SANTA SUSANNA 28>29 OCTUBRE2010 · 2011. 11. 3. · X!REUNIÓN!DELA...

X REUNIÓN DE LA RED NACIONAL DE MICROORGANISMOS EXTREMÓFILOS (X MEETING OF THE SPANISH SCIENTIFIC NETWORK ON EXTREMOPHILIC MICROORGANISMS)

PROGRAMA Y ABSTRACTS

SANTA SUSANNA, 28-‐29 OCTUBRE 2010

Responsable de organización

Casamayor, Emilio Ortega Dep. Continental Ecology-‐Limnology Centre d’Estudis Avançats de Blanes CEAB-‐CSIC Acces Cala St Francesc, 14 17300 Blanes [email protected] Diseño y logotipo: Empar Rossello. www.artega.net

X REUNIÓN DE LA RED NACIONAL DE MICROORGANISMOS EXTREMÓFILOS (REDEX 2010) 28-‐29 OCTUBRE DE 2010, SANTA SUSANNA (CATALUNYA)

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ÍNDICE

BIENVENIDA_______________________________________________________ 4 AGRADECIMIENTOS __________________________________________________ 5 INFORMACIÓN GENERAL_______________________________________________ 6 PROGRAMA _______________________________________________________ 7 RESÚMENES (PONENCIAS INVITADAS) _____________________________________ 11 RESÚMENES (CONTRIBUCIONES REDEX) ___________________________________ 17 LISTA DE ASISTENTES ________________________________________________ 41 NOTAS _________________________________________________________ 48


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BIENVENIDA La X Reunión de la Red Nacional de Microorganismos Extremófilos se organiza desde el Centro de Estudios Avanzados de Blanes (CSIC) con el apoyo del Ministerio de Ciencia y Tecnología (MICINN, BIO2008-‐04954-‐E) y el Comité Diversitas, dentro de las actividades 2010 Año Internacional de la Biodiversidad. La reunión tendrá lugar los días 28-‐29 de Octubre de 2010 en las instalaciones del hotel AquaHotel Ona Brava en Santa Susanna, a escasos kilómetros de Blanes (http://www.aquahotel.com/es/hotel/onabrava-‐santa-‐susanna/). Los ambientes extremos tienen a niveles hostiles para el desarrollo de formas de vida compleja uno o varios de los parámetros ambientales de mayor relevancia para los seres vivos (temperatura, acidez, salinidad, presión o radiación). Los organismos que viven en estos ambientes se denominan extremófilos, son básicamente microorganismos y están perfectamente adaptados a estas duras condiciones. Los termófilos toleran temperaturas de hasta 115ºC, los halófilos se desarrollan en salmueras, los piezófilos crecen a presiones de hasta 1100 atmósferas, los psicrófilos se reproducen a temperaturas inferiores a 5ºC, los acidófilos viven a pH inferior a 5, y los alcalófilos pueden desarrollarse a pH superior a 9. A lo largo de la evolución los microorganismos han sabido adaptarse y sobrevivir en estas condiciones, ocupando una gran diversidad de ambientes que el hombre ha considerado tradicionalmente como exentos de vida. En este proceso de adaptación han modificado sus componentes macromoleculares para obtener el mayor grado de eficiencia, produciéndose como consecuencia una extraordinaria diversificación, que en sus aspectos moleculares es, sin lugar a dudas, mucho más extensa que la observada en formas de vida aparentemente más complejas. Los organismos extremófilos y sus proteínas y polímeros tienen aplicación en biotransformaciones industriales o en procesos de biorremediación, constituyen modelos terrestres de posibles formas de vida en otros planetas (exobiología) y son dianas buscadas en bioprospección. El encuentro Redex 2010 reúne a varios de los mejores especialistas nacionales en algunos de estos aspectos.


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AGRADECIMIENTOS El X encuentro de la Red Nacional de Microorganismos Extremófilos ha sido financiado por la Acción Complementaria del MICINN BIO2008-‐04954-‐E, por el Comité Nacional de Diversitas y por el Centro de Estudios Avanzados de Blanes del CSIC. En esta ocasión hemos querido potenciar el contacto con la empresa privada y con la investigación que se realiza en algunas de ellas ligada al desarrollo de productos de interés farmacológico y biotecnológico. Agradecemos, por tanto, las contribuciones de PharmaMar, Esteve, Arquebio y Roche Diagnostics, y su disponibilidad para facilitar el intercambio y la interacción entre el mundo académico y el empresarial, entre ciencia básica y aplicada, en la búsqueda de sinergias y objetivos comunes que beneficien a ambas partes. También queremos destacar la contribución de Roche Applied Science en la preparación de esta reunión. Finalmente agradecemos especialmente a Empar Rosselló-‐Móra la creación y cesión del logo Redex.


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INFORMACIÓN GENERAL LOCALIZACIÓN AquaHotel Ona Brava Avinguda del Mar, 6 08398 Santa Susanna, Barcelona Tel: 937 678 370 Fax: 937 677 050 El hotel se encuentra a 1 minuto a pie de la parada Sta. Susanna de Cercanías Renfe Línea C1 (Barcelona-‐Blanes-‐Maçanet/Massanes), en pleno paseo marítimo. La estancia y manutención de todos los participantes estarán cubiertas por la organización, así como los actos sociales previstos. Los horarios de desayuno (8-‐10h), comida (13-‐14.30h) y cena (19-‐21.30h) son estrictos y con servicios de buffet libre que serán retirados al cumplirse el límite horario. Las habitaciones deberán dejarse libres el día de partida entre las 10 y las 11 de la mañana. Existe un servicio de guarda-‐maletas en recepción. DESPLAZAMIENTO Para llegar al hotel tenéis dos opciones: (i) Llegada a Barcelona y viaje en cercanías Renfe (Rodalies R1) hasta Santa Susanna (línea Blanes-‐Maçanet). El trayecto dura aprox. 1 hora y la parada Sta. Susanna se encuentra poco antes de llegar a Blanes. El hotel está a 50 metros de la parada. Si llegáis al aeropuerto del Prat tenéis un tren hasta Sants (frecuencia cada 30 min) o bien un servicio de autocares (Aerobus) hasta Plaça Catalunya (5 euros por trayecto). Tanto en Sants como en Plaça Catalunya hay acceso a la línea R1. (ii) Llegada al aeropuerto de Girona (Vilobí d’Onyar). En este caso es necesario coger un bus (Buses Sagalés Línea 3 Aeroport de Girona-‐Costa Brava -‐ Costa Brava Sur/Maresme Norte, horarios disponibles en http://www.sagales.com) o transfer hasta Sta Susanna (trayecto 40 min por carretera). Podéis contratar el transfer hasta el hotel por unos 15 euros contactando directamente con Aquatel (93 767 80 78). ESTRUCTURA DE LAS COMUNICACIONES ORALES En esta ocasión las charlas de los miembros de la red no están organizadas en sesiones temáticas sino que se encuentran mezcladas para mantener un buen tono de atención en la audiencia. Tenemos además 5 ponencias invitadas de aspectos muy variados. Continuando con la política de la red, el resto de charlas están reservadas para estudiantes. El idioma de la comunicación será indistinto a escoger entre inglés o castellano. La sala que utilizaremos se encuentra junto a recepción enfrente del hall y contará con ordenador portátil, proyector, servicio de megafonía y otros complementos. Los tiempos asignados totales serán 30 min (25 exposición+5 preguntas) las ponencias y 15 min (12+3) las charlas. Debido a lo ajustado de la programación el cumplimiento de los horarios será estricto.


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PROGRAMA MIÉRCOLES, 27 OCTUBRE 2010

Llegada al hotel por la tarde/noche a vuestra conveniencia. Entrega de la documentación del congreso a partir de las 19h. Cena (retirada del servicio a las 21.30h) JUEVES, 28 OCTUBRE 2010 8:30h Entrega de documentación. Depósito de las charlas sesiones I y II por parte de

los conferenciantes (disponible Microsoft Office 2007) 8:45-‐9:00h Acto de presentación (Sala Rosa dels Vents) 9:00-‐10:00h Ponencias invitadas 9:00h Microorganismos marinos como fuente de nuevos fármacos para la salud humana. ¿Pueden ser rentables? Fernando de la Calle PharmaMar, S.A. 9:30h Bioprospección del potencial microbiano para el desarrollo y escalado de procesos industriales innovadores y sostenibles Caterina Gómez, Jaume Mir Arquebio, S.L. 10:00-‐11:00h Charlas sesión I (Moderador: Ricardo Amils) 10:00-‐10:15h Felipe Gómez. “The permafrost in the Imuruk lake basaltic field (Alaska) as a

Martian permafrost analogue: microbiological diversity” 10:15-‐10:30h Paulina Corral. “A novel haloarchaeal species isolated from hypersaline lakes

of Inner Mongolia, China” 10:30-‐10:45h Alba Cuecas. “Distribution of microbial communities along a temperature

gradient in Mae Fang Hot Springs (Thailand)” 10:45-‐11:00h Tomàs Llorens-‐Marès. “Activity, composition and dynamics of psychrophiles

in the slush of Lake Redon (Limnological Observatory of the Pyrenees)”


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11:00-‐11:15h Breve pausa 11:15-‐12:30h Charlas sesión II (Moderadora: Victoria Béjar) 11:15-‐11:30h Carlos Bricio. “Accumulation of gaseous forms of nitrogen during

denitrification in Thermus thermophilus” 11:30-‐11:45h Elena González-‐Toril. “Microbial diversity of two acidic pit lakes from the

Iberian pyritic belt (SW Spain)” 11:45-‐12:00h Arantxa Peña. “Horizontal gene transfer and their role in Salinibacter ruber

evolution” 12:00-‐12:15h Javier Rodríguez-‐Moya. “Clues of signal transduction pathways involved in

osmo-‐ and heat-‐ stress response in the halophilic bacterium Chromohalobacter salexigens”

12:15-‐12:30h Raúl Muñoz. “Evaluation of MALDI-‐TOF MS whole cell profiles to assess the

culturable diversity of aerobic and moderately halophilic prokaryotes thriving in solar saltern sediments”

12:30-‐12:45 Sergio Valea. “Endoevaporitic heterotrophic microorganisms from halite crust

of the Atacama Desert, Chile” 13:00-‐14:00h Comida Tipo buffet libre. Café en el bar (zona del hall) 14:00h Encuentro en recepción y salida conjunta en autocar. Se ruega puntualidad. 15:30-‐17:45h Actividades y Visita guiada 18:00-‐19:00h Regreso al hotel 19:15-‐20:00h Ponencia invitada La conferencia científica como teatro Carlos Pedrós-‐Alió Institut de Ciències del Mar, CSIC, Barcelona 20:45h Cena del congreso y acto social. Se ruega puntualidad.


