Holistic Approach to the Restoration of a Vandalized Monument: The Cross of the Inquisition, Seville City Hall, Spain

The Cross of the Inquisition, sculpted in 1903 and raised on a column with a fluted shaft and ornamented with vegetable garlands, is located in a corner of the Plateresque façade of the Seville City Hall. The Cross was vandalized in September 2019 and the restoration concluded in September 2021. A geological and microbiological study was carried out in a few small fragments. The data are consistent with the exposure of the Cross of the Inquisition to an urban environment for more than 100 years. During that time, a lichen community colonized the Cross and the nearby City Hall façades. The lichens, bryophytes and fungi colonizing the limestone surface composed an urban community, regenerated from the remains of the original communities, after superficial cleaning of the limestone between 2008 and 2010. This biological activity was detrimental to the integrity of the limestone, as showed by the pitting and channels, which evidence the lytic activity of organisms on the stone surface. Stone consolidation was achieved with Estel 1000. Preventol RI80, a biocide able to penetrate the porous limestone and active against bacteria, fungi, lichens, and bryophytes, was applied in the restoration.The research was funded by Atelier Samthiago and the restoration of the Cross of the Inquisition was supported by the Seville City Hall

Role of subterranean microbiota in the carbon cycle and greenhouse gas dynamics

Subterranean ecosystems play an active role in the global carbon cycle, yet only a few studies using indirect methods have focused on the role of the cave microbiota in this critical cycle. Here we present pioneering research based on in situ real-time monitoring of CO2 and CH4 diffusive fluxes and concurrent δ13C geochemical tracing in caves, combined with 16S microbiome analysis. Our findings show that cave sediments are promoting continuous CH4 consumption from cave atmosphere, resulting in a significant removal of 65% to 90%. This research reveals the most effective taxa and metabolic pathways in consumption and uptake of greenhouse gases. Methanotrophic bacteria were the most effective group involved in CH4 consumption, namely within the families Methylomonaceae, Methylomirabilaceae and Methylacidiphilaceae. In addition, Crossiella and Nitrosococcaceae wb1-P19 could be one of the main responsible of CO2 uptake, which occurs via the Calvin-Benson-Bassham cycle and reversible hydration of CO2. Thus, syntrophic relationships exist between Crossiella and nitrifying bacteria that capture CO2, consume inorganic N produced by heterotrophic ammonification in the surface of sediments, and induce moonmilk formation. Moonmilk is found as the most evolved phase of the microbial processes in cave sediments that fixes CO2 as calcite and intensifies CH4 oxidation. From an ecological perspective, cave sediments act qualitatively as soils, providing fundamental ecosystem services (e.g. nutrient cycling and carbon sequestration) with direct influence on greenhouse gas emissions.This work was supported by the Spanish Ministry of Science, Innovation through project PID2019-110603RB-I00, MCIN/AEI/FEDER, UE/10.13039/501100011033 and with collaboration of projects RTI2018-099052-B-I00 and PID2020-114978GB-I00. This research has also received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 844535 — MIFLUKE

Microbial Activity in Subterranean Ecosystems: Recent Advances

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

Crossiella, a Rare Actinomycetota Genus, Abundant in the Environment

The genus Crossiella contains two species, C. equi, causing nocardioform placentitis in horses, and C. cryophila, an environmental bacterium. Apart from C. equi, which is not discussed here, environmental Crossiella is rarely reported in the literature; thus, it has not been included among “rare actinobacteria”, whose isolation frequency is very low. After C. cryophila, only five reports cover the isolation of Crossiella strains. However, the frequency of published papers on environmental Crossiella has increased significantly in recent years due to the extensive use of next-generation sequencing (NGS) and a huge cascade of data that has improved our understanding of how bacteria occur in the environment. In the last five years, Crossiella has been found in different environments (caves, soils, plant rhizospheres, building stones, etc.). The high abundance of Crossiella in cave moonmilk indicates that this genus may have an active role in moonmilk formation, as evidenced by the precipitation of calcite, witherite, and struvite in different culture media. This review provides an overview of environmental Crossiella, particularly in caves, and discusses its role in biomineralization processes and bioactive compound production.This research was funded by the projects PID2020-114978GB-I00 and PID2019-110603RB-I00. The Malaga City Council financed data from the archaeological basement of the Thyssen Museum of Malaga through a conservation contract for this Roman site

The deterioration of Circular Mausoleum, Roman Necropolis of Carmona, Spain

The Circular Mausoleum tomb in the Roman Necropolis of Carmona was carved on a calcarenite sequence in an ancient quarry located in the town of Carmona, Southern Spain. This rock-cut tomb, representative of Roman burial practices, currently suffers from serious deterioration. A detailed survey over several years permitted the identification of the main tomb's pathologies and damaging processes, which include loss of material (scaling, flaking, granular disintegration), surface modifications (efflorescences, crusts and deposits) and extensive biological colonization. The results obtained in this study indicated that anthropogenic changes were largely responsible and enhanced the main alteration mechanisms observed in the Circular Mausoleum. Based on the deterioration diagnosis, effective corrective actions were proposed. This study shows that any conservative intervention in the interior of the tomb should be preceded by accurate in situ measurements and laboratory analyses to ascribe the source of the deterioration damages and thus designing effective treatments.This research was funded by the Consejeria de Cultura, Junta de Andalucia, the Spanish Ministry of Science and Innovation project CGL2010-17183, and the program Torres Quevedo (PTQ 13-06296 and PTQ 12-05601)

Deterioro asociado a películas microbianas en la Iglesia de San Roque, Campeche, México

