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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

16 páginas.- 5 graficas.- 6 tablas.- 91 referencias.- The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2019.02588/full#supplementary-material, A PAHs-contaminated industrial soil was analyzed using PCR amplification of the gene 16S ribosomal RNA for the detection and identification of different isolated bacterial strains potentially capable of degrading PAHs. Novel degrader strains were isolated and identified as Achromobacter xylosoxidans 2BC8 and Stenotrophomonas maltophilia JR62, which were able to degrade PYR in solution, achieving a mineralization rate of about 1% day–1. A. xylosoxidans was also able to mineralize PYR in slurry systems using three selected soils, and the total extent of mineralization (once a plateau was reached) increased 4.5, 21, and 57.5% for soils LT, TM and CR, respectively, regarding the mineralization observed in the absence of the bacterial degrader. Soil TM contaminated with PYR was aged for 80 days and total extent of mineralization was reduced (from 46 to 35% after 180 days), and the acclimation period increased (from 49 to 79 days). Hydroxypropyl-ß-cyclodextrin (HPBCD) was used as a bioavailability enhancer of PYR in this aged soil, provoking a significant decrease in the acclimation period (from 79 to 54 days) due to an increase in PYR bioavailable fraction just from the beginning of the assay. However, a similar global extension of mineralization was obtained. A. xylosoxidans was then added together with HPBCD to this aged TM soil contaminated with PYR, and the total extent of mineralization decreased to 25% after 180 days, possibly due to the competitive effect of endogenous microbiota and the higher concentration of PYR in the soil solution provoked by the addition of HPBCD, which could have a toxic effect on the A. xylosoxidans strain. © Copyright © 2019 Villaverde, Láiz, Lara-Moreno, González-Pimentel and Morillo., This work was supported by the Spanish Ministry of Economy and Competitiveness under the research project CTM2O13-42599-R (AEI/FEDER, UE) and Junta de Andalucía Proyect RNM 894. AL-M acknowledges the Spanish Ministry of Education. Culture and Sports for her FPU fellowship (FPU15/03740).

2 páginas.- 17 referencias.- Data availability. This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under the accession number VSZQ00000000. The version described in this paper is the first version, VSZQ01000000. BioProject and raw data are available at the accession numbers PRJNA562075 and SRR10059654, respectively., Streptomyces parvus strain C05 was isolated from the walls of a Roman tomb located in Carmona, Seville, Spain. Subsequent studies determined the capability of this strain for producing granaticins. Here, we report the 8.7-Mbp draft genome sequence for this bacterium., Financial support was obtained through project 0483_PROBIOMA_5_E, cofinanced by the European Regional Development Fund within the framework of the Interreg V-A Spain-Portugal program (POCTEP) 2014 –2020.

Póster presentado en EGU 2019, Viena (Austria) 7 al 12 de abril de 2019. Puede ver el Abstract en https://meetingorganizer.copernicus.org/EGU2019/EGU2019-9604.pdf, 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 howed 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.

Póster presentado en EGU 2019, Viena (Austria) 7 al 12 de abril de 2019. Puede ver el Abstract en https://meetingorganizer.copernicus.org/EGU2019/EGU2019-8949.pdf, 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 (DominguezMoñino 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., References - Agarossi, 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.