THE MURAL EXECUTION TECHNIQUE OF THE CUBICLE “DEI FORNAI” REVEALED BY LASER CLEANING (p.75-77)

WORKSHOP ON THE CONSERVATION OF THE

SUBTERRANEAN CULTURAL HERITAGE

Seville, Spain

25-27 March 2014

ORGANIZED BY TECHNOHERITAGE

SPANISH NETWORK OF SCIENCE AND TECHNOLOGY FOR THE CONSERVATION OF CULTURAL HERITAGE

with the collaboration of Andalusian Historical Heritage Institute

The Conservation of Subterranean Cultural Heritage. Programme and Abstracts Cover image: Burial Chamber of Leone, Commodilla Catacomb, Rome © Red de Ciencia y Tecnología para la Conservación del Patrimonio Cultural, 2014 The copyright of the texts belong to their respective authors Typesetting: M.A. Rogerio Edited by Instituto de Recursos Naturales y Agrobiología de Sevilla, CSIC Printed by Coria Grafica, S.L. Impreso en España --- Printed in Spain

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Last Circular and Programme

On behalf of the Organizing Committee, we are pleased to invite you to attend the Workshop on the Conservation of the Subterranean Cultural Heritage, to be held in Seville, Spain. The workshop is organized by the Spanish Network of Science and Technology for the Conservation of Cultural Heritage (TechnoHeritage). The objective of the workshop is to promote an interdisciplinary forum for discussion on all aspects on the conservation of the subterranean cultural heritage, providing at the same time an up-to-date and comprehensive picture of the state-of-the-art of most recent investigations. The ultimate aims is to strengthen the foundation of scientific and technological research, overcoming the barriers of exchanging information that not always is published in scientific journals. The participation of researchers and professionals from the archeology-history-conservation-maintenance-restoration areas and the architectural-sciences-engineering areas will stimulate cooperation and integration between otherwise heterogeneous fields. The official languages of the Congress are English/Spanish with simultaneous translation. In addition to the invited lectures a poster session will be scheduled. Poster size: 100 cm x 120 cm. A Workshop proceeding (English) will be published and a copy of the book will be sent to full delegates around eight months after the Workshop. The book will include the invited lectures and the posters presented at the Workshop. The publication of communications cannot be guaranteed if the deadline is not met and the registration fee paid. The manuscripts should be sent to the address: [email protected] before the workshop commencement or directly delivered at the

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Secretary during the Workshop. The Workshop will be printed by CRC Press/Balkema, Taylor and Francis Group, a premier publisher of scientific and technical books and will be included in their international catalogue. The venue will be at Hotel Sevilla Palmera, 28 Cardenal Ilundain street, 41013 Seville, a four stars hotel. All sessions, coffee breaks, and lunches will be organized at the hotel. Registration Fees* Full delegate: € 150 (including congress program, coffee breaks, lunches, social dinner, workshop proceedings and visit to the Roman necropolis of Carmona) Local organization Cesáreo Sáiz Jiménez, Instituto de Recursos Naturales y Agrobiología de Sevilla, CSIC. Miguel Angel Rogerio Candelera, Instituto de Recursos Naturales y Agrobiología de Sevilla, CSIC.

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Programme and lectures

THE CONSERVATION OF THE SUBTERRANEAN CULTURAL HERITAGE

Hotel Sevilla Palmera, Seville, Spain, March 25-27, 2014

Monday 24 March 2014 18:30-20:30 Registration and Welcome

Tuesday 25 March 2014 9:30 -10:00 Workshop opening

Session 1. Tombs and catacombs

Chairman: José Manuel Galán, Centro de Ciencias Humanas y Sociales, CSIC, Madrid

10:00-11:00 Plenary Lecture: José Manuel Galán, Centro de

Ciencias Humanas y Sociales, CSIC, Madrid. The rock-cut tomb-chapel of Djehuty (ca. 1470 a. C.) on the West Bank of Luxor, Egypt.

11:00-