TY - JOUR A1 - Stohl, Leonie A1 - Tonon, Chiara A1 - Cook, Jake A1 - Gorbushina, Anna A1 - Dehn, Frank A1 - von Werder, Julia A1 - Manninger, Tanja T1 - Understanding bioreceptivity of concrete: material design and characterization N2 - The climate crisis is driving an increasing demand for ecologically oriented concepts. In the building sector, this demand includes not only the use of environmentally friendly materials but also the greening of urban areas. One promising approach is the development of bioreceptive concrete façades, which support the growth of green biofilms directly on their surfaces. These innovative façades are anticipated to deliver benefits comparable to those of macroscopically greened façades, such as enhanced biodiversity and improved air quality, while offering the advantages of being more self-sustaining and stable systems once fully established. However, the development of bioreceptive concrete presents substantial challenges. Due to the interdisciplinarity and novelty of this field, standardized methods for material characterization and bioreceptivity assessment are currently lacking. This study proposes an approach for evaluating surface properties crucial for bioreceptivity, developed on differently structured samples of ultra-high-performance concrete (UHPC). Existing methods and standards from concrete technology are critically reviewed and, where necessary, modified to meet the unique requirements of measuring bioreceptive material properties. Special attention is given to the surface pH value and water retention characteristics, as these are essential for promoting microbial growth and ensuring the long-term stability of green biofilms. The observed surface characteristics vary according to the imprinted surface structures, offering a spectrum of material properties and enabling the evaluation of their impact on bioreceptivity. The findings presented form the foundation for subsequent laboratory weathering experiments, which will be discussed in a complementary publication. KW - Novel methods KW - Bioreceptivity testing PY - 2025 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-655023 DO - https://doi.org/10.1617/s11527-025-02863-y SN - 1359-5997 VL - 58 SP - 1 EP - 18 PB - Springer Science and Business Media LLC AN - OPUS4-65502 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 - TY - JOUR A1 - Vogel, Christian A1 - Urban, Klaus A1 - Ackerhans, C. A1 - Gorbushina, Anna T1 - Analysis of Carbon and Nitrogen from Atmospheric Sources by Bulk Deposition Sampling at various locations in Germany N2 - Atmospheric deposition of particulate matter is an important indicator of air pollution and a significant factor in material surface fouling. The elemental composition of this nutrient-containing dust depends largely on the exposure region and time as well as on climate. Therefore, in this paper we report an analysis of atmospheric pollutions with a self-made low-cost bulk deposition sampler directed at sampling deposition via air transport and rainfall. We used the device in diverse environments - thus comparing an urban region, an area surrounded by forest and an area mainly dominated by agriculture. The total organic carbon (TOC) and total nitrogen (TN) amounts were selected as indicator parameters and analyzed in a biweekly rhythm for three and a half and two years, respectively. The TOC value responded to particulate matter in the urban area, especially significant were the influences of the New Year's firework in urban and pollen in the rural forest area. In contrast, the TN value was more under the influence of the nitrogen emissions in the agriculture-dominated area. However, the TN value did not correlate with the NOx values in the urban area because the atmospheric nitrogen emissions in the city might originate from various emission sources. Summarizing, the TOC and TN values of the self-made low-cost bulk deposition sampler were in good agreement with environmental events of their immediate surrounding. Moreover, the selected containers and sampling procedures are universally applicable to monitor and analyze organic as well as inorganic parameters (e.g. metal ions) of atmospheric deposition. KW - Passive sampling KW - Biomonitoring KW - Air Analysis PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-609373 DO - https://doi.org/10.1016/j.envadv.2024.100583 SN - 2666-7657 VL - 17 SP - 1 EP - 9 PB - Elsevier Ltd. AN - OPUS4-60937 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 - Wen, Keqing A1 - Gorbushina, Anna A1 - Schwibbert, Karin A1 - Bell, Jérémy T1 - Microfluidic platform with precisely controlled hydrodynamic parameters and integrated features for generation of microvortices to accurately form and monitor biofilms in flow N2 - Microorganisms often live in habitats characterized by fluid flow, and their adhesion to surfaces in industrial systems or clinical settings may lead to pipe clogging, microbially influenced corrosion, material deterioration, food spoilage, infections, and human illness. Here, a novel microfluidic platform was developed to investigate biofilm formation under precisely controlled (i) cell concentration, (ii) temperature, and (iii) flow conditions. The developed platform central unit is a single-channel microfluidic flow cell designed to ensure ultrahomogeneous flow and condition in its central area, where features, e.g., with trapping properties, can be incorporated. In comparison to static and macroflow chamber assays for biofilm studies, microfluidic chips allow in situ monitoring of biofilm formation under various flow regimes and have better environment control and smaller sample requirements. Flow simulations and experiments with fluorescent particles were used to simulate bacteria flow in the platform cell for calculating flow velocity and direction at the microscale level. The combination of flow analysis and fluorescent strain injection in the cell showed that microtraps placed at the center of the channel were efficient in capturing bacteria at determined positions and to study how flow conditions, especially microvortices, can affect biofilm formation. The microfluidic platform exhibited improved performances in terms of homogeneity and robustness for in vitro biofilm formation. We anticipate the presented platform to be suitable for broad, versatile, and high-throughput biofilm studies at the microscale level. KW - Topographical pattern KW - E. coli KW - Fluorescence KW - Bacteria trapping KW - Particle velocimetry PY - 2024 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-610450 DO - https://doi.org/10.1021/acsbiomaterials.4c00101 SN - 2373-9878 VL - 10 IS - 7 SP - 4626 EP - 4634 PB - ACS Publ. CY - Washington, DC AN - OPUS4-61045 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -