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12 páginas.- 5 figuras.- 2 tablas.- 39 referencias.- Comunicación oral presentada en el VIII Congreso Español sobre Cuevas y Minas Turísticas "Minas y Cuevas: Patrimonio Geológico y Turístico". J.M. Calaforra y J.J. Durán Valsero (eds.), Pulpí (Almería) 19-22 octubre 2022.- Enlace al libro de Actas completo https://www.cuevasturisticas.es/actas-cuevatur-2022, Las cuevas vienen siendo estudiadas desde diversos puntos de vistas: ambiental, geológico, geoquímico, microbiológico, turístico, etc. Sin embargo, las cuevas y otros ambientes subterráneos han sido poco explorados desde el punto de vista biotecnológico, aunque representan nichos ecológicos indispensables que podrían ofrecer nuevas moléculas de interés para la medicina y la farmacología. Se han investigado las bacterias de diversas cuevas de la Península Ibérica, así como de las Islas Canarias para comprobar su bioactividad. Entre ellas, han mostrado un interesante potencial antimicrobiano algunas de las bacterias aisladas de la Cueva de Altamira. Los análisis in vitro e in silico mostraron la inhibición de bacterias patógenas Gram- positivas, Gram-negativas y hongos. El interés de esta investigación se apoya en la conocida resistencia de las bacterias patógenas a los antimicrobianos, lo que se ha convertido en un problema mundial en las últimas décadas. Nuestros datos muestran la presencia de bacterias productoras de compuestos antimicrobianos en ambientes subterráneos; y el estudio de sus genomas indicarían la existencia de genes implicados en la síntesis de nuevos compuestos. Palabras clave: Cueva de Altamira, compuestos antimicrobianos, Crossiella, Pseudomonas, predicción de genes., Esta publicación es parte del proyecto de investigación 0483_PROBIOMA_5_E.

9 páginas.- 5 figuras.- 3 tablas.- referencias.-. The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb. 2022.982933/full#supplementary-material .- This article is part of the Research Topic Microbial Roles in Caves, The present study reports a sudden fungal outbreak that occurred in the corridor near the entrance of the Catacombs of SS. Marcellino and Pietro in Rome (Italy) observed after 1 year of a restoration treatment that interested the walls of the entrance of the Catacombs and some artifacts placed in situ. The colonization was observed on the vault at the entrance and in correspondence with the restored marble pieces displayed on the left side of the corridor. No growth was observed on the right side where similarly treated marble slabs were placed. Samples taken in correspondence with fungal biofilm were analyzed through the combined use of microscopical, cultural, and molecular tools and showed that the vault and the left side of the corridor entrance were colonized by a complex fungal biofilm consisting mainly of Coniophora sp. and other genera, such as Hypomyces, Purpureocillium, Acremonium, Penicillium, and Alternaria, many of which are well known as responsible of biodeterioration of stone surfaces. Regarding the brown-rot basidiomycete Coniophora, it was able to form very large colonies on the substrata with a diameter of up to 57 cm. Although the direct observation under a light microscope evidenced the presence of abundant brown fungal conidia, several attempts to cultivate the microorganism failed, therefore only through DNA sequencing analyses, it was possible to identify and characterize this fungus. There is very little literature on the genus Coniophora which is reported as one of the causes of wet-rot decay of wood in buildings. A connection with calcium-containing materials such as bricks and mortars was demonstrated, but no data were available about the possible role of this species in the biodeterioration of stones. This study features the first finding of a strain related to the basidiomycetous genus of Coniophora in the order Boletales in association with evident phenomena of biodeterioration., This project was supported thanks to the special funds provided by the University of Messina (approved on 21 February 2022 by the Academic Senatus) (Italy), the Spanish Ministry of Science and Innovation, and project PID2020-114978GB-I00 and project CIACCO DTC TE1 - FASE II - PROGETTI RSI8.