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VIERNES 29 OCTUBRE 2010 8:45h Depósito de las charlas sesiones III y IV 9:00-‐10:00h Ponencias invitadas (Sala Rosa dels Vents) 9:00h 454 Sequencing y su utilidad en genómica microbiana Miguel Alvarez Tejado Roche Applied Science 9:30h Proceso de I+D de medicamentos Antonio Párraga Laboratorios Esteve 10:00-‐11:15h Charlas sesión III (Moderadora: Pepa Antón) 10:00-‐10:15h Angeles Aguilera. “Photosynthetic performance of phototrophic biofilms in

extreme acidic environments” 10:15-‐10:30h Yamal Al Ramahi. “Thermoestable fluorescent protein palette for the study of

thermophilic metabolism and its regulation” 10:30-‐10:45h Laura Alvarez. “Lateral transfer of the denitrification pathway” 10:45-‐11:00h Hakima Amjres. “Halomonas rifensis sp. nov., an exopolysaccharide-‐

producing, halophilic bacterium isolated from brikcha, a solar saltern in Chefchaouen, Morocco”

11:00-‐11:15h Ana B. Fernández. “A novel species of the genus Bacillus isolated from a saline

lake in Iran” 11:15-‐11:45h Pausa café 11:45-‐13:45h Charlas sesión IV (Moderador: Antonio Ventosa) 11:45-‐12:00h Ali Tahrioui. “The HanRI quorum-‐sensing system of Halomonas anticariensis, a

moderately halophilic bacterium” 12:00-‐12:15h Nahid Oueriaghli. “Diversity of archaea in saline and non-‐saline soils” 12:15-‐12:30h Anna Vegara. “Bioinformatics and experimental analysis of the GS/GOGAT

pathway and its possible regulatory proteins in the genome from Haloferax mediterranei”


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12:30-‐12:45h Mercedes Reina-‐Bueno. “Transcriptional regulation of the ectD and ectE genes, encoding two copies of the enzyme ectoine hydroxylase of the halophilic bacterium Chromohalobacter salexigens”

12:45-‐13:00h Rafael Ruiz de la Haba. “Salimicrobium salexigens sp. nov., a moderately

halophilic bacterium isolated from salted hides” 13:00-‐13:15h Irene Sánchez-‐Andrea. “Attenuattion of acid mine drainage (AMD) in Rio Tinto

sediments through sulfate reduction” 13:15-‐13:30h Judith Villamor. “Isolation, conservation and characterization of virus of

Salinibacter rubber” 13:30-‐13:45 Roy Mackenzie. “Bacterial diversity in thermal environments studied by DGGE

and electronic microscopy” 13:45-‐14:00h Acto de despedida 14:00h Comida Tipo buffet libre. Café en el bar (zona del hall)


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RESÚMENES (PONENCIAS INVITADAS)


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MARINE MICROBIOLOGY AS A SOURCE OF NEW DRUGS FOR HUMAN HEALTH. REALLY A BUSINESS OPPORTUNITY? Fernando De La Calle

PharmaMar. Avda. Los Reyes, 1. Colmenar Viejo 28770. Madrid. Spain The marine ecosystem represents 95% of the biosphere. Evolution has equipped marine organisms with the appropriate tools to survive, developing exquisitely complex biological and chemical mechanisms for defence, attack, signalling and other still unknown purposes. Such novel chemical structures often result in new modes of action and open up the potential of new ways to treat cancer and other diseases. However, the conversion of a bioactive molecule into medicine is a long and risky process. Currently, the classical process of drug discovery requires an enormous financial investment. High failure rates, the isolation of non patentable compounds and the low chance of finally reaching market are major challenges for R&D within big-‐pharma. However, in the area of natural products for human health new business opportunities are being created with the advent of innovative strategies based on biotechnology tools and efficient methods for exploring biodiversity and analysis of metabolite profiles. In general, marine microorganisms can play two important roles in the drug discovery process:

As sources of new chemical structures As vehicles to supply marine natural products

The use of “culturable” marine microorganisms for drug discovery involves many steps including collection and pre-‐treatment of marine samples, development of isolation methods for specific microbes, molecular de-‐replication to remove repeated strains, molecular taxonomy, culture, screening, chemical de-‐replication, structural elucidation, optimization of culture conditions, scale-‐up and lead optimization. However, new bio-‐technologies are required to increase the output of new compounds for clinical development and include innovative strategies to induce the “silent genes for unknown metabolites” (mainly PKS and NRPS) and efficient methods for chemical de-‐replications in order to avoid redundancies. The use of microbial metabolites in the supply of marine natural products can be illustrated by several real examples, where biotechnology and fermentation are more feasible options than other manufacturing strategies such as total chemical synthesis or aquaculture. In conclusion, the enormous potential of the marine micro-‐diversity remains highly unexplored and new biotechnology tools are emerging to increase the potential to cultivate non-‐easily cultivable microorganisms and to advance in the knowledge of genomic pathways to relate secondary metabolites and bioactivity.


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EXPLORING MICROBIAL POTENTIAL TO DEVELOP AND SCALE-‐UP INNOVATIVE AND SUSTAINABLE INDUSTRIAL BIOSOLUTIONS Caterina Gómez, Jaume Mir Arquebio, SL. Edifici Eureka. Campus Universitat Autònoma de Barcelona, Bellaterra Microbes, such as bacteria and fungi, can be found everywhere on the planet. They have adopted a wide variety of strategies for surviving under all temperature conditions, extreme pressure and high salt or hydrocarbon concentrations. They are responsible for recycling dead material and maintaining the balance of elements. There is no limit to their capability to “bioprocess” organic material in the environment. We use these bioprocessing capabilities to carry out innovative processes with commercial applications in a wide range of industrial and medical fields. Increasing knowledge on microbiology, biotechnology, biochemistry and genetic engineering enables us to find, develop and implement innovative biosolutions to a continuously expanding wide range of applications within the Pharmaceutical, Fine-‐Chemical, Cosmetic, Agro-‐food and Environmental areas. We have developed a wide range of services to achieve successful bioprocess development and scaling-‐up and industrial implementation among them, genetic and biochemical diversity searching and screening, strain development and improvement, protein design, bioprocess development, bioproduct upstream and downstream processes design, technology transfer and strategic consultancy. We also look to improve industrial performance by implementing clean and sustainable bioprocesses as innovative solutions to better safeguard the natural environment and the world’s resources.


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THE SCIENTIFIC PRESENTATION AS THEATER Carles Pedrós-‐Alió Institut de Ciències del Mar-‐CSIC, Barcelona Giving a lecture is certainly a privilege for the speaker. For more than one hour the audience will fix their eyes on the speaker and their ears on his or her words. In fact, the audience wants to learn interesting things along the talk and feel motivated by the speaker but also wants to enjoy with the discoveries presented. The speaker should be aware on the theater performance skills to know how to successfully deal with this great opportunity. Personal background. My scientific interest is to understand the ecology of aquatic microorganisms. Recently, I have started to use genomics as a tool to generate hypotheses that can later be tested experimentally. I also like to study extreme or unusual environments to throw light on the functioning of more conventional ones. For example, hypersaline systems, karstic lakes, or the Polar waters illuminate particular aspects of the ecology of microorganisms that are more difficult to identify in temperate marine waters. Another interest is in understanding the diversity of microorganisms, in particular the mechanisms maintaining a large number of rare bacteria in aquatic ecosystems.


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454 SEQUENCING Y SU UTILIDAD EN GENÓMICA MICROBIANA Miguel Alvarez Tejado Roche Diagnostics SL. Roche Applied Science En 2005 las tecnologías de secuenciación masiva de ADN dieron su primer paso a la aplicación real en la comunidad científica. Este primer avance lo dio la empresa de biotecnología 454 Life Sciences (Branford, CT USA). Desde entonces la tecnología de esta compañía, ahora propiedad de Roche, ha conseguido enormes avances tecnológicos, y ha posibilitado la realización de proyectos que antes eran económica o técnicamente inviables. Esta tecnología, denominada 454 sequencing, está en su tercera generación de desarrollo químico y es capaz de generar cientos de miles de secuencias de más de 400 pares de bases de media, con una gran exactitud y en un tiempo de 10 horas aproximadamente. Con esta capacidad de secuenciación, se posibilita el desarrollo de proyectos genómicos de gran complejidad, como pueden ser la secuenciación de genomas y transcriptomas microbianos, y de muestras de ADN de origen ambiental, facilitando un abordaje metagenómico. Esta aproximación metagenómica en microbiología está permitiendo secuenciar muestras muy complejas, en las que el organismo de estudio no se puede crecer en condiciones de laboratorio, o donde la diversidad microbiana hace muy difícil su análisis por métodos convencionales. Durante esta ponencia se hablará del fundamento de la tecnología de secuenciación 454, y se presentarán diferentes ejemplos de aplicación en campos de utilidad a la comunidad de microbiólogos.


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PROCESO DE I+D DE MEDICAMENTOS Antonio Párraga Laboratorios Dr Esteve, Barcelona El desarrollo de fármacos es un proceso largo, complejo y costoso por varias razones: (i) sólo un candidato a fármaco sobre 10.000 llegará al mercado o al paciente; (ii) se tarda hasta 10 -‐13 años para desarrollar un medicamento y llevarlo al mercado, y (iii) se requiere una inversión de alrededor de mil millones de euros en promedio para cada nuevo medicamento. El momento actual de la Industria Farmacéutica es muy delicado. Mientras que los costes de I+D de nuevos medicamentos y los tiempos para obtenerlos se han incrementado notoriamente en los últimos años, el número de fármacos que llega al mercado es cada vez inferior. Este fenómeno se ha acuñado con el término brecha de innovación. La Industria Farmacéutica tiene como desafío continuar con el desarrollo y lanzamiento de nuevos productos diferenciados. Para esto es necesario obtener nuevos medicamentos más eficaces y más seguros en la lucha contra las enfermedades. ¿Cómo superar este reto? Aprovechando los puntos fuertes de la Industria Farmacéutica, la Industria Biotecnológica y la Academia, para transformar las invenciones en innovaciones a través de un proceso de innovación abierta. Las compañías como ESTEVE no pueden competir con los grandes programas de las multinacionales, pero pueden enfocarse en áreas específicas y utilizar las sinergias en su entorno para tener éxitos en la obtención de nuevos medicamentos innovadores. ESTEVE es un grupo químico-‐farmacéutico líder en España en el ámbito de la salud, que desarrolla su actividad en el área farmacéutica mediante la investigación y desarrollo de nuevos fármacos; comercializa un amplio portafolio de productos de prescripción, medicamentos publicitarios, vacunas, genéricos y para uso veterinario; y también está presente en el segmento de los principios activos farmacéuticos (APIs), ofreciendo de este modo una solución global en el sector Salud. A través de diversas alianzas estratégicas también tiene actividad en el segmento de los dermocosméticos (ISDIN, alianza con el Grupo Puig) y en terapia respiratoria domiciliaria (ESTEVE-‐Teijin Healthcare, alianza con la empresa japonesa Teijin Pharma).


