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The detection of fecal viral pathogens in water is hampered by their great variety and complex analysis. As traditional bacterial indicators are poor viral indicators, there is a need for alternative methods, such as the use of somatic coliphages, which have been included in water safety regulations in recent years. Some researchers have also recommended the use of reference viral pathogens such as noroviruses or other enteric viruses to improve the prediction of fecal viral pollution of human origin. In this work, phages previously tested in microbial source tracking studies were compared with norovirus and adenovirus for their suitability as indicators of human fecal viruses. The phages, namely those infecting human-associated Bacteroides thetaiotaomicron strain GA17 (GA17PH) and porcine-associated Bacteroides strain PG76 (PGPH), and the human-associated crAssphage marker (crAssPH), were evaluated in sewage samples and fecal mixtures obtained from different animals in five European countries, along with norovirus GI + GII (NoV) and human adenovirus (HAdV). GA17PH had an overall sensitivity of ≥83% and the highest specificity (>88%) for human pollution source detection. crAssPH showed the highest sensitivity (100%) and specificity (100%) in northern European countries but a much lower specificity in Spain and Portugal (10 and 30%, respectively), being detected in animal wastewater samples with a high concentration of fecal indicators. The correlations between GA17PH, crAssPH, or the sum of both (BACPH) and HAdV or NoV were higher than between the two human viruses, indicating that bacteriophages are feasible indicators of human viral pathogens of fecal origin and constitute a promising, easy to use and affordable alternative to human viruses for routine water safety monitoring.

This article considers current trends in antimicrobial resistance (AMR) research and knowledge gaps relevant to policymaking in the water sector. Specifically, biological indicators of AMR (antibiotic-resistant bacteria and their resistance genes) and detection methods that have been used so far are identified and discussed, as well as the problems with and solutions to the collection of AMR data, sewage surveillance lessons from the COVID-19 pandemic, and the financial burden caused by AMR, which could be synergically used to improve advocacy on AMR issues in the water sector. Finally, this article proposes solutions to overcoming existing hurdles and shortening the time it will take to have an impact on policymaking and regulation in the sector This work was supported by the European Union’s Horizon 2020 Research and Innovation Programme under the Marie Skłodowska-Curie grant agreement no. 792686, acronym ENVIROSTOME. This work was also supported by the Generalitat de Catalunya (ICRA-ENV 2017 SGR 1124) and by a New Lecturer Research Grant from the Society for Applied Microbiology. EGV thanks the National Council for Science and Technology (CONACYT, Mexico) for the postdoctoral fellowship (CVU no. 441304)

Programa de Doctorat: Ciències Experimentals i Tecnologies Les activitats mineres han estat crucials per al desenvolupament econòmic i social d’Europa al llarg dels segles. No obstant, avui en dia a Europa hi ha més mines abandonades que operatives que causen greus impactes sobre els ecosistemes aquàtics, principalment mitjançant l’abocament incontrolat, i encara no regulat, d’efluents contaminants, essent d’especial preocupació els efluents hipersalins i metàl·lics. En la present tesis, els biofilms aquàtics s’han fet servir per avaluar l’eficiència de diferents tecnologies de tractament (en el cas dels efluents metàl·lics) i diferents dilucions (per als efluents salins), per reduir els seus impactes ens els ecosistemes aquàtics. Els resultats demostren la viabilitat d’utilitzar els biofilms com a bioindicador, especialment l’ús de les variables funcionals del biofilm, essent les que van respondre més ràpidament a l’estrès i la recuperació. Per altra banda, aquesta tesi proporciona noves evidencies per a determinar els llindars de salinitat a partir dels quals els impactes sobre les els ecosistemes aquàtics poden assolir un punt de no retorn. Mining activities have been crucial for the economic and social development of Europe over the centuries. However, nowadays there are significantly more abandoned than operational mines in Europe, which cause serious impacts on freshwater ecosystems mostly through the uncontrolled and still non-regulated discharge of polluted effluents being of special concern hypersaline and metal effluents. In the present thesis, aquatic biofilms have been used to evaluate the efficiency of different treatment technologies (in the case of metal effluents) and different dilutions (for saline effluents), to reduce their impacts on the aquatic ecosystems. The results demonstrated the feasibility of using biofilms as a bioindicator, especially the use of the functional variables of the biofilm, being the ones that responded faster to stress and recovery. On the other hand, this thesis provides new evidence to determine the salinity thresholds from which impacts on aquatic ecosystems can reach a point of no return.

[spa] El control de la transmisión de patógenos víricos a través de la ruta fecal-oral ha adquirido recientemente relevancia e interés público. Para mantener este control, es necesario determinar indicadores apropiados para la predicción de comportamiento y transmisión de los virus en las matrices ambientales. Los bacteriófagos han sido propuestos como indicadores víricos de contaminación fecal desde hace varias décadas y están siendo incorporados gradualmente como organismos indicadores en numerosas normativas de gestión del agua, como las nuevas directivas de la Unión Europea para el control de calidad de aguas de consumo y aguas regeneradas. Como consecuencia, será necesario disponer de métodos que permitan detectar bacteriófagos de manera sencilla, rápida, robusta y eficiente. Esta necesidad ha impulsado la mejora y el desarrollo de nuevos métodos de detección, ya que los métodos tradicionales requieren más de un día de trabajo para proporcionar resultados y pueden ser difíciles de implementar en laboratorios de análisis rutinario. Bajo este contexto se ha desarrollado el método Bluephage, un método colorimétrico que permite la detección cualitativa de los diferentes grupos de colifagos mediante la liberación de la enzima β-glucuronidasa al exterior celular por la lisis bacteriana. Este método es posible gracias a una serie de modificaciones genéticas realizadas en las cepas huéspedes empleadas para la detección de colifagos, bloqueando el transporte de los sustratos de la enzima al interior celular y sobreexpresando la enzima intracelularmente. Sólo después de la lisis mediada por fagos, la enzima sobreexpresada puede salir de la célula, encontrar el sustrato en el medio, degradarlo y producir un cambio colorimétrico del medio de amarillo a azul. Este método supone una mejora respecto a los métodos tradicionales, ya que permite la detección en menos de 6 horas de hasta 1 colifago en volúmenes de muestra de hasta 100 ml. Presenta una sensibilidad, especificidad y precisión equivalentes a los métodos estandarizados y es adaptable a volúmenes de muestra grandes y pequeños, a muestras de diferentes orígenes, a la detección de todos los grupos de colifagos y a resultados cuantitativos mediante la aplicación de la técnica del Número Más Probable sobre los resultados cualitativos. En definitiva, Bluephage es el método microbiológico más rápido descrito hasta la fecha para la detección de contaminación fecal utilizando microorganismos cultivables y puede proveer resultados cuando la información rápida de contaminación fecal es esencial., [eng] Relevance and public interest in controlling fecal transmission of viral pathogens has been recently increasing worldwide. Consequently, this has created a need for establishing suitable indicators for predicting viral behavior and transmission in the environment. Bacteriophages have been proposed as viral fecal indicators for decades and they are currently being incorporated into multiple guidelines and regulations of water management, such as the new European Union directives for maintaining water quality of drinking water and reclaimed water. Thus, the development and improvement of methods that allow a fast, user-friendly, robust, and efficient bacteriophage detection is required, as traditional methods entail more than a day of work to provide results and their implementation can be cumbersome for routine analysis laboratories. Under these global circumstances, Bluephage has been developed as a possible solution for these difficulties. Bluephage is a colorimetric methodology qualitatively capable of detecting all groups of coliphages thanks to the release into the external medium of the enzyme β- glucuronidase during lytic processes. This is attainable thanks to the genetic modifications performed on the host bacteria used to detect these coliphages, which blocks the import of the substrate into the cells and overexpresses this intracellular enzyme. Only after phage-mediated lysis, the overexpressed enzyme can be released from the cells, encounter the substrate in the medium, cleave it and produce a colorimetric change of the medium from yellow to blue. This method is an improvement over traditional methods, allowing the detection of up to 1 coliphage in samples up to 100 ml in less than 6 hours. Its sensitivity, specificity and precision are equivalent to standardized methods and it can be scaled to different sample volumes, adapted for analysis of samples from different origins and detection of different types of coliphages. It is also possible to achieve quantitative results from the qualitative data using the Most Probable Number approach. Overall, Bluephage is the fastest microbiological culture- based method for detecting viral fecal pollution described to date and can provide results where rapid information about fecal pollution is essential.

The selection of the many biological indicators described in scientific literature is rarely based on systematic or clear-cut processes, and often takes into account only a single or very few taxa, or even disregards the complex interactions that exist between the components of biodiversity. In certain cases, the particular context of a site–for example in the Mediterranean Basin–makes it difficult to apply the choice of indicators to other regions proposed in the literature. Therefore, the selection of appropriate methodologies for generating relevant indicators for a particular site is of crucial importance. Here, we present a simple quantitative methodology capable of incorporating multidisciplinary information for assessing and selecting appropriate methods and indicators for monitoring local biodiversity. The methodology combines several ecological levels (species, habitats, processes, and ecosystem disturbances), and embraces biological interactions and common functional guilds (detritivores, producers, herbivores, and carnivores). We followed an iterative selection procedure consisting of five phases: 1) collection focal area useful information; 2) classification of this information into interrelated datasets; 3) assessment and selection of the relevant components using a quantitative relevance index; 4) the adding of taxonomic, physiognomic and functional similarities to the relevant components; and 5) the quantitative selection of the priority indicators in the study area. To demonstrate the potential of this methodology, we took as a case study the biodiversity components and their ecological interactions present in a protected area. We show that our methodology can help select appropriate local and long-term indicators, reduce the number of components required for thorough biodiversity monitoring, and underline the importance of ecological processes This work has been carried out through the Biodiversity Monitoring Centre of Mediterranean Mountains (CMBMM) thanks a convention of Diputació de Barcelona and Universitat de Barcelona

Urine from patients with prostate cancer (PCa) contains gene transcripts that have been used for PCa diagnosis and prognosis. Historically, patient urine samples have been collected after a digital rectal examination of the prostate, which was thought necessary to boost the levels of prostatic secretions in the urine. We herein describe methodology that allows urine to be collected by patients at home and then posted to a laboratory for analysis. RNA yields and quality were comparable to those for post digital rectal examination urine, and there was improved sensitivity for the detection of TMPRSS2:ERG transcripts by RT-PCR. The At-Home collection protocol has opened up the potential to perform large-scale PCa studies without the inconvenience, cost, discomfort and expense of patients having to visit the clinic.

Plant traits—the morphological, anatomical, physiological, biochemical and phenological characteristics of plants—determine how plants respond to environmental factors, affect other trophic levels, and influence ecosystem properties and their benefits and detriments to people. Plant trait data thus represent the basis for a vast area of research spanning from evolutionary biology, community and functional ecology, to biodiversity conservation, ecosystem and landscape management, restoration, biogeography and earth system modelling. Since its foundation in 2007, the TRY database of plant traits has grown continuously. It now provides unprecedented data coverage under an open access data policy and is the main plant trait database used by the research community worldwide. Increasingly, the TRY database also supports new frontiers of trait-based plant research, including the identification of data gaps and the subsequent mobilization or measurement of new data. To support this development, in this article we evaluate the extent of the trait data compiled in TRY and analyse emerging patterns of data coverage and representativeness. Best species coverage is achieved for categorical traits—almost complete coverage for ‘plant growth form’. However, most traits relevant for ecology and vegetation modelling are characterized by continuous intraspecific variation and trait–environmental relationships. These traits have to be measured on individual plants in their respective environment. Despite unprecedented data coverage, we observe a humbling lack of completeness and representativeness of these continuous traits in many aspects. We, therefore, conclude that reducing data gaps and biases in the TRY database remains a key challenge and requires a coordinated approach to data mobilization and trait measurements. This can only be achieved in collaboration with other initiatives

Altres ajuts: The study was supported by grants AECC-JB-2013-02 of Asociación Española Contra el Cáncer, 7th Framework Programe, IRSES (PROTBIOFLUID −269285) - Belgium, pre-doctoral fellowship of Vall d'Hebron Research Institute (VHIR) and cofinanced by the European Regional Development Fund(ERDF). Rapid and reliable diagnosis of prostate cancer (PCa) is highly desirable as current used methods lack specificity. In addition, identification of PCa biomarkers that can classify patients into high- and low-risk groups for disease progression at early stage will improve treatment decision-making. Here, we describe a set of protein-combination panels in urinary extracellular vesicles (EVs), defined by targeted proteomics and immunoblotting techniques that improve early non-invasive detection and stratification of PCa patients.We report a two-protein combination in urinary EVs that classifies benign and PCa patients (ADSV-TGM4), and a combination of five proteins able to significantly distinguish between high- and low-grade PCa patients (CD63-GLPK5-SPHM-PSA-PAPP). Proteins composing the panels were validated by immunohistochemistry assays in tissue microarrays (TMAs) confirming a strong link between the urinary EVs proteome and alterations in PCa tissues. Moreover, ADSV and TGM4 abundance yielded a high diagnostic potential in tissue and promising TGM4 prognostic power. These results suggest that the proteins identified in urinary EVs distinguishing high- and low grade PCa are a reflection of histological changes that may be a consequence of their functional involvement in PCa development. In conclusion, our study resulted in the identification of protein-combination panels present in urinary EVs that exhibit high sensitivity and specificity for PCa detection and patient stratification. Moreover, our study highlights the potential of targeted proteomic approaches-such as selected reaction monitoring (SRM)-as diagnostic assay for liquid biopsies via urinary EVs to improve diagnosis and prognosis of suspected PCa patients.

