and in vitro functional tests were used to validate their pathogenicity. Results: Four novel NCSS variants have been identified. They are located in intron 33 and 34 of ABCA4 (c.4774-9G>A and c.4849-8C>G, respectively), intron 2 of POC1B (c.101-3T>G) and intron 3 of RP2 (c.884-14G>A). Functional analysis detected different aberrant splicing events, including intron retention, exon skipping and intronic nucleotide addition, whose molecular effect was either the disruption or the elongation of the open reading frame of the corresponding gene. Conclusions: Our data increase the genetic diagnostic yield of IRD patients and expand the landscape of pathogenic variants, which will have an impact on the genotype-phenotype correlations and allow patients to opt for the emerging gene and cell therapies

Gut microbiome signatures linked to HIV-1 reservoir size and viremia control

Background The potential role of the gut microbiome as a predictor of immune-mediated HIV-1 control in the absence of antiretroviral therapy (ART) is still unknown. In the BCN02 clinical trial, which combined the MVA.HIVconsv immunogen with the latency-reversing agent romidepsin in early-ART treated HIV-1 infected individuals, 23% (3/13) of participants showed sustained low-levels of plasma viremia during 32 weeks of a monitored ART pause (MAP). Here, we present a multi-omics analysis to identify compositional and functional gut microbiome patterns associated with HIV-1 control in the BCN02 trial. Results Viremic controllers during the MAP (controllers) exhibited higher Bacteroidales/Clostridiales ratio and lower microbial gene richness before vaccination and throughout the study intervention when compared to non-controllers. Longitudinal assessment indicated that the gut microbiome of controllers was enriched in pro-inflammatory bacteria and depleted in butyrate-producing bacteria and methanogenic archaea. Functional profiling also showed that metabolic pathways related to fatty acid and lipid biosynthesis were significantly increased in controllers. Fecal metaproteome analyses confirmed that baseline functional differences were mainly driven by Clostridiales. Participants with high baseline Bacteroidales/Clostridiales ratio had increased pre-existing immune activation-related transcripts. The Bacteroidales/Clostridiales ratio as well as host immune-activation signatures inversely correlated with HIV-1 reservoir size. Conclusions The present proof-of-concept study suggests the Bacteroidales/Clostridiales ratio as a novel gut microbiome signature associated with HIV-1 reservoir size and immune-mediated viral control after ART interruption.Peer ReviewedObjectius de Desenvolupament Sostenible::3 - Salut i BenestarObjectius de Desenvolupament Sostenible::10 - Reducció de les DesigualtatsPostprint (published version

Transcriptomic, biochemical and individual markers in transplanted Daphnia magna to characterize impacts in the field.

Daphnia magna individuals were transplanted across 12 sites from three Spanish river basins (Llobregat, Ebro, Jucar) showing different sources of pollution. Gene transcription, feeding and biochemical responses in the field were assessed and compared with those obtained in re-constituted water treatments spiked with organic eluates obtained from water samples collected at the same locations and sampling periods. Up to 166 trace contaminants were detected in water and classified by their mode of action into 45 groups that included metals, pharmaceuticals, pesticides, illicit drugs, and other industrial compounds. Physicochemical water parameters differentiated the three river basins with Llobregat having the highest levels of conductivity, metals and pharmaceuticals, followed by Ebro, whereas the Jucar river had the greatest levels of illicit drugs. D. magna grazing rates and cholinesterase activity responded similarly than the diversity of riparian benthic communities. Transcription patterns of 13 different genes encoding for general stress, metabolism and energy processes, molting and xenobiotic transporters corroborate phenotypic responses differentiated sites within and across river basins. Principal Component Analysis and Partial Least Square Projections to Latent Structures regression analyses indicated that measured in situ responses of most genes and biomarkers and that of benthic macroinvertebrate diversity indexes were affected by distinct environmental factors. Conductivity, suspended solids and fungicides were negatively related with the diversity of macroinvertebrates cholinesterase, and feeding responses. Gene transcripts of heat shock protein and metallothionein were positively related with 11 classes of organic contaminants and 6 metals. Gene transcripts related with signaling paths of molting and reproduction, sugar, protein and xenobiotic metabolism responded similarly in field and lab exposures and were related with high residue concentrations of analgesics, diuretics, psychiatric drugs, β blockers, illicit drugs, trizoles, bisphenol A, caffeine and pesticides. These results indicate that application of omic technologies in the field is a promising subject in water management

Controles locales y regionales del metabolismo en ríos de cabecera: aportaciones del primer proyecto colaborativo de la AIL