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RESÚMENES (CONTRIBUCIONES REDEX)


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THE PERMAFROST IN THE IMURUK LAKE BASALTIC FIELD (ALASKA) AS A MARTIAN PERMAFROST

ANALOGUE: MICROBIOLOGICAL DIVERSITY M. T. Gómez (1), O. Prieto-‐Ballesteros (1), D.C. Fernandez-‐Remolar (1), J. A. Rodríguez-‐Manfredi (1), D. Gómez Ortíz (2), J.S. Kargel (3), J. Gómez-‐Elvira (1), R. Amils (1,4), Felipe Gómez (1) (1) Centro de Astrobiología-‐INTA, 28850 Torrejón de Ardoz, and (2) Rey Juan Carlos University, 28933 Móstoles, Spain. (3) University of Arizona, Tucson AZ 85721, USA. (4) Centro de Biología Molecular Severo Ochoa, Universidad Autonoma de Madrid. 28049 Madrid, Spain. We are studying the permafrost in the Imuruk lake volcanic field area (Alaska) from an Astrobiological perspective, in order to reach three main objectives: 1) Define preservation patterns of biosignatures in cold environments that may be used in future space exploration missions; 2) develop new instrumentation for detecting life in situ or remotely, and 3) develop new instrumentation for detection and mapping of permafrost niches where life (or biochemical tracers of past life) may be preserved. These aims will be achieved by permafrost characterization using geophysical sounding and drilling, sampling different levels of the rock cores and analyzing their mineralogy, geochemistry and microbiology. In order to map the permafrost underground, electric tomography sounding was performed. Resulting tomographic data indicate that the permafrost of the studied area is at a mean depth of 0.50 meter from the surface, sometimes even shallower. Drilling points were selected depending on the permafrost depth known from the tomographic data analysis. Three perforations were done all along the hill. Samples were collected at several depths in the three holes for mineralogical, geochemical and biological analysis. They were in situ fixed with formaldehyde in order to be maintained till laboratory analysis was developed. Several growth fresh media were inoculated with samples from different depths in the field for microorganisms enrichment. First results report enrichment in several inoculated media including some specific for heterotrophic aerobic bacteria, anaerobic chemolithotrophic and methanogen bacteria. Two different molecular methods are being used for microbial determination: “In situ” hybridizitation (was used for microbial determination and cell counting also) and 16S rRNA genes amplification, cloning and sequencing. First results in cell counting determined a population density gradient vs. depth. Acknowledgments: Centro de Astrobiologia-‐INTA (Spain) supported the 2005 expedition to Imuruk Lake. This expedition was in part supported with the Grant ESP 2004-‐05008 “Detección de biomoléculas en exploración planetaria” from the Spanish Government.


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A NOVEL HALOARCHAEAL SPECIES ISOLATED FROM HYPERSALINE LAKES OF INNER MONGOLIA, CHINA Paulina Corrala, Carmen Gutiérreza, Ana Castilloa, Masahiro Kamekurab and Antonio Ventosaa (a) Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville, Spain (b) Halophiles Research Institute, 677-‐1 Shimizu, Noda-‐shi, Chiba-‐ken 278-‐0043, Japan The family Halobacteriaceae has long been identified as the most abundant microorganisms in hypersaline environments (Oren, 1994). Currently this family comprises 27 genera (Oren et al., 2009). The genus Halorubrum was established by McGenity and Grant (1995) and currently contains 23 species, including four haloalkaliphilic species, Hrr. alkaliphilum, Hrr. luteum, Hrr. tibetense, Hrr. vacuolatum and 19 neutrophilic species: Hrr. aidingense, Hrr. arcis, Hrr. californiense, Hrr. cibi, Hrr. coriense, Hrr. distributum, Hrr. ejinorense, Hrr. ezzemoulense, Hrr. kocurii, Hrr. lacusprofundi, Hrr. lipolyticum, Hrr. litoreum, Hrr. orientale, Hrr. saccharovorum, Hrr. sodomense, Hrr. tebenquichense, Hrr. terrestre, Hrr. trapanicum and Hrr. xinjiangense. In the present study, two halophilic archaea, strains EN-‐2T and SH-‐4, were isolated from the saline lakes Erliannor and Shangmatala, respectively, located in Inner Mongolia, China. The aim of this study is to describe the features of these strains which are proposed as a new species of the genus Halorubrum. The characterization was performed according to the proposed minimal standards for the description of new taxa of the order Halobacteriales (Oren et al., 1997). Cells of the two strains were non-‐motile rods and strictly aerobic. Colonies were red-‐pigmented. Strains EN-‐2T and SH-‐4, grow under optimal conditions at 35-‐40 ºC, at 3.4 M NaCl and at pH 7.2-‐7.5. MgCl2 was not required for growth. Cells lysed in destilled water and minimal NaCl concentration to prevent cell-‐lysis is 12 % (w/v). On the basis of 16S rRNA gene sequence analysis, strains EN-‐2T and SH-‐4 were closely related to Halorubrum cibi B31T (97.9 and 98.0 % similarity, respectively), Halorubrum tibetense 8W8T (97.6 and 97.3 %), Halorubrum alkaliphilum DZ1T (96.8 and 97.1 %), Halorubrum luteum CGSA15T (96.8 and 97.0 %) and Halorubrum lipolyticum 9-‐3T (96.8 and 97.0 %). DNA-‐DNA hybridization experiments showed that strains EN-‐2T and SH-‐4 were not related to these species, with levels of DNA-‐DNA relatedness equal to or below 45 %. The DNA-‐DNA hybridization between strains EN-‐2T and SH-‐4 was higher than 70 % showing that both strains are members of the same species. Polar lipid analysis revealed that strains EN-‐2T and SH-‐4 contained phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-‐Me), sulfated diglycosyl diether (S-‐DGA-‐3) and two unidentified glycolipids. The DNA G+C content of both strains was 62.1 mol%. It was concluded that strains EN-‐2T and SH-‐4 represent a novel species of the genus Halorubrum, for which the name Halorubrum aquaticum sp. nov. is proposed. The type strain is EN-‐2T.


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DISTRIBUTION OF MICROBIAL COMMUNITIES ALONG A TEMPERATURE GRADIENT IN MAE FANG HOT SPRINGS (THAILAND) A. Cuecas1, M.C. Portillo1, P. Pasomsup2, W. Kanoksilapatham2, J.M. González1 1 Institute of Natural Resources and Agrobiology, IRNAS-‐CSIC, Seville, Spain 2 Department of Microbiology, Faculty of Science, Silpakorn University, Nakhon Pathom, Thailand The distribution of bacteria along a wide range (39-‐90ºC) temperature gradient was studied in Mae Fang hot springs (Northern Thailand). A wide variety of microorganisms has been reported in terrestrial hot springs [1] including some hot springs in Thailand [2-‐4] but scarce studies are available on the distribution of thermophilic microorganisms along a wide range of temperatures and the relationship among the different bacterial groups and their influence on baterial community structure. The spatial distribution of thermophilic cyanobacteria and other thermophiles is a current topic of interest [5]. The microbial communities were detected in base to molecular methods based on DNA, PCR amplification of 16S rRNA genes, DGGE, cloning and sequencing. At the highest temperatures, Aquificales, Dictyoglomi, Thermodesulfobacteria and Thermotogae were the major components of the bacterial community. In the mid temperature range (70-‐50ºC) microbial mats were dominated by Chloroflexi and Cyanobacteria. A number of candidate divisions and scarcely known bacterial groups were detected along the gradient with a maximum of phylum diversity at 50ºC. The warm stream below 50ºC was represented by mesophilic bacterial clades and the development of a wide variety of heterotrophic bacteria, including Proteobacteria. The distribution, displacement, and coexistence of the major bacterial components of the communities stablished along the study temperature gradient will be analyzed. This work was supported by projects PA1001993 (JMG) and PA1002058 (MCP) from the CSIC and BIO288 from the regional Government of Andalusia. [1] Ward, D.M., M.J. Ferris, S.C. Nold, M.M. Bateson. 1998. A natural view of microbial biodiversity hot spring cyanobacterial mat communities. Microbiology and Molecular Biology Reviews 62: 1353-‐1370. [2] Portillo, M.C., V. Sririn, W. Kanoksilapatham & J.M. Gonzalez. 2009. Differential microbial communities in hot spring mats from Western Thailand. Extremophiles 13: 321-‐331. [3] Kanokratana, P., S. Chanapan, K. Pootanakit & L. Eurwilaichitr. 2004. Diversity and abundance of Bacteria and Archaea in the Bor Khlueng Hot Spring in Thailand. Journal of Basic Microbiology 44: 430-‐444. [4] Sompong, U., P.R. Hawkins, C. Besley & Y. Peerapornpisal. 2005. The distribution of cyanobacteria across physical and chemical gradients in hot springs in Northen Thailand. FEMS Microbiology Ecology 52: 365-‐376. [5] Papke, R.T., N.B. Ramsing, .M. Bateson, D.M. Ward. 2003. Geographical isolation in hot spring cyanobacteria. Environmental Microbiology 5: 650-‐659.


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ACTIVITY, COMPOSITION AND DYNAMICS OF BACTERIAL PSYCHROPHILES IN THE SLUSH OF LAKE REDON (LIMNOLOGICAL OBSERVATORY OF THE PYRENEES) Tomàs Llorens-‐Marès, JC Auguet, EO Casamayor Limnological Observatory of the Pyreenes-‐Centre d’Estudis Avançats de Blanes, CEAB-‐CSIC, Accés Cala Sant Francesc, 14, 17300 Blanes, Girona, Spain High altitude lakes are covered by ice and snow for several months of the year, and a rich and very active community of psychrophilic microorganisms naturally develops in the slush layers. These slush layers are formed between different ice sheets and contain a mixture of snow and liquid water at temperatures near 0 C°. Our study was carried out in the Pyrenees (Lake Redon, 2240 m above mean sea level) on late winter (March) and spring (May). We compared microbes present in the slush layers and those inhabiting the water column studying chemical parameters, bacterial activity, CARDFISH counts, and cloning and sequencing of the bacterial 16S rRNA gene. One additional ice-‐melt pond (May) was added to the comparative analyses. Dissolved organic matter (DOC) concentration was similar among samples but nitrogen and phosphorous did not. In spite of the low temperature and the seasonal occurrence of the habitat, microbial biomass and activity were very high, and always greater in the slush than in the plankton and much higher in May. All the clones identified were closely related to other clones found worldwide. The most represented phyla were Bacteroidetes, Betaproteobacteria, a a few Actinobacteria and Verrucomicrobia (only detected in March). Unexpectedly, Bacteroidetes were potentially responsible for the large increase in bacterial activity from March to May, probably due to the increase of nitrogen sources during the ablation and the potential to quickly degrade at very low temperatures a large spectrum of organic compounds provided by the algal bloom. Bacteroidetes are unveiled as key organisms in these environments offering exciting biotechnological and ecological perspectives.