Altamira, Lascaux and many other caves house some of the world¿s most prominent Paleolithicrock art. Open to the visitors for long time, some of these caves had to be closed to the public after decades of visits due to microbial outbreaks and severe deterioration of the paintings. Once closed to visits, the tourists were rerouted to replicas of Altamira and Lascaux paintings, however politics and public pressures still exist for a reopening, and Altamira still allows the visits on Friday. In September 2002, Altamira Cave had to be closed to the public because of the presence of phototrophic microorganisms on the famous bisons decorating the ceiling of the Polychrome Hall, a phenomenon similar to that suffered by Lascaux 50 years before. Colonization by phototrophic microorganisms was a consequence of decades of use of artificial lighting in the Polychrome Hall and was accompanied by the development of white microbial colonizations, directly on the red paintings. Lascaux Cave was closed in 1963, and after 2001 suffered several fungal outbreaks due to a wrong management and the use of benzalkonium chloride as biocide for controlling the fungi. A research carried out by the CSIC showed that the walls and ceiling of Altamira Cave were colonized by bacteria, but this contrasted with the absence of visible fungal development. It was suggested that the microbial communities colonizing the walls plays an important role in the ecosystem and the presence of antifungal compounds produced by the bacteria would protect the cave rock surfaces from fungal colonization. In fact, some species of actinobacteria found in Altamira Cave, such as Actinomadura, Amycolatopsis, Nocardia, Micromonospora, Rhodococcus, Streptomyces, etc., have antifungal activities.

The first tomb of a Roman Necropolis in the area of Carmona (Seville, Spain) was accidentally discovered in 1830. Few more tombs were exhumed between 1868 and 1869. These tombs were plundered until the beginning of the archaeological excavations in 1882. This necropolis, located in the town of Carmona, represents one of the most significant Roman burial sites in Southern Spain, and was in use during the 1st and 2nd centuries AD. Initially, the necropolis was used as quarry from which stone blocks were extracted and used for building construction. The quarry was then abandoned and reused as necropolis due to the workability of the rock. The Necropolis has suffered numerous and extensive interventions since its discovery. The first one was the adaptation of the necropolis for visits in 1885, including the construction of a trail, design of gardens around the tombs and plantation of trees. Nowadays, the gardens are considered one of the most deteriorating factors affecting the conservation of the tombs due to irrigation and consequent percolation of water inside the tombs, in addition to the damage caused by roots. Taking into account the cultural, artistic and religious importance of the Necropolis of Carmona, its preservation and conservation are a major issue, both from cultural and economic points of view. This importance emphasizes the need to define accurate and sustainable intervention protocols based on a deep understanding of the environmental and/or anthropogenic-induced deterioration processes. The design of effective preservation and/or conservation strategies should be based on exhaustive in situ surveys and laboratory investigations. Here we review 10 years of research in the Necropolis and we focus on the Circular mausoleum, as a case study, including an assessment of its conservation state, and the identification of the main deteriorating agents. In addition, an intervention proposal for the preservation of the Circular Mausoleum was presented.