Este trabajo se ha llevado a cabo en el marco del proyecto FOMIX CONACYT-Gobierno del Estado de Campeche: "Influencia del entorno urbano en los procesos de degradación de edificios militares y religiosos de la época colonial en la ciudad de Campeche", clave: CAMP2005-C01-028

Combining stable isotope (δ13C) of trace gases and aerobiological data to monitor the entry and dispersion of microorganisms in caves

Altamira Cave (north of Spain) contains one of the world's most prominent Paleolithic rock art paintings, which are threatened by a massive microbial colonization of ceiling and walls. Previous studies revealed that exchange rates between the cave and the external atmosphere through the entrance door play a decisive role in the entry and transport of microorganisms (bacteria and fungi) and nutrients to the interior of the cave. A spatial-distributed sampling and measurement of carrier (CO2) and trace (CH4) gases and isotopic signal of CO2 (δ13C) inside the cave supports the existence of a second connection (active gas exchange processes) with the external atmosphere at or near the Well Hall, the innermost and deepest area of the cave. A parallel aerobiological study also showed that, in addition to the entrance door, there is another connection with the external atmosphere, which favors the transport and increases microorganism concentrations in the Well Hall. This double approach provides a more complete knowledge on cave ventilation and revealed the existence of unknown passageways in the cave, a fact that should be taken into account in future cave management.This research was supported by the Spanish Ministry of Sciences and Innovation, project CGL2010-17108/BTE. EG-A is supported by a CSIC JAE-Predoctoral grant. SC benefits of a postdoctoral fellowship from the Spanish Ministry of Science and Innovation, research programme Juan de la Cierva. AF-C was funded by a postdoctoral fellowship the JAE-Doc Program (CSIC). AZM was supported by FCT grant SFRH/BPD/63836/2009. This is a TCP-CSD 2007–00058 paper

A study on the state of conservation of the Roman Necropolis of Carmona (Sevilla, Spain)

The Roman Necropolis of Carmona (Sevilla, Spain) has experienced a severe deterioration since the accidental discovery in 1868 and subsequent archaeological excavations starting in 1882. To this deterioration contributed the location in a soft calcarenite rock quarry, the adverse environmental conditions and the numerous and extensive wrong archaeological and managing interventions along more than a century. The cultural, artistic and religious importance of this Necropolis converts the protection and conservation of this archaeological site in a major issue. This work present the data obtained in a multidisciplinary research were geology, geomorphology, micro-environmental and climatic monitoring, rock petrophysical characterization, description of weathering forms and biological colonization were considered in order to propose corrective measures to minimize deterioration.This research was funded by the Consejería de Cultura, Junta de Andalucía, the Spanish Ministry of Science and Innovation, project CGL2010-17183 and the CSIC project 201230E125

Recolonization of mortars by endolithic organisms on the walls of San Roque church in Campeche (Mexico): A case of tertiary bioreceptivity

San Roque church (Campeche, Mexico) was built at the end of the 17th century with a micritic limestone and lime mortar in baroque style. In 2005 the church exhibited heavy biodeterioration associated with the development of extensive dark green phototrophic-based biofilms. Several cyanobacteria belonging to the order Chroococcales and lichenized fungi (Toninia nordlandica, Lobaria quercizans, Lecanora subcarnea, Cystocoleus ebeneus) were predominant in the dark biofilm samples, as revealed by DNA-based molecular techniques. In 2009, a cleaning and restoration intervention was adopted; however, after few months, microbial recolonization started to be noticeable on the painted church walls, representing an early phototrophic-based recolonization. According to molecular analysis, scanning electron microscopy observations and digital image analysis of cross sections, new phototrophic-based colonization, composed of cyanobacteria and bryophytes, developed mainly beneath the restored mortars. The intrinsic properties of the mortars, the tropical climate of Campeche and the absence of a biocide treatment in the restoration protocol influenced the recolonization of the church façades and enhanced the overall rate of deterioration in a short-term period.This work was carried out under the Mexico Research Project “Influencia del entorno urbano en los procesos de degradación de edificios militares y religiosos de la época colonial en la ciudad de Campeche” (FOMIX CONACYT, CAMP2005-C01-028) and Junta de Andalucía through funding to research team RMN 201. This is a TCP CSD2007-00058 paper. SC benefited from a postdoctoral fellowship from the Spanish Ministry of Science and Innovation, Juan de la Cierva research program. This work was also supported by Fundação para a Ciência e a Tecnologia (Portugal) with a postdoctoral grant to AZM (SFRH/BPD/63836/2009)

Microclimate, airborne particles, and microbiological monitoring protocol for conservation of rock-art caves: The case of the world-heritage site La Garma cave (Spain)

Cave heritage is often threatened by tourism or even scientific activities, which can lead to irreversible deterioration. We present a preventive conservation monitoring protocol to protect caves with rock art, focusing on La Garma Cave (Spain), a World Heritage Site with valuable archaeological materials and Palaeolithic paintings. This study assessed the suitability of the cave for tourist use through continuous microclimate and airborne particles monitoring, biofilm analysis, aerobiological monitoring and experimental visits. Our findings indicate several factors that make it inadvisable to adapt the cave for tourist use. Human presence and transit within the cave cause cumulative effects on the temperature of environmentally very stable and fragile sectors and significant resuspension of particles from the cave sediments. These environmental perturbations represent severe impacts as they affect the natural aerodynamic control of airborne particles and determine bacterial dispersal throughout the cave. This monitoring protocol provides part of the evidence to design strategies for sustainable cave management.This research was supported by the Consejeria de Universidades, Igualdad, Cultura y Deporte del Gobierno de Cantabria and the Spanish Ministry of Science and Innovation through projects PID2019-110603RB-I00 and PID2020-114978 GB-I00, funded by MCIN/AEI/10.13039/501100011033