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ACCUMULATION OF GASEOUS FORMS OF NITROGEN DURING DENITRIFICATION IN THERMUS

THERMOPHILUS Carlos Bricioa, Laura Alvareza, Andrew J. Gatesb, David J. Richardsonb, José Berenguera

(a) Centro de Biología Molecular Severo Ochoa (UAM-‐CSIC). Universidad Autónoma de Madrid. Cantoblanco, 28049, MADRID. (b) University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ, UK Several strains of Thermus thermophilus exist that are able to grow anaerobically through nitrate respiration or through an apparently complete denitrification pathway, with the accumulation of gaseous forms of nitrogen oxides (1). The first step of the process is catalyzed by a tetrameric nitrate reductase (Nar) encoded within a Laterally transferable Genetic element called NCE. This NCE also encodes the nitrate/nitrite transporters, a NADH dehydrogenase, and the transcription regulators and signaling systems required for the induction by nitrate of the system under anaerobic conditions (2). Further steps of the dinitrification require a nitrite reductase similar in sequence to those known as to cd1 type (Nir), and a putative cytochrome c-‐containing nitric oxide reductase (Nor). Genes encoding both enzymes are apparently clustered in the genome of the denitrificant strains (see communication by L. Alvarez et al.). Whether or not the pathway ends in N2 has been controversial due to the fact that any homologue to known nitrous oxide reductases (NosZ) has been found encoded within the genome of the denitrificant strain PRQ25. In an attempt to solve this, we analyzed the formation of nitrous oxide from nitrate and nitrite by cultures and protein extracts of wild type and norC mutant strains. The results obtained showed in the wild type strains lower levels of nitrous oxide accumulation than it could be expected if all the nitrate used at the start of the culture was reduced to this gas, supporting that a relevant amount of dinitrogen (N2) is being produced in the process. Therefore, the denitrification in T. thermophilus PRQ25 is apparently complete, being the last step of the process likely catalyzed by an enzyme unrelated to conventional NosZ. Putative candidates will be discussed. 1. Cava el al. 2008. Environmental Microbiology, 10: 522-‐533 2. Cava et al. 2007. Molecular Microbiology, 64: 630-‐646.


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MICROBIAL DIVERSITY OF TWO ACIDIC PIT LAKES FROM THE IBERIAN PYRITIC BELT (SW SPAIN) Elena González-‐Torila, María Sánchez-‐Seguraa, Enrique López Pamob, Esther Santofimiab, Ricardo Amilsa,c, Angeles Aguileraa (a) Centro de Astrobiología (INTA-‐CSIC). Carretera de Ajalvir Km 4, Torrejón de Ardoz. 28850 Madrid. Spain. (b) Instituto Geológico y Minero de España, 28003, Madrid, Spain. (c) Centro de Biología Molecular (UAM-‐CSIC). Universidad Autónoma de Madrid. Canto Blanco. 28049 Madrid. Open cast mining often produces acidic pit lakes. In the provinces of Huelva and Sevilla (SW Spain) the intensive mining carried out in the Iberian Pyritic Belt (one of the most outstanding massive sulphide bodies in the world) has left a legacy of mine pits with considerable size. This study analyzed two of these pit lakes, Nuestra Señora del Carmen (NSC, 0.68 Ha and 32 m deep) and Concepción (CC, 1,2 Ha and 15 m deep). Depth profiles of both lakes were analyzed. The hydrochemical characteristics were explored by different analytical techniques, in situ and in laboratory. Both cases showed a clear chemical and thermal stratification with well defined chemoclines and thermoclines. Microbial community composition of the depth profiles was analyzed by 16S and 18S rRNA gene cloning and sequencing. Water samples were collected in May 2009. Both pit lakes resulted acidic and with high concentration of heavy metals, but NSC resulted more extreme in both aspects. pH ranged between 2.08 and 2.38 in NSC and 2.7 and 3.5 in CC. Heavy metal concentration was higher in NSC specially iron (597 mg/L in the up layer and 703 mg/L in the deep in NSC and 9.45 mg/L in the up layer and 395 mg/L in the deep in the case of CC). Thus the hydrochemical presented by NSC is the characteristic in acid mine drainage (AMD) and Río Tinto waters, while CC’s waters resulted relatively lighter. Microorganisms detected in NSC were characteristics of AMD and Río Tinto, including obligate oxidising bacteria (Leptospirillum) and the facultative iron reducing bacteria (At. ferroxidans, Actinobacteria, Acidimicobiales, Thermoplasmales) detected in the deep layer. Diversity in CC was much higher. Microorganisms related to AMD and Río Tinto (Acidiphilium, Acidobacteria and Ferrovum) and also microorganisms never detected in these systems were identified. Taking into consideration the hydrochemical characteristics of these pit lakes and the spatial distribution of the identified microorganisms, a model explaining their geomicrobiology will be advanced.


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HORIZONTAL GENE TRANSFER AND THEIR ROLE IN SALINIBACTER RUBER EVOLUTION Arantxa Peñaa, Hanno Teelingb, Jaime Huerta-‐Cepasc, Cristina Lópeza, Toni Gabaldónc, Rudi Amannb y Josefa Antóna aDepartamento de Fisiología, Genética y Microbiología, and IMEM, Universidad de Alicante, Apartado 99, 03080 Alicante, Spain. bMax Planck Institute for Marine Microbiology, Celsiusstrasse 1, 28359 Bremen, Germany. cBioinformatics and Genomics Programme. Center for Genomic Regulation (CRG). Dr Aiguader, 88. Barcelona 08003, Spain In the “genomic era”, some authors have proposed that Lateral Gene Transfer (LGT) is the major force in the evolution of prokaryotes that can accelerate the adaptation to new environments. From the overwhelming surge of genome sequence information, a lot of candidates for HGT are being identified among prokaryotes.. Recently, S. ruber strain M31 DSM13588 and S. ruber M8 have been sequenced, annotated and compared. S. ruber genomes harbour 34 genes located outside hypervariable regions that are transcribed during standard growth and probably derive from lateral gene transfers with Archaea preceding the strains divergence. These genes present a good codon usage adaptation index, indicating that the LGT was not recent and the codon usage of these genes has been adapted to the new genome. When reverse transcriptase-‐PCR analysis was carried out, the results indicated a diffent expression profile for 6 of them. On the other hand, we detected 6 genes present only in M8, but not in M31. These specific genes had rather poor codon usage indices, suggesting that these 6 genes are transferred more recently, most likely after sepatation of the species. The results of RT-‐PCR for these specific genes showed that only 3 genes had expression in, almost, one point of the growth curve. We amplified these genes in all the strains from a collection previously obtained from differente isolation sources in an attempt to clarify the evolution of the specie through geographical diferences.


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CLUES OF SIGNAL TRANSDUCTION PATHWAYS INVOLVED IN OSMO-‐ AND HEAT-‐ STRESS RESPONSE IN THE HALOPHILIC BACTERIUM CHROMOHALOBACTER SALEXIGENS J. Rodríguez de Moya, M. Argandoña, C. Vargas and J. J. Nieto Department of Microbiology and Parasitology, University of Seville, Spain Chromohalobacter salexigens is a moderately halophilic bacterium able to grow in a wide range of salinities (0.5 to 3 M NaCl) and up to 45 ºC. Its main osmo-‐ and thermo-‐ adaptation strategy is the cytoplasmic accumulation of the biostabilizers ectoine and hydroxyectoine, two compounds with great industrial interest. In order to coordinate its processes of osmo and thermoregulation, cells are equipped with different systems and mechanisms of sensing physical stimuli correlated to changes in the external medium (sensing), with pathways to transduce these stimuli into useful signals which can be processed in the cell (signal transduction). In addition, there should be mechanisms of regulating proper responses in the cell to recover from the environmental stress and to maintain all necessary physiological functions (regulation). These different signal transduction pathways have not yet well been elucidated in halophilic bacteria. Here we report the characterization of a response regulator of a two component system, EupR, involved in the transcriptional regulation of ectoine(s) transport, together with the prediction of its putative histidine kinase sensor. This finding is the first example of the involvement of a two-‐component response regulator in the osmoadaptation of a true halophilic bacterium. We have also carried out global proteomic analyses by two dimension differential gel electrophoresis (2D-‐DIGE) in order to identify proteins with a differential expression at low salt conditions (0.6 M NaCl, 37 ºC), high salt conditions (2.5 M NaCl, 37 ºC) and high salt and high temperature conditions (2.5 M NaCl, 45 ºC). By using peptide-‐mass fingerprinting data and/or by TOF-‐TOF techniques, three proteins were identified. One of them was induced by salinity, and two others were overexpressed by heat stress. A preliminary in silico analysis related these proteins to different signal transduction mechanisms as chemotaxis or the synthesis of the second messenger c-‐di-‐GMP. These data pave the way to further elucidation of the signal transduction pathways involved in the osmo-‐ and heat-‐ stress responses in C. salexigens.


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EVALUATION OF MALDI-‐TOF MS WHOLE CELL PROFILES TO ASSESS THE CULTURABLE DIVERSITY OF AEROBIC AND MODERATELY HALOPHILIC PROKARYOTES THRIVING IN SOLAR SALTERN SEDIMENTS Raul MuñozA, Arantxa López-‐LópezA, Mercedes UrdiainA, Josefa AntónB, Edward R.B. MooreC and Ramon Rosselló-‐MóraA A Marine Microbiology Group, Department of Ecology and Marine Resources, Institut Mediterrani d'Estudis Avançats (CSIC-‐UIB), E-‐07190 Esporles, Mallorca, Spain. B Departamento de Fisiología, Genética y Microbiología and INEM, Universidad de Alicante, Apartado 99, Alicante, Spain. C Culture Collection of the University of Gothenburg (CCUG) Department of Clinical Bacteriology, Sahlgrenska University Hospital and Sahlgrenska Academy of the University of Gothenburg, P.O. Box 7193, SE-‐402 34 Gothenburg, Sweden. Matrix Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-‐TOF MS) has been widely used in organic chemistry since de 1980s. Reportedly, this technique has been adapted to the analysis of whole cell extracts and used for the identification of prokaryotes, mostly in the clinical field, since it delivers profiles based mainly on their ribosomal proteins. It seemed appealing to bring this approach into the ecological scope. We have tested MALDI-‐TOF MS onto a large set of isolates from an environmental sample taken from a crystallizer pond in a solar saltern in Mallorca. For this purpose we brought to pure culture a total of 374 strains isolated from different growth conditions. All cultures were analyzed by MALDI-‐TOF MS, and their profiles were clustered by means of their overall similarity. Altogether, we observed a total of 25 different clusters at 52% similarity, and two of them embraced over 65% of the total isolates. In order to check whether the different phenotypic groups represented potential independent species we further characterized the strains by 16S rRNA sequencing, DNA-‐DNA hybridization (DDH) and RAPD. The selection of two to four strains belonging to each individual cluster always affiliated together in the same 16S rRNA clade. Besides, the preliminary DDH results indicated that members of different clusters always hybridized below the species threshold, thus can be considered different genospecies. On the other hand, those strains belonging to the same cluster showed DDH values within what is considered the species boundary. The intra-‐cluster RAPD analyses indicated that in most cases different populations of the same species coexisted in the sample, and rarely clonal variants were isolated. Finally, the results also indicated that most of the isolates belonged to the phylum Firmicutes (about 87%) with Thalassobacillus sp. being the most retrieved species. Besides, Rhodovibrio sp. was the mostly retrieved Gram-‐negative organisms in our experiment.


