TY - CONF A1 - Fuentes, Elsa A1 - Prieto, Beatriz T1 - Effect of climate change on phototrophic biofilms colonizing granitic rocks and their biodeteriogenic activity. N2 - Material cultural heritage from NW of the Iberian Peninsula is linked to the use or presence of granite, whether in historical buildings, sculptures, archaeological sites or any other artistic manifestation. This resistant material has the ideal physical characteristics that favor its longevity, but, like any material exposed to the environment, it is susceptible to deterioration. To this respect, environmental changes due to climate change and global change may modify biological colonization-substrate interactions. The main objective of the present research was to analyse the effects of Climate Change, more specifically changes in water availability, increased temperature, increased CO2 concentrations and changes in UV-B radiation patterns, on the survival and biodeteriogenic activity of phototrophic biofilms on granite heritage and the consequences for the built heritage. Both field and laboratory work were carried out. Field studies were conducted on three rural churches to characterise the current biological colonization. A higher proportion of algae than cyanobacteria was identified, with the genera Trentepohlia and Desmococcus standing out among the former. The most common current fungi are lichenised fungi, but black fungal genera such as Catenulostroma, Rhinocladiella and Knufia were also identified. Bacterial genera related to the production of sphinganes, EPS, carotenoids and some causing the so-called pink discolouration are also present. The effect of climate change on the growth and physiological state of the organisms was analysed in the laboratory. For this purpose, mixed biofilms were developed in the laboratory and exposed to changing conditions of temperature, water availability, CO2 and UV-B. A reduction in growth was observed with increasing water restriction, although in the initial colonization process, rock bioreceptivity was shown to be a more important aspect in favoring anchorage and retention of organisms. Temperature was shown to have a growth-enhancing effect when water availability was low, whereas an increase in CO2 only resulted in increased growth under conditions of high-water availability. Increased UV-B produced a reduction in growth, especially at the highest dose (equivalent to the current dose of a west-facing wall in southern Portugal, which is considered as high), while little difference in the physiological state of the organisms was observed at the current doses in Galicia (considered as medium) and southern Ireland (considered as low). Clear changes in microbial composition were observed, with a trend towards increased or greater resistance of cyanobacteria to higher temperature, lower water availability and higher UV-B doses. In addition, all these changes in development, physiological and microbial composition had an effect on the predominant pigments, tending towards more yellowish-brownish colourations, and on the ability of the organisms to generate biodeterioration by affecting ion concentrations on water solutions by enhancing their consumption or adhesion which lead to granite deterioration. Future work should take into account the combination of a greater number of climatic parameters simultaneously and also the possible resistances associated with the nature of the SAB community studied. T2 - 19th International Biodeterioration and Biodegradation Symposium CY - Berlin, Germany DA - 09.09.2024 KW - Biofilm KW - Granite KW - Climate change KW - Biodeterioration KW - Cultural heritage PY - 2024 AN - OPUS4-62271 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Knisz, J. A1 - Eckert, R. A1 - Gieg, L. A1 - Koerdt, Andrea A1 - Lee, J. A1 - Silva, E. A1 - Skovhus, T. L. A1 - An Stepec, Biwen Annie A1 - Wade, S. A. T1 - Microbiologically Influenced Corrosion - More than just Microorganisms N2 - Microbiologically influenced corrosion (MIC) is a phenomenon of increasing concern which affects various materials and sectors of society. MIC describes the effects, often negative, that a material can experience due to the presence of microorganisms. Unfortunately, although several research groups and industrial actors worldwide have already addressed MIC, discussions are fragmented, while information sharing and willingness to reach out to other disciplines is limited. A truly interdisciplinary approach, that would be logical for this material/biology/chemistry-related challenge, is rarely taken. In this review we highlight critical non-biological aspects of MIC that can sometimes be overlooked by microbiologists working on MIC but are highly relevant for an overall understanding of this phenomenon. Here, we identify gaps, methods and approaches to help solve MIC related challenges, with an emphasis on the MIC of metals. We also discuss the application of existing tools and approaches for managing MIC and propose ideas to promote an improved understanding of MIC. Furthermore, we highlight areas where the insights and expertise of microbiologists are needed to help progress this field. KW - MIC KW - Biodeterioration KW - Biocorrosion KW - Interdisciplinarity KW - Multiple lines of evidence PY - 2023 DO - https://doi.org/10.1093/femsre/fuad041 SN - 0168-6445 VL - 47 IS - 5 SP - 1 EP - 70 PB - FEMS Microbiology Reviews AN - OPUS4-58066 