Podeu consultar la versió en castellà i anglès a recurs relacionat, Manual d’utilització de la aplicació RiuNet per telèfons mòbils i tauletes, en el que es mostra tots els continguts (textos, imatges, metodologies i algoritmes) que conté l’app. RiuNet és un projecte de ciència ciutadana que permet avaluar l’estat hidrològic i ecològic dels rius de la Península Ibèrica per qualsevol persona seguint una adaptació de la metodologia oficial que s’utilitza a nivell de tot l’estat. A la part final del document s’hi ha inclòs les fulles de camp que poden ser utilitzades si no es disposa d’un telèfon o tauleta.

Objectives: to develop a risk classifier using urine-derived extracellular vesicle RNA (UEV-RNA) capable of providing diagnostic information of disease status prior to biopsy, and prognostic information for men on active surveillance (AS). Patients and methods: post-digital rectal examination UEV-RNA expression profiles from urine (n = 535, multiple centres) were interrogated with a curated NanoString panel. A LASSO-based Continuation-Ratio model was built to generate four Prostate-Urine-Risk (PUR) signatures for predicting the probability of normal tissue (PUR-1), D'Amico Low-risk (PUR-2), Intermediate-risk (PUR-3), and High-risk (PUR-4) PCa. This model was applied to a test cohort (n = 177) for diagnostic evaluation, and to an AS sub-cohort (n = 87) for prognostic evaluation. Results: each PUR signature was significantly associated with its corresponding clinical category (p

Podeu consultar la versió en català i anglès a recurs relacionat, Manual de utilización de la aplicación RiuNet para teléfonos móviles y tabletas, en el que se muestra todos los contenidos (textos, imágenes, metodologías y algoritmos) que contiene la app. RiuNet es un proyecto de ciencia ciudadana que permite evaluar el estado hidrológico y ecológico de los ríos de la Península Ibérica por cualquier persona siguiendo una adaptación de la metodología oficial que se utiliza a nivel de todo el estado. En la parte final del documento se ha incluido las hojas de campo que pueden ser utilizadas si no se dispone de un teléfono o tableta.

An assessment of the physico-chemical and the biological quality of surface waters through the use of macroinvertebrates as bioindicators was conducted in two rivers in Algeria, located in semi-arid and arid regions. These are Wadi M’zi (Laghouat region) and Wadi Djedir (Djelfa region). The sampling strategy developed in this work is based on the analysis of the upstream and downstream waters of each watercourse. Eleven physico-chemical parameters (T°C, pH, CE, OD, Cl-, SO4 2-, NO3 -, Salinity, Ca2+, Mg2+ and HCO3 -) were measured to establish a diagnosis of the state of health of these aquatic ecosystems. Macroinvertebrates were studied using kick-net sampling at eight study sites. The faunistic inventory of benthic macroinvertebrates identified 37 families, mostly represented by insect larvae (96%). Among these, Diptera and Ephemeroptera were the most dominant orders, with other groups being relatively poorly represented. The results obtained show that the physico-chemical quality of the water is slightly deteriorated, but the quality is still evaluated as ‘good’ according to the biotic indices IBMWP (Iberian Biological Monitoring Working Party) and IBGN (Standardised global biological index). Keywords: surface waters, Algeria, macro-invertebrates, physico-chemical quality, IBMWP, IBGN. L’évaluation de la qualité physico-chimique et biologique des eaux superficielles par l’utilisation des macroinvertébrés comme bio-indicateur est menée dans deux cours d’eau de l’Algérie situés en région semi-aride et aride. Il s’agit de l’oued M’zi (région de Laghouat) et de l’oued Djedir (région de Djelfa). La stratégie d’échantillonnage développée dans ce travail repose sur l’analyse des eaux en amont et en aval de chaque cours d'eau. Onze paramètres physico-chimiques (T°C, pH, CE, OD, Cl-, SO4 2-, NO3 -, Salinité, Ca2+, Mg2+ and HCO3 -) ont été mesurés afin d’établir un diagnostique de l’état de santé de ces écosystèmes aquatiques. Les macroinvertébrés ont été étudiés à partir de prélèvements réalisés à l’aide d’un filet troubleau au niveau de huit stations d’étude. L’inventaire faunistique des macroinvertébrés benthiques a permis de recenser 37 familles, représentées essentiellement par les larves d’insectes avec un pourcentage de 96 %. Parmi ces insectes, les Diptères et les Éphéméroptères sont les ordres les plus dominants par rapport aux autres groupes qui sont faiblement représentés. Les résultats obtenus montrent que la qualité physico-chimique des eaux est légèrement détériorée, tandis que la qualité biologique évaluée par les deux indices biotiques IBMWP « Iberian Biological Monitoring Working Party » et IBGN « Indice biologique global normalisé » révèle une bonne qualité de l’eau. Mots-clés : eaux superficielles, Algérie, macro-invertébrés, qualité physico-chimique, IBMWP, IBGN.

Background Prevention of unnecessary biopsies and overtreatment of indolent disease remains a challenge in the management of prostate cancer. Novel non-invasive tests that can identify clinically significant (intermediate-risk and high-risk) diseases are needed to improve risk stratification and monitoring of prostate cancer patients. Here, we investigated a panel of six DNA methylation biomarkers in urine samples collected post-digital rectal exam from patients undergoing prostate biopsy, for their utility to guide decision making for diagnostic biopsy and early detection of aggressive prostate cancer. Results We recruited 408 patients in risk categories ranging from benign to low-, intermediate-, and high-risk prostate cancer from three international cohorts. Patients were separated into 2/3 training and 1/3 validation cohorts. Methylation biomarkers were analyzed in post-digital rectal exam urinary sediment DNA by quantitative MethyLight assay and investigated for their association with any or aggressive prostate cancers. We developed a Prostate Cancer Urinary Epigenetic (ProCUrE) assay based on an optimal two-gene (HOXD3 and GSTP1) LASSO model, derived from methylation values in the training cohort, and assessed ProCUrE’s diagnostic and prognostic ability for prostate cancer in both the training and validation cohorts. ProCUrE demonstrated improved prostate cancer diagnosis and identification of patients with clinically significant disease in both the training and validation cohorts. Using three different risk stratification criteria (Gleason score, D’Amico criteria, and CAPRA score), we found that the positive predictive value for ProCUrE was higher (59.4–78%) than prostate specific antigen (PSA) (38.2–72.1%) for all risk category comparisons. ProCUrE also demonstrated additive value to PSA in identifying GS ≥ 7 PCa compared to PSA alone (DeLong’s test p = 0.039), as well as additive value to the PCPT risk calculator for identifying any PCa and GS ≥ 7 PCa (DeLong’s test p = 0.011 and 0.022, respectively). Conclusions ProCUrE is a promising non-invasive urinary methylation assay for the early detection and prognostication of prostate cancer. ProCUrE has the potential to supplement PSA testing to identify patients with clinically significant prostate cancer. Electronic supplementary material The online version of this article (10.1186/s13148-018-0575-z) contains supplementary material, which is available to authorized users.

In the last years, the practice of removing all the deadwood after wildfire and use it with energetic purposes has increased. With the objective to evaluate and decrease the impacts of these practices upon biodiversity, from the University of Girona the project Anifog has been created, conducted by the Pecat research team, where a Handbook of Good Management Practices in burned forests has been done to guide managers and forest workers. In my Final Grade Work has been evaluated how two different management practices following the handbook recommendations affect the diversity of saproxylic beetles. The first management consists on logging dead trees or those damaged by the wildfire leaving part of the wood on-site (Sustainable logging), and the second one consists on leaving the forest unaltered after the fire (Non-intervention). Saproxylic beetles have an important ecological role in the recycling of nutrients found in deadwood and, therefore, are important for the recovering of the ecosystem after a wildfire. Furthermore, they are used in our study as bioindicators of the deadwood and its associated biodiversity

The Amazon jungle is a zone of great importance in regards of its diversity; due to its climatic characteristics it offers optimal conditions to a big number of species, animal and vegetal. In particular the chiropters, present an elevated diversity and abundance in the neotropic zones and their populations are fundamental to the good functioning of the communities and even the ecosystem. Especially those that belong to the family Phyllostomidae, the majority frugivorous who have a huge importance in the seed dispersion and, with that, they intervene in the regeneration and succession of the forest. The population of chiropters it’s in decline due to being subjugated to a combination of menaces occasioned mainly by human activity, the majority of said menaces are related to the alteration of nature, and so, directly related with the bat’s habitat perturbation. The main objective of this study is to determine the diversity of the chiropters of two sampled zones in San Rafael, the Community and its communal Reserve. To be able to determine if there are differences between both studied zones and analyze if said differences have been occasioned by the anthropization of the area

The capability of clostridia spores to act as pathogen indicators in sewage sludge treatment was investigated. Sulfite-reducing clostridia and E. coli levels were monitored during waste activated sludge pre-treatments (alkali and ultrasound) and its subsequent mesophilic anaerobic digestion. E. coli was maintained or reduced depending on treatment type and intensity. However, alkali pre-treatment (35.3 gNaOH/kg TS) by itself and alkali (157 gNaOH/kg TS) and ultrasound (27,000 kJ/kg TS) pre-treatments followed by anaerobic digestion provoked reproducible clostridia increases. Specifically, up to 2.7 log 10 after 35.3 gNaOH/kg TS pre-treatment and up to 1.9 and 1.1 log 10 after digesting the 157 gNaOH/kg TS and 27,000 kJ/kg TS pre-treated sludge, respectively. Having rejected the hypotheses of sporulation and floc dissipation, the most plausible explanation for these clostridia increases is re-growth. These results question the suitability of clostridia spores as indicators of sludge treatment and other biological treatments where clostridia may have a role.

