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Póster presentado en la S7. Biodeterioration: fundamentals, present and future perspectives. Session in honour of Prof. Cesáreo Sáiz Jiménez del 3rd International Congress Science and Technology for the Conservation of Cultural Heritage.-- Technoheritage 2017 21-24th May 2017, Cádiz, Spain, Caves are considered experimental locations of remarkable geological, ecological and microbiological interest, with diverse compartments that differ in physical, chemical and microbial composition. The study of the microbial diversity is important for many different reasons, but perhaps one of the most important is that caves may be potential sources of novel microorganisms producing bioactive compounds of medical interest. In the last few years Cueva del Tesoro has been investigated regarding the need to control phototrophic biofilms colonizing cave speleothems. Cleaning of caves and speleothems should be preceded by an inventory of species and testing of treatment protocols due to the fact that cleaning with biocides revealed important negative effects in some caves. We observed that the green phototrophic biofilms in this cave were uncommon and presented some characteristics worthy of study. These communities were triggered by the artificial lighting which is located in selected places along the visitor´s trail. The assemblages were mainly composed of Cyanobacteria (Friedmannia sp., Nostoc sp., Phormidium sp., Gleocapsa-like members, etc.) Chlorophyta (Friedmannia sp., Chlorella sp, Choricystis sp.), Rhodophyta (Cyanidium sp.), Bacillariophyta (Diadesmis contenta). Some locations develop abundant populations of the bryophyte Eucladium verticillatum. Some of the Chlorophyta and Rhodophyta are parasitized by unknown virus-like structures located on the cell walls, a topic of potential scientific interest. Regarding the bacteria, three novel species are being studied and will be described: Acinetobacter thesauricus, Bacillus thesauricus and Paracoccus speluncae. In addition, two new species of the fungal genus Aspergillus were described: Aspergillus thesauricus and Aspergillus baeticus. The species name thesauricus refers to the place where the bacterial and fungal species were first isolated and described. We tested three cleaning methods: i) Mechanical cleaning with liquid nitrogen, ii) Cleaning with sodium hypochlorite and iii) Cleaning with hydrogen peroxide. The most effective method was the use of hydrogen peroxide which was used by the restoration company carrying out the works. In addition, a remodelling of the lighting system and installation of new LED lamps in more appropriate places were suggested. Cueva del Tesoro revealed to harbour a rich bacterial, fungal and algal diversity. The removal and cleaning of the biofilms was focused on biodeteriorated walls and speleothems. However, the analysis of the microbial communities revealed that this cave seems to be an important reservoir of microbial diversity, which suggests that in addition to cleaning and control of the biofilms, some actions are needed to preserve this cave and its microbial diversity. In fact, it was proposed and accepted by the managing cave authority to leave some reservoirs of biofilms in the cave, distant from the rock art galleries, that does not represent a threat for the cultural heritage, in order to preserve biodiversity.