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Voigt, Oliver A1 - Knabe, Nicole A1 - Nitsche, Sarah A1 - Erdmann, Eileen A1 - Schumacher, Julia A1 - Gorbushina, Anna T1 - An advanced genetic toolkit for exploring the biology of the rock‑inhabiting black fungus Knufia petricola N2 - Microcolonial black fungi are a group of ascomycetes that exhibit high stress tolerance, yeast-like growth and constitutive melanin formation. They dominate a range of hostile natural and man-made environments, from desert rocks and salterns to dishwashers, roofs and solar panels. Due to their slow growth and a lack of genetic tools, the underlying mechanisms of black fungi’s phenotypic traits have remained largely unexplored. We chose to address this gap by genetically engineering the rock-inhabiting fungus Knufia petricola (Eurotiomycetes, Chaetothyriales), a species that exhibits all characteristics of black fungi. A cell biological approach was taken by generating K. petricola strains expressing green or red fluorescent protein variants. By applying: (1) traditional gene replacement; (2) gene editing and replacement via plasmid-based or ribonucleoprotein (RNP)-based CRISPR/Cas9, and (3) silencing by RNA interference (RNAi), we constructed mutants in the pathways leading to melanin, carotenoids, uracil and adenine. Stable single and double mutants were generated with homologous recombination (HR) rates up to 100%. Efficient, partially cloning-free strategies to mutate multiple genes with or without resistance cassettes were developed. This state-of-the-art genetic toolkit, together with the annotated genome sequence of strain A95, firmly established K. petricola as a model for exploring microcolonial black fungi. KW - Subaerial biofilms KW - Biodeterioration KW - Fluorescent microscopy KW - CRISPR/Cas9 KW - RNA interference PY - 2020 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-518601 DO - https://doi.org/10.1038/s41598-020-79120-5 VL - 10 IS - 1 SP - 22021 PB - Springer Nature AN - OPUS4-51860 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Tonon, C. A1 - Breitenbach, Romy A1 - Voigt, Oliver A1 - Turci, F. A1 - Gorbushina, Anna A1 - Favero-Longo, S. E. T1 - Hyphal morphology and substrate porosity -rather than melanization- drive penetration of black fungi into carbonate substrates N2 - Due to their ability to penetrate, deteriorate and discolour stone surfaces, rock-inhabiting black fungi represent a remarkable issue for cultural heritage conservation. Black microcolonial fungi (MCF) can also adapt to different environmental conditions, by converting from yeast-like morphology to a peculiar meristematic development with swollen cells (torulose hyphae, TH), to extremely thin structures (filamentous hyphae, FH). Furthermore, black MCF produce protective pigments: melanin, dark pigment particularly evident on light stone surfaces, and carotenoids. Black fungi produce melanin in critical, oligotrophic conditions as well as constitutively. Melanin function is mostly related to stress resistance and the ability of fungi to generate appressorial turgor to actively penetrate plant cells in pathogenic species. An involvement of melanins in stone surface penetration has been suggested, but not experimentally proved. In this work, we tested the role of hyphal melanisation in penetration mechanisms on the model black fungus Knufia petricola A95 in lab conditions. The wild-type and three mutants with introduced targeted mutations of polyketide-synthases (melanin production) and/or phytoene dehydrogenase (carotenoid synthesis) were inoculated on artificial carbonate pellets (pressed Carrara marble powder) of different porosity. After 5, 10, 17 and 27 weeks, hyphal penetration depth and spread were quantified on periodic acid Schiff-stained cross-sections of the pellets, collecting measurements separately for TH and FH. Droplet assay of the mutants on different media were conducted to determine the role of nutrients in the development of different fungal morphologies. In our in vitro study, the hyphal penetration depth, never exceeding 200 μm, was proven to be consistent with observed penetration patterns on stone heritage carbonate substrates. Pellet porosity affected penetration patterns of TH, which developed in voids of the more porous pellets, instead than actively opening new passageways. Oppositely, the thin diameter of FH allowed their penetration independently of substrate porosity. Instead, the long-hypothesized crucial role of melanin in black MCF hyphal penetration should be rejected. TH were developed within the pellets also by melanin deficient strains, and melanized strains showed an endolithic component of non-melanized TH. FH were non-melanized for all the strains, but deeply penetrated all pellet types, with higher penetration depth probably related to their potential exploratory (nutrient-seeking) role, while TH may be more related to a resistance to surface stress factors. In the melanin deficient strains, the absence of melanin caused an increased penetration rate of FH, hypothetically related to an earlier necessity to search for organic nutrients. KW - Biodeterioration KW - Bioreceptivity KW - Black microcolonial fungi KW - Marble KW - Stone cultural heritage KW - Stress tolerance PY - 2020 DO - https://doi.org/10.1016/j.culher.2020.11.003 VL - 48 SP - 244 EP - 253 PB - Elsevier Masson SAS CY - Paris, Amsterdam AN - OPUS4-51933 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -