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Microbial diversity in lava tubes from Canary Islands (Spain) has never been explored thus far offering a unique opportunity to study subsurface microbiology. Abundant yellow coloured mats developing on coralloid speleothems in a lava tube from La Palma Islands were studied by next-generation sequencing and DNA/RNA clone library analyses for investigating both total and metabolically active bacteria. In addition, morphological and mineralogical characterization was performed by field emission scanning electron microscopy (FESEM), micro-computed tomography, X-ray diffraction and infrared spectroscopy to contextualize sequence data. This approach showed that the coralloid speleothems consist of banded siliceous stalactites composed of opal-A and hydrated halloysite. Analytical pyrolysis was also conducted to infer the possible origin of cave wall pigmentation, revealing that lignin degradation compounds can contribute to speleothem colour. Our RNA-based study showed for the first time that members of the phylum Actinobacteria, with 55% of the clones belonging to Euzebyales order, were metabolically active components of yellow mats. In contrast, the DNA clone library revealed that around 45% of clones were affiliated to Proteobacteria. Composition of microbial phyla obtained by NGS reinforced the DNA clone library data at the phylum level, in which Proteobacteria was the most abundant phylum followed by Actinobacteria.

Three actinobacteria strains isolated from a green biofilm covering the biodeteriorated interior walls of Vilar de Frades Church (Portugal) were studied using a polyphasic approach. The three strains were aerobic, non-spore forming and Gram-positive. Phylogenetically, the most closely related described species was Rubrobacter radiotolerans (94.2-94.3% and 81.9-82.5% similarities for the 16S rRNA and rpoB gene sequences, respectively). The fatty acid profile was dominated by anteiso-C(17:1) ω9c, and MK-8 was the only menaquinone present. These data clearly showed that the three strains could represent a new species, for which we propose the name Rubrobacter bracarensis sp. nov., with strain VF70612_S1(T) (=CECT 7924=DSMZ 24908) as the type strain., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

While microbial communities in limestone caves across the world are relatively understood, knowledge of the microbial composition in lava tubes is lagging behind. These caves are found in volcanic regions worldwide and are typically lined with multicolored microbial mats on their walls and ceilings. The Mount Etna (Sicily, S-Italy) represents one of the most active volcanos in the world. Due to its outstanding biodiversity and geological features, it was declared Natural Heritage of Humanity by the UNESCO in 2013. Despite the presence of more than 200 basaltic lava tubes, the microbial diversity of these hypogean systems has never been investigated so far. Here, we investigated bacterial communities in four lava tubes of Mount Etna volcano. Field emission scanning electron microscopy (FESEM) was carried out for the morphological characterization and detection of microbial features. We documented an abundant presence of microbial cells with different morphotypes including rod-shaped, filamentous, and coccoidal cells with surface appendages, resembling actinobacteria reported in other lava tubes across the world. Based on 16S rRNA gene analysis, the colored microbial mats collected were mostly composed of bacteria belonging to the phyla Actinomycetota, Pseudomonadota, Acidobacteriota, Chloroflexota, and Cyanobacteria. At the genus level, the analysis revealed a dominance of the genus Crossiella, which is actively involved in biomineralization processes, followed by Pseudomonas, Bacillus, Chujaibacter, and Sphingomonas. The presence of these taxa is associated with the carbon, nitrogen, and ammonia cycles, and some are possibly related to the anthropic disturbance of these caves. This study provides the first insight into the microbial diversity of the Etna volcano lava tubes, and expands on previous research on microbiology of volcanic caves across the world. [ABSTRACT FROM AUTHOR]

A number of mural paintings and building materials from monuments located in central and south Europe are characterized by the presence of an intriguing rosy discolouration phenomenon. Although some similarities were observed among the bacterial and archaeal microbiota detected in these monuments, their origin and nature is still unknown. In order to get a complete overview of this biodeterioration process, we investigated the microbial communities in saline environments causing the rosy discolouration of mural paintings in three Austrian historical buildings using a combination of culture-dependent and -independent techniques as well as microscopic techniques. The bacterial communities were dominated by halophilic members of Actinobacteria, mainly of the genus Rubrobacter. Representatives of the Archaea were also detected with the predominating genera Halobacterium, Halococcus and Halalkalicoccus. Furthermore, halophilic bacterial strains, mainly of the phylum Firmicutes, could be retrieved from two monuments using special culture media. Inoculation of building materials (limestone and gypsum plaster) with selected isolates reproduced the unaesthetic rosy effect and biodeterioration in the laboratory. [ABSTRACT FROM AUTHOR]