Comunicación oral presentada en la S5.-Biodeterioration: Fundamentals, Present and Future Perspectives. Session in Honour of Prof. Cesáreo Sáiz Jiménez V del 3rd International Congress Science and Technology for the Conservation of Cultural Heritage.-- Technoheritage 2017 21-24th May 2017, Cádiz, Spain, Cave and catacombs microbiota and especially fungi are sensitive to organic input changes in their close environment. New input of organic sources (soil, dead animals and microorganisms, etc.) associated to favorable microclimatic parameters often lead to the overgrowth of fungi that may cover (colonize) a wide extension of available surfaces. The present research reports of a sudden fungal outbreak occurred in the corridor near the entrance of the Catacombs of SS. Marcellino and Pietro in Rome. A heavy fungal growth was observed one year after a restoration treatment that interested the walls of the entrance of the Catacombs and some artifacts placed in situ. Some marble slabs in fact, were restored and placed back into the walls. The colonization was observed only on the left side of the corridor around the marble slabs and on the vault. No growth was observed in the right side were similarly treated marbles slabs were placed. However, a high circulation of airborne fungal spores was observed. Mycological analysis were done on samples taken on the catacombs vault and side, as well as from soil and tree roots coming from upper ground above the catacombs. The combined use of microscopical, cultural and molecular tools showed that the vault and the left side of the corridor entrance were colonized by the brown Basydiomycetes Coniophora marmorata, that was able to form colonies with a diameter up to 57 cm, while secondary colonizations were due to different microfungi belonging to the genera Hypomyces chlorinigenus, Purpureocillium ilacinum, Acremonium persicinum, Penicillium spp. and Alternaria sp. The comparative analysis of roots and soils showed that fungi were present in all samples but a different distribution and diversity was observed. Due to the fast rate of growth of the fungi on the walls, a three steps emergency treatment was done with intervention on the environment, on the biodeteriogens and further by a close monitoring on monthly basis. Despite the successful intervention, it was clear that several causes could have created this fungal outbreak and for this reason, a continuous monitoring of the catacomb’s surfaces was planned. * Both Authors contributed equally in the isolation and characterization of fungal isolates.

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

11 páginas.-- 2 figuras.-- 40 referencias., [EN]: Gruta de las Maravillas (Aracena, Huelva) is a cavity o( geological interest, with an important annual regime o( visits. We carried out a seasonal study o( the cave's aerobiology with the aim o( knowing the concentration and distriblltion o( bacterial and (ungal communities, and determining their seasonal pattern and visitors' impacto The analysis showed that while some species o(bacteria are present along the (our seasons, such as Micrococcus yunnanensis, the (ungi present a higher variability, which means that there is not a predominant species. This study ollows us to know the air quality and to compare these results with the ones previollsly obtained in other touristic caves., [ES]: La Gruta de las Maravillas (Aracena, Hu elva) es una cavidad de interés geológico, que mantiene un rég imen anual de visitantes importante. Se consideró de interés realizar un estudio estacional de su aerobiología con el objetivo de conocer la concentración y distribución de las comunidades bacterianas y fúngicas presentes en la cueva, determinar su pauta estacional y el impacto de las visitas. Los análisis realizados indican que mientras algunas especies están presentes a lo largo de las cuatro estaciones del año, como la bacteria Microcaccus yunnanensis, los hongos presentan mayor variabilidad no existiendo una especie predominante. Este estudio permite conocer la calidad del aire y comparar los resultados con los obtenidos previamente en otras cuevas visitables, Este proyecto ha sido cofinanciado por el programa operativo FEDER de Andalucía 2007-2013, y por el Proyecto de Excelencia de la Junta de Andalucía, código RNM5137. La participación de CSJ y VJ es a través del proyecto CSIC 201230E125