Streams play a key role in the global biogeochemical cycles, processing material from adjacent terrestrial systems and transporting it downstream. However, the drivers of stream metabolism, especially those acting at broad spatial scales, are still not well understood. Moreover, stream metabolism can be affected by hydrological changes associated with seasonality, and thus, assessing the temporality of metabolic rates is a key question to understand stream function. This study aims to analyse the geographical and temporal patterns in stream metabolism and to identify the main drivers regulating the wholeecosystem metabolic rates at local and regional scales. Using a coordinated distributed experiment, we studied ten headwaters streams located across five European ecoregions during summer and fall 2014.We characterized the magnitude and variability of gross primary production (GPP) and ecosystem respiration (ER) with the open-channel method. Moreover, we examined several climatic, geographical, hydrological, morphological, and physicochemical variables that can potentially control stream metabolic rates. Daily rates of stream metabolism varied considerately across streams, with GPP and ER ranging from 0.06 to 4.33 g O2 m?2 d?1 and from 0.72 to 14.20 g O2 m?2 d?1, respectively. All streams were highly heterotrophic (P/R < 1), except the southernmost one. We found that the drier climates tended to have the highest GPP, while humid regions presented the highest ER. Between the sampling periods no statistical differences were found. Partial-least squares models (PLS) explained ?80% of the variance in GPP and ER rates across headwater streams and included both local and regional variables. Rates of GPP varied primarily in response to the local variables, such as streambed substrate and stream water temperature. In contrast, regional variables, such as the mean annual temperature or the land use of the catchment, had more relevance to explain ER. Overall, our results highlight that stream metabolism depends on both local and regional drivers and show the positive experience of a young network of researchers to assess scientific challenges across large-scale geographic areas.Los ecosistemas fluviales tienen un papel relevante en los ciclos biogeoquímicos globales, ya que procesan el material de la cuenca y lo transportan río abajo. Sin embargo, los factores que afectan al metabolismo, especialmente aquellos que actúan en una escala espacial más amplia, todavía no se conocen completamente. Además, el metabolismo fluvial puede variar a causa de cambios hidrológicos relacionados con la estacionalidad; lo que también hace importante incluir la variabilidad temporal para entender el funcionamiento de los ríos. Mediante el uso de un experimento distribuido y coordinado, estudiamos diez ríos de cabecera localizados a lo largo de cinco ecoregiones europeas durante verano y otoño del 2014. Se midió la magnitud y la variabilidad de la producción primaria bruta (PPB) y la respiración ecosistémica (RE) mediante el método de canal abierto. Así mismo, se examinaron los factores climáticos, hidrológicos y físico-químicos que potencialmente pueden afectar las tasas metabólicas fluviales. Las tasas diarias de metabolismo fluvial variaron considerablemente entre ríos, con valores de PPB de entre 0.06 a 4.33 g O2 m-2 día-1, y de RE de entre 0.72 a 14.20 g O2 m-2 día-1. Todos los ríos fueron altamente heterotróficos (P/R < 1), excepto el río más meridional. Se observó que los climas secos tendían a tener mayor PPB y las regiones húmedas tendían a tener una mayor RE. No se encontraron diferencias estadísticas entre los períodos de estudio. Los modelos de regresión de mínimos cuadrados parciales (PLS) explicaron --80% de la varianza de PPB y RE para los ríos estudiados. Las tasas de PPB variaron principalmente con factores locales como el sustrato del lecho del río y la temperatura del agua. En cambio, las tasas de RE estuvieron estrechamente relacionadas con factores regionales, como la temperatura media anual y los impactos en la cuenca. En conjunto, nuestros resultados resaltan la necesidad de incluir la escala local y regional en el estudio del metabolismo fluvial y muestran la experiencia positiva de una red coordinada de jóvenes investigadores para lograr retos científicos de amplia escala geográfica.We thank the support of the AIL board throughout the development of the project and all the affiliated institutions for the accessibility of laboratory premises, especially the Limnology Department of Uppsala University and the Department d’Ecologia of the Universitat de Barcelona. We greatly thank all the extended team of participants for their valuable contribution in DOMIPEX project. The authors were supported by the following founding: NC by Wenner-Gren foundation stipend (Sweden, 2014-2016) and a Juan de la Cierva grant (FJCI-2014-23064), EE by a pre-doctoral grant from the Basque Government (2014-2017), AGB by a Swedish Research Council grant (SMAREF 2014-2016), PRL by a Ramón Areces Foundation Postdoctoral Scholarship AM by the Univ. of the Basque Country, AMGF and SP by pre-doctoral research grants from the Spanish Ministry of Economy and Competitiveness (Ref: BES-2013-065770, Ref: BES-2012-059743, respectively), and RR by the IMDEA-Water Institute. DOMIPEX project was founded by the First Call of Collaborative Projects among Young Researchers of the Iberian Association of Limnology (AIL; 2013-2015)

