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Trabajo presentado en la XIX Conference of the Iberian Association of Limnology (Inland waters and XXI century challenges: from scientific knowledge to environmental management), celebrada en Coimbra del 24 al 29 de junio de 2018., Invasive species are one of the most important threats to biodiversity globally, with freshwaters being one of the most susceptible ecosystems. Freshwater crayfish are among the world's most successful aquatic invaders. Crayfish play a key role within trophic networks and are often important ecosystem engineers. In recent years, different invasive crayfish species have expanded rapidly throughout Europe, and their distributions overlap. Thus, complex interactions among invasive crayfish may be occurring, which could have consequences for the fitness and/or the invasion success of the different species. To date, most of the research on interactions among crayfish species has focused on ethological studies of agonistic struggles or competition for resources. However, studies exploring the mechanisms underlying such interactions remain very limited. In this study, we combine an ethological approach with the study of the gut microbiota and the proteome to explore proximal mechanisms of species interactions. There is increasing evidence that gut microbiota can modulate the host behaviour and can be altered by biotic stress (e.g. presence of a competitor). Stressful factors can also cause changes in protein expression, which can result in behaviour modifications. To study these mechanisms we used two of the most widespread invasive crayfish species in Europe: signal crayfish (Pacifastacus leniusculus) and red swamp crayfish (Procambarus clarkii). We registered inter- and intraspecific interactions during one-day experiments. Upon completing the experiment, we extracted digestive tract, hepatopancreas and gills from each crayfish to analyse the proteome and to quantify bacteria and viruses from the gut microbiota by flow cytometer. Here, we present the results of: (1) a quantitative description of agonistic interactions; (2) gut microbiota characteristics (quantification of virus and bacteria) and its relationship with host-behaviour; and (3) proteome under different stress scenarios caused by the presence of competitors where statistical descriptive analyses (gel analysis, cluster and PCA) on 1D gels showed discrimination between species, sexes and treatments. Overall, the results of this study provide a better understanding of the underlying mechanisms explaining the competitive interactions between invasive species.

Trabajo presentado en el Congreso Internacional de la Sociedad Española de Etología y Ecología Evolutiva. (ECO-ETOLOGÍA 2016), celebrado en Granada del 20 al 23 de septiembre de 2016.-- Gregorio Moreno-Rueda, Fabián Casas, Eloísa Collantes-Martín, Mar Comas, Rodrigo Megía-Palma, David Ochoa, Eliana Pintus, Manuel Pizarro, Senda Reguera, José M. Rivas, José L. Ros-Santaella, Francisco J. Zamora-Camacho (Editores)., Invasive species are often able to cope with variety environmental conditions enabling them to expand to new habitats. Once introduced and successfully established those invasive species, often threaten native species in their ecosystems. New environmental conditions may act as selective pressures that drive organismal adaptation and evolution. However, alongside genetic adaptation, invasive organisms also show a high degree of plasticity in response to environmental challenge. To elucidate whether epigenetic mechanisms allow an organism to respond to the environment, we study how individuals of two invasive crayfish species respond to environmental challenges by means of a cutting-edge proteomic approach. Our objective is to investigate which types of proteins and how many of each type are expressed when individuals are exposed to different environmental conditions. Our target species are two invasive crayfish, i.e. the red swamp crayfish, Procambarus clarkii; and the parthenogenetic marbled crayfish, Procambarus fallax f. virginalis. The use of a parthenogenetic species will allow us to assess whether changes in protein expression are uniquely due to plasticity (as individuals are genetically identical). We conduct an experiment in which we expose the two crayfish species to treatments consisting of different combinations of temperature and water velocity for several days. After exposure, individuals will be collected and immediately subjected to tissue extraction from the muscle, gills and hepatopancreas to examine the proteome responses. Here, we will present the details of the experimental setup and hypotheses tested as well as any available results.

Trabajo presentado en el XXI Symposium of the InternationalAssociation of Astacology (IAA21), celebrado en Madrid del 5 al 8 de septiembre de 2016., Invasive species are often able to cope with variety environmental conditions enabling them to expand to new habitats. Once introduced and successfully established those invasive species, often threaten native species in their ecosystems. New environmental conditions may act as selective pressures that drive organismal adaptation and evolution. However, alongside genetic adaptation, invasive organisms also show a high degree of plasticity in response to environmental challenge. To elucidate whether epigenetic mechanisms allow an organism to respond to the environment, we study how individuals of two invasive crayfish species respond to environmental challenges by means of a cutting-edge proteomic approach. Our objective is to investigate which types of proteins and how many of each type are expressed when individuals are exposed to different environmental conditions. Our target species are two invasive crayfish, i.e. the red swamp crayfish, Procambarus clarkii ; and the parthenogenetic marbled crayfish, Procambarus fallax f. virginalis. The use of a parthenogenetic species will allow us to assess whether changes in protein expression are uniquely due to plasticity (as individuals are genetically identical). We conduct an experiment in which we expose the two crayfish species to treatments consisting of different combinations of temperature and water velocity for several days. After exposure, individuals will be collected and immediately subjected to tissue extraction from the muscle, gills and hepatopancreas to examine the proteome responses. Here, we will present the details of the experimental setup and hypotheses tested as well as any available results.

Trabajo presentado en el Congreso Internacional de la Sociedad Española de Etología y Ecología Evolutiva. (ECO-ETOLOGÍA 2016), celebrado en Granada del 20 al 23 de septiembre de 2016.-- Gregorio Moreno-Rueda, Fabián Casas, Eloísa Collantes-Martín, Mar Comas, Rodrigo Megía-Palma, David Ochoa, Eliana Pintus, Manuel Pizarro, Senda Reguera, José M. Rivas, José L. Ros-Santaella, Francisco J. Zamora-Camacho (Editores)., Las especies invasoras son consideradas una de las más importantes amenazas para la biodiversidad a nivel global, siendo los ecosistemas de agua dulce de los más susceptibles. Los cangrejos de río juegan un papel clave en las interacciones con otras especies y en las redes tróficas, además de ser importantes ingenieros del ecosistema. En los últimos años, diferentes especies invasoras de cangrejo se han expandido rápidamente por Europa dando lugar a zonas de solapamiento, donde establecen complejas interacciones en las que cada invasor podría sufrir diferentes consecuencias sobre su fitness, recursos o su capacidad de invasión. Hasta la fecha, la mayoría de la investigación se ha centrado en estudios etológicos de luchas agonísticas o competencia por recursos y refugio. Sin embargo, en este estudio vamos más allá y combinamos un enfoque etológico con el estudio del proteoma para abordar los mecanismos proximales de la interacción, utilizando dos de las especies invasoras de cangrejo más extendidas en Europa: el cangrejo señal (Pacifastacus leniusculus) y el cangrejo rojo americano (Procambarus clarkii). El estudio del proteoma nos permitirá analizar la expresión de proteínas que tiene lugar en un individuo o en un tejido durante un tiempo concreto. Para ello, llevamos a cabo un experimento durante un periodo de 30 días en el que evaluamos los resultados de la interacción intra- e inter-específica de ambas especies; tanto por contacto directo (individuos tenían posibilidad de contacto físico entre ellos) como indirecto (sin contacto directo ya que los individuos permanecieron separados en jaulas). Los individuos fueron sexados, medidos y se les evaluaron las heridas tanto antes como después del experimento, además de extraer tejido del sistema digestivo, hepatopáncreas y branquias; a partir de los cuales, se identificarán las proteínas expresadas. Aquí, presentaremos los detalles del experimento, hipótesis testadas así como resultados disponibles hasta la fecha.