9 páginas, 1 figura, 2 tablas, 16 referencias.-- El libro consta de 448 páginas.-- Trabajo presentado al IV Congreso español sobre cuevas turísticas. CUEVATUR 2012, celebrado del 18-20, de octubre, 2012, en Aguilar de Campoo, Palencia, España., [ES]: La Aerobiología es una ciencia de reciente aplicación en cuevas. Los estudios del aire de las cuevas son escasos y no hay información respecto a la concentración de esporas fúngicas en el aire que permitan una clasificacción de la atmósfera de una cueva como no peligrosa para la conservación de las pinturas rupestres o incluso para la salud humana. Con el objetivo de conocer los problemas derivados del uso turístico de las cuevas y su impacto sobre el arte rupestre, recientemente se ha propuesto una clasificación con cinco categorías que van desde una cueva que no presenta problemas a una que está en peligro: la categoría 1 identifica a una cueva sin problemas de hongos, la categoría 2 representa una señal de alarma para las cuevas, la categoría 3 incluye a cuevas amenazadas por los hongos, la categoría 4 la componen las cuevas muy afectada por los hongos, y la categoría 5 es una cueva con daños ecológicos irreversibles y potencialmente muy peligrosa para la conservación de las pinturas. En este trabajo aplicamos esta clasificación a tres cuevas andaluzas (cueva del Tesoro, El Rincón de la Victoria, Málaga; cueva de Ardales, Málaga y Gruta de las Maravillas, Aracena, Huelva), con diferente régimen de visitas. Asimismo, se consideró de interés el estudio estacional de la aerobiología de las cuevas, con el fin de observar si existían pautas estacionales y/o la concentración y tipos de microorganismos presentes en el aire sufrían modificaciones a lo largo del año., [EN]: Aerobiology of caves is rarely studied. At present, no clear information has been generated on the limits of acceptance of fungal spores in air, which would permit a classification of the cave atmosphere as not dangerous for the conservation of rock-art paintings and for the health of visistors. We should rely on the preventive conservation and the early detection of microbial outbreaks and with this aim we are using an index of fungal hazard in show caves. This index is based on data on the concentration of fungal spores in the cave's air, knowledge of the cave's history and management, and a detailed survey of the different halls of the caves. The index classifies caves into five risk categories: category 1 designates a cave without a fungal problem, category 2 is a warning sign for caves, category 3 includes caves threatened by fungi, category 4 is assigned to caves already affected by fungi, and category 5 designates caves with an irreversible ecological impact. We have applied this index to three Andaluslian show caves: Treasure Cave (El Rincón de la Victoria, Málaga), Ardales Cave (Ardales, Málaga) and Wonders Grotto (Aracena, Huelva). In addition, we considered of interest to study if there were seasonal patterns in the aerobiology of some caves or the microorganisms present in the air suffer modification throughout the year., Este proyecto ha sido cofinanciado por el programa operativo FEDER de Andalucía 2007- 2013, Y del proyecto de Excelencia de la Junta de Andalucía, código RNM5137. La participación de CSJ y VJ es a través del "Programa de investigación en tecnologías para la valoración y conservación del patrimonio cultural", TCP CSD2007-00058.

Trabajo presentado como póster al 22nd IUBMB & 37th FEBS Congress, celebrado en Sevilla (España) del 4 al 9 de septiembre de 2012., [Backgroun and Purpose]: MELAS (mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes) is a mitochondrial disease most usually caused by point mutations in tRNA genes encoded by mitochondrial DNA (mtDNA). Approximately 80% of cases of MELAS syndrome are associated with a m.3243A > G mutation in the MT-TL1 gene, which encodes the mitochondrial tRNALeu (UUR). Currently, no effective treatments are available for this chronic progressive disorder. Treatment strategies in MELAS and other mitochondrial diseases consist of several drugs that diminish the deleterious effects of the abnormal respiratory chain function, reduce the presence of toxic agents or correct deficiencies in essential cofactors., [Experimental Approach]: We evaluated the effectiveness of some common pharmacological agents that have been utilized in the treatment of MELAS, in yeast, fibroblast and cybrid models of the disease. The yeast model harbouring the A14G mutation in the mitochondrial tRNALeu(UUR) gene, which is equivalent to the A3243G mutation in humans, was used in the initial screening. Next, the most effective drugs that were able to rescue the respiratory deficiency in MELAS yeast mutants were tested in fibroblasts and cybrid models of MELAS disease., [Key Results]: According to our results, supplementation with riboflavin or coenzyme Q10 effectively reversed the respiratory defect in MELAS yeast and improved the pathologic alterations in MELAS fibroblast and cybrid cell models., [Conclusions and Implications]: Our results indicate that cell models have great potential for screening and validating the effects of novel drug candidates for MELAS treatment and presumably also for other diseases with mitochondrial impairment., This work was supported by FIS PI10/00543 grant, FIS EC08/00076 grant, Ministerio de Sanidad, Spain and Fondo Europeo de Desarrollo Regional (FEDER-Unión Europea), SAS111242 grant, Servicio Andaluz de Salud-Junta de Andalucía, Proyecto de Investigación de Excelencia de la Junta de Andalucía CTS-5725, and by AEPMI (Asociación de Enfermos de Patología Mitocondrial), FEEL (Fundación Española de Enfermedades Lisosomales) and Federación Andaluza de Fibromialgia y Fatiga Crónica (ALBA Andalucía). MM receiveda fellowship from Colegio Oficial de Farmacéuticos de Sevilla. This group was founded by the Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER),ISCIII.