Analysis of monitoring programmes and their suitability for ecotoxicological risk assessment in four Spanish basins

Data from four Spanish basin management authorities were analysed. Chemical and biological data from four Spanish basin management authorities were analysed, focusing on three consecutive years. Aims were to i) determine the chemicals most likely responsible for the environmental toxicological risk in the four Spanish basins and ii) investigate the relationships between toxicological risk and biological status in these catchments. The toxicological risk of chemicals was evaluated using the toxic unit (TU) concept. With these data we considered if the potential risk properly reflects the risk to the community or, alternatively, if new criteria should be developed to improve risk assessment. Data study revealed inadequacies in processing and monitoring that should be improved (e.g., site coincidence for chemical and biological sampling). Analysis of the chemical data revealed high potential toxicological risk in the majority of sampling points, to which metals were the main contributors to this risk. However, clear relationships between biological quality and chemical risk were found only in one river. Further investigation of metal toxicity may be necessary, and future analyses are necessary to accurately estimate the risk to the environment.The present work was funded by the Spanish Ministry of Economics and Competitiveness through the Consolider-Ingenio 2010 Program (project Scarce CSD2009-00065). The authors would like to acknowledge the Confederacion Hidrografica del Ebro (C. Duran and V. Sanchez-Tello), Confederacion Hidrografica del Guadalquivir (V. Cifuentes), Agenda Andaluza del Agua, Agenda Catalana de l'Aigua (A Munne, L Tirapu) and Confederacion Hidrografica del Jucar (MA Pinon) which kindly provided the monitoring data. Nuria De Castro-Catala holds a predoctoral grant from the University of Barcelona. We would like to thank two anonymous reviewers for their suggestions which have improved this manuscript.López Doval, JC.; De Castro Catala, N.; Andrés Doménech, I.; Blasco, J.; Ginebreda, A.; Muñoz, I. (2012). Analysis of monitoring programmes and their suitability for ecotoxicological risk assessment in four Spanish basins. Science of the Total Environment. 440:194-203. https://doi.org/10.1016/j.scitotenv.2012.07.035S19420344

Early-Career Coordinated Distributed Experiments: Empowerment Through Collaboration

Este artículo contiene 7 páginas, 1 tabla, 3 figuras.Coordinated distributed experiments (CDEs) enable the study of large-scale ecological patterns in geographically dispersed areas, while simultaneously providing broad academic and personal benefits for the participants. However, the effective involvement of early-career researchers (ECRs) presents major challenges. Here, we analyze the benefits and challenges of the first CDE exclusively led and conducted by ECRs (i.e. ECR-CDE), which sets a baseline for similar CDEs, and we provide recommendations for successful CDE execution. ECR-CDEs achieve most of the outcomes identified in conventional CDEs as well as extensive benefits for the young cohort of researchers, including: (i) receiving scientific credit, (ii) peer-training in new concepts and methods, (iii) developing leadership and communication skills, (iv) promoting a peer network among ECRs, and (v) building on individual engagement and independence. We also discuss the challenges of ECR-CDEs, which are mainly derived from the lack of independence and instability of the participants, and we suggest mechanisms to address them, such as resource re-allocation and communication strategies. We conclude that ECR-CDEs can be a relevant tool to empower ECRs across disciplines by fostering their training, networking and personal well-being.The authors were supported by the following founding: NC the support of the Beatriu de Pinós postdoctoral program of the Government of Catalonia’s Secretariat for Universities and Research of the Ministry of Economy and Knowledge (BP2016- 00215), EE by a predoctoral grant from the Basque Government (2014-2017), AB by a Generalitat de Catalunya—Beatriu de Pinós (BP-00385-2016), AMG-F by a predoctoral research grant (BES-2013-065770) from the Spanish Ministry of Economy and Competitiveness, MAr by a postdoctoral grant from the Basque Government, MIA by a Juan de la Cierva postdoctoral grant (FJCI-2015-26192), PR-L by a Margalida Comas postdoctoral contract (PD/031/2018) funded by the Government of the Balearic Islands and the European Social Fund, AP by a Ramón Areces Foundation Postdoctoral Scholarship, and AL by a Kempe Foundation stipend. DOMIPEX project was founded by the First Call of Collaborative Projects among Young Researchers of the Iberian Association of Limnology (AIL; 2013-2015).Peer reviewe