Persepolis, a UNESCO World Heritage Site in south-western Iran, dates back to more than 2500 years ago, and is colonized by a great diversity of lichen-forming fungi. A survey of the lichen-forming fungi revealed a species abundant in different areas of the cultural site, which turned out to be a new species of the genus Circinaria. The new species, Circinaria persepolitana, is introduced and described on the basis of morphological and molecular data. Circinaria persepolitana is characterized by having a crustose thallus, rimose to areolate, usually with bullate areoles, with an olive green to olive-brown surface and angular to elongate areoles in the marginal zone. Phylogenetic analyses including other species of the genus showed that the new species is phylogenetically close to C. mansourii, C. ochracea and C. reptans. We propose a new combination of Circinaria reptans (Looman) Sohrabi, Owe-Larsson & Paukov. The bioweathering capacity of the new species was also analyzed by scanning electron microscopy, examining the interface between the lichen thallus and the lithic substratum to assess its potential threat to the conservation of heritage surfaces. We found this species to be a potential biodeteriogenic agent, as thalli were closely attached to the lithic substratum and biogeophysical and biogeochemical changes at the rock surface could be associated with the colonization.

Environmental control strategies are commonly practised to limit biodeterioration issues threatening indoor cultural heritage objects, while they are still poorly exploited for the conservation of outdoor stone heritage surfaces, including rock art. In this study, we evaluated the environmental factors driving the diversity and abundance of lithobiontic communities in the Rock Engravings National Park of Naquane (UNESCO WHS n. 94, Italy). The survey considered 23 rocks that had been cleaned in the last three (3YC) or twelve (12YC) years or more than 40 years ago (NRC). A cyanobacteria-dominated biofilm and lichens (37 taxa) were the most widespread and abundant lithobiontic components, prevailing on 3YC–12YC and NRC rocks, respectively. On the latter, a turnover of xerophytic and meso-hygrophytic lichen communities was observed. On 3YC–12YC rocks lichen colonization, if present, was limited to nitrophytic species, including common epiphytes from surrounding trees and a small number of meso-hygrophytic species, with a prevalence of asexual reproductive strategies. Multivariate analyses including environmental parameters (canonical correspondence analyses) indicated that tree cover and the presence of bare or vegetated ground upstream of the rocks, probably prolonging wetness and providing nutrients by water transport, are the factors mostly related to the microbial and lichen recolonization of 3YC–12YC surfaces. On this basis, an experiment on preventive conservation was conducted, consisting of a new cleaning of a strongly recolonized 3YC surface combined with the building of a small wall to protect part of the rock from prolonged water fluxes. The fluorimetric and colorimetric monitoring of the rock surface, carried out 40 months after this new cleaning intervention, displayed recolonization on the unprotected area only, indicating the potential of preventive conservation strategies in outdoor environments.

Elemental behaviour, during the process of weathering of glazed sekishu roof-tiles affected by Lecidea s.lat. sp. (a crustose lichen), was investigated using optical and fluorescence microscopy, field emission scanning electron microscopy (FE-SEM) and transmission electron microscopy. Sekishu roof tiles have an opaque reddish brown glaze on their surfaces which consist of an alkali feldspar-type X-ray amorphous glass recrystallized at 1200°C. Optical and fluorescence microscopy revealed the presence of corrosion pits (at a depth of ∼50 μm) at the lichen-glaze interface. Elemental mapping by FE-SEM identified the concentrations of Ti and Fe in the section of the glazed tile analysed. The behaviour of C was correlated with those elements, suggesting the possibility of biomineralization.

Fungal deterioration of frescoes was studied in situ on a selected Serbian church, and on a laboratory model, utilizing standard and newly implemented microscopy techniques. Scanning electron microscopy (SEM) with energy-dispersive X-ray confirmed the limestone components of the plaster. Pigments used were identified as carbon black, green earth, iron oxide, ocher, and an ocher/cinnabar mixture. In situ microscopy, applied via a portable microscope ShuttlePix P-400R, proved very useful for detection of invisible micro-impairments and hidden, symptomless, microbial growth. SEM and optical microscopy established that observed deterioration symptoms, predominantly discoloration and pulverization of painted layers, were due to bacterial filaments and fungal hyphal penetration, and formation of a wide range of fungal structures (i.e., melanized hyphae, chlamydospores, microcolonial clusters, Cladosporium-like conidia, and Chaetomium perithecia and ascospores). The all year-round monitoring of spontaneous and induced fungal colonization of a “mock painting” in controlled laboratory conditions confirmed the decisive role of humidity level (70.18±6.91% RH) in efficient colonization of painted surfaces, as well as demonstrated increased bioreceptivity of painted surfaces to fungal colonization when plant-based adhesives (ilinocopie, murdent), compared with organic adhesives of animal origin (bone glue, egg white), are used for pigment sizing.

The suitability of image analysis by colour-based pixel classification to quantify lichen colonization on the surface of and within marble, travertine and mortar stonework has been investigated. High resolution images of lichenized stonework surfaces were acquired at different field sites using a scanner, thus avoiding invasive surveys, and the percentage cover of lichen species was subsequently measured in the laboratory using dedicated software. Furthermore, microphotographs of polished cross-sections of lichenized marble, travertine and mortar, stained using the periodic acid-Schiff (PAS) method to visualize hyphae, were produced by the same software to quantify hyphal spread within the substratum, a parameter which can be used more successfully than the commonly used depth of hyphal penetration to quantify how much the lichen has affected the conservation of a stone substratum. Significant statistical differences in hue, saturation and intensity (HSI) of the lichen thalli and PAS-stained hyphae, with respect to the lithic substrata, allowed the software to discriminate and quantify the lichen species cover on, and hyphal spread within, the three investigated lithotypes. Since such a quantitative approach highlights the volume of influence of lichens on stonework, where bioweathering processes are likely to develop, it could be used to support decisions on the preservation of our stone cultural heritage.

The anatomy of the endolithic, calcicolous lichen Verrucaria rubrocincta Breuss is described using optical and scanning electron microscopy. This lichen is locally abundant in caliche plates of open desert pavements in the Sonoran Desert of south-western Arizona. The endolithic growth of V. rubrocincta is distinctly layered. The upper layer is a fine-grained calcite (micrite). This layer is sparsely penetrated by hyphae and therefore cannot be interpreted as a lithocortex sensu stricto. Beneath the micrite is the photobiont layer. Below this photobiont layer hyphae form a pseudomedulla penetrating up to 1 cm into the caliche. Calcium oxalates occur in the pseudomedulla but are absent from uninhabited caliche. The analogy of a greenhouse describes the ecophysiological adaptations of this endolithic growth to the environmental extremes of the Sonoran Desert. The micrite acts as a roof and the photobiont layer and pseudomedulla represent the greenhouse interior. Verrucaria rubrocincta has thus evolved a strategy to successfully establish and survive within an extreme environment. Our study illustrates biodeterioration and biomineralization processes acting simultaneously within a single lichen species. Mineralogical evidence suggests that the micrite on the thallus surface is biologically induced. The hyphae of the lichen biodeteriorate the caliche thus forming the pseudomedulla. Simultaneously with this process micrite forms at the surface protecting the thallus from exposure and counter-balancing rock degradation. These combined effects of biodeterioration and biomineralization do not markedly accelerate erosion because inhabited and uninhabited areas of the same plate show similar surface heights.