Bacterial diversity on rock surface of the ruined part of a French historic monument: The Chaalis abbey

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Highlights

  • Whatever the degree of deterioration stone is always covered by a microbial biofilm.

  • Presence of bacteria well adapted to oligotrophic conditions.

  • Presence of bacteria that may damage the stone support.

Abstract

The aim of the present study was to analyze the epilithic bacterial diversity of the ruins of the Chaalis Abbey, France. To do this, stone samples were collected at the surface of three areas presenting different macroscopic characteristics: a Discolored Area (DCA), a Damaged Area (DMA) and an Undamaged Area (UDMA). The bacterial diversity investigation after DNA extraction and amplified ribosomal DNA restriction analysis targeting the 16S rRNA genes revealed that clone libraries were dominated by Alphaproteobacteria and Actinobacteria. The main part of recovered phylotypes belonged to genera previously associated with stone colonization and/or biodeterioration. Indeed most abundant Alphaproteobacteria phylotypes belonged to the genus Sphingomonas and to the species Bosea thiooxidans, and most abundant Actinobacteria phylotypes belonged to the genera Rubrobacter and Arthrobacter. Members of Cyanobacteria, Bacteroidetes, TM7, Betaproteobacteria, Deinococcus, Acidobacteria and Chloroflexi were also recovered but to a lesser extent. In conclusion, these results suggest that the ruins of the Chaalis Abbey are covered by bacterial communities mainly composed of Alphaproteobacteria and Actinobacteria able to colonize and, for some of them, to deteriorate stone.

Introduction

The surface of stone monuments exposed to the outside is continually subjected to physicochemical environmental factors that act as weathering agents: wind, temperature, rain, relative humidity, condensation, and air pollution. These environmental factors can more or less alter the rocky substrate in accordance to its mineralogical composition and structure (Miller et al., 2012). Biological factors also impact stone deterioration processes. Biodeterioration is defined as “any undesirable change in a material brought about by vital activities of organisms” (Hueck, 2001). Microorganisms that play a potential role in biodeteriorative processes are autotrophic and heterotrophic bacteria, fungi, algae, lichens and protozoa (Gómez-Alarcón et al., 1995, Tomaselli et al., 2000, Gaylarde and Gaylarde, 2005, Sterflinger, 2010). This consortium of heterogeneous microbial species forms a biofilm where microbial cells are embedded in extracellular polymeric substances (EPS) (Gorbushina, 2007, Di Martino, 2016). The interaction of microorganisms and building stone is complex and has been reviewed extensively (Gaylarde and Morton, 1999, Griffin et al., 1991, Kumar and Kumar, 1999, Mihajlovski et al., 2015, Saiz-Jimenez, 1999, Saiz-Jimenez, 2001, Sand and Bock, 1991, Scheerer et al., 2009, Steiger et al., 2011, Warscheid and Braams, 2000, Urzì, 2004). Indeed, the stone surface of monuments is a common habitat for a wide range of microorganisms. Microbial communities development occurs at the interface between the stone and the atmosphere, on top of or inside the rock (de los Rios et al. 2004). Microorganisms can cause various damages on stone surfaces. The biodeterioration impact of microorganisms can be classified into three categories that may occur separately or simultaneously, namely: (i) biophysical (ii) biochemical and (iii) aesthetic. These different categories have been described and reviewed by several authors (e.g. Allsopp et al., 2004, Gaylarde et al., 2003, Griffin et al., 1991, Kumar and Kumar, 1999, Saiz-Jimenez, 2001, Scheerer et al., 2009, Warscheid and Braams, 2000). Biophysical deteriorations refer to actions that directly affect the material components and its mechanical properties, such as the fungal hyphal growth into the mineral support (Gadd, 2007, Hoffland et al., 2004). Biochemical deteriorations can be divided into: assimilatory and dissimilatory processes. Assimilatory process occurs when microorganisms use the mineral support as a source of nutrients and energy. This can results in the modifications of the material properties. In dissimilatory processes, microbial metabolism products can chemically react with the mineral support and thus alter the latter. Finally, even if biophysical and biochemical biodeteriorative processes can lead to aesthetic deterioration, the latter is often associated to the discoloration of the support. These discoloration can be caused by pigments released from, or contained within, the microorganisms and also from the biofilm EPS that facilitate entrapment of airborne particles, aerosols, minerals, and organic compounds and thus increase the dirty appearance of the substrate (Kemmling et al., 2004). Even if the materials properties are not initially affected, as time passes the fouling biofilm impact may exceed the purely aesthetic consideration and may cause physicochemical damages to the support. The concept of bioreceptivity was defined by Guillitte (1995) as “the aptitude of a material to be colonized by one or several groups of living organisms without necessarily undergoing any biodeterioration” or as “the totality of material properties that contribute to the establishment, anchorage and development of fauna and/or flora” (Guillitte, 1995). Thus, the intrinsic properties of the stone material itself, i.e., surface roughness, porosity and chemical composition also influence microbial colonization and deterioration processes (Miller et al., 2012). In addition, as the intrinsic characteristics of the stone materials change over time, bioreceptivity is not considered as a static property and several types of bioreceptivity exit (Guillitte, 1995). Thus, the state of conservation of building materials can also affect its bioreceptivity (Ortega-Calvo et al., 1995) as well as the accumulation of exogenous deposits such as soil, dust or organic particles (Guillitte, 1995). Carbonate stones such as marble and limestones are particularly sensitive to deterioration by physical and chemical weathering and biodeterioration associated to the presence of macro-and microorganisms (Lamenti et al., 2000, Saiz-Jimenez, 2001). As a large number of the most important cultural heritage structures are built with carbonate stones, their conservation for long period of times is at risk (Gauri and Bandyopadhyay, 1999). Finally, the colonization of the outdoor surface of monuments is heterogeneous depending on the weathering level of the studied area (de Los Ríos et al., 2009, Qi-Wang et al., 2011). This suggests that these variable weathering levels may be due to different aggressive colonizers or that they may lead to the colonization by various kinds of microorganisms.