11 páginas.-- 2 figuras.-- 41 referencias.-- Conferencia presentada en el Sexto Congreso Español sobre Cuevas Turísticas, celebrado en Nerja (Málaga), en septiembre/octubre de 2016.-- Descarga de las Actas en http://www.cuevasturisticas.es/cuevatur/docs/2016/pdf/LibroCuevastur2016.pdf, [ES]: La geomorfología del medio subterráneo depende de su historia geológica, y de los fenómenos que lo modelan, como la erosión y otros agentes externos. Además, sobre los sedimentos, las paredes o los espeleotemas de cuevas se desarrollan poblaciones de microorganismos que modifican la superficie mineral. Así, la colonización de este medio estará en función de las condiciones ambientales, del grupo de microorganismos en cuestión y de la biorreceptividad de los sustratos. Las cuevas se consideran espacios especializados que se caracterizan por una temperatura uniforme, elevada humedad y escasos recursos nutricionales y energéticos. Las cuevas turísticas presentan mayor disponibilidad de energía, generada por la luz artificial y la entrada de materia orgánica, y favorecen la presencia de un mayor número de organismos fotosintéticos y heterótrofos. El desarrollo de microorganismos sobre las paredes y los espeleotemas de las cuevas pueden producir procesos de biodeterioro, es decir, cambios indeseables en el sustrato. En este trabajo se presenta la información obtenida sobre la microbiota que coloniza espeleotemas iluminados de dos cuevas turísticas, la Cueva de Nerja y la Cueva del Tesoro (Málaga), mediante el empleo combinado de la microscopía y del análisis molecular. Los resultados ponen de manifiesto que los microorganismos se organizan en comunidades estructurales complejas denominadas biofilms. El empleo de la microscopía ha permitido conocer la composición del sustrato y su relación con el biofilm, así como observar procesos de biodeterioro relacionados con cambios físicos y químicos del sustrato tales como decoloración, pérdida de consistencia, ruptura y disolución. En relación con la composición de los biofilms, se ha observado una biodiversidad dominada por cianobacterias y microalgas, junto con actinobacterias, proteobacterias, arqueas, hongos e incluso protozoos y pequeños artrópodos. El análisis molecular revela la presencia de nuevas especies de microorganismos en ambas cuevas. El conocimiento de estas comunidades, así como sus mecanismos de biodeterioro permite el diseño de medidas preventivas y correctoras destinadas al control y a una adecuada conservación del patrimonio natural y cultural de ambas cuevas., [EN]: The geomorphology of the subterranean environment depends on its geological history, and phenomena such as erosion and other external agents. Moreover, in the caves the development of microorganisms can change the surface of sediments, walls and speleothems. Therefore, the colonization of a cave will depend on the environmental conditions, the type of microorganisms involved and the substrata bioreceptivity. Caves are considered specialized areas characterized by a uniform temperature, high humidity and low nutritional and energy resources. Visited caves have more available energy, produced by artificial lighting and the input of organic matter, and these conditions allow the growth of photosynthetic and heterotrophic organisms. The development of microorganisms on the cave walls and speleothems may produce biodeterioration, i.e., undesirable changes in the substratum. This work provides information on the microbiota that colonizes illuminated speleothems in two tourist caves, Nerja and Tesoro (Málaga), through the combined use of optical and electron microscopy, together with molecular analysis. The results showed that, in both caves, microorganisms are organized into complex structural communities called biofilms. The microscopic approach provides data on the substratum composition and its relationship with the biofilm. Biodeterioration processes related to physical and chemical changes of the substratum such as discoloration, loss of consistency, breaking and dissolution, were also observed. The biofilms were dominated by cyanobacteria and microalgae together with actinobacteria, proteobacteria, archaea, protozoa, fungi and even small arthropods. Moreover, molecular analysis revealed the presence of new species of microorganisms in both caves. Knowledge about these communities and their mechanisms of biodeterioration allows the design of preventive and corrective actions, according with a proper conservation of natural and cultural heritage of both caves.

4 páginas.-- 3 figuras..- 18 referencias.-- Comunicación presentada en el Congreso Estudio y Conservación del Patrimonio Cultural. Actas 16-19 de noviembre 2015, Málaga, Las cuevas albergan variados grupos de microorganismos, entre los que se encuentran los de metabolismo fotosintético, que se desarrollan en las zonas iluminadas.1,2 En general, estos microorganismos forman parte de comunidades estructurales complejas denominadas biofilms, que también pueden estar formados por microorganismos heterótrofos e incluso pequeños invertebrados. De forma natural, los organismos fotosintéticos colonizan las zonas más externas de las cuevas pero en las cuevas turísticas, el sistema de iluminación eléctrica permite su desarrollo en las zonas más profundas, alrededor de los focos de luz. En estos medios es común la presencia de una flora que se relaciona con la otros ambientes aerofíticos, cuyo desarrollo depende de la biorreceptividad del sustrato y de las condiciones ambientales disponibles. La disponibilidad de algunos requisitos como agua, radiación fotosintética activa (PAR) o propiedades del sustrato determinan el tipo de microorganismo presente y su abundancia.