[cat] La present tesi té com a objectiu l’estudi de la comunitat microeucariota que es desenvolupa en dos sistemes avançats de tractament d’aigües residuals amb alta càrrega amoniacal. La comunitat microeucariota, especialment els ciliats, són una part essencial per al correcte funcionament dels sistemes biològics de tractament d’aigües residuals, on serveixen, a més a més, com a organismes indicadors del funcionament d’aquests processos. Concretament, el principal objectiu de la tesi doctoral és la caracterització de les comunitats que es desenvolupa en aquests sistemes i la determinació d’aquelles espècies amb potencial com a bioindicadores dels processos, per tal de donar eines de gestió complementàries als paràmetres fisicoquímics. Els sistemes estudiats han estat el procés Shortcut Biological Nitrogen Removal (SBNR) i la fase de nitritació parcial (Partial Nitritation; PN) del procés combinat PN-Anammox (Anaerobic Ammonium Oxidation). Ambdós processos, en comparació amb el procés convencional d’eliminació de nitrogen, són capaços d’eliminar del medi grans concentracions de nitrogen (en forma amoniacal) amb un estalvi de fins el 25 i el 50-60% d’oxigen dissolt i del 40 i el 100% de matèria orgànica en el sistema SBNR i el PN-Anammox, respectivament. Aquest fet els fa especialment adequats per al tractament d’aigües residuals amb ratios de carboni/nitrogen molt baixes, com la que presenta l’efluent dels processos de digestió anaeròbia de fangs. Ambdós sistemes han operat amb tecnologia de biofilm, concretament la tecnologia Moving-Bed Biofilm Reactor (MBBR), basada en la presència de suports plàstics en suspensió (carriers) on s’hi desenvolupa, adherida, la biomassa. Per l’estudi d’aquests sistemes, doncs, s’ha implementat un protocol efectiu que permet determinar, de forma independent, la comunitat microeucariota que es desenvolupa al biofilm i la que es manté en suspensió al líquid mescla. El protocol permet, a més a més, la comparació en termes d’abundància entre ambdues comunitats, al utilitzar la mateixa unitat de mesura (ind/ml) en els dos hàbitats (biofilm i licor mescla). En la memòria es caracteritzen les comunitats microeucariotes associades al sistema SBNR i al reactor PN, i es descriuen aquelles espècies de ciliats amb potencial com a bioindicadores del seu funcionament. Entre aquestes espècies destaca Epistylis camprubii, una nova espècie de ciliat sessílid descrita arrel de la seva abundant presència al biofilm dels carriers dels processos estudiats. A nivell ecològic, s’han aportat noves dades sobre l’ecologia de les espècies que han colonitzat els dos sistemes, establint (o eixamplant) els rangs de tolerància d’aquests organismes al nitrogen amoniacal, nitrit, nitrat, amoníac lliure i àcid nitrós, entre altres paràmetres abiòtics. Per últim, en un context actual de clara expansió de les tècniques moleculars com a eina d’estudi de la biodiversitat, s’ha comparat el potencial de detecció de la tècnica de Next Generation Sequencing Ilumina amb els resultats obtinguts mitjançant microscòpia òptica., [eng] The main objective of the present thesis is to study the microeukaryote community that develops in two advanced wastewater treatment systems treating high ammonium loaded wastewater, in order to characterise their communities and determine the microeukaryote species that can serve as bioindicators of the process performance. The studied wastewater treatment systems are the Shortcut Biological Nitrogen Removal (SBNR) and the Partial Nitritation (PN) reactor of the combined PN-Anammox (Anaerobic Ammonium Oxidation) process. Both processes are designed for nitrogen removal of high ammonium loaded wastewater in response to the need to reduce energy costs and improve treatment efficiency. Concretely, the SBNR process can reduce oxygen demand and organic source demand due to nitrification by up to 25% and 40%, respectively, while the combined PN-Anammox process implies a 50%-60% reduction in oxygen demand, does not require an external organic carbon source for denitrification and reduces sludge production by about 90%. Both systems operated using Moving-Bed Biofilm Reactor (MBBR) technology, and the reactors were filled with plastics supports for the biofilm development (carriers). To achieve the goals of the thesis, an effective method to study the microeukaryote communities that remain in suspension in the liquid phase and the ones that develop adhered to the fixed substrate has been proposed. The thesis also describes the microeukaryote communities related to the SBNR process and the PN reactor, and determine the species that can be useful as bioindicators of the processes performance. Moreover, a new peritrich species, Epistylis camprubii, has been described. The unusual physicochemical characteristics of both SBNR and PN processes provided new data on tolerance ranges of protists species, specifically of ciliates, to ammonium, nitrites, nitrates, free ammonia and nitrous acide, among other physicochemical parameters. Finally, a V4 18S rDNA molecular survey (Illumina) has been carried out as a complement to the microscopic results in order to verify the feasibility of the microscopic and molecular analyses.

Este artículo contiene 13 páginas, 9 figuras, 3 tablas., We analysed with multigene (18S and COI) metabarcoding the effects of the proliferation of invasive seaweeds on rocky littoral communities in two Spanish Marine Protected Areas. The invasive algae studied were Caulerpa cylindracea, Lophocladia lallemandii and Asparagopsis armata. They are canopy-forming, landscape-dominant seaweeds, and we were interested in their effects on the underlying communities of meiobenthos and macrobenthos, separated in two size fractions through sieving. A new semiquantitative treatment of metabarcoding data is introduced. The results for both markers showed that the presence of the invasive seaweed had a significant effect on the understory communities for Lophocladia lallemandii and Asparagopsis armata but not for Caulerpa cylindracea. Likewise, changes in MOTU richness and diversity with invasion status varied in magnitude and direction depending on the alga considered. Our results showed that metabarcoding allows monitoring of the less conspicuous, but not least important, effects of the presence of dominant invasive seaweeds., This research was funded by projects Metabarpark and CORCLIM from the Spanish National Parks Autonomous Agency (OAPN 1036/ 2013 and 766S/2012), and ChallenGen CTM2013-48163 and ANIMA CGL2016-76341-R projects from the Spanish Government. OSW is currently funded by project SeaDNA, NE/N005759/1 grant from the Natural Environment Research Council (NERC).

Antioxidant enzymes are involved in important processes of cell detoxification during oxidative stress and have, therefore, been used as biomarkers in algae. Nevertheless, their limited use in fluvial biofilms may be due to the complexity of such communities. Here, a comparison between different extraction methods was performed to obtain a reliable method for catalase extraction from fluvial biofilms. Homogenization followed by glass bead disruption appeared to be the best compromise for catalase extraction. This method was then applied to a field study in a metal-polluted stream (Riou Mort, France). The most polluted sites were characterized by a catalase activity 4-6 times lower than in the low-polluted site. Results of the comparison process and its application are promising for the use of catalase activity as an early warning biomarker of toxicity using biofilms in the laboratory and in the field.

The ecological status of streams depends on an equilibrium between hydrological processes and biological dynamics. Water discharge is the main requisite for a wealthy riparian habitat. Nevertheless, human practices severely affect water availability through stream water derivation and groundwater withdrawal. In this sense, impacts upon aquifer water storage and its effects on base flow generation have a significant effect on stream biology. Consequently, biological indicators will point out poor conditions resulting from such human impacts. In this paper, the effects of groundwater exploitation on stream discharge and surface water quality are evaluated and compared to biological indicators in a Mediterranean catchment. The stream-aquifer relationship is investigated by considering the hydrological context of each river reach, including human pressures, and the hydraulic head in the contiguous alluvial aquifer, where it exists. These data allow us to differentiate distinct types of reaches that are defined according to a "Stream-Aquifer relationship and chemical Pressure" (SAP) classification, which is used later on to standardize the different hydrochemical and biological features of the sampling points. Stream water and groundwater hydrochemistry are compared to depict the hydraulic behavior of the sampled watercourses during wet and dry periods. Specific elements are used as tracers of groundwater inputs, wastewater influence, or even stagnant conditions during the dry season. This dataset defines a framework to interpret the biological status of each reach based on the Iberian biological monitoring working party indicator (IBMWP). Affinities between hydrological and hydrochemical conditions with biological indicator values allow the causal effects of groundwater exploitation on stream ecology to be defined. The use of multivariate principal component analysis shows that the dataset variance is distributed according to the SAP classification, and that variable grouping is in agreement with the observed hydrological processes and their effects on biological indicator values. This work provides evidence of the importance of groundwater dynamics on biological indicators in a human-modified environment. When using the SAP classification, biological indicators acquire a broader meaning as they reveal the status of biological processes and the causal references. Such information is relevant for water management assessment within the context of the European Water Framework Directive, as it emphasizes the control of groundwater exploitation as a key parameter in the preservation of stream ecological status and the achievement of the objectives of the directive This project was funded by grants from Spanish Government research projects CICYT CGL-2005-08019-C04-02 and CGL2008-06373-C03-03/BTE, and from the Patronat Francesc Eiximenis grant (Diputació de Girona)

[eng] Nearshore marine ecosystems like seagrass meadows face a wide range of anthropogenic influences, impacting the system at different spatial and temporal scales. Managing these systems in the face of these pressures requires detailed knowledge of how seagrass habitats respond to these various threats. A plethora of useful indicators have been developed to help managers and policy makers track seagrass meadow health and status, detect environmental impacts or measure the effectiveness of management interventions. However, choosing between these indicators can often be a daunting task since they vary considerably in their overall behaviour in relation to ecosystem and environmental changes. This thesis assesses the most commonly employed seagrass indicators to determine if they are adequate and appropriate to the specific needs of coastal ecosystem management. This assessment is based on evaluating three fundamental characteristics of each indicator – the robustness of its response, the specificity or generality of its response, and the time of response. We use a variety of complementary approaches to explore indicator behaviour. In Chapter 3, we use field-based studies to assess how seagrass indicators respond to the construction of a breakwater in the vicinity of a Posidonia oceanica seagrass meadow. Chapters 4 and 5 examine long-term trends in seagrass indicators to improving water quality conditions after significant regional management interventions. In addition, in Chapter 6, we comprehensively review seagrass indicator responses to multiple stressors. Chapters 3, 4 and 5 focus largely on the Catalan Coast in the Mediterranean with Posidonia oceanica as a target species. Chapter 6 in contrast is a broad review of a wide range of indicators used across several seagrass species worldwide. A central learning across these studies was that the level of biological organisation of the indicator (i.e. Physiological, biochemical, growth, morphological, structural or demographic) is critical in determining the specificity or generality of response: the lower the level (e.g. biochemical), the most specific the response, while the higher the level (e.g. population, community), the wider the response. Thus, biochemical indicators are ideal to determine the identity or even the origin of a pressure while structural indicators, in contrast, are useful as generic indicators of declining conditions. Response times are also heavily determined by the level of organisation, particularly in the detection of improving environmental quality along the Catalan coast. Biochemical indicators responded unequivocally to water quality improvements observed both in the experimental field study (Chapter 3) as well as in the analysis of the long-term data series (Chapters 4 and 5). The meta-analysis confirmed that these trends in specificity and response time were not unique to Posidonia oceanica or the Catalan coast and highlighted the critical role of plant size in determining indicator time responses. Large species take considerably longer to register a response to environmental degradation as well as improvement – a critical factor that needs to be accounted for in designing monitoring programmes and interpreting ecosystem trends. Taken together, these results suggest that differences in the behaviour of seagrass indicators require that they should be carefully selected to match the objectives of management. Based on the results reported in this thesis, where different sets of indicators have been proven successful for given management objectives, we finally develop a simple decision tree to help managers chose the most reliable sets of indicators matching their objectives. Understanding the diversity of responses that seagrass indicators display can make them a powerful set of tools in the ecosystem manager’s toolkit. Carefully employed, they can serve as bespoke solutions to a wide range of management objectives as we seek to monitor and protect these vital ecosystems and coastal water quality in the face of increasing coastal pressures., [cat] Entendre com responen les fanerògames marines a les pressions, és clau per a poder gestionar tant els herbeis com les aigües costaneres. Actualment, disposem d’un gran nombre d’indicadors basats en fanerògames marines. Però, la manca d’informació sobre com responen als canvis ambientals, fa que no sigui fàcil escollir quins indicadors són els més adients per a cada tipus i objectiu de gestió. Aquesta tesi avalua les tres característiques bàsiques de la resposta dels indicadors més utilitzats als canvis ambientals: la robustesa de la resposta, la especificitat dels indicadors a diferents pressions i el temps de resposta. Per analitzar aquestes tres característiques, fem servir diferents aproximacions complementàries. Al capítol 3, analitzem la resposta de diferents indicadors a les obres d’ampliació del port de Blanes, situat just al costat d’un herbei de Posidonia oceanica. Als capítols 4 i 5, estudiem com responen els indicadors a la millora de la qualitat de l’aigua a la costa catalana. Finalment, al capítol 6, presentem una metaanàlisi que estudia com responen els indicadors a diferents factors d’estrès. De totes tres aproximacions, hem pogut comprovar que el nivell d’organització dels indicadors (i.e. bioquímic, estructural) és clau a l’hora de determinar el grau d’especificitat de la resposta dels indicadors a les pressions: generalment, a més baix nivell d’organització (e.g. bioquímic), més específica és la resposta i com més alt (e.g. demogràfic), més ampli és el rang de pressions que un indicador pot detectar. El temps de resposta dels indicadors varia també en funció del nivell d’organització dels indicadors, especialment, quan es tracta de la detecció de millores ambientals. A més a més, la metaanàlisi destaca la importància de la mida de les plantes per determinar el temps de resposta. Les espècies grans triguen més a detectar la degradació de les condicions ambientals i, molt més, a detectar la millora, especialment, si s’utilitzen indicadors estructurals o demogràfics. Basant-nos en els resultats d’aquesta tesi, hem elaborat un esquema per ajudar els gestors a escollir el conjunt d’indicadors que més s’ajusti a cada objectiu de gestió. Utilitzats correctament, aquests indicadors són molt útils per fer el seguiment, tant de l’estat de salut dels herbeis, com de la qualitat del medi.

