A. Dionísio

Biodiversity of cyanobacteria and green algae on monuments in the Mediterranean Basin: an overview

The presence and deteriorating action of micro-organisms on monuments and stone works of art have received considerable attention in the last few years. Knowledge of the microbial populations living on stone materials is the starting point for successful conservation treatment and control. This paper reviews the literature on cyanobacteria and chlorophyta that cause deterioration of stone cultural heritage (outdoor monuments and stone works of art) in European countries of the Mediterranean Basin. Some 45 case studies from 32 scientific papers published between 1976 and 2009 were analysed. Six lithotypes were considered: marble, limestone, travertine, dolomite, sandstone and granite. A wide range of stone monuments in the Mediterranean Basin support considerable colonization of cyanobacteria and chlorophyta, showing notable biodiversity. About 172 taxa have been described by different authors, including 37 genera of cyanobacteria and 48 genera of chlorophyta. The most widespread and commonly reported taxa on the stone cultural heritage in the Mediterranean Basin are, among cyanobacteria, Gloeocapsa, Phormidium and Chroococcus and, among chlorophyta, Chlorella, Stichococcus and Chlorococcum. The results suggest that cyanobacteria and chlorophyta colonize a wide variety of substrata and that this is related primarily to the physical characteristics of the stone surface, microclimate and environmental conditions and secondarily to the lithotype.

Bioreceptivity of building stones: a review.

In 1995, Guillitte defined bioreceptivity, a new term in ecology, as the ability of a material to be colonized by living organisms. Information about the bioreceptivity of stone is of great importance since it will help us to understand the material properties which influence the development of biological colonization in the built environment, and will also provide useful information as regards selecting stones for the conservation of heritage monuments and construction of new buildings. Studies of the bioreceptivity of stone materials are reviewed here with the aim of providing a clear set of conclusions on the topic. Definitions of bioreceptivity are given, stone bioreceptivity experiments are described, and finally the stone properties related to bioreceptivity are discussed. We suggest that a standardized laboratory protocol for evaluating stone bioreceptivity and definition of a stone bioreceptivity index are required to enable creation of a database on the primary bioreceptivity of stone materials.

Isolation of five Rubrobacter strains from biodeteriorated monuments

In the last few years, the microbial colonisation of mural paintings in ancient monuments has been attracting the attention of microbiologists and conservators. The genus Rubrobacter is commonly found in biodeteriorated monuments, where it has been reported to cause rosy discolouration. However, to date, only three species of this genus have been isolated, all from thermophilic environments. In this paper, we studied three monuments: the Servilia and Postumio tombs in the Roman Necropolis of Carmona (Spain), and Vilar de Frades church (Portugal), in search of Rubrobacter strains. In all cases, biodeterioration and the formation of efflorescences were observed, and five Rubrobacter strains were isolated. These isolates showed different physiology and migration in denaturing gradient gel electrophoresis, suggesting they might represent new species within this genus. The isolates reproduced some biodeterioration processes in the laboratory and revealed their biomediation in crystal formation.

Main geophysical techniques used for non-destructive evaluation in cultural built heritage: a review

Geophysical methodologies have been implemented, tested and validated as diagnostic and / or monitoring tools in artworks or historical monuments. They are non-destructive and can give an image of internal structure of investigated medium. This paper is a review about the main geophysical techniques applied to the study of cultural built heritage (excluding the archaeology field). A brief description of the used methodologies is presented, the main investigations done in this field are showed, the method or methods most appropriate to answer each problem (moisture detection, characterization of the materials, study of the structural continuity of the material, assessment of intervention’s effectiveness) are indicated and the main advances and gaps and future developments are also pointed out.

Reproducing stone monument photosynthetic-based colonization under laboratory conditions

In order to understand the biodeterioration process occurring on stone monuments, we analyzed the microbial communities involved in these processes and studied their ability to colonize stones under controlled laboratory experiments. In this study, a natural green biofilm from a limestone monument was cultivated, inoculated on stone probes of the same lithotype and incubated in a laboratory chamber. This incubation system, which exposes stone samples to intermittently sprinkling water, allowed the development of photosynthetic biofilms similar to those occurring on stone monuments. Denaturing gradient gel electrophoresis (DGGE) analysis was used to evaluate the major microbial components of the laboratory biofilms. Cyanobacteria, green microalgae, bacteria and fungi were identified by DNA-based molecular analysis targeting the 16S and 18S ribosomal RNA genes. The natural green biofilm was mainly composed by the Chlorophyta Chlorella, Stichococcus, and Trebouxia, and by Cyanobacteria belonging to the genera Leptolyngbya and Pleurocapsa. A number of bacteria belonging to Alphaproteobacteria, Bacteroidetes and Verrucomicrobia were identified, as well as fungi from the Ascomycota. The laboratory colonization experiment on stone probes showed a colonization pattern similar to that occurring on stone monuments. The methodology described in this paper allowed to reproduce a colonization equivalent to the natural biodeteriorating process.