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ENDOEVAPORITIC HETEROTROPHIC MICROORGANISMS FROM HALITE CRUST OF THE ATACAMA

DESERT, CHILE Sergio Valea1, Asunción de los Rios1, Alfonso F. Davila2, Carmen Ascaso1, Jacek Wierzchos1

(1) Institute of Natural Resources, Centre for Environmental Science (CSIC), 28006 Madrid, Spain. (2) SETI Institute – NASA Ames Research Center; Moffet Field, CA 94035 (USA). Relatively abundant cyanobacterial endolithic colonization has been previously described in halite (NaCl) crusts in the hyper-‐arid core of the Atacama Desert [1]. Mineral deliquescence of halite has been proposed as the main physical factor facilitating this endoevaporitic colonization [2]. Halite crusts also provide natural shelter against harmful radiation, allowing the infiltration of photosynthetic active radiation inside the salt crust at the same time. Analyses with several microscopy techniques revealed that microorganisms form small aggregates composed by cyanobacterial cells, accompanied by groups of heterotrophic microorganisms [3]. Molecular characterization by isolation of genomic DNA and PCR-‐DGGE analysis of 16S rRNA gene fragments of the associated heterotrophic microorganisms revealed significant differences in the community structure between three Salares sampled: Yungay, Salar Grande and Salar de Llamará, probably relating to a response to different environmental conditions. The highly restrictive environment of Yungay, showed the lowest phylotypes richness. Phylogenetic approaches based on Bayesian inference methods revealed that heterotrophic microorganisms belong to Archaea and Bacteria domains. Most of the sequences clustered together with uncultured microorganisms from diverse hypersaline environments. None of phylotypes showed similarity with species found in other habitats within Atacama region. The closest GenBank relatives of the majority of the Archaea sequences were different genera of Haloarchaea. Predominant Bacteria sequences were closely related to Bacteriodetes. We suggest that heterotrophic Archaea and Bacteria could play an important role in the dynamic of these endoevaporitic communities. Maintenance of specific microbial loops may help the survival of this unique microbial ecosystem in this extremely dry, oligotrophic and saline environment. [1] Wierzchos J, Ascaso C, McKay CP (2006) Endolithic cyanobacteria in halite rocks from the hyperarid core of the Atacama Desert. Astrobiology 6:415-‐422 [2] Davila AF, Gómez-‐Silva B, De los Ríos A, Ascaso C, Olivares H, McKay C, Wierzchos J (2008) Facilitation of endolithic microbial survival in the hyperarid core of the Atacama Desert by mineral deliquescence. J Geophys Res 113 G01028, 9 pp [3] De los Ríos A, Valea S, Ascaso C, Davila AF, Kastovsky J, McKay CP, Gómez-‐Silva B, Wierzchos J (2010) Comparative analysis of the microbial communities inhabiting halite evaporates of the Atacama Desert. Int Microbiol 13:79-‐89


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PHOTOSYNTHETIC PERFORMANCE OF PHOTOTROPHIC BIOFILMS IN EXTREME ACIDIC ENVIRONMENTS Virginia Souza-‐Egipsya, Elena González-‐Torilb, Ricardo Amilsb,c, Angeles Aguilerab (a) Departamento de Biología Vegetal, Facultad de Ciencias, Universidad de Málaga. Campus Universitario de Teatinos s/n, 29071 Málaga. Spain. (b) Centro de Astrobiología (INTA-‐CSIC). Carretera de Ajalvir Km 4, Torrejón de Ardoz. 28850 Madrid. Spain. (c) Centro de Biología Molecular (UAM-‐CSIC). Universidad Autónoma de Madrid. Canto Blanco. 28049 Madrid. Spain. Although low pH and high metals concentrations are restrictive to most aquatic life, large phototrophic biofilms and mats composed of filamentous green algae such Zygnemopsis or phototrophic protists such Euglena are often observed to thrive in extreme acidic environments. On the other hand, photosynthesis is known to be particularly sensitive to stressful environmental conditions, such as salinity, pH or presence of toxicants. There are relatively few reports regarding photosynthesis in acidic environments in the literature, and most have focussed on primary productivity measurements in acidic lakes. The aim of the present study was to compare the photosynthetic characteristics (photosynthetic capacity and efficiency, light compensation point and light saturation parameter) as well as dark respiration of different phototrophic biofilms isolated from Río Tinto and to relate them to their species composition and the physicochemical characteristics of their respective sampling locations. In order to do this, rates of dark respiration (DR) and light-‐saturated net photosynthesis (Pmax) were determined as oxygen exchange using an oxygen electrode in eight different phototrophic biofilms taken from the riverbed surface at six different sampling locations. All the biofilms analyzed showed photoinhibition over the light intensity of 0 to 100 mmol photon m-‐2 s-‐1 except in the case of the filamentous algae Zygnemopsis that showed a light-‐saturated photosynthesis model. Diatom biofilms showed the lowest compensating irradiance (Ic) with 5 mmol photon m-‐2 s-‐1, followed by Euglenas biofilms with ca. 20 mmol photon m-‐2 s-‐1, Zygnemopsis was the species with highest values of Ic ca. 40 mmol photon m-‐

2 s-‐1. Statistical analysis showed that Pmax and DR is clearly influenced by the environmental conditions. This suggests a great physiological plasticity of these acidophilic species.


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THERMOESTABLE FLUORESCENT PROTEIN PALETTE FOR THE STUDY OF THERMOPHILIC

METABOLISM AND ITS REGULATION Yamal Al-‐Ramahi, Leticia Torres, José Berenguer, Aurelio Hidalgo Centro de Biología Molecular Severo Ochoa (UAM-‐CSIC). Universidad Autónoma de Madrid. Cantoblanco, 28049, MADRID.

Thermus thermophilus is an excellent model organism for basic and applied research (1). Unlike other organisms Thermus thermophilus grows easily in the laboratory, can acquire exogenous DNA by means of an extraordinarily efficient mechanism of natural competence, and usually is the organism of choice for structural biology thanks to the ease of its enzymes and multiproteinic complexes to crystallize. Thermus thermophilus is suitable host for the production of thermozymes, for the thermoestabilisation of thermolabile proteins and it is an excellent model for the study of the respiratory metabolism and its regulation. Although some tools such as antibiotic resistance genes are being developed as molecular markers, there is still a shortage of tools to study the physiology of thermophiles and its regulation, especially in comparison to other models such as Escherichia coli. Fluorescent proteins constitute the basis of powerful tools providing solutions for the study of physiological processes and the development of biotechnological applications. Since the discovery of the green fluorescent protein from Aqueorea victoria by Shimomura almost fifty years ago, great knowledge has been achieved regarding the three dimensional structure and the mechanisms for fluorescence emission, a broad color palette has been developed and the number of successful applications is rapidly increasing. Fluorescent proteins have been shown to be suitable as nontoxic and noninvasive, easily diffusing probes to study different biological models ranging from bacterial organisms to mammals. There were no existing examples of the applicability of these techniques in thermophiles until recently (2). The results achieved by protein engineering of superfolder green fluorescent protein (the first termoestable variant efficiently used in Thermus thermophilus) evidence potential to broaden the spectrum of the thermoestable fluorescent protein palette and its future applications.

1. Cava et al. 2009. Extremophiles, 13 : 213-‐231 2. Cava et al. 2008. Environmental Microbiology, 10: 605-‐ 610.


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LATERAL TRANSFER OF THE DENITRIFICATION PATHWAY Laura Alvareza, Carlos Bricioa, Derek Lithauerb, Estariette Van Heerdenb, José Berenguera (a) Centro de Biología Molecular Severo Ochoa (UAM-‐CSIC). Universidad Autónoma de Madrid. Cantoblanco, 28049, MADRID. (b) University of the Free State, Bloemfontein, South Africa. The denitrification pathway consists of four consecutive steps that reduce nitrate to N2 [1]. Denitrification has been proposed to be an ancient pathway by phylogenetic studies [2] because the corresponding genes have been detected in the genomes of ancient bacterial and archaeal lineages, such as extreme thermophiles and hyperthermophiles. Our model organism for the study of denitrification at high temperatures is Thermus thermophilus. Nitrate respiration is a common and strain-‐specific property in Thermus thermophilus, encoded by a gene cluster (NCE) that can be laterally transferred by conjugation [3]. In contrast, the nitrite respiration and further denitrification steps are restricted to a few isolates of this species, and the proteins involved are unknown. In order to test if the whole pathway could also be transferred, we have carried out transformation and conjugation assays, managing to obtain derivatives from the aerobic strain HB27 that are able to grow under anaerobic conditions using different nitrogen oxides as electrons donors and producing gas in the process. Thanks to massive sequencing methods, the complete genome sequences of a partial denitrificant strain, NAR1, and of a complete denitrificant strain, PRQ25, have been obtained.The analysis of PRQ25’s genome has allowed us to identify a homolog to nitrite reductase of the cytochrome cd1 (NirS) type and a classical nitric oxide reductase most likely dependent on a cytochrome c (cNor). The operons encoding these complexes were identified 7 kbp upstream of the NCE within a region that likely belongs to a megaplasmid (pTT27) present in most isolates of this species. The presence of transposases and synteny analysis reveal this region as a hot spot for the insertion of new genes. Genetic comparisons between these sequences and the draft genome of Thermus scotoductus SA1, isolated from a deep mine in South Africa, show high homology of the corresponding denitrification islands, supporting the idea of a frequent mobilization and transference of the denitrification genes even between different species. 1. Zumft, W.G., Microbiol Mol Biol Rev, 1997. 61(4): p. 533-‐616. 2. Philippot, L., Biochim Biophys Acta, 2002. 1577(3): p. 355-‐76. 3. Ramírez-‐Arcos, S et al. Biochim Biophys Acta, 1998. 1396(2): p. 215-‐27.


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HALOMONAS RIFENSIS SP. NOV., AN EXOPOLYSACCHARIDE-‐PRODUCING, HALOPHILIC BACTERIUM

ISOLATED FROM BRIKCHA, A SOLAR SALTERN IN CHEFCHAOUEN, MOROCCO Hakima Amjresa,b, Inmaculada Llamasa, Jamal Abrinib, Victoria Béjara and Emilia Quesadaa (a) Microbial Exopolysaccharide Research Group, Department of Microbiology, Faculty of Pharmacy, Cartuja Campus, University of Granada, 18071 Granada, Spain. (b) Biotechnology and Applied Microbiology Research Group, Department of Biology, Faculty of Sciences, University Abdelmalek Essâadi, BP2121, 93002 Tetouan, Morocco. We have conducted a polyphasic taxonomic study of strain HK31T, a moderately halophilic bacterium isolated from a solar saltern in Chefchaouen, Morocco. The strain is a Gram-‐negative, oxidase-‐positive rod, motile by peritrichous flagella. It requires NaCl for growth and grows best at salt concentrations (mixture of sea salts) of 0.5-‐20% w/v (optimum 5-‐7.5%), temperatures from 25ºC to 45ºC (optimum 32ºC) and a pH range of 5 to 10 (optimum pH 6-‐9). It does not produce acids from sugars and its metabolism is respiratory with oxygen as terminal electron acceptor. It accumulates poly-‐β-‐hydroxyalkanoate granules and forms mucoid colonies due to its excretion of an exopolysaccharide. Its G+C content is 61.5 mol%. Its 16S rRNA gene sequence indicates that it belongs to the genus Halomonas in the class Gammaproteobacteria. The most phylogenetically related species is Halomonas anticariensis, with which HK31T shows 16S rRNA identity values of 96.48%. Its major fatty acids are C18:1ω7c (38.25%), C16:0 (26.73%), C19:0 cyclo ω8c (14.85%), C16:1ω7c/iso-‐C15:0 2-‐OH (6.63%) and C12:0 3-‐OH (5.85%) and the predominant respiratory lipoquinone is ubiquinone, with nine isoprene units (Q-‐9). The proposed name for the new species is Halomonas rifensis. The type strain is strain HK31T (=CECT 7698 =LMG 25695)


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A NOVEL SPECIES OF THE GENUS BACILLUS ISOLATED FROM A SALINE LAKE IN IRAN Ana B. Fernándeza, Cristina Sánchez-‐Porroa, Mohammad A. Amoozegarb, H. Babavalian Fardb, M. Ramezanib and Antonio Ventosaa

(a) Dept of Microbiology and Parasitology, Faculty of Pharmacy, University of Sevilla, 41012 Sevilla, Spain (b) Dept of Microbiology, Extremophiles Lab, Faculty of Biology, College of Science, University of Tehran, Iran In 2008 we started a study on the microbial diversity of Lake Aran-‐Bidgol, located in central area of Iran, allowing the isolation of several new strains that were studied in detail. Strain Amb33 was selected for further study. This strain is Gram-‐positive with rod-‐shaped and motile cells, its colonies are round, convex, smooth, with entire margins and have a slightly orange pigmentation. It can grow in a salt concentration range between 0.5 and 20% NaCl (optimum between 5.0-‐7.5% NaCl), in a range of pH values between 5.5 and 9.0 (optimum at 7.5), and an optimum growth temperature of 35ºC. Amb33 is catalase and oxidase positive, reduces nitrate to nitrite, is able to hydrolyze esculin, starch, casein, gelatin and DNA, but does not hydrolyse Tween 80 or urea. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain Amb33 exhibited higher phylogenetic relationships with species of the genus Bacillus, specifically B. hwajinpoensis SW-‐72T (98.5%) and B. algicola KMM 3737T (96.8%). Also, the DNA G+C content of strain Amb33 is 47 mol%. DNA-‐DNA hybridization studies between strain Amb33 with the most closely related species Bacillus hwajinpoensis SW-‐72T and Bacillus algicola KMM 3737T showed hybridization percentages of 33 and 36%, respectively. Currently, we are completing the chemotaxonomic study of strain Amb33, although phylogenetic analysis and phenotypic and genotypic characteristics carried out so far indicate that strain Amb33 can constitute a novel species within the genus Bacillus.


33

THE HANRI QUORUM-‐SENSING SYSTEM OF HALOMONAS ANTICARIENSIS, A MODERATELY

HALOPHILIC BACTERIUM Ali Tahrioui, Inmaculada Llamas, Emilia Quesada Department of Microbiology. Faculty of Pharmacy. University of Granada. Campus Universitario Cartuja s/n, 18071 Granada, Spain. Quorum sensing is a cell-‐density-‐dependent gene-‐expression mechanism that involves the production of signal molecules such as N-‐acyl homoserine lactones (AHLs) in Gram-‐negative bacteria. Quorum-‐sensing mechanisms are known to regulate genes involved in expression of virulence factors and exoenzymes, conjugal DNA transfer and plasmid copy number control, production of antibiotics, iron acquisition, biofilm formation and exopolysaccharide production (González and Keshavan, 2008; Whitehead et al., 2001; De Kievit and Iglewski, 2000). One significant group of organisms, for which quorum sensing has not been previously studied are the moderate halophiles. We have reported that Halomonas anticariensis FP35T synthesizes four AHLs, N-‐butyryl-‐DL-‐homoserine lactone (C4-‐HL), N-‐hexanoyl-‐DL-‐homoserine lactone (C6-‐HL), N-‐octanoyl-‐DL-‐homoserine lactone (C8-‐HL) and N-‐dodecanoyl-‐DL-‐homoserine lactone (C12-‐HL). We have also found that the production of these compounds is growth-‐phase dependent and that maximum activity is reached during the late-‐exponential to stationary phases (Llamas et al., 2005).

We have recently identified a quorum-‐sensing system in Halomonas anticariensis FP35T that is composed of luxR/luxI homologs: hanR, the transcriptional regulator gene and hanI, the autoinducer synthase gene. To address how the hanR/hanI system is regulated we have carried out insertional mutagenesis experiments, RT-‐PCR, transcriptional lacZ fusion analysis and betagalactosidase assays. Our results indicate that the hanR/hanI system constitutes an operon which expression is regulated by a promoter located upstream of hanR gene and a secondary promoter upstream of hanI gene. HanI protein is responsible for the production of the AHLs and HanR protein has a slightly effect on the synthesis of AHLs.

In order to understand the role of cell-‐to-‐cell communication in Halomonas anticariensis FP35T we have carried out a Phenotype MicroArrays (PM) analysis to assay 1536 phenotypes. Preliminary results indicate that a cell-‐density-‐dependent gene-‐expression mechanism is involved in carbohydrate utilization, nitrogen and amino acid metabolism, transport of small molecules, as well as in other phenotypes. References -‐De Kievit and Iglewski. 2000. Infect. Immun. 68: 4839-‐4849. -‐González and Keshavan, N. 2006. Mol. Biol. Rev. 70: 859-‐875. -‐Llamas et al. 2005. Extremophiles 9: 333-‐341. -‐Whitehead et al., 2001. FEMS Microbiol. Rev. 25: 365-‐404.


34

DIVERSITY OF ARCHAEA IN SALINE AND NON-‐SALINE SOILS Nahid Oueriaghli, Rocío Luque, Fernando Martínez-‐Checa, Inmaculada Llamas, Emilia Quesada, Victoria Béjar Microbial Exopolysaccharides Research Group, Department of Microbiology, Faculty of Pharmacy, University of Granada, Granada 18071, Spain. The Archaea were originally described as microorganisms that thrive in extreme environments. Nevertheless, recent studies have shown that they coexist with the members of the Domains Bacteria and Eukarya in most environments. In this work we have studied the diversity of archaea in two different habitats, a saline habitat in Rambla Salada (Murcia) and an agricultural non-‐saline soil in Motril (Granada). These soils have been analysed using molecular (DGGE and FISH) and classical techniques. Molecular techniques: The DNA from a total of 24 samples was extracted and analyzed by denaturing gradient gel electrophoresis (DGGE). The sequences obtained from DGGE profiles were compared with those existing in the data bases (BLAST). The majority of the Archaea found in Rambla Salada soils belonged to the genus Halorhabdus, Haloarcula and Methanobrevibacter. In the agricultural soils of Motril, however, the archaeal community was essentially consisted of “Candidatus Nitrososphaera”, “Candidatus Aciduliprofundum”, Thermoplasma, and Methanobrevibacter. Our results so far indicate that the communities of archaea are quite different in the habitats studied. Thus, uncultured archaea predominated in the argricultural soils, while in Rambla Salada soils we found that the haloarchaea was the dominant group. Classical techniques: We studied a collection of 50 archaeal strains isolated from Rambla Salada. Most of them were related to halophilic archaea, such as Natrinema, Haloferax, Halostagnicola, Haloarcula, and Halorubrum. We also found a novel archaeon classified into the genus Natronomonas. It is noteworthy that the members of Haloarcula were identified in hypersaline habitats using both molecular and classical techniques. Classical studies in agricultural soils of Motril are in progress.


35

BIOINFORMATICS AND EXPERIMENTAL ANALYSIS OF THE GS/GOGAT PATHWAY AND ITS POSSIBLE REGULATORY PROTEINS IN THE GENOME OF HALOFERAX MEDITERRANEI Carmen Pire Galiana, Julia Mª Esclapez Espliego, Francisco Pérez Pomares, Laia Pedro Roig, Vanesa Bautista Saiz, Rosa Mª Martínez Espinosa, Basilio Zafrilla Requena, Anna Vegara Luque, Francisco Llorca Alcaraz, Gloria Bravo Barrales, Susana Díaz González, Mónica Camacho Carrasco, Mª José Bonete Pérez Extremophilic Proteins Group, Universidad de Alicante Nitrogen (N) is one of the basic elements of all organisms and their availability limits primary productivity in both natural environments and agriculture [1]. In the nature, N can be found in different redox states, from +5 in the most oxidized compound (nitrate) to -‐3 in the most reduced compound (ammonia). Haloferax mediterranei is a haloarchaea able to grow in minimal media using nitrate and/or nitrite as sole nitrogen source. Under these conditions, ammonium assimilation occurs by GS/GOGAT pathway. This ammonia assimilation pathway, that implies glutamine synthetase (GS) and glutamate synthase (GOGAT), is of crucial importance since the products L-‐glutamine and L-‐glutamate play a key role as nitrogen donors in the biosynthetic reactions. The GS/GOGAT pathway is particularly important because it allows ammonia assimilation into L-‐Glu at low intracellular ammonia concentrations and it seems that efficiently substitutes the other glutamate biosynthetic reaction (that of glutamate dehydrogenase, GDH) in these conditions. Glutamine synthetase, the first enzyme of the pathway GS/GOGAT, seems to act universally in ammonia assimilation [2]. GS/GOGAT pathway from Hfx. mediterranei could allow the assimilation of the ammonium produced by assimilatory nitrite reductase, while glutamate dehydrogenase would allow the assimilation of ammonium when this nitrogen source is present in the culture media at high concentration [3]. In bacteria, the catalytic activity of GS is regulated by adenylylation, and covalent binding of AMP to a specific tyrosine residue of GS occurs in response to an increase in intracellular nitrogen availability. This modification is catalyzed by adenylyl transferase, which is controlled by the PII protein [4]. The regulation of GS from Hfx. mediterranei is being studied in our Group; the halophilic protein gene has been expressed and, by sequence comparison with other GS, we have seen that also it has the tyrosine residue capable of covalent modification; two genes whose products ( GlnK1 and GlnK2) belong to the family of PII proteins have been expressed and, by to the genome of this organism that our Group has sequenced, and the bioinformatic analysis done, we have identified an adenylyl transferase that could act in the halophilic glutamine synthetase regulation. Work funded by the Project BIO2008-‐00082 from Ministerio de Educación y Ciencia (Programa Biotecnología) (Spain). References: [1] Moreno-‐Vivián, C., Cabello, P., Martínez-‐Luque, M., Blasco, R. and Castillo, F. (1999) J. Bacteriol. 181, 6573. [2] Martinez-‐Espinosa, R.M., Esclapez, J., Bautista, V. and Bonete, M.J. (2006) FEMS Microbiol. Lett. 264, 110. [3] Bonete, M.J., Camacho, M., Martínez-‐Espinosa, R.M., Esclapez, J., Bautista, V., Pire, C., Zafrilla, B., Díaz, S., Pérez-‐Pomares, F. and Llorca, F. (2007) En: “Communicating Current Research and Educational Topics and Trends in Applied Microbiology” (Méndez-‐Vilas, A., ed.), pp. 170-‐183, Formatex. [4] Stadtman, E.R. (2001) J. Biol. Chem. 276, 44357.