[spa] En esta tesis, se evaluó la vulnerabilidad y la capacidad de aclimatación al estrés ambiental, relacionada al cambio climático, en las especies Cystoseira tamariscifolia, Ellisolandia elongata, Cystoseira compressa y Padina pavonica. En estas especies, se evaluaron los efectos de los factores de estrés físico, como la irradiancia elevada de PAR (lambda = 400-700 nm) y UVR (lambda = 280-400 nm), la temperatura y factores de estrés químico, como nutrientes, metales pesados y CO2. Se realizaron seis estudios experimentales variando irradiancia, temperatura, condiciones de nutrientes, acidificación y metales pesados. El enfoque común en estos estudios fue el uso bioindicadores funcionales como; rendimiento cuántico máximo de PSII (Fv/Fm) como un indicador de estado fisiológico de macroalgas y de la fotoinhibición, la tasa de transporte de electrones (ETR) como un indicador de la capacidad fotosintética. Como indicadores bioquímicos se utilizaron; las relaciones estequiometrias (C:N) como indicador del estado nutricional y el contenido de compuestos fenólicos y micosporinas, tales como sustancias fotoprotectoras y antioxidantes. Con el objetivo de evaluar el estado fisiológico de estas especies de macroalgas en el Mar Mediterráneo (Mar de Alborán). La disminución del rendimiento cuántico y la tasa máxima de transporte de electrones, el aumento de compuestos fenólicos y actividad antioxidante o el aumento de la relación C: N se producen en condiciones de estrés y por lo tanto, estas variables se validan como indicadores de estrés. Sin embargo, se encontró un aumento de los compuestos fenólicos y de la actividad fotosintética, que muestra una relación entre la actividad antioxidante y la producción algal. Este no es un resultado extraño ya que una alta actividad fotosintética se relaciona con una producción alta de oxígeno lo que puede producir estrés oxidativo. La acumulación de compuestos fenólicos en condiciones de aumento de los niveles de nitrato y de CO2 o la liberación de fenoles bajo mayor radiación solar en C. tamariscifolia, nos muestra que esta especie tiene mecanismos bioquímicos eficaces para aclimatarse a las variaciones esperadas en los factores del cambio climático aunque esté limitado por la temperatura., [eng] In this thesis, the vulnerability and capacity of acclimation to environmental stress related to Climate change mainly in Cystoseira tamariscifolia and other species as Ellisolandia elongata, Cystoseira compressa and Padina pavonica are evaluated. The effects of physical stressors as elevated irradiance of PAR (lambda=400-700 nm) and UVR (lambda=280-400 nm) and temperature and chemical stressors as nutrient, heavy metals and CO2, separately and in interaction with different levels were evaluated. Six experimental studies were conducted under varying irradiance, temperature, nutrient conditions, acidification and heavy metals. The common approach in these studies was the use functional bioindicators to evaluate the physiological state macroalgal species of Mediterranean Sea (Alboran Sea) in studies conducted both in situ in ultra-oligotrophic (Cabo de Gata-Níjar Natural Park, Almeria) and oligotrophic waters (La Araña beach, Malaga) and with algae transported to controlled experimental systems under out-door conditions. In addition, an indoor experiment study was performed in Cystoseira tamariscifolia collected in the North Atlantic Ocean, the northern limit of distribution of this species. In this study, the decrease of maximal quantum yield and electron transport rate , the increase of phenolic compounds and antioxidant activity or the increase of C:N ratio are produced in stress conditions and thus they are validated as stress indicator. In addition, it is possible to evaluate the direction of the physiological response i.e. positive or negative to expected changes under climate change factors or other anthropogenic impacts, as eutrophication (increased nitrate levels in the water column) or pollution by heavy metals. However, on the other hand, the increase of phenolic compounds is also produced under increased photosynthetic activity showing a link between antioxidant and algal production. This not a strange result since a high photosynthetic activity is related to a high oxygen production which can be produced oxidative stress. Non-photochemical quenching, oxygen consumption through Mehler reaction and increased antioxidant activities are down regulation mechanisms to survive under promoted oxygenic scenario. Phenolic accumulation under increased nitrate and CO2 levels or the release of phenols under increased irradiance in C. tamariscifolia shows us that this species has effective biochemical mechanisms to acclimate for the expected variations in climate change factors although this is limited by temperature. Phenolic compounds are related to secondary metabolism but in C. tamariscifolia but the direct positive relation found with photosynthetic activity and internal nitrogen in all experiments seem to link the phenols to primary metabolism. In summary, increased CO2 under high irradiance, but not photoinhibitory, conditions will be favorable growth and physiological responses. The nitrate enrichment reduced stress provoked by irradiance or pollution by Cooper due to the photoprotection mechanisms are favored by nitrate increase. However, the positive effect of CO2 and nitrate is dependent on temperature, summer temperature in the field or 4oC increased temperature in outdoor experiments provoked physiological stress. Consequently, ocean acidification will be favorable for C. tamariscifolia only under no very high increase of temperature, less 2-3ºC and without nutrient limitation. The oligotrophication produced in certain areas of Mediterranean Sea will be unfavorable for Cystoseira tamariscifolia communities in a climate change scenario. The data on vulnerability and acclimation to climate change factors of Cystoseira tamariscifolia, Ellisolandia elongata and Padina pavonica presented in this study can help the management of macroalgal communities, mainly in protected areas. In addition, the physiological and biochemical data will help to predict the effects of climate change on bioactive compounds with antioxidant capacity and their potential biotechnological uses as phenolic compounds, mycosporine like aminoacids and carotenoids.

INTRODUCTION High-grade prostatic intraepithelial neoplasia (HGPIN) is a recognized precursor stage of PCa. Men who present HGPIN in a first prostate biopsy face years of active surveillance including repeat biopsies. This study aimed to identify non-invasive prognostic biomarkers that differentiate early on between indolent HGPIN cases and those that will transform into actual PCa. METHODS We measured the expression of 21 candidate mRNA biomarkers using quantitative PCR in urine sediment samples from a cohort of 90 patients with initial diagnosis of HGPIN and a posterior follow up of at least two years. Uni- and multivariate statistical analyses were applied to analyze the candidate biomarkers and multiplex models using combinations of these biomarkers. RESULTS PSMA, PCA3, PSGR, GOLM, KLK3, CDH1, and SPINK1 behaved as predictors for PCa presence in repeat biopsies. Multiplex models outperformed (AUC = 0.81–0.86) the predictive power of single genes, including the FDA-approved PCA3 (AUC = 0.70). With a fixed sensitivity of 95%, the specificity of our multiplex models was of 41–58%, compared to the 30% of PCA3. The PPV of our models (30–38%) was also higher than the PPV of PCA3 (27%), suggesting that benign cases could be more accurately identified. Applying statistical models, we estimated that 33% to 47% of repeat biopsies could be prevented with a multiplex PCR model, representing an easy applicable and significant advantage over the current gold standard in urine sediment. DISCUSSION Using multiplex RTqPCR-based models in urine sediment it is possible to improve the current diagnostic method of choice (PCA3) to differentiate between benign HGPIN and PCa cases. Prostate 75:1102–1113, 2015. © 2015 Wiley Periodicals, Inc.

[cat]La península Ibèrica és un territori molt divers pel que fa als ecosistemes aquàtics. No obstant, són escasses les publicacions referents a la flora que els ocupa. En aquesta tesi es presenta un catàleg florístic de 846 mostres d’algues caràcies recol•lectades a 456 masses d’aigua diferents. S’han identificat 28 espècies o varietats pertanyents als cinc gèneres citats a Espanya, essent Chara el més freqüent, i s’amplia l’àrea de distribució de 18 espècies o varietats i l’ecologia de 8 d’elles, algunes vulnerables o en perill d’extinció. D’aquesta manera s’incrementa el coneixement que es té sobre aquest grup algal i s’apunten algunes de les causes principals que han ocasionat el descens d’espècies al llarg del temps a la conca del riu Duero, relacionades principalment amb l’agricultura i la ramaderia. Per altra banda, les caràcies són organismes amb una elevada plasticitat morfològica en relació a les condicions ambientals i existeixen complexes d’espècies que fan que la seva identificació no sempre resulti una tasca fàcil. La parella conflictiva Chara aspera-Chara galioides es diferencia bàsicament pel diàmetre dels anteridiòfors madurs, però aquest caràcter podria no tenir significació sistemàtica. Amb l’objectiu de conèixer l’efecte de la llum, la temperatura i la salinitat sobre la morfometria de diverses estructures, s’han cultivat a una cambra i es proposen nous caràcters vegetatius que farien factible la seva separació com a dues espècies diferents, tals com el diàmetre de l’eix principal. Per acabar, els ecosistemes aquàtics lòtics i lenítics de la península Ibèrica són sotmesos a gran quantitat de pressions antropogèniques que posen en seriós compromís la seva continuïtat i la supervivència de la interessant flora que sovint els ocupa. Desgraciadament, Espanya no compta actualment amb índexs adequats per conèixer l’estat d’aquests ecosistemes. Tanmateix, són molts els estudis i normatives com la Directiva Marc de l’Aigua que posen de manifest el paper bioindicador dels macròfits, en general i de les caràcies, en particular. En aquesta tesi es proposen dues eines per avaluar l’estat ecològic dels llacs i llacunes de la conca del riu Duero (INEQUAL) i de rius d’àmbit mediterrani (IMF). Aquests dos índexs podrien ajudar a l’hora de prendre mesures de gestió, conservació i restauració dels ecosistemes aquàtics ibèrics., [eng] The Iberian Peninsula is a very diverse country in terms of aquatic ecosystems. However, there are few publications on the flora that occupies these bodies of water. This thesis presents a floristic catalogue of 846 charophyte samples collected from 456 different localities. We have identified 28 species or varieties of the five genera currently present in Spain, being Chara the most frequently sampled, and we have provided new data on the area of distribution of 18 species or varieties and the ecology of 8 of them, some vulnerable or endangered. Thus, the knowledge we have on this algal group increases and we have also proposed some of the main causes that have led to the decline of species over time in the Douro river basin, which are mainly related to agriculture and farming. Moreover, the charophytes are organisms with high morphological plasticity in relation to the environment conditions and there also exist complexes of species that make their identification not always a straightforward matter. The pair of taxa Chara aspera-Chara galioides was traditionally discriminated by the diameter of the mature antheridia, but this character could not have any systematic significance. In order to determine the effect of light, temperature and salinity on the morphometry of different structures, they have been grown in a culture chamber and new vegetative characteristics, such as the diameter of the main axis, are proposed to split them as two different species. Finally, lotic and lentic aquatic ecosystems of the Iberian Peninsula are subjected to many anthropogenic pressures that seriously compromise their continuity and the survival of their interesting flora. Unfortunately, Spain does not currently have any suitable index for the assessment of the ecological status of these ecosystems. However, many studies and the Water Framework Directive highlight the bioindicator role of macrophytes, in general and charophytes, in particular. In this thesis, we propose two indices to assess the ecological status of the lakes and lagoons of the Duero river basin (INEQUAL) and the Mediterranean rivers (IMF). With this information it would be possible to apply management, conservation and, if necessary, restoration measures in these aquatic habitats., [spa] La Península Ibérica es un territorio muy diverso en lo que se refiere a ecosistemas acuáticos. No obstante, son escasas las publicaciones referentes a la flora que los ocupa. En esta tesis se presenta un catálogo florístico de 846 muestras de algas caráceas recolectadas en 456 masas de agua diferentes. Se han identificado 28 especies o variedades pertenecientes a los cinco géneros citados en España, siendo Chara el más frecuente, y se amplía el área de distribución de 18 especies o variedades y la ecología de 8 de ellas, algunas vulnerables o en peligro de extinción. De esta manera se incrementa el conocimiento que se tiene sobre este grupo algal y se apuntan algunas de las causas principales que han ocasionado el descenso de especies a lo largo del tiempo en la cuenca del río Duero, relacionadas principalmente con la agricultura y la ganadería. Por otra parte, las caráceas son organismos con una elevada plasticidad morfológica en relación a las condiciones ambientales y existen complejos de especies que hacen que su identificación no siempre resulte una tarea fácil. La pareja conflictiva Chara aspera-Chara galioides se diferencia básicamente por el diámetro de los anteriodióforos maduros, pero este carácter podría no tener significación sistemática. Con el objetivo de conocer el efecto de la luz, la temperatura y la salinidad sobre la morfometría de diversas estructuras, se han cultivado en una cámara y se proponen nuevos caracteres vegetativos que harían factible su separación como dos especies diferentes, tales como el diámetro del eje principal. Para acabar, los ecosistemas acuáticos lóticos y leníticos de la Península Ibérica están sometidos a multitud de presiones antropogénicas que ponen en serio compromiso su continuidad y la supervivencia de la interesante flora que a menudo los ocupa. Desgraciadamente, España no cuenta actualmente con ningún índice adecuado para conocer el estado de estos ecosistemas. Sin embargo, son muchos los estudios y normativas como la Directiva Marco del Agua que ponen de manifiesto el papel bioindicador de los macrófitos, en general y de las caráceas, en particular. En esta tesis se proponen dos herramientas para evaluar el estado ecológico de lagos y lagunas de la cuenca del río Duero (INEQUAL) y de ríos de ámbito mediterráneo (IMF). Estos dos índices podrían ayudar a tomar medidas de gestión, conservación y restauración de los ecosistemas acuáticos ibéricos.