9 páginas.-- 6 figuras.-- 20 referencias, [EN]: In recent years caves hove revealed to be a source of new microbial species. The great adaptability o( microorganisms al/ow them establishing themselves in these subterranean environments where they develop different cammunities. Cueva del Tesoro (Rincón de la Victoria, Mólaga) harbours a great biodiversity barely explored. Recently, we hove described two new fungal species of the genus Aspergil/us. Three novel bacterial species of the genera Acinetabader, Bacil/us and Paracaccus are under research. The colomzatians of cyanobacteria and algae do requiere (urther investigation. These data support that the high biodiversity of Cueva del Tesoro should be preserved if it does not represent a threot for the conservation of the Cultural Heritage. [ES]: En los últimos años las cuevas se han revelado como una fuente de nuevas especies de microorganismos. La gran adaptación de los microorganismos a todo tipo de hábltats les permite vivir en estos ambientes donde se desarrollan formando diferentes comunidades. La Cueva del Tesoro (Rincón de la Victoria, Málaga) alberga una gran biodiversidad apenas explorada . Recientemente, hemos descrito dos nuevas especies de hongos del género Aspergil/us, y tres posibles nuevas especies de bacterias de los géneros Acinetobacter, Bacil/us y Paracoccus están siendo investigadas. Por otra parte, las colonizaciones de cianobacterias y algas requieren una investigación más detallada. Estos datos demuestran que la diversidad biológica de la Cueva del Tesoro es muy alta, por lo que debería ser preservada, siempre que no represente una amenaza para la conservación del Patrimonio Cultural., Este proyecto ha sido financiado por el proyecto de Excelencia de la Junta de Andalucía, código RNM5137 yun contrato del Excmo. Ayuntamiento del Rincón de la Victori a, Málaga

The type of branching is a morphological character of great significance, separating taxa within the true-branched cyanobacteria. In this paper, true-branched morphospecies of cyanobacteria belonging to genus Iphinoe Lamprinou et Pantazidou, from different and geographically isolated caves of Greece, were investigated by coupling classical approach i.e. microscopy analysis (LM, SEM, TEM, CLSM) and molecular analysis (16S rRNA gene diversity). The morphological study revealed interspecific differences which could be attributed to adaptations to local conditions: Iphinoe cf. spelaeobios from the cave 'Diktaion Andron' (Crete island) exhibit different type of branching (Y- and T-type) and smaller size compared to the type species Iphinoe spelaeobios (T- and V-type of branching) from the caves 'Kastria' and 'Koutouki' (Peloponnese, Attica). Despite their phenotypic differences and geographical isolation molecular analysis showed identical 16S rRNA sequences which indicate that both populations belong to the same genus. © Czech Phycological Society (2013).

Caves have generally been found to host phototrophic micro-organisms from various taxonomic groups, with cyanobacteria comprising an important group that have adapted to these stable and highly specific environments. A polyphasic study based on aspects of classical morphology and molecular data revealed two new monospecific genera from fresh material of Greek and Spanish caves. Both taxa are characterized by obligatory true branching (T-type, V-type and false branching), the presence of heterocysts, and reproduction by hormocysts and akinetes. They shared some similarities in their morphological characteristics as revealed by light, scanning electron and transmission electron microscopy, but phylogenetic analysis based on 16S rRNA gene sequences showed that the two phylotypes were different (89.8% similarity); this represents an example of shared morphology in genetically different strains of cave-adapted species. Phenotypic and genetic traits strongly support classification of the phylotypes as independent taxa in the order Stigonematales (the most differentiated and complicated group of cyanobacteria), family Loriellaceae Geitl 1925. Hence, the names Iphinoe spelaeobios Lamprinou and Pantazidou gen. nov., sp. nov. and Loriellopsis cavernicola Herna ́ndez-Marine ́and Canals gen. nov., sp. nov. are proposed. © 2011 IUMS.