Due to their extraordinary properties, Actinobacteria can thrive in extreme environments, such as limestone caves, lava tubes and stone monuments. They grow forming microbial mats and speleothems on the walls and ceilings of caves, ranging from extensive coatings to small colonies (Riquelme et al. 2015). Their colour includes yellow, tan, orange, grey, pink and white. Recently, we have found abundant yellow and white-coloured bacterial mats coating the cave walls and secondary mineral deposits (speleothems) of lava tubes from La Palma Island, Canary Islands, Spain (Gonzalez-Pimentel et al. 2018) and Mount Etna in Catania (Sicily, Italy). Field Emission Scanning Electron Microscopy (FESEM) of the coloured microbial mats revealed abundant Actinobacteria-like cells, including a variety of filaments and spore structures with smooth surface ornamentation or profuse surface appendages. The DNA-/RNA-based analyses confirmed that these microbial mats are mainly composed of metabolically active Actinobacteria (Gonzalez-Pimentel et al. 2018).It is well known that Actinobacteria, mainly isolated from marine and soil ecosystems, are an important source of bioactive compounds, with Streptomyces ranking first with a huge number of bioactive secondary metabolites (Guo et al. 2015). These compounds, not only produced by Streptomyces but also by Bacillus, are very important to the industrial sector, such as pharmacology, biofuel and food industries, as well as to the conservation of stone cultural heritage, due to their antimicrobial properties (Silva et al. 2017). In the last decades, these sectors have intensified demands for exploring novel eco-friendly bioactive compounds, which stresses the need to investigate new groups of Actinobacteria from underexplored habitats. Yet, Actinobacteria from caves have not been the target of intensive screening for bioactive secondary metabolites. Hence, Actinobacterial-like microbial mats were collected and isolated from lava tubes in La Palma and Mount Etna to investigate their biotechnological potential. The screening of antimicrobial activity was based both on culture-dependent techniques using the agar diffusion assay and on metagenomics. Our study has showed that the strain Streptomyces sp. MZ0467C isolated from La Palma lava tube has antimicrobial activity against Microbacterium sp., Rhodococcus sp., Arthrobacter sp., Kocuria sp., Sphingomonas sp. and Paenibacillus sp. due to its ingenious adaptations and metabolic strategies to survive under extreme environmental conditions. This demonstrates that Actinobacteria from subterranean environments are promising sources of antibacterial compounds with interest for cultural heritage conservation.References:Gonzalez-Pimentel JL. et al. 2018. Yellow coloured mats from lava tubes of La Palma (Canary Islands, Spain) are dominated by metabolically active Actinobacteria. Sci. Rep. 8, 1944.Guo X. et al. 2015. Red soils harbor diverse culturable actinomycetes that are promising sources of novel secondary metabolites. Appl. Environ. Microbiol. 81, 3086-3103.Riquelme C. et al. 2015. Actinobacterial diversity in volcanic caves and associated geomicrobiological interactions. Front. Microbiol. 6, 1342.Silva M. et al. 2017. Green mitigation strategy for cultural heritage: Bacterial potential for biocide production. Environ. Sci. Pollut. Res. Int. 24, 4871-4881. [ABSTRACT FROM AUTHOR]

Etruscan and Roman necropolises are cemeteries with different types of burial practices, which are remarkable for their magnificent mural paintings. The conservation of these paintings is difficult, mainly derived from environmental factors and the opening of the tombs to public visits. One of the most complex phenomena observed on these sites is biodeterioration. The Circular Mausoleum tomb (Roman Necropolis of Carmona, Seville, Spain) is characterized by a heavy colonization of phototrophic microorganisms on the walls and ceiling. In addition, some areas near the ceiling exhibited an important number of violet stains of unknown origin. Previously, Agarossi (1994) found similar violet stains, attributed to streptomycetes, in two Etruscan tombs. However, no data on the species involved or the chemical structure of the pigments were reported. Here we show that the violet stains observed in the Circular Mausoleum are produced by a strain of the actinobacterium Streptomyces isolated from the mortar walls, and able to synthesize the same violet pigment in the laboratory. Streptomyces parvus MC05 was identified after whole genome study by means of Next Generation Sequencing methods. Pairwise comparisons carried out for genomes of the type strain of S. parvus DSM40348T and the MC05 strain were performed using Jspecies service. While results observed for ANIb, ANIm and Tetra indexes indicated that both strains belong to the same species, the S. parvus MC05 strain differs from the DSM20348T type strain in genome size and secondary metabolites production. AntiSMASH analysis implemented for both strains showed that the S. parvus MC05, isolated from the tomb, presents the capacity of synthesize bioactive compounds which were absents in the type strain.HPLC-MS of the culture extracts from S. parvus MC05 showed the production of three main granaticin derivatives (dihydrogranaticin A, granaticin A and granaticin B) in addition to minor products of other granaticin analogues. Granaticin pigments exhibited antibacterial activity, which justified the low number of clones of Gram positive bacteria found in the whole microbial community study. Gram negative bacteria were not affected (Dominguez-Moñino et al. 2017).To conclude, the origin of the violet stains in the walls of the Circular Mausoleum is the presence of S. parvus MC05, a member of the complex microbial community thriving on the tomb. In growing periods (rainy season) the bacterium excretes the soluble violet granaticins, compounds with a benzoisochromanequinone structure, which diffuses to the mortar and surrounding substrata in wetting periods.ReferencesAgarossi, G. 1994. In Studi e Ricerche sulla Conservazione delle Opere d'Arte Dedicati alla Memoria di Marcello Paribeni. F. Guidobaldi (ed.), CNRS, pp. 1−11. Dominguez-Moñino, I. et al. 2017. Science of the Total Environment 598, 889-899. [ABSTRACT FROM AUTHOR]