Score-based biotic indices are widely used to evaluate the water quality of streams and rivers. Few adaptations of these indices have been done for South America because there is a lack of knowledge on macroinvertebrate taxonomy, distribution and tolerance to pollution in the region. Several areas in the Andes are densely populated and there is need for methods to assess the impact of increasing human pressures on aquatic ecosystems. Considering the unique ecological and geographical features of the Andes, macroinvertebrate indices used in other regions must be adapted with caution. Here we present a review of the literature on macroinvertebrate distribution and tolerance to pollution in Andean areas above 2 000masl. Using these data, we propose an Andean Biotic Index (ABI), which is based on the BMWP index. In general, ABI includes fewer macroinvertebrate families than in other regions of the world where the BMWP index has been applied because altitude restricts the distribution of several families. Our review shows that in the high Andes, the tolerance of several macroinvertebrate families to pollution differs from those reported in other areas. We tested the ABI index in two basins in Ecuador and Peru, and compared it to other BMWP adaptations using the reference condition approach. The ABI index is extremely useful for detecting the general impairment of rivers but class quality boundaries should be defined independently for each basin because reference conditions may be different. The ABI is widely used in Ecuador and Peru, with high correlations with land-use pressures in several studies. The ABI index is an integral part of the new multimetric index designed for high Andean streams (IMEERA). Rev. Biol. Trop. 62 (Suppl. 2): 249-273. Epub 2014 April 01. Los índices bióticos basados en puntuación son ampliamente utilizados para evaluar la calidad del agua de los arroyos y ríos. Varias áreas de los Andes están densamente pobladas y hay necesidad de métodos para evaluar el impacto de la creciente presión humana sobre los ecosistemas acuáticos. Dadas las características ecológicas y geográficas únicas de los Andes, los índices de macroinvertebrados utilizados en otras regiones deben adaptarse con cautela. Aquí se presenta una revisión de la literatura sobre distribución de macroinvertebrados y la tolerancia a la contaminación en las zonas andinas por encima de 2 000msnm. Usando estos datos, se propone un Índice Biolótico Andino (ABI), que se basa en el índice de BMWP. En general, ABI incluye un menor número de familias de macroinvertebrados que en otras regiones del mundo donde se ha aplicado el índice BMWP porque la altitud restringe la distribución de varias de ellas. Nuestra revisión muestra que la tolerancia de varias familias a la contaminación en los ríos altoandinos difiere de lo reportado en otras áreas. Probamos el índice ABI en dos cuencas en Ecuador y Perú, y comparamos con otras adaptaciones BMWP utilizando el enfoque de condición de referencia. Nuestros resultados muestran que el índice de ABI es extremadamente útil para detectar el deterioro general de los ríos, pero que los límites entre las clases de calidad deben ser definidos independientemente para cada cuenca debido a que las condiciones de referencia pueden ser diferentes. El ABI es ampliamente utilizado en Ecuador y Perú, y es parte integral del nuevo índice multimétrico diseñado para corrientes altas andinas (IMEERA).

[spa] El agua bajo el enfoque ecosistémico es un recurso clave para el desarrollo socioeconómico y en Chile presenta problemas de escasez, como lo es para la cuenca del Río Limarí, IV Región (semiárida), localizada en una zona de transición climática, con precipitaciones en disminución desde 30 años. Proyecciones futuras, indican un aumento de su la demanda, siendo imprescindible compatibilizar los procesos de desarrollo de estas con la protección hídrica, bajo una gestión integrada. Se propone una metodología de evaluación en el marco de la norma secundaria de calidad de agua continental superficial, a través del Plan de Vigilancia Ambiental para aportar a la toma de decisiones para una gestión preventiva. El río posee una dinámica temporal y espacial, en su estructura hidromorfológica, física y química con consecuencias biológicas, dado por su propia naturaleza y las actividades antrópicas de la cuenca. Los ríos están conformados por parches heterogéneos que interactúan, y por si solos constituyen un paisaje fluvial, por lo que las escalas espaciales y temporales son cruciales de definir al evaluar sus estados ecológicos. Se evaluó la calidad del agua en seis subcuencas, elaborando previamente un modelo conceptual, basado en bioindicación, el cual se puso a prueba y se ajustó en dos campañas de terreno. Se levantaron datos del componente hidromorfógico (índice QBR e IHF), físico y químico (metodología estandarizada APHA.) y biológico (índices bióticos). Los resultados indicaron la influencia de actividades agrícolas y mineras en la calidad del agua, un aumento de perturbaciones hacia los tramos aguas abajo y bajo embalse. Los hábitat mas heterogéneos fueron los tramos de mayor altura, con valores de IHF (> 75 ) y QBR (buena calidad). Las estaciones aguas abajo presentaron pH más básicos (> 7.5), altas DQO > 40 mg/L y tendencia a eutrofización. El índice Ch SIGNAL reflejó aguas de mala calidad en los tramos más bajos, comparado con los de mayor altura, y correlación significativa positiva con índices hidromorfológicos, como criterio de validación. El modelo propuesto fue difundido y consensuado con actores de interés en la gestión. Se elaboró un protocolo de monitoreo, y se recomendó el índice biótico Ch SIGNAL para el Programa de Vigilancia. Se concluye que, los macroinvertebrados bentónicas, sirven de bioindicadores al constituir variables de estado, de cambios físicos y químicos del sistema, pueden ser una herramienta para evaluar el estado ecológico de los ríos, ante la norma secundaria de calidad de aguas continentales superficiales., [eng] According to the ecosystem concept approach, water is a key factor for the socioeconomic development and, in Chile it represents scarcity problems such as the case of the river Limarí, IV semi-arid Región (30°15’ - 31°20’ south latitude), located at an area of climatic transition with decreasing rainfall since the last 30 years indicating an increase in demand and showing that it is indispensable to find integral compatibility between the development and hydrological availability protection. An evaluation methodology under the frame of the secondary standard of surface freshwater quality through an Environmental Vigilance Plan that should help as prevention management determinations is proposed. This river presents a temporal and spatial dynamics in its hydromorphological, physic and chemical structure with biological consequences due to its own structure and anthropic activities at the basin. Rivers are structured as heterogeneous patchs, that interact and by themselves they conform a fluvial landscape, so that spatial and temporal characteristics are crucial to be assessed in evaluating river ecological state. Water quality was evaluated at six sub-basins, with a previous design of a conceptual model based on bioindication that was tested and adjusted in two field trips. Data on hydromorphological (QBR and IHF indexes) physical and chemical data (standard APHA methodology) and biology (biotic indexes). Results indicated the influence of agriculture and mining activities in water quality, such as an increase of the disturbances towards low waters and down a reservoir. The most heterogeneous habitats were the higher altitude sites, with high IHF values (> 75) and QBR (good river border quality). The lower sampling stations showed more basic pH (> 7.5), high DQO (>40 mg/L) and a tendency to eutrophication. The Ch SIGNAL index showed bad quality waters at lower sites compared to higher altitude and a positive correlation with hydromorphological indixes as a validation criteria. The proposed model was publicized and approved by interested management persons. A monitoring protocol was elaborated and the biotic Index Ch SIGNAL was recommended to be used in the Vigilance Program. Conclusions state that benthic macro invertebrates are systems useful bio indicators of physic and chemical state variables as a tool to evaluate the river ecological state, in relation with the secondary standard of surface freshwater quality.

Major oil spills can have long-term impacts since oil pollution does not only result in acute mortality of marine organisms, but also affects productivity levels, predator-prey dynamics, and damages habitats that support marine communities. However, despite the conservation implications of oil accidents, the monitoring and assessment of its lasting impacts still remains a difficult and daunting task. Here, we used European shags to evaluate the overall, lasting effects of the Prestige oil spill (2002) on the affected marine ecosystem. Using δ 15N and Hg analysis, we trace temporal changes in feeding ecology potentially related to alterations of the food web due to the spill. Using climatic and oceanic data, we also investigate the influence of North Atlantic Oscillation (NAO) index, the sea surface temperature (SST) and the chlorophyll a (Chl a) on the observed changes. Analysis of δ 15N and Hg concentrations revealed that after the Prestige oil spill, shag chicks abruptly switched their trophic level from a diet based on a high percentage of demersal-benthic fish to a higher proportion of pelagic/semi-pelagic species. There was no evidence that Chl a, SST and NAO reflected any particular changes or severity in environmental conditions for any year or season that may explain the sudden change observed in trophic level. Thus, this study highlighted an impact on the marine food web for at least three years. Our results provide the best evidence to date of the long-term consequences of the Prestige oil spill. They also show how, regardless of wider oceanographic variability, lasting impacts on predator-prey dynamics can be assessed using biochemical markers. This is particularly useful if larger scale and longer term monitoring of all trophic levels is unfeasible due to limited funding or high ecosystem complexity.