The purpose of this study was to analyze the diversity of bacteria living at the surface of the ruins of the royal abbey of Chaalis north of Paris, France. In order to get a global characterization of the epilithic bacterial diversity present on the outdoor face-wall of the monument, stone samples were collected from areas presenting various degrees of deterioration (Fig. 1 and Fig. 2).

Section snippets

Sampling site and samples

The royal abbey of Chaalis is located in the north of Paris near Senlis. The domain, classified as historic monument since September 9, 1965, contains the ruins of the old abbey, the old abbey chapel and its frescoes of the Renaissance and a park with a rose garden (Fig. 1). Monuments of the abbey are built with Lutetian limestone, a material widely used in the construction of monuments in France, which has characteristics of porosity and roughness conducive to bacterial growth (Vázquez et al.,

Phylotype richness and its reliability

Analysis of ARDRA patterns allowed grouping 100 DCA clones into 50 ribotypes, 97 DMA clones into 49 ribotypes and 76 UDMA clones into 28 ribotypes. At least one clone per ARDRA pattern was then partially sequenced (655–985 bp). A total of 9 and 6 clones contained chimeric sequences for DCA and DMA libraries respectively, and were thus discarded from analysis. Two clones corresponding to Chloroplast sequences in the UDMA library were also discarded from analysis. Pairwise alignments of 16S rDNA

Discussion

Many monuments suffer from problems of deterioration that results from the action of biological and non-biological factors. Describing the diversity of microorganisms that colonize the stone of historical monuments is an essential first step in the understanding of biodeterioration phenomena. The aim of the present study was to analyze the diversity of bacteria colonizing the surface of the ruins of a French Historical monument, the Abbey of Chaalis, using a molecular approach. In order to get

Conclusion

A large number of historical monuments are made of carbonate stones such as limestone. However limestone undergoes significant deterioration phenomena over time. These deterioration phenomena are due in part to the presence and activity of microorganisms that colonize the stony substrate. Moreover, the state of conservation of building stone also affects microbial colonization. Analyzing the diversity of microorganisms colonizing the stone and understanding their involvement in biodeterioration

Acknowledgment

The authors thank the site administrator of the royal abbey of Chaalis, Aymar de Virieu, who has made it possible for us to conduct research on the ruins. The work was supported in part by “la Fondation des Sciences du Patrimoine“ PATRIMA.

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