5 pages, 4 figures, 20 references. El resumen corresponde al trabajo publicado en el libro: Heritage, Weathering and Conservation volumen 1, páginas 323-327, ISBN 0-415-41272-2. Consta de 6 páginas, 3 figuras, 20 referencias. Derechos de copyright: Taylor & Francis. International Conference, Madrid, 21-24 June, 2006. 8th Thematic Network on Cultural and Historic Heritage Scientific Meeting Stone, Weathering and Atmospheric Pollution Network Scientific Meeting., Molecular surveys of microbial communities in Roman catacombs of Saint Callixtus (Rome, Italy) have revealed the presence of Phyllobacterium 16S rRNA gene sequences. In these environment, Phyllobacterium appears to be linked to the development of cyanobacterial colonization of catacomb walls. This finding represents a novel niche for members of this genus since previously described Phyllobacterium has always been associated to plant nodules or the rhizosphera., This work was supported by the Spanish Ministry of Education and Science (MEC), project BTE2002-04492-C02-01 Social Fund fellowships and JMG from a MEC contract, Ramón y Cajal programme.

The biocidal effect of Algophase has been tested against the sheated cyanobacterium Gloethece membranacea CCAP 1430/3. The maximum effect of Algophase was observed at the recommended concentration (3%) and in the presence of gypsum in the cultures. Transmission electron microscopy confirmed the biocidal effect. The effect was fast, producing damage after an hour of contact. Main changes were rims of black accumulations of mucopolysaccharides in the cell wall and sheath thinness.

Las algas y cianobacterias de ambientes terrestres no han sido tan estudiadas como las acuáticas, a pesar de encontrarse muy difundidas y existir un gran número de especies. Viven en medios diversos: suelos, rocas, cortezas de árboles y arbustos, hielo, animales, etc. La biología y taxonomía de las algas edáficas han sido extensamente revisadas (METTING, 1981; HOFFMANN, 1989; ARCHIBALD, 1990); las de los otros biotopos terrestres han sido menos consideradas. El objetivo del presente trabajo es el estudio florístico de las algas y cianobacterias de ambientes terrestres -suelos, cortezas- y de charcos temporales en la Punta de la Mora (Tarragona).

Uno de los aspectos más interesantes que ofrece la biotecnología relacionada con las algas es la depuración de las aguas residuales. Las lagunas de alta carga, ideadas por Oswald en 1957 (OSWALD & GOLUEKE, 1960), difieren de las lagunas convencionales (o de estabilización) por ser menos profundas (30-40 cm) y por la continua agitación del agua que se obtiene generalmente mediante unas aspas. Este tipo de lagunas combinan la producción de biomasa con la depuración (ABELIOVICH, 1986). En este trabajo se presentan resultados parciales de la flora algal y de su periodicidad anual en dos lagunas de alta carga piloto.

15 pages, 41 figures, 2 tables, 40 references., The chlorophytes and the cyanobacteria present in samples from the outer zones of the cathedrals of Salamanca and Toledo (Spain) were isolaled and determined. A total of 21 taxa were studied, 8 belonging to Cyanobacteria and 13 to Chlorophyta. The most common species within the samples studied are the chlorophytes Klebsormidium flaccidum and Muriella terrestris, and the cyanobacteria Microcoleus vaginatus and Phormidium autumnale. Species distribution is discussed in relation to microclimatic conditions. It is found that in some cases, samples taken near ground level are characterized by the absence of cyanobacteria, which are however present in samples taken from places exposed to sunlight, at 50 m height. The possible biodeterioration mechanisms in which these chlorophytes and cyanobacteria could be involved are discussed., This research has been supported by the C.E.C. through contract EV4V 0061 E and C.I.C.Y.T. (PAT89-0767-C04), Spain.

The literature on cyanobacteria and algae in relation to the colonization of historic buildings and monuments, together with probable biodeterioration mechanisms and control methods, is reviewed. Because the organisms concerned are the same as those colonizing natural habitats, attention is also given to colonization and distribution of similar organisms in nature. Epilithic and endolithic cyanobacteria and algae are discussed. Directions for future research are also suggested.

3 pages, 1 figure, 1 reference.-- La monografía citada se publica con motivo del Symposium celebrado del 26-31, agosto, 1990, en Ontario, Canadá., The weathering of building stone is a natural process in which physical, chemical and biological agents are involved, similar to soil formation. Among the biological agents the atmophytic cyanobacleria and algae are primary colonizers due to their phototrophism and low nutrient requirements. Their presence on and in the stone may represent a contribution to its microbial deterioration., This research has been supported by the CEC through contract EV4E 0061 E and CICYT, Spain.