[cat]En aquesta memòria de tesi es detalla la quantificació de biomarcadors d’exposició (acumulació de Pb, Hg, Cd, Tl, Fe, Mg, Zn, Cu, Mn, Mo, Co, Ni, Cr, Sr, Ba i B a teixits tous i durs) i d’efecte (morfometria, hematologia, activitat enzimàtica, genotoxicitat i histopatologia) en un total de 243 exemplars de musaranya comuna, Crocidura russula (Hermann, 1780). En el present estudi, es va tenir en compte la importància de l’edat i el sexe en els paràmetres quantificats ja que són dos factors biòtics que expliquen part de la variabilitat observada. Les zones d’estudi van ser 4 zones contaminades i 4 zones de referència triades per la seva importància tant mediambiental (6 de les 8 zones estudiades tenen estatus de protecció parcial o total) com d’interès per la societat: l’abocador de Garraf, el més gran de la península Ibèrica, la mina abandonada d’Aljustrel a la zona portuguesa de la faixa piritosa ibèrica, el Parc Nacional de Doñana afectat pel vessament de la mina de pirita d’Aznalcóllar, situada a la part espanyola de la faixa piritosa ibèrica i el Delta de l’Ebre afectat diverses activitats humanes incloent caça, industria i residus domèstics. Dels exemplars analitzats, 138 van ser capturats pròpiament per estudis ecotoxicològics a 3 de les zones contaminades (Garraf, Aljustrel i Doñana) i 105 eren mostres de col•lecció procedents d’altres estudis ecològics (Delta de l’Ebre). Especialment en les zones contaminades estudiades, es van observar marcades diferències en la bioacumulació entre elements essencials i no essencials per diferències en biodisponibilitat, toxicitat, i capacitat de transferència i biomagnificació al llarg de les cadenes tròfiques entre ambdós grups d'elements. Els majors augments detectats als teixits de les musaranyes de les zones contaminades es van donar entre els metalls pesants no essencials (Pb, Cd, Hg, Tl i Ni), els nivells tissulars dels quals no estan metabòlicament regulats a mamífers, donant com a resultat alts increments a teixits durs i tous quan augmenta la biodisponibilitat al medi ambient. Dels elements quantificats només Pb, Hg, Cd, i, possiblement Tl, Ni, B, Ba i Sr, van acumular-se fins a nivells que poden ser tòxics en mamífers. Ja que hi ha escassa informació sobre com afecta l’exposició crònica a les poblacions naturals d’insectívors, les dades conjuntes dels biomarcadors d’efecte obtingudes serveixen com valors de referència per a l’espècie i poden contribuir a estimar els efectes tòxics sobre els organismes. Es van detectar disminucions en la massa corporal i l'activitat de GST, i increments de massa i patologies atribuïbles a la contaminació a fetge i ronyó i freqüència de micronuclis en les musaranyes de les zones contaminades. Dels dos factors biòtics estudiats, l'edat va ser el més important ja que 10 dels elements, amb especial rellevància en el cas del Cd, van presentar patrons d’acumulació diferencials entre joves i adults. Per contra, mascles i femelles de musaranya comuna només van presentar petites diferències en l'acumulació de Pb, Hg, Ni, Mo, Mn, Co i Fe. Com amb els marcadors d’exposició, la rellevància de edat en els biomarcadors d’efecte va ser també major que la del sexe per explicar la variabilitat observada. L’ús de diversos biomarcadors d’exposició d’efecte permeten d’una banda obtenir una millor idea del conjunt d'efectes tòxics de la contaminació sobre el normal desenvolupament dels sistemes biològics, així com indiquen la idoneïtat de la musaranya comuna com a bioindicador de contaminació ambiental especialment per l’avaluació de risc ambiental a zones protegides i per les poblacions humanes., [eng] In this report, a quantification of biomarkers of exposition (accumulation of Pb, Hg, Cd, Tl, Fe, Mg, Zn, Cu, Mn, Mo, Co, Ni, Cr, Sr, Ba and B in soft and hard tissues) and effect (morfometry, haematology, enzymatic activity, genotoxicity and histopathology) in a total of 243 specimens of the greater white-toothed shrew, Crocidura russula (Hermann, 1780) was found. In the present study, the age- and sex-dependent variations were also considered in order to explain the variability due to these important biotic factors. The study sites were 4 polluted sites and 4 reference sites selected both for their environmental relevance (6 out the 8 sites have granted partial or total protection status) and for the interest for the society: the landfill of Garraf, the bigger dump site in the Iberian Peninsula; the abandoned mine of Aljustrel in the Portuguese part of the Iberian Pyrite Belt (IPB); the National Park of Doñana affected by the wastes of the mine of Aznalcóllar, sited in the Spanish part of the IPB; and the Ebro Delta disturbed during decades by several human activities including hunting, industries and domestic wastes. A total of 138 shrews were captured for ecotoxicological studies in 3 polluted sites (Garraf, Aljustrel, and Doñana) and 105 specimens were captured for other ecological studies and stored in a zoological collection (Ebro Delta). Especially in the polluted sites studied, there were marked differences between the bioaccumulation patterns of essential and non-essential elements due to differences in the bioavailability, toxicity, and transfer and bioaccumulation throughout the food chains between the two groups of elements. The highest increases in the tissues of shrews from the polluted sites were on non-essential heavy metals (Pb, Cd, Hg, Tl, and Ni), without metabolic regulation of tissular levels in mammals that ends in high burdens in soft and hard tissues when bioavailability increase in the environment. Among elements quantified, only Pb, Hg, Cd, and, probably Tl, Ni, B, Ba and Sr, were accumulated till levels that may produce toxic effects in mammals. The scarce information of the effects of chronic exposure in natural populations of insectivores render more relevant the use of a battery of biomarkers of effect, that serve as reference values for the species and as assessment of toxic effects in organism. Decreases of body mass and GST activity and increases of mass and pathologies attributable to pollution in liver and kidneys were detected in shrews from the polluted sites. Among biotic factors, age was important because 10 of the elements quantified, with special remark on Cd, showed differential bioaccumulation patterns in juveniles and adults. In contrast, only slight differences in bioaccumulation of Pb, Hg, Ni, Mo, Mn, Co, and Fe were found between males and females. The same pattern of importance of age and sex as in the biomarker of exposure was found in the biomarkers of effect. The use of several biomarkers of exposure and effect provide information of the whole toxic effects of pollution in the normal function of biological systems and point out C. russula as a suitable bioindicator of environmental pollution, specially to evaluate environmental risk both in protected sites and human populations.

Prostate cancer (PCa) is the most frequently diagnosed type of cancer in developed countries. The decisive method of diagnosis is based on the results of biopsies, morphologically evaluated to determine the presence or absence of cancer. Although this approach leads to a confident diagnosis in most cases, it can be improved by using the molecular markers present in the tissue. Both miRNAs and proteins are considered excellent candidates for biomarkers in formalin-fixed paraffin-embedded (FFPE) tissues, due to their stability over long periods of time. In the last few years, a concerted effort has been made to develop the necessary tools for their reliable measurement in these types of samples. Furthermore, the use of these kinds of markers may also help in establishing tumor grade and aggressiveness, as well as predicting the possible outcomes in each particular case for the different treatments available. This would aid clinicians in the decision-making process. In this review, we attempt to summarize and discuss the potential use of microRNA and protein profiles in FFPE tissue samples as markers to better predict PCa diagnosis, progression, and response to therapy.

Altres ajuts: Asociación Española Contra el Cáncer Junta Provincial de Barcelona; Red de Genómica del Cáncer y Genotipado de tumores C03/10; Movember Foundation 2012, Valor 2010/00220, programa INNPACTO and Lisa Piccione for reviewing the document. In order to successfully cure patients with prostate cancer (PCa), it is important to detect the disease at an early stage. The existing clinical biomarkers for PCa are not ideal, since they cannot specifically differentiate between those patients who should be treated immediately and those who should avoid over-treatment. Current screening techniques lack specificity, and a decisive diagnosis of PCa is based on prostate biopsy. Although PCa screening is widely utilized nowadays, two thirds of the biopsies performed are still unnecessary. Thus the discovery of non-invasive PCa biomarkers remains urgent. In recent years, the utilization of urine has emerged as an attractive option for the non-invasive detection of PCa. Moreover, a great improvement in high-throughput "omic" techniques has presented considerable opportunities for the identification of new biomarkers. Herein, we will review the most significant urine biomarkers described in recent years, as well as some future prospects in that field.

Bioassessments are used to measure system health and assess disturbance. While fish-based freshwater bioassessments are cost-effective and perform well in speciose systems, such bioassessments remain difficult to implement in species-poor Mediterranean regions. Population size structure metrics may provide meaningful biological information where depauperate communities preclude the richness and composition measures generally used. We focus our assessments of population size structure responses to anthropogenic perturbation on one of the most widespread native stream fish (Squalius laietanus). We explore a number of population size statistics as metrics for a Mediterranean region, where current bioassessments perform poorly. Our sampling encompassed 311 sites across Catalonia (NE Spain) where we characterized anthropogenic perturbation using a summary of impacts, including local data on stream condition and landscape indicators of degradation, via a principal component analysis. Anthropogenic perturbation in streams was collinear with altitudinal gradients and highlights the importance of appropriate statistical techniques. Of the population size structure metrics explored, average length was the most sensitive to anthropogenic perturbation and generally increased along the disturbance gradient. Although we expected to find consistent changes in variance, kurtosis, and skewness, the observed relationships were weak. River basin mediated responses suggest the importance of environmental landscape factors. The unexpected increases of mean S. laietanus body size with anthropogenic perturbation, strong effects of river basin, collinearity with spatial gradients and the species-specific nature of responses preclude the direct application of size structure in freshwater bioassessments. Although its application in fish-based freshwater bioassessments appears difficult, population size structure can provide insights in species-specific applications and management Additional financial support was provided by theSpanish Ministry of Science (projects CGL2009-12877-C02-01 andConsolider-Ingenio 2010 CSD2009-00065)

[eng] Birds have been widely used as bioindicators. In this study we face the use of birds as bioindicators of metal pollution in two different scenarios of contamination: one that takes place in an aquatic environment, the Ebro river basin, and a second that occurs in a terrestrial environment, the Bolivian Andes. In the case of the Ebro river basin, the pollution threat is a factory located at the river bend, close to Flix, that due to its long operational activity and along with the construction of a dam next to the plant around 1960, resulted in the accumulation of 200,000–360,000 tons of industrial wastes in the riverbed, occupying an area of 700 m of length and 60 m of width. In this study case we evaluated whether aquatic birds such as the Purple Heron reflect the potential pollutant exposure from Flix Reservoir among different riverine and deltaic areas, and assess their usefulness as bioindicators. Also we examined if the polluted wastes of Flix reservoir affect the levels of pollution of the habitats where waterbird populations of the Ebro Delta (situated 90 km downstream) forage and feed. As results stable isotopes shown the high nitrification and lower carbon signatures in the river, and together with the niche width metrics, that Common and Sandwich Tern behave as strict specialists at the Ebro Delta, with narrow niche widths, while in the opposite way, Little Tern, Little Egret, Purple Heron, Night Heron and the Moorhen behave like generalists foragers, with broad niche widths. Mercury levels in nestlings of Purple Heron at Flix site and eggs of Audouin’s Gull, Little Tern and Common Tern ‘Banya’ at the Ebro Delta are high enough to be of special concern. Arsenic (specially used in combination with carbon signatures) discriminates outstandingly between marine and freshwater species. Both eggs and nestling feathers of Purple Herons are adequate bioindicators for trace element pollution, but nestling feathers present certain advantages. The six selected blood parameters (TOSC ROO*, TOSC OH*, BhCE, CbE, LDH and MN) provide the first evidence of an association of biological responses with pollutants in heron species. The second study case takes place in Bolivia. Anthropogenic mining has taken place in the Bolivian Andes since at least the fifteenth century. Particularly the East of Oruro Department in the Eastern Andean Cordillera is affected by a long term history of mining. These mining activities were characterized by the deposition of large tailing piles, which accumulated in abandoned and active mines, where trace metals such as lead, arsenic, cadmium, and antimony could reach surface waters and soils, and so the biota. In this case we evaluated the potential of Tinamou species as sentinels of exposure to local trace metal pollution and studied different routes of lead accumulation into the sentinel species, in order to assess the usefulness of feather levels as a measure of the exposure to this pollutant. As results we found that the detected small home ranges detected in Ornate Tinamou (lower than 1 km2) and their sedentary habits make of this species an adequate bioindicator of local pollution. We recommend feathers for future monitoring programs as they are a non invasive sample. Higher rates of histopathological damage were found in kidney at the polluted sites. We detected high levels of trace elements detected at the polluted sites in the two studied species of Tinamou (Ornate and Darwin Tinamou), many of them trespassing hazard levels. The calamus is the most suitable section for assessing the original endogenous lead levels in feather samples., [spa] En este estudio las aves serán utilizadas como bioindicadoras de contaminación por metales pesados en dos escenarios diferentes: el primero en un ambiente acuático, la cuenca del rio Ebro, mientras que el segundo tiene lugar en un ambiente terrestre, los Andes bolivianos. En el primer caso la fuente de contaminación es una fábrica localizada cerca de Flix, que produjo cientos de toneladas de lodos tóxicos. Así evaluamos si aves acuáticas como la Garza imperial reflejan la exposición potencial a los contaminantes de Flix entre zonas fluviales y deltaicas, comprobando su utilidad como bioindicadoras, y también examinamos si dichos residuos afectan a la contaminación de los hábitats donde la avifauna del Delta del Ebro se alimenta. Como resultados los isótopos estables mostraron la alta nitrificación y signaturas más bajas de carbono en el río y observamos que el Charrán común y patinegro actúan como especialistas en el Delta, mientras que Charrancito, Garceta común, Garza Imperial, Martinete y Gallineta de agua son generalistas, presentando amplios nichos tróficos. El mercurio en volantones de Garza Imperial en Flix y huevos de Gaviota de Audouin, Charrancito y Charrán común “Banya” en el Delta presentan valores alarmantes. El arsénico discrimina claramente entre especies marinas y dulceacuícolas. Ambos huevos y plumas son bioindicadores adecuados de contaminación, pero las plumas presentan más ventajas. El segundo estudio tiene lugar en los Andes Bolivianos, donde la actividad minera ha dado lugar a la acumulación de residuos. Evaluamos el potencial de las especies de Tinamúes como bioindicadoras de la exposición a dicha contaminación, incluyendo las diferentes vías de acumulación de plomo en las especies bioindicadoras. Como resultados, los pequeños territorios detectados y hábitos sedentarios hacen del Tinamú Pisacca un bioindicador adecuado de la contaminación local. Se recomienda el uso de plumas al son una muestra no invasiva. El mayor daño histopatológico se encontraron en muestras de riñón de las zonas contaminadas. La contaminación minera ha quedado de manifiesto con los altos niveles detectados en las dos especies estudiadas de Tinamú. El cálamo es la sección más adecuada para la evaluación de los niveles originales endógenos de plomo.