3 páginas.- 1 tabla.- 14 referencis, Pseudomonas sp. strain L5B5 is an antimicrobial-producing bacterium isolated from an air sample collected in a pyrite mine in Lousal, Portugal. Genomic analyses predicted genes involved in virulence factors. Here, we report the complete genome sequence of this bacterium, which consists of a circular chromosome with a length of 6,811,662 bp. Copyright © 2021 Gonzalez-Pimentel et al., Financial support was obtained through project 0483_PROBIOMA_5_E, cofinanced by the European Regional Development Fund within the framework of the INTERREG VA Spain-Portugal program (POCTEP), 2014 to 2020.

16 páginas.- 4 figuras.- 2 tablas.- 81 referencias.- : The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/microorganisms10081575/s1 .- (This article belongs to the Section Environmental Microbiology), Antimicrobial resistance has become a global problem in recent decades. A gradual reduction in drug discoveries has led to the current antimicrobial resistance crisis. Caves and other subsurface environments are underexplored thus far, and they represent indispensable ecological niches that could offer new molecules of interest to medicine and biotechnology. We explored Spanish show caves to test the bioactivity of the bacteria dwelling in the walls and ceilings, as well as airborne bacteria. We reported the isolation of two strains of the genus Crossiella, likely representing a new species, isolated from Altamira Cave, Spain. In vitro and in silico analyses showed the inhibition of pathogenic Gram-positive and Gram-negative bacteria, and fungi, as well as the taxonomical distance of both strains from their closest relative, Crossiella cryophila. The presence of an exclusive combination of gene clusters involved in the synthesis of lanthipeptides, lasso peptides, nonribosomal peptides and polyketides indicates that species of this genus could represent a source of new compounds. Overall, there is promising evidence for antimicrobial discovery in subterranean environments, which increases the possibility of identifying new bioactive molecules. Crossiella; predicted gene clusters; Spanish show caves, Financial support for research in caves and mines was obtained through project 0483_PROBIOMA_5_E, co-financed by the European Regional Development Fund within the framework of the Interreg V-A Spain-Portugal program (POCTEP) 2014–2020

18 páginas._ 3 figuras.- 1 tabla.- 102 referencias, Of the several critical challenges present in environmental microbiology today, one is the assessment of the contribution of microorganisms in the carbon cycle in the Earth-climate system. Karstic subterranean ecosystems have been overlooked until recently. Covering up to 25% of the land surface and acting as a rapid CH4 sink and alternately as a CO2 source or sink, karstic subterranean ecosystems play a decisive role in the carbon cycle in terms of their contribution to the global balance of greenhouse gases. Recent data indicate that microbiota must play a significant ecological role in the biogeochemical processes that control the composition of the subterranean atmosphere, as well as in the availability of nutrients for the ecosystem. Nevertheless, there are still essential gaps in our knowledge concerning the budgets of greenhouse gases at the ecosystem scale and the possible feedback mechanisms between environmental-microclimatic conditions and the rates and type of activity of microbial communities in subterranean ecosystems. Another challenge is searching for bioactive compounds (antibiotics) used for treating human diseases. At present, there is a global health emergency and a strong need for novel biomolecules. In recent decades, great research efforts have been made to extract antibiotics from marine organisms. More recently, caves have been receiving considerable attention in search of novel antibiotics. Cave methanotrophic and heterotrophic bacteria are producers of bioactive compounds and may be potential sources of metabolites with antibacterial, antifungal or anticancer activities of interest in pharmacological and medical research, as well as enzymes with a further biotechnological use. Here we also show that bacteria isolated from mines, a still unexplored niche for scientists in search of novel compounds, can be a source of novel secondary metabolites, Financial support was obtained through project 0483_PROBIOMA_5_E, co-financed by the European Regional Development Fund within the framework of the Interreg V-A Spain-Portugal program (POCTEP) 2014–2020. This work was also supported by the Spanish Ministry of Economy and Competitiveness through projects CGL2016-75590-P and PID2019-110603RB-I00, AEI/FEDER, UE.

