Leonila Laiz

On the origin of fiber calcite crystals in moonmilk deposits

In this study, we show that moonmilk subaerial speleothems in Altamira Cave (Spain) consist of a network of fiber calcite crystals and active microbial structures. In Altamira moonmilks, the study of the typology and distribution of fiber crystals, extracellular polymeric substances, and microorganisms allowed us to define the initial stages of fiber crystal formation in recent samples as well as the variations in the microstructural arrangement in more evolved stages. Thus, we have been able to show the existence of a relationship among the different types of fiber crystals and their origins. This allowed us to outline a model that illustrates the different stages of formation of the moonmilk, developed on different substrata, concluding that microbes influence physicochemical precipitation, resulting in a variety of fiber crystal morphologies and sizes.

Microbiological study of the dripping waters in Altamira cave (Santillana del Mar, Spain).

The culturable microbial populations in dripping waters from Altamira cave were studied and compared with those of the ceiling rock. Water communities have low proportions of gram-positive bacteria, and are mainly composed of gram-negative rods and cocci (Enterobacteriaceae and Vibrionaceae), while those of ceiling rocks are mainly Streptomyces spp. The community differences are probably related to environmental cave conditions: high humidity, relatively low and stable temperature, water pH close to neutrality and nature of the organic matter. All these factors seem to favor colonization and long-term growth of actinomycetes over other heterotrophic bacteria on ceiling rocks.

Geomicrobiological Study of the Grotta dei Cervi, Porto Badisco, Italy

Speleothems (active stalactites, wall concretions), rock walls, ceiling, and soils from the galleries of Grotta dei Cervi, Porto Badisco, Italy, were sampled to investigate the culturable heterotrophic microbial communities present in this cave. Sampling was carried out in a transect of about 230 m from the entrance to the central gallery where numerous Gram-positive bacteria were isolated from all studied sites. Members of the genera Agromyces, Arthrobacter, Rhodococcus , and particularly Streptomyces , of the order Actinomycetales, are spread throughout the whole cave. The ability of actinomycetes, and particularly of Nocardiopsis , to colonize salt-stressed environments is favored by the presence of ectoine, a compatible solute for osmotic adaptation. Selected actinomycete isolates were tested for the formation of crystals. Strains from all tested genera, except isolates of Gordonia and Nocardia , produced vaterite and/or calcite. Production of Mgcalcite was restricted to strains of Brachybacterium, Rhodococcus , and Streptomyces , whereas struvite was only precipitated by an unidentified isolate. These findings indicate that actinomycetes may play a role in the formation of mineral deposits in caves.Speleothems (active stalactites, wall concretions), rock walls, ceiling, and soils from the galleries of Grotta dei Cervi, Porto Badisco, Italy, were sampled to investigate the culturable heterotrophic microbial communities present in this cave. Sampling was carried out in a transect of about 230 m from the entrance to the central gallery where numerous Gram-positive bacteria were isolated from all studied sites. Members of the genera Agromyces, Arthrobacter, Rhodococcus , and particularly Streptomyces , of the order Actinomycetales, are spread throughout the whole cave. The ability of actinomycetes, and particularly of Nocardiopsis , to colonize salt-stressed environments is favored by the presence of ectoine, a compatible solute for osmotic adaptation. Selected actinomycete isolates were tested for the formation of crystals. Strains from all tested genera, except isolates of Gordonia and Nocardia , produced vaterite and/or calcite. Production of Mgcalcite was restricted to strains of Brachybacterium, Rh...

Biomediated Precipitation of Calcium Carbonate Metastable Phases in Hypogean Environments: A Short Review

Natural precipitates of metastable polymorphs of CaCO 3 , such as vaterite, are rarely found in nature however, they have been widely synthesized in laboratory under particular conditions (ie, supersaturated solutions, relative high temperatures, etc.). By SEM and XRD we recognize vaterite spherulites from culturable microbial colonies isolated from hypogean environments. Spherical bodies (∽10μin diameter), probably composed of vaterite, occur in submilimetric microbial mats and biofilms on volcanic substrates (Saint Callixtus Catacombs, Rome, Italy) and karstic caves (Altamira, Candamo, and Tito Bustillo caves, Spain, and Grotta dei Cervi, Italy) where cyanobacteria and actinomycetes are the major microbial components. These particles form beneath dense biofilms, where particular physicochemical conditions are developed by the microbial activity. Natural biofilms seems to generate microenvironments favoring the formation and preservation of metastable CaCO 3 polymorphs. This also shows a major role of microbes in processes of low-temperature alteration of different hypogean rock-substrates.

Occurrence of halotolerant/halophilic bacterial communities in deteriorated monuments

The use of traditional microbiological methods provides the isolation of a large number of heterotrophic bacteria on deteriorated monuments. However, the spectrum of isolated bacteria changed when the protocols used in studies of halophilic bacteria were applied to mural paintings, efflorescences or mineral deposits. In fact, enumeration of the heterotrophic viable bacteria indicates that the higher counts were, generally, obtained in media with 10% of salt concentration. Media with magnesium sulphate always yielded higher counts than sodium chloride, particularly in environments where magnesium salts were abundant. The predominance of bacilli communities was remarkable. Four cases, in search of halophilic bacteria, are described.

Microbial Communities Associated With Hydromagnesite and Needle-Fiber Aragonite Deposits in a Karstic Cave (Altamira, Northern Spain)

Microbial communities, where Streptomyces species predominate, were found in association with hydromagnesite, Mg5(CO3)4(OH)2.4H2O, and needle-fiber aragonite deposits in an Altamira cave. The ability to precipitate calcium carbonate in laboratory cultures suggests that these and other bacteria present in the cave may play a role in the formation of moonmilk deposits.

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.

Pathogenic and opportunistic microorganisms in caves

Funding from the project RNM-5137, Consejeria de Innovacion, Junta de Andalucia, is acknowledged. This is also a TCP CSD2007-00058 paper.

The Actinobacterial Colonization of Etruscan Paintings

The paintings from Tomba della Scimmia, in Tuscany, are representative of the heavy bacterial colonization experienced in most Etruscan necropolises. The tomb remained open until the late 70′s when it was closed because of severe deterioration of the walls, ceiling and paintings after decades of visits. The deterioration is the result of environmental changes and impacts suffered since its discovery in 1846. We show scanning electron microscopy and molecular studies that reveal the extent and nature of the biodeterioration. Actinobacteria, mainly Nocardia and Pseudonocardia colonize and grow on the tomb walls and this process is linked to the availability of organic matter, phyllosilicates (e.g. clay minerals) and iron oxides. Nocardia is found metabolically active in the paintings. The data confirm the specialization of the genera Nocardia and Pseudonocardia in the colonization of subterranean niches.

Bacteria isolated from rock art paintings: the case of Atlanterra shelter (south Spain).

The Sierra de la Plata is an Aljibe yellow sandstone formation from the Acheulian period. There are a few shelters, some of them with rock art paintings. The most representative one, and subjected to anthropogenic pressure, is that of Atlanterra, situated in a residential area. This shelter contains some rock art paintings made with iron oxides. The bacteria present in these paintings were isolated and identified using an automatic method: fatty acid methyl esters profiling. Most of the bacteria belong to the Bacillus genus, B. megaterium being the most abundant species. The isolated strains are able to reduce hematite. This is significant due to the fact that Fe(III)-(hydr)oxides are the most abundant pigments in rock art.