Podeu consultar el llibre complet a: http://hdl.handle.net/2445/32393, The relevancy of parasites as potential indicators of environmental quality has been increasing over the last years, mostly due to the variety of ways in which they respond to anthropogenic pollution. The use of fish parasites as bioindicators of heavy metal pollution in aquatic ecosystems has been widely studied. However, little information concerning terrestrial habitats is presently available. In fact, in the last two decades several studies have been performed worldwide in different habitats and/or conditions (theoretically both in polluted and unpolluted terrestrial ecosystems, but mainly in aquatic ecosystems) in order to investigate heavy metal pollution using parasitological models. Different groups of vertebrates (mainly fish, mammals and birds) and several parasitological models have been tested involving acanthocephalans mostly, but also cestodes and nematodes. It is not the aim of this chapter to do a complete revision of the available data concerning this subject. Instead, we emphasize some general aspects and compile a mini-review of the work performed in this field by our research group. The results obtained until now allow confirming several parasitic models as promising bioindicator systems to evaluate environmental cadmium and mainly lead pollution in terrestrial non-urban habitats, as it was already demonstrated for aquatic ecosystems. The present knowledge also allows confirming that parasites can reveal environmental impact. Environmental parasitology is an interdisciplinary field, which needs simultaneous expertise from toxicology, environmental chemistry and parasitology. Furthermore, environmental parasitology should be taken into account in order to increase the efficiency of environmental monitoring programs.

[cat] Els resultats que es presenten en aquesta tesi estan centrats bàsicament en l'estudi de la biodiversitat de les diatomees epilítiques presents en els rius de la conca de l'Ebre i en la seva aplicació com a indicadores de l'estat ecològic de les masses d'aigua. Durant els mesos d'estiu del 2005 i 2006 es va realitzar el mostreig de la flora de diatomees epilítiques en un total de 197 i 211 estacions, respectivament, pertanyents a la Xarxa de Control de l'Estat de les Masses d'Aigües Superficials (CEMAS) de la Confederació Hidrogràfica de l'Ebre (CHE). Aquests mostrejos van ser impulsats per la CHE amb l'objectiu d'establir xarxes de control biològic, basades en bioindicadors del fitobentos per tal de conèixer l'estat ecològic de les aigües fluvials d'acord amb el que estableix la Directiva Marc de l'Aigua (DMA). La recollida de les mostres, el seu tractament químic i, la quantificació de les espècies per localitat, s'han basat en les normatives estandarditzades europees CEN/TC 230 EN 13946:2003, norma prEN 14407: 2004 i, en els protocols de la CHE per l'avaluació de la qualitat biològica en rius mitjançant diatomees. L'examen de les mostres recollides ha permès la identificació de 499 tàxons, repartits en 91 gèneres, dels quals 492 han estat a nivell específic o infra-específic. Dels tàxons identificats, 361 estan representats en 751 fotografies realitzades al microscopi òptic, agrupats en 25 làmines. Com a particularitats de la flora s'esmenta d'una banda la presència de 7 espècies al•lòctones segons les consideracions principalment de Coste & Ector 2000 Achnanthidium catenatum, A. druartii, Diadesmis confervacea, Didymosphenia geminata, Gomphoneis minuta, Navicula kotschyi y Reimeria uniseriata) i per l'altre, la presència en algunes localitats de diverses formes teratològiques que podrien indicar condicions adverses del medi (estrès químic o contaminació), encara que en alguns casos podrien ser originades simplement per causes naturals. Com a resultat de la determinació de l’estat ecològic de les masses d’aigua s'ha comprovat l'aplicació i funcionament òptim dels índexs de diatomees (Index de Pol•lució Específica IPS, Index Biològic de Diatomees IBD i Index Europeu CEE), sent l'IPS l'índex més convenient per la bioindicació de l'aigua a la conca de l'Ebre. No obstant, aquests índexs no reflecteixen prou bé les diferències entre algunes ecoregions de la conca de l'Ebre, com ara les capçaleres calcàries. Tenint en compte els resultats obtinguts amb l'IPS, la proporció de localitats estudiades que complirien amb la DMA en aquests 2 anys d'estudi són bastant similars, seria del, [eng] "Diatoms of the Ebro basin: Biodiversity and ecological state of water" TEXT: The aim of this PhD thesis is to contribute to the taxonomic knowledge of the epilithic diatom flora from the rivers of the Ebro basin and to determinate the water quality status in the studied area using benthic diatoms as indicators. The studied area is located inside the Water Quality Surveillance Network of the Ebro Water Agency, CHE (NE Spain). It includes the Ebro and Garona rivers and all the rivers and streams which run into them in the Spanish territory, mostly over a calcareous and sedimentary substrata. A total of 408 epilithic diatoms samples were collected from different environments and ecoregions within the limits of the network of the Ebro Water Agency, CHE (NE Spain) during the summers of 2005 (197 sites) and 2006 (211 sites). Epilithic diatom were sampled, and then treated to prepare permanent diatoms slides following the standard methods (European Committee for Standardization 2003, 2004). The number of taxa identified in the material studied amounted to 499 (492 a specific or infra specific level), belonging to 91 different genera. 361 taxa are presented in 751 light microscope micrographs regrouped in 25 plates. We noticed the presence of species considered rare or exotic for European freshwaters (Coste & Ector, 2000) such as Diadesmis confervacea; Achnanthidium catenatum, Reimeria uniseriata, Navicula kotschyi, Gomphoneis minuta and Didymosphenia geminata. A few diatoms species presented teratological forms have been detected in some localities. However, we could not find a direct relationship of these forms occurrence and any particular cause. In order to evaluate the water quality assessment, three diatoms indices were applied (Specific Pollution Index IPS, Biological Diatom Index BDI and European Index CEC). All of them correlated significantly with water physical and chemical variables, but SPI achieved better correlations, and is therefore to be the best diatom index to be applied in the Ebro basin, although in some cases it did not properly reflected special situations in this basin as calcareous headwaters. According to the cluster and canonical correspondence analyses, nutrient enrichment (from the headwaters to the mouth), environmental variables and impacts were to be the most important variables structuring diatom communities in the Ebro basin.

Informe sobre l'ajust de l’Índex d’Integritat Biòtica (IBICAT) basat en l’ús dels peixos com a indicadors de la qualitat ambiental als rius de Catalunya

[eng] The effects of the main anthropogenic impacts in the fluvial ecosystem of the lower Ebro River are evaluated along the five chapters of the present thesis, using the benthic macroinvertebrate community as a bioindicator. Different approaches are integrated in function of the type of stressor, considering mainly heavy metal and organochlorine pollution and hydrological alterations. These approaches focus on the taxonomy and biological traits of the community, and on the bioaccumulation of pollutants, population and life history studies on keystone species such as Ephoron virgo (Ephemeroptera: Polymitarcyidae). The results evidence a strong ecological response, and demonstrate that these effects act at different levels of organization, including communities, populations and individuals., [cat] Els efectes dels principals impactes antropogènics de l'ecosistema fluvial del tram baix del riu Ebre s'estudien al llarg dels cinc capítols d'aquesta tesi, utilitzant la comunitat de macroinvertebrats bentònics com a bioindicadors. Depenent del tipus d'estrès, tenint en compte principalment la contaminació per metalls pesants i organoclorats i les alteracions hidrològiques, s'han considerat diferents aproximacions. Concretament, l'estudi se centra en la taxonomia i els trets biològics de la comunitat, la bioacumulació de contaminants, i els estudis poblacionals i relacionats amb el cicle biològic d'espècies clau com l'Ephoron virgo (Ephemeroptera: Polymitarcyidae). Els resultats evidencien una resposta ecològica important, i demostren que aquests efectes actuen a diferents nivells d'organització, incloent comunitats, poblacions i individus.

[spa] La exposición a sustancias químicas altamente tóxicas como los contaminantes orgánicos persistentes (COP), representa una seria amenaza para los depredadores marinos y sus redes tróficas. Dicha exposición depende en gran medida de la ecología, distribución y áreas de alimentación de la fauna marina. Sin embargo, la contribución relativa de estas características biológicas para explicar los niveles de contaminación que se encuentran en los organismos es aún desconocida para la mayoría de los depredadores, incluyendo las aves marinas. Metodologías novedosas como el análisis de isótopos estables y los sistemas de geolocalización proporcionan información esencial para poder entender la influencia de los distintos factores que determinan la exposición a los contaminantes marinos. En la presente tesis se evalúa la influencia de factores ecológicos clave, principalmente posición trófica, áreas de forrajeo, distribución y movimientos, sobre las relaciones isotópicas de carbono y nitrógeno (δ13C y δ15N) y los niveles de contaminación encontrados en la mayor parte de las especies de Procellariiformes que crían en el Atlántico Nordeste y en el mar Mediterráneo. En particular, se determinaron los niveles de hidrocarburos policíclicos aromáticos (HAP), bifenilos policlorados (PCB) y pesticidas organoclorados como el DDT. En este trabajo, en primer lugar, se evalúa la influencia relativa de la distribución y la ecología trófica de las aves marinas sobre las firmas de isótopos estables y las cargas de contaminantes encontradas. A continuación se exponen los patrones geográficos encontrados para δ13C y δ15N y los contaminantes estudiados en el NE Atlántico y el Mediterráneo. En segundo lugar, se expone la marcada relación encontrada entre la ecología trófica, incluyendo posición trófica y áreas de forrajeo, de las aves estudiadas y los niveles de contaminación que presentan. En conjunto, esta tesis pone de manifiesto la utilidad de combinar los avances y metodologías propios de la química ambiental con las nuevas metodologías utilizadas para el estudio de la ecología de la fauna salvaje, así como el gran valor de las aves marinas pelágicas como centinelas del estado de contaminación que sufren nuestros océanos., [eng] Exposure to contaminants, such as Persistent Organic Pollutants (POPs), is currently considered a serious anthropogenic threat to marine predators and their food webs. Exposure to contaminants by marine wildlife is mainly related to their trophic ecology, distribution and movements. However, the contribution of these biological features to pollutant levels remains poorly understood in most marine predators, including seabirds. New methodologies, including dietary and geographic tracers, such as stable isotope analyses, and a wide array of devices to track movement at different spatial and temporal scales, can provide new light into this issue. Although stable isotope signatures in marine wildlife provide valuable information about their trophic ecology, isotopic baseline levels also show geographical differences. Therefore, to understand isotopic differences among separate wildlife populations we first need to evaluate the influence of spatial variability in stable isotope signatures. In the present dissertation, some ecological factors, mainly feeding ecology, breeding locality and movements, and their influence shaping the isotopic signatures and the contaminant burdens, were examined in most Procellariiformes breeding in the Northeast Atlantic Ocean and the Mediterranean Sea. In particular, the isotopic signatures of Carbon and Nitrogen (δ13C and δ15N) as well as burdens of polycyclic aromatic hydrocarbons (PAHs) and organochlorinated compounds (OCs), e.g., polychlorinated biphenyls (PCBs) and pesticides such as DDT, in seabirds were determined. In this study, firstly, we evaluate the relative influence of geographic origin, movements and trophic ecology in shaping stable isotope signatures and contaminant burdens in pelagic seabirds. Then, we show the spatial patterns in the isotopic signatures, OCs and PAHs that emerge among Mediterranean and northeast Atlantic seabirds and evaluate pollutant sources and the influence of long-range transport mechanisms on these basins. Secondly, we provide evidences that show no marked differences in the trophic ecology of the studied seabird species throughout their breeding distributions, but clearly differ among species, pointing out the marked relationship of seabird trophic ecology with their PCB, DDT and PAH burdens. Overall, this study underlines the usefulness of combining environmental chemistry methodologies with new approaches in the study of animal ecology, showing the value of pelagic seabirds in marine contamination monitoring

Among increasingly used pharmaceutical products, beta-blockers have been commonly reported at low concentrations in rivers and littoral waters of Europe and North America. Little is known about the toxicity of these chemicals in freshwater ecosystems while their presence may lead to chronic pollution. Hence, in this study the acute toxicity of 3 beta-blockers: metoprolol, propranolol and atenolol on fluvial biofilms was assessed by using several biomarkers. Some were indicative of potential alterations in biofilm algae (photosynthetic efficiency), and others in biofilm bacteria (peptidase activity, bacterial mortality). Propranolol was the most toxic beta-blocker, mostly affecting the algal photosynthetic process. The exposure to 531microg/L of propranolol caused 85% of inhibition of photosynthesis after 24h. Metoprolol was particularly toxic for bacteria. Though estimated No-Effect Concentrations (NEC) were similar to environmental concentrations, higher concentrations of the toxic (503microg/L metoprolol) caused an increase of 50% in bacterial mortality. Atenolol was the least toxic of the three tested beta-blockers. Effects superior to 50% were only observed at very high concentration (707mg/L). Higher toxicity of metoprolol and propranolol might be due to better absorption within biofilms of these two chemicals. Since beta-blockers are mainly found in mixtures in rivers, their differential toxicity could have potential relevant consequences on the interactions between algae and bacteria within river biofilms.