14 páginas.- 5 firuras.- 57 referencias.- Supplementary Materials: The following are available online at https://www.mdpi.com/article/10.3390/app11115027/s1, The aerobiology of caves in Southern Spain possesses special characteristics, different from caves located in Northern Spain. Previous studies demonstrated the influence of outdoor air on caves in the north and the existence of two different patterns, depending on the season. In summer there is an abundance of Ascomycota, whereas in winter Basidiomycota predominates, which are related to the periods of stagnation and ventilation, respectively. In caves in Southern Spain the presence of airborne Basidiomycota is scarce and Ascomycota represents the main group of fungi widely distributed across the caves in all seasons. The most characteristic features were the abundant presence of entomopathogenic fungi (Beauveria bassiana, Parengyodontium album, Pochonia chlamydosporia, Leptobacillium symbioticum, Leptobacillium leptobactrum) and Cladosporium cladosporioides in Cueva del Tesoro, Cueva de Ardales and Gruta de las Maravillas. However, the presence of yeasts of the genera Cutaneotrichosporon, Trichosporon, Cryptococcus, Naganishia, Cystobasidium, Microstroma and Phragmotaenium was exclusive to Gruta de las Maravillas. Fungal hazard in the three show caves were determined using an ecological indicator based on the concentration of spores in cave air. © 2021 by the authors. Licensee MDPI, Basel, Switzerland., Funding: This research was funded by Junta de Andalucia, grant number RNM-5137. I.D.-M. acknowledges the pre-doctoral grant associated to project RNM-5137.

21 páginas.- 9 figuras.- 2 tablas.- 96 referencias, Mines, like other subterranean environments, have ecological conditions which allow the thriving of microorganisms. Prokaryotes and fungi are common inhabitants of mines, developing a metabolism suitable for growing in such inhospitable environments. The mine of Lousal, Portugal, is an interesting site for the study of microorganisms present in their galleries. Aerobiological studies resulted in the isolation of a Pseudomonas sp., strain L5B5, closely related to the opportunistic fish pathogen P. piscis MC042T, and to the soil bacteria P. protegens CHA0T, P. protegens Cab57, and P. protegens Pf-5. Strain L5B5 was able to inhibit the growth of the pathogenic bacteria Bacillus cereus, Staphylococcus aureus, and Acinetobacter baumanii, as well as the cave fungi Aspergillus versicolor, Penicillium chrysogenum, Cladosporium cladosporioides, Fusarium solani, and Ochroconis lascauxensis. In silico analyses based on de novo genome hybrid assembly and RNA-Seq, performing seven conditions based on culture and phases of growth resulted in the prediction and detection of genetic mechanisms involved in secondary metabolites, with the presence of a possible new gene cluster transcribed under the tested conditions, as well as feasible virulence factors and antimicrobial resistance mechanisms, The research was supported by the European Union’s project 0483_PROBIOMA_5_E, co-financed by the European Regional Development Fund within the framework of the Interreg V-A Spain-Portugal program (POCTEP) 2014–2020.