The influence of inherent properties of building limestones on their bioreceptivity to phototrophic microorganisms.

The influence of open porosity, water absorption capillarity, water vapour permeability, surface roughness, stone pH and chemical composition on stone bioreceptivity to phototrophic microorganisms was assessed by means of a thorough stone characterisation with subsequent artificially inoculation of limestone samples with a multi-species phototrophic culture and placing them inside a growth chamber for 90 days. A principal component analysis and an analysis of variance (ANOVA) were carried out in order to evaluate the direct relationships between stone bioreceptivity and petrophysical properties. From the principal component analysis, two main components were obtained and assigned a petrophysical/photosynthetic biomass meaning. Stone bioreceptivity, quantified by the amount of chlorophyll a and intensity of chlorophyll a fluorescence present on the stone samples after 90 days-incubation, was included in both principal components. The first component was linked to the amount of chlorophyll a and was highly and linearly associated to capillarity and roughness, and less associated with open porosity and water vapour permeability. The second component, linked to the intensity of chlorophyll a fluorescence measured on the stone surfaces, was not linearly associated with the petrophysycal properties, showing the fallibility of this in vivo chlorophyll quantification technique on the estimation of photosynthetic biomass growing on stone materials, particularly when endolithic growth occurs.

Enigmatic reticulated filaments in subsurface granite.

In the last few years, geomicrobiologists have focused their researches on the nature and origin of enigmatic reticulated filaments reported in modern and fossil samples from limestone caves and basalt lava tubes. Researchers have posed questions on these filaments concerning their nature, origin, chemistry, morphology, mode of formation and growth. A tentative microbial origin has been elusive since these filaments are found as hollow tubular sheaths and could not be affiliated to any known microorganism. We describe the presence of similar structures in a 16th century granite tunnel in Porto, Northwest Portugal. The reticulated filaments we identify exhibit fine geometry surface ornamentation formed by cross-linked Mn-rich nanofibres, surrounded by a large amount of extracellular polymeric substances. Within these Mn-rich filaments we report for the first time the occurrence of microbial cells.

Siliceous Speleothems and Associated Microbe-Mineral Interactions from Ana Heva Lava Tube in Easter Island (Chile)

Coralloid-type speleothems were recorded on the ceiling of the Ana Heva lava tube in Easter Island (Chile). These speleothems were morphologically, geochemically and mineralogically characterized using a wide variety of microscopy and analytical techniques. They consist dominantly of amorphous Mg silicate and opal-A. Field emission scanning electron microscopy revealed a variety of filamentous and bacillary bacteria on the surface of the Ana Heva coralloid speleothems, including silicified filamentous microorganisms. Among them, intriguing reticulated filaments resemble those filaments documented earlier in limestone caves and lava tubes. The identification of silicified microorganisms on the coralloid speleothems from the Ana Heva lava tube suggests a possible role of these microorganisms in silica deposition.

Primary bioreceptivity of limestones used in southern European monuments

Abstract Different Mediterranean Basin limestones, like Calcário Ançã (Portugal), Calcário Lioz (Portugal), Piedra San Cristobal (Spain), Piedra Escúzar (Spain) and Pietra di Lecce (Italy), have been widely used as building materials in the European architecture. The aim of this study was focused on biodeterioration, mainly on evaluation of the primary bioreceptivity of those materials. A set of samples was inoculated with a cultured photosynthetic biofilm under laboratory conditions. Several assessment tools were applied to monitor the colonization overtime of the different lithotypes. After 3 months of incubation the colonization occurred endolithically in some lithotypes, namely Piedra San Cristobal and Piedra Escúzar. Spectrophotometric determination of chlorophyll a was a useful analytical technique to achieve the total amount of photosynthetic biomass on rock substrates, demonstrating that Piedra Escúzar and Calcário Lioz were the highest and lowest bioreceptive lithotypes, respectively. Microscopic and image analyses were essential to understand the stone colonization process and its pattern of distribution. Physical stone parameters and exposure conditions were shown to play an important role in the establishment and development of photosynthetic colonization.

Stone decay induced by fire on historic buildings: the case of the cloister of Lisbon Cathedral (Portugal)

Abstract Lisbon Cathedral, built in Late Romanesque style, is one of the most ancient Portuguese monuments. Its cloister was severely damaged by a fire that occurred in 1755 right after an earthquake. The aim of this investigation is to study stone thermal damage through the application of in situ and laboratory techniques. With this study it is possible to identify and characterize (chemically and mineralogically) the main thermal decay forms. Special attention is given to colour modification and granular disintegration. Through the application of an indirect ultrasound method it is verified that only a small number of stone blocks are relatively sound (11%). In terms of chromatic alteration, two factors are considered to explain heat-induced colour modification: the transformation of goethite into hematite and an increase in hematite single crystalline domains. It is also established that the most probable high-temperature range to which the cloister stones were subjected during the fire was 300–350 °C.