36

TRANSCRIPTIONAL REGULATION OF THE ECTD AND ECTE GENES, ENCODING TWO COPIES OF THE ENZYME ECTOINE HYDROXYLASE OF THE HALOPHILIC BACTERIUM CHROMOHALOBACTER

SALEXIGENS Mercedes Reina-‐Bueno, Montserrat Argandoña, Fernando Iglesias-‐Guerra, Carmen Vargas, Joaquín J. Nieto Department of Microbiology and Parasitology. Faculty of Pharmacy. University of Seville. C/ Profesor García González Nº 2, 41012 SEVILLA The obligately halophilic γ-‐proteobacterium Chromohalobacter salexigens synthesizes de novo and accumulates the compatible solutes ectoine, hydroxyectoine, and trehalose in response to abiotic stress such as high salinity and temperature. The synthesis of hydroxyectoine from ectoine is catalysed by a Fe2+ and 2-‐oxoglutarate-‐dependent ectoine hydroxylase enzyme, which is encoded by the ectD and ectE paralogs. We have shown that EctD is the main responsible for hydroxyectoine synthesis via ectoine hydroxylation. Our previous analyses by using 13C-‐NMR and HPLC techniques revealed that hydroxyectoine accumulation is up-‐regulated by salinity and high temperature. In this study, we present data on the transcriptional control of hydroxyectoine synthesis in C. salexigens. First, we used qPCR to quantify the expression of the ectD and ectE genes under different combinations of salt and temperature stress. Second, two promoters upstream of ectD were mapped by primer extension. The first one was predicted to be dependent on the vegetative factor σ70, whereas the second was activated by temperature and showed -‐10 and -‐35 regions that matched the consensus sequences of promoters dependent on heat stress sigma factor σ32. In agreement with this finding, expression of an ectDp::lacZ transcriptional fusion was drastically reduced in an E. coli rpoH (rpoH encoding σ32) background. Expression from this σ32-‐dependent promoter was also dependent on the transcriptional activator EctZ, whose gene is located upstream of ectD in the C. salexigens genome.


37

SALIMICROBIUM SALEXIGENS SP. NOV., A MODERATELY HALOPHILIC BACTERIUM ISOLATED FROM

SALTED HIDES Rafael R. de la Habaa, Pinar Yilmaza,b, Cristina Sánchez-‐Porroa, Meral Birbirb and Antonio Ventosaa aDepartment of Microbiology and Parasitology, Faculty of Pharmacy, University of Sevilla, 41012 Sevilla, Spain bDivision of Plant Diseases and Microbiology, Department of Biology, Faculty of Arts and Sciences, Marmara University, 34722 Istanbul, Turkey During the course of studies focused on the determination of the bacterial diversity of salted hides a great number of strains were isolated and some of them have been studied in detail. Strains 29CMIT and 53CMI are novel moderately halophilic bacteria closely related to the genus Salimicrobium. This genus was proposed to accommodate a new species, S. luteum, as well as to transfer the species Marinococcus albus and Bacillus halophilus to the new genus Salimicrobium, as S. album and S. halophilum, respectively. Recently, another species within this genus has been described, S. flavidum, isolated from a marine solar saltern of the Yellow Sea, Korea. The polyphasic characterization of strains 29CMIT and 53CMI was performed following the recommendations of the proposed Minimal Standards for describing new taxa of aerobic, endospore-‐forming bacteria. Both strains were Gram-‐positive, moderately halophilic and endospore-‐forming cocci. Cells were non-‐motile, strictly aerobic, growing in the presence of 3-‐25 % (w/v) NaCl (optimum at 7.5-‐12.5 % [w/v] NaCl), at pH 5.0-‐9.0 (optimum at pH 7.0-‐8.0) and at temperatures between 20-‐40 ºC (optimum at 37 ºC). Phylogenetic analysis based on 16S rRNA gene sequence comparison showed that both strains were closely related to species of the genus Salimicrobium, within the phylum Firmicutes. Strain 29CMIT and 53CMI exhibited 16S rRNA gene sequence similarity values of 97.8 % and 97.7 % with Salimicrobium halophilum DSM 4771T, 97.5 % and 98.1 % with Salimicrobium album DSM 20748T, 97.4 % and 98.1 % with Salimicrobium flavidum ISL-‐25T and 97.3 % and 97.9 % with Salimicrobium luteum BY-‐5T. The DNA G+C content was 50.7 and 51.5 mol% for strains 29CMIT and 53CMI, respectively. The DNA-‐DNA hybridization between both strains was 98 %, whereas the values between strain 29CMIT and the species Salimicrobium album CCM 3517T, Salimicrobium luteum BY-‐5T, Salimicrobium flavidum ISL-‐25T and Salimicrobium halophilum CCM 4074T were 45 %, 28 %, 15 % and 10 %, respectively, showing unequivocally that strains 29CMIT and 53CMI constitute a new genospecies. The major cellular fatty acids were anteiso-‐C15:0, anteiso-‐C17:0, iso-‐C15:0 and iso-‐C14:0. The main respiratory isoprenoid quinone was MK-‐7, although small amounts of MK-‐6 were also found. On the basis of the phylogenetic analysis and phenotypic, genotypic and chemotaxonomic characteristics, we propose strains 29CMIT and 53CMI as a novel species of the genus Salimicrobium, with the name Salimicrobium salexigens sp. nov.


38

ATTENUATTION OF ACID MINE DRAINAGE (AMD) IN RIO TINTO SEDIMENTS THROUGH SULFATE REDUCTION Irene Sánchez-‐Andreaa, Nuria Rodriguezb, Ricardo Amilsa,B, Jose Luis Sanza (a) Dpt. of Molecular Biology, Universidad Autónoma de Madrid, 28049 Madrid, Spain (b) Extremophile group, Centre of Astrobiology, INTA-‐CSIC, 28850 Ajalvir, Spain. The Tinto River in southwestern Spain is the focus of an increasing amount of research including geochemistry, microbiology and Mars-‐analogue studies. This basin shows the characteristics of Acid Mine Drainage (AMD) (low pH and high amounts of dissolved metals) where the aerobic, acidophilic, chemolithotrophic Bacteria and Archaea dissolve metal sulfides by oxidizing Fe(II) and sulfur compounds. Products resulting from these oxidation processes can be used by Fe(III) and sulfate reducing prokaryotes under anaerobic conditions. To go into the knowledge of the processes taken place in the underlying sediments in depth, cores of two sampling sites (SN and JL dams) were analyzed in this study through a physical-‐chemical (pH, redox and ICP-‐MS) and quantitative microbial community analysis (CARD-‐FISH). Depth profiles of cell numbers showed that: (i) the microbial communities are greatly different at both samples sites: 5,4E+08 cells/wet weight of sediment in JL respect 9,7E+06 in SN dam, and (ii) strongly drop with the depth: from 1,01E+09 in the superficial layer to 2,1E+07 in the depth layer in JL damn and from 1,2E+07 to 3,2E+06 in SN damn. Hybridization with domain-‐probes showed that in all cases Bacteria (98,6%) dominated over Archaea (1,4%) and Eukarya (ND). Class and phylum-‐probes hybridization showed that bacteria belonging to Delta-‐proteobacteria (13,7%) were the most abundant, followed by Gamma-‐proteobacteria (5,5%) ones. Genera probes showed that Syntrophobacter spp (6,1%), Desulfurella spp. (9,3%) and Desulfosporosinus spp. (5,6%) -‐all of them sulphate-‐reducing bacteria – were predominant in the sediments. Fe-‐reducing microorganisms (Acidithiobacillus spp. and Acidiphilium spp.) were also detected although in lower numbers. In the layers where sulphate-‐reducing bacteria showed the highest numbers there was an increased in the pH value and a decrease of the redox potential, dissolved metals and iron. These results suggest that the attenuation of AMD is biologically driven by reducing sulphate and through sulphide-‐mediated iron reduction with the subsequent metals and iron precipitation as sulfides.


39

ISOLATION, CONSERVATION AND CHARACTERIZATION OF VIRUS OF SALINIBACTER RUBER J. Villamor1,2, I. Meseguer1, F. Santos2, J. Antón2 (1) Departamento de Producción Vegetal y Microbiología, Universidad Miguel Hernández de Elche. (2) Departamento de Fisiología, Genética y Microbiología, Universidad de Alicante The sequencing of the genome of two strains of the extremely halophilic bacterium S. ruber has revealed than 10% of the genes are strain specific, are mostly located in areas coding for a high number of transposases, and are related to the cell envelopes synthesis and extracellular compounds. This fact points to a significative influence of phages in the genomic differentiation between strains. For this reason, and in order to study their role in the intraspecific diversification of S. ruber, we have undertaken the systematic isolation of viruses infecting this bacterium. So far, there are available 21 isolated viruses infecting extremely halophilic microorganisms: 20 of them infecting different genera of haloarchaea and one infecting the extremely halophilic bacterium Salicola spp. However, to date, there no isolated Salinibacter viruses. In this study we have developed the methodology for the isolation and quantification of viruses for S. ruber using viruses water samples from hypersaline environments to infect M8 and M31 cultures. The isolated viruses have been further characterized by transmission electron microscopy (TEM) and their genome sizes estimated using pulsed file gel electrophoresis (PFGE). Our results indicate that M8 and M31 strains have different sensitivity to phages. So far, we have isolated over 20 phages infecting S. ruber. Two of them infecting strain M8, ΦM8CR-‐1 y ΦM8CR-‐2, that were isolated from crystallizer ponds from “Bras del Port” brines, are now being studied in more detail.


40

BACTERIAL DIVERSITY IN THERMAL ENVIRONMENTS STUDIED BY DGGE AND ELECTRONIC MICROSCOPY Roy Mackenzie1, Leslie Abarzua, Homero Urrutia Departamento de Microbiología, Facultad de Ciencias Biológicas, Universidad de Concepción, Chile 1 Present address: Instituto de Ciencias del Mar, CSIC, Barcelona Currently, there is an increasing interest in the study of biodiversity in extreme environments, since they are the only type of ecosystems in which the existence of microbial endemism has been demonstrated. These microbial communities are a unique component of biodiversity, both from the ecological point of view, and from the possibility to isolate microorganisms that help to clarify physiological and evolutionary processes in microbes. The bacterial diversity of three hot springs from the Chilean Patagonia was compared (Porcelana’s hot spring, Cahuelmó’s hot spring and Porcelana’s Fumarole), along transects in a thermal gradient with a variation of ±20ºC ( ̴60-‐40ºC) in each hot spring. Results show that a 40% of the bacterial community is shared among all three hot springs. Moreover, the bacterial community at higher temperature is always different from the cooler points, however, mesophilic communities exhibited greater similarities between them. Despite the existence of a shared proportion among the hot springs in this region, our results suggests that a geographic isolation may exist, because each hydrothermal system showed specific environmental conditions that selected different bacterial groups.


41

LISTA DE ASISTENTES


42

A

Abrusci, Concha

Departamento de Biologia Molecular

Universidad Autonoma de Madrid

Cantoblanco, 28049 Madrid

[email protected]

Aguilera Bazán, Ángeles

Centro de Astrobiología

Instituto Nacional de Técnica Aeroespacial

CAB-‐INTA-‐CSIC

Ctra. de Torrejón a Ajalvir, km 4

28850 Torrejón de Ardoz, Madrid

[email protected]

Al-‐Ramahi, Yamal

Grupo de Biotecnología y Genética de bacterias

termófilas

Centro de Biología Molecular Severo Ochoa

CBM-‐UAM-‐CSIC

C/Nicolás Cabrera 1

28049 Madrid

Alvarez, Laura


termófilas


CBM-‐UAM-‐CSIC


28049 Madrid

[email protected]

Alvarez Tejado, Miguel

Product Manager

Roche Diagnostics SL

Roche Applied Science

Av. de la Generalitat, 171-‐173

08174 Sant Cugat del Vallès

miguel.alvarez-‐[email protected]

Amils i Pibernat, Ricardo

Unidad de Microbiología Aplicada


CSIC-‐UAM

c/ Nicolás Cabera nº 1, Cantoblanco

28049 Madrid

&

Departamento de Habitabilidad, Laboratorio de

Extremofilia


CSIC-‐INTA

Carretera Ajalvir km 4


[email protected]

Amjres, Hakima

Grupo Exopolisacáridos microbianos

Facultad de Farmacia

Universidad de Granada

Campus Universitario de Cartuja s/n

18071 Granada

Antón Botella, Josefa Departamento de Fisiología, Genética y Microbiología Universidad de Alicante Apartado 99 03080 Alicante [email protected]

B

Béjar, Victoria

Grupo Exopolisacáridos Microbianos




18071 Granada

[email protected]


43

Berenguer, José


termófilas


CBM-‐UAM-‐CSIC


28049 Madrid

[email protected]

Bricio, Carlos


termófilas


CBM-‐UAM-‐CSIC


28049 Madrid

[email protected]

C

Casamayor, Emilio Ortega

Microbial Community Ecology

Centre d’Estudis Avançats de Blanes

CEAB-‐CSIC

Acces Cala St Francesc, 14

17300 Blanes

[email protected]

Cifuentes, Ana Marine Microbiology Group Institut Mediterrani d’Estudis Avançats IMEDEA-‐CSIC-‐UIB C/Miquel Marqués 21 07190 Esporles

Corral, Paulina

Dept Microbiology and Parasitology

Faculty of Pharmacy

University of Sevilla

C/. Prof. García González, 2

41012 Sevilla

[email protected]

Cuecas, Alba

Instituto de Recursos Naturales y Agrobiología,

IRNAS-‐CSIC

Avda. Reina Mercedes 10,

41012 Sevilla, Spain

Tel. +34 95 462 4711

Fax +34 95 462 4002

[email protected]

D

De la Calle, Fernando

Head of Microbiology R&D Dept.

PharmaMar SA

Avda. los Reyes, 1.

28770 Colmenar Viejo (Madrid)

[email protected]

De los Ríos Murillo, Asunción Instituto de Recursos Naturales Centro de Ciencias Medioambientales CCMA-‐CSIC Serrano 115 dpdo Madrid 28006 [email protected]

E

Esclapez Espliego, Julia Extremophilic Proteins Group Universidad de Alicante Facultad de Ciencias División de Bioquímica y Biología Molecular Ap. 99 03080 Alicante [email protected] F

Fernández González, Ana Beatriz


Faculty of Pharmacy



41012 Sevilla

[email protected]


44

G

Gómez, Caterina

Jefa de proyectos

Arquebio SL

Edifici Eureka -‐ Campus de la UAB 08193 Bellaterra (Cerdanyola del Vallès)

[email protected]

Gómez, Felipe

Departamento de Habitabilidad, Laboratorio de

Extremofilia


CSIC-‐INTA

Carretera Ajalvir km 4


[email protected]

González Grau, Juan Miguel

Instituto de Recursos Naturales y Agrobiología,

IRNAS-‐CSIC

Avda. Reina Mercedes 10,

41012 Sevilla, Spain

Tel. +34 95 462 4711

Fax +34 95 462 4002

[email protected]

González, Pedro Departamento de Fisiología, Genética y Microbiología Universidad de Alicante Apartado 99 03080 Alicante

González Toril, Elena


Instituto Nacional de Técnica Aeroespacial

CAB-‐INTA-‐CSIC

Ctra. de Torrejón a Ajalvir, km 4


[email protected]

H

Hidalgo, Aurelio


termófilas


CBM-‐UAM-‐CSIC


28049 Madrid

[email protected]

L

Llamas Company, Inmaculada Grupo exopolisacáridos microbianos

Dpto. Microbiologia




18071-‐Granada

[email protected]

Llorens Marès, Tomàs Microbial Community Ecology Centre d’Estudis Avançats de Blanes CEAB-‐CSIC Accés Cala St Francesc, 14 17300 Blanes [email protected] [email protected] López López, Arantxa Marine Microbiology Group Institut Mediterrani d’Estudis Avançats IMEDEA-‐CSIC-‐UIB C/Miquel Marqués 21 07190 Esporles [email protected]

Luque Aznar, Mª Rocío

Grupo “Exopolisacáridos microbianos”

Departamento de Microbiología



18071 Granada

[email protected]


45

M

Mackenzie, Roy Institut de Ciències del Mar ICM-‐CSIC Ps Marítim de la Barceloneta 37-‐49 08003 Barcelona [email protected]

Marin Palma, Irma

Departamento de Biologia Molecular

Universidad Autonoma de Madrid

Cantoblanco, 28049 Madrid

[email protected] Martínez-‐Checa, Fernando

Grupo de Exopolisacáridos Microbianos




18071 Granada

[email protected] Muñoz, Raúl Marine Microbiology Group Institut Mediterrani d’Estudis Avançats IMEDEA-‐CSIC-‐UIB C/Miquel Marqués 21 07190 Esporles [email protected]‐csic.es N

Nieto Gutiérrez, Joaquín J.

Department of Microbiology and Parasitology

University of Seville

C/Profesor García González 2

41012 Sevilla

[email protected] O Oueriaghli, Nahid Grupo de exopolosacaridos microbianos Facultad de Farmacia Universidad de Granada Campus Universitario de Cartuja s/n 18071 Granada [email protected]

P Párraga, Antonio

TPP & TFP Management Head

Target Product Profile &

Target Formulation Profile

Laboratorios Dr. Esteve

Barcelona

[email protected] Pedrós-‐Alió, Carlos Institut de Ciències del Mar ICM-‐CSIC Ps Marítim de la Barceloneta 37-‐49 08003 Barcelona [email protected]

Peña-‐Pardo, Arantxa

Departamento de Fisiología, Genética y

Microbiología

Universidad de Alicante

Apartado 99

03080 Alicante

[email protected]

Pire Galiana, Carmen

Extremophilic Proteins Group


Facultad de Ciencias

División de Bioquímica y Biología Molecular

Ap. 99

03080 Alicante

[email protected]

R

Reina Bueno, Mercedes




41012 Sevilla

[email protected]


46

Rodríguez de Moya Vera, Javier




41012 Sevilla

[email protected]

Rodríguez, Nuria Lab. of Extremophiles Centre of Astrobiology INTA-‐CSIC 28850 Torrejón de Ardoz [email protected]

Rosselló-‐Móra, Ramon Marine Microbiology Group Institut Mediterrani d’Estudis Avançats IMEDEA-‐CSIC-‐UIB C/Miquel Marqués 21 07190 Esporles rossello-‐[email protected]

Ruiz de la Haba, Rafael Dept Microbiology and Parasitology Faculty of Pharmacy University of Sevilla C/. Prof. García González, 2 41012 Sevilla [email protected] S

Sánchez-‐Andrea, Irene

Department of Molecular Biology,

Applied Microbiology Unit

Autonoma University of Madrid

C/ Darwin, 2

28049 Madrid

[email protected]

Sanz, Jose Luis

Department of Molecular Biology,

Applied Microbiology Unit

Autonoma University of Madrid

C/ Darwin, 2

28049 Madrid

[email protected]

T

Tahrioui, Ali

Grupo exopolisacáridos microbianos

Dpto. Microbiologia




18071 Granada

[email protected]

Torres, Leticia


termófilas


CBM-‐UAM-‐CSIC


28049 Madrid

Torres, Marta

Grupo Exopolisacáridos microbianos




18071 Granada

Triadó Margarit, Xavier Microbial Community Ecology Centre d’Estudis Avançats de Blanes CEAB-‐CSIC Accés Cala St Francesc, 14 17300 Blanes [email protected]

V

Valea, Sergio Instituto de Recursos Naturales Centro de Ciencias Medioambientales CCMA-‐CSIC Serrano 115 dpdo Madrid 28006


47

Vegara, Anna

Extremophilic Proteins Group


Facultad de Ciencias

División de Bioquímica y Biología Molecular

Ap. 99

03080 Alicante

Ventosa, Antonio


Faculty of Pharmacy



41012 Sevilla

[email protected]

Villamor, Judith

Departamento de Fisiología, Genética y

Microbiología


Apartado 99

03080 Alicante


48

NOTAS


49

NOTAS

SANTA SUSANNA 28>29 OCTUBRE2010 · 2011. 11. 3. · X!REUNIÓN!DELA...

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