[cat] La detecció de l’orgien de la contaminació fecal a l’aigua perment millorar la gestió del medi i mitigar la contaminació en l’origen recuperant amb més eficiència les zones afectades; a més a més permet estimar el risc sanitari que porta associat; prendre les mesures legals oportunes i establir responsabilitat econòmiques. Fins a l’actualitat s’han descrit un elevat número de tècniques i marcadors que indiquen la presència de contaminació i permeten conèixer el seu origen. Abans de la implementació d’aquestes metodologies cal conèixer altres factors. Aquests mètodes s’han definit en una determinada àrea demogràfica i per una determinada població, per tant, cal conèixer si existeix variació geogràfica i temporal, saber-ne la prevalència i persistència al medi, valorar la distribució entre espècies, materialitzar procediments stàndars que puguin ser utilitzats en qualsevol laboratori. A més a més, estudis previs han suggerit que cap dels indicadors de l’origen de la contaminació fecal presenta un 100% d’eficiència i especifitat, així que és necessària la combinació de varis d’ells per tal d’incrementar la discriminació entre diferents oríigens. Aquesta treball consisteix en tres parts diferenciades que volen aprofundir ens les mancances dels mètodes per a determinar l’origen de la contaminació. En la primera part (Capítol 2) s’ha valorat 10 marcadors diferents descrits prèviament: dues espècies del gènere Bifidobacterium marcadores de contaminació humana: Bif. adolescentis i Bif. dentium, dos marcadors específics d’humans (HF183 i HF134) i dos de rumiants (CF128 i CF193) que detecten espècies no cultivades del grup Bacteroidetes, un marcador del gen esp de soques d’Enterococcus faecium específiques d’humans i 3 marcadors d’ADN mitocondrial de cèl•lules humanes, bovines i porcines. Tot i que alguns dels marcadors moleculars analitzats presenten una elevada sensibilitat i especificitat, cap d’ells assegura un 100% d’encert. Això ha fer que es valorés la possibilitat de desenvolupar models predictius amb combinacions d’ells. D’aquesta manera s’ha obtingut una discriminació del 79,6% entre 4 oríigens diferents: humà, boví, porcí i avícola i d’un 90,1% distingint entre contaminació d’origen humà o no humà. A la segona part, veient la importància de Bif. adolescentis i Bif. dentium com a marcadors de contaminació fecal humana, s’ha utilitzat la tècnica de DGGE per tal d’aprofundir en l’ecologia del gènere. S’ha analitzat la distribució d’espècies en mostres d’aigua residual de diferents orígens. Bif. adolescentis s’ha confirmat com una de les espècies majoritàries en mostres d’origen humà. A la vegada, s’han definit espècies majoritàries en mostres d’origen boví com Bif. pseudolongum ssp. pseudolongum i Bif. thermophilum. Aquesta espècie també ha estat majoritària en porcs juntament amb una espècie que no identificada prèviament. En aus s’ha detectat una sola espècie identificada com Bif. saeculare. En els últims anys s’han dissenyat un elevat nombre de PCRs i q-PCRs específiques d’espècies no cultivades del grup Bacteroidetes per discriminar entre diferents hostes. En la tercera part, s’ha volgut aprofundir en el coneixement de la persistència del gènere Bacteroides i conèixer els factors ambientals que afecten la seva supervivència. Mitjançant experiments in situ al riu Llobregat s’ha observat la persistència de les soques tipus Bact. fragilis i Bact. thetaiotaomicron, majoritàries en aparell digestiu humà i espècies ambientals de l’aigua residual. S’ha realitzat el seguiment de la supervivència mitjançant tècniques de cultiu i tècniques moleculars. S’ha observat una major supervivència de Bact. fragilis a baixes temperatures que altes. Bact. thetaiotaomicron ha mostrat el contrari, major supervivència a l’estiu quan s’ha registrat una menor concentració d’oxigen dissolt a l’aigua. En aquesta espècie s’ha descrit prèviament una menor tolerància a l’oxigen. La presència de predadors a l’aigua també afecta de manera important la supervivència del gènere., [eng] Faecal pollution is one of the main causes of degradation of surface and coastal water quality. Faecal pollution can be attributed to different sources, for example urban sewage, sewer misconnections, slaughterhouse wastewaters, manure run-off and solid disposals. The principal faecal bacterial indicators E. coli and Enterococcus are used to show the presence of pollution as well as the faecal load in water. However, these indicators do not discriminate between the possible sources of pollution. During the last decade many authors have described different methodologies based on molecular tools to detect different Microbial Source Tracking (MST) markers. However there is a lack of knowledge about the behaviour of the markers when they appear in the environment. Usually these markers have been described in a specific geographical area. For this reason their geographical and temporal stability has to be tested. Other aspects such as their prevalence and persistence in the aquatic environment must be known and standard operating procedures must be determined. The current project aimed to address these questions. 144 samples of different human and animal sources around Catalonia were collected in order to evaluate the applicability of some described MST markers, e.g. Bifidobacterium adolescentis, Bif. dentium, Bacteroides species human and ruminant specifics, esp gene of Ent. faecium and mitochondrial DNA of human, bovine and swine cells. The specificity and sensitivity of the markers was determined. Each marker alone does not give 100% source discrimination and for that reason predictive models were developed. These models increased the capacity of source discrimination using the lowest possible number of markers. Bif. adolescentis and Bif. dentium have been described as human specific species. Denaturant gradient gel electrophoresis technique was used to broaden the knowledge of Bifidobacterium distribution among different animal sources and to describe new species specificity to enable their uses as new MST markers. In addition, Bacteroidetes species have shown themselves to be capable MST markers. How environmental factors such as temperature and dissolved oxygen affect their survival in the environment has been studied in order to predict the durability of the markers in these environments. For this purpose, molecular techniques such as PCR and Q-PCR and cultivable methodologies combined with colony hybridization were used.

Several methods and approaches for measuring parameters to determine fecal sources of pollution in water have been developed in recent years. No single microbial or chemical parameter has proved sufficient to determine the source of fecal pollution. Combinations of parameters involving at least one discriminating indicator and one universal fecal indicator offer the most promising solutions for qualitative and quantitative analyses. The universal (nondiscriminating) fecal indicator provides quantitative information regarding the fecal load. The discriminating indicator contributes to the identification of a specific source. The relative values of the parameters derived from both kinds of indicators could provide information regarding the contribution to the total fecal load from each origin. It is also essential that both parameters characteristically persist in the environment for similar periods. Numerical analysis, such as inductive learning methods, could be used to select the most suitable and the lowest number of parameters to develop predictive models. These combinations of parameters provide information on factors affecting the models, such as dilution, specific types of animal source, persistence of microbial tracers, and complex mixtures from different sources. The combined use of the enumeration of somatic coliphages and the enumeration of Bacteroides-phages using different host specific strains (one from humans and another from pigs), both selected using the suggested approach, provides a feasible model for quantitative and qualitative analyses of fecal source identification.

Durant el període 1995-2002 Catalunya va patir una sequera prolongada que va afectar, en diferent grau, als sistemes fluvials de les Conques Internes Catalanes, especialment per la seva durada que va comprendre des d'un mínim de 2 anys en les conques del nord fins a 6 anys en les del sud. Dins aquest marc, aquesta tesi pretén estudiar l'efecte d'aquesta sequera sobre la qualitat físico-química, les comunitats de macroinvertebrats i els índexs biòtics emprats en el seguiment de la qualitat ecològica dels nostres rius, en concret en l'índex BMWPC, l'ASPT, el nombre de tàxons i l'índex EPT. També s'hi ha fet una actualització del BMWPC, i la predicció mitjançant descriptors ambientals del seu valor amb l'objectiu d'avaluar la integritat de les comunitats de macroinvertebrats en els punts de la xarxa de seguiment de les Conques Internes Catalanes (CIC). S'hi inclou també l'estudi de la variació interanual dels principals descriptors físics, químics i biològics analitzats per conques i separant-hi els paràmetres biològics de tots els altres, per la qual cosa s'ha empleat l'anàlisi de PCA i les distàncies de pertorbació obtingudes a partir d'aquestes anàlisis, d'acord amb la metodologia formulada per Sabater et al. (1991). Els resultats obtinguts mostren que els descriptors que s'han utilitzat són els adequats i aquests són, en part, els mateixos que recomana la Directiva Marc de l'Aigua. Es pot dir que l'efecte de sequera és més important especialment per les comunitats menys alterades i que les reduccions de cabal afecten tan a les estructures de les comunitats de macroinvertebrats (efecte parcialment apaivagat als rius regulats) com a la qualitat físico-química dels rius de les CIC. Pel que fa a l'índex BMWPC no ha mostrat cap davallada de seu valor en resposta a la sequera, ja que s'han mesurat durant tot el període uns valors iguals o un xic millors en funció de l'equilibri que s'obté entre els tàxons sensibles que desapareixen i els lenítics que s'incorporen a cada comunitat, la qual cosa fa que sigui un índex robust en front a la sequera. Per avaluar els canvis en l'estructura de les comunitats de macroinvertebrats és precís l'ús de l'índex ASPT, que valora la qualitat relativa de tota la comunitat i s'ha mostrat com a l'òptim per observar l'efecte de la llarga sequera soferta. En relació amb les distàncies de pertorbació s'ha confirmat que augmenten quan minven els cabals i decreixen quan aquests s'incrementen., During the period between 1995 and 2002 Catalonia suffered a long lasting drought that affected in varying degrees the river system of the Catalan Internal Basins (CIB), especially in which refers to the time length, ranging from a minimum of two years in the north basins to six years in the southern ones. Within this framework, this thesis aims to study the effects of this drought on the physical-chemical quality, the macroinvertebrate communities and the biotic indices used in monitoring the ecological quality of our rivers. Particularly the BMWPC, EPT, ASPT and richness (number of taxa) indexes were used.The update of the BMWPC, and the prediction of the index changes through environmental descriptors of its value in order to evaluate the integrity of the macroinvertebrate communities in the different points of the control network monitoring the CIB have also been made. The interannual variation of the main physical, chemical and biological descriptors by basins was performed, and analysed by separating the biological parameters from all others. For it, the PCA analysis has been used and the disturbance distances in accordance with the methodology formulated by Sabater et al. (1991) were established.The results obtained show that the descriptors used are suitable and are partially the same ones that the Water Framework Directive recommends. The drought effect is important especially for the less altered communities and the flow reductions affect both the structures of the macroinvertebrates communities (effect partially appeased in the regulated rivers) and the physico-chemical quality of the rivers and the CIB.The BMWPC index has not been showing any decrease of its value in relation to the drought. It is to note that it has been measured during periods with equal or slightly better values in terms of the balance achieved between disappearing sensitive taxa and lentic taxa being incorporated into each community, Then the index, proved to be robust in drought conditions. To assess changes in the structure of the macroinvertebrate communities it is necessary to use the ASPT index which establishes the relative quality of the whole community. It has also been proved to be the optimum index to use in order to study the effect of the drought suffered. In relation to the distances from disturbance, it has been confirmed that they have increased when the flows diminished and decreased when flow increased.

A key issue in the implementation of the Water Framework Directive is the classification of streams and rivers using biological quality parameters and type-specific reference conditions. Four groups of stream types were defined in NE Spain on the basis of 152 diatom samples by means of detrended correspondence analysis and classification techniques. Diatom analysis was restricted to epilithic taxa, and the sites included gradients ranging from near-natural streams to sites with poor ecological quality. The main gradient shows a clear separation of sites in relation to the degree of human influence: polluted streams (mainly located in the lowlands) differ from streams in mountainous areas and in the Pyrenees. A second gradient is related to physiographical features. Headwater streams can be distinguished by their catchment geology. The type-specific diatom taxa for the stream types studied were determined by using indicator species analysis (IndVal). The type-specific taxa from near-natural streams are coincident with the indicator taxa for high ecological status. Human impact reduced the typological heterogeneity of the diatom community composition. Overall, the diatom communities in NE Spain exhibit a regional distribution pattern that closely corresponds with that observed in river systems elsewhere. Physiographical differences are only evident in undisturbed sites, while nutrient enrichment and other human disturbances may mask the regional differences in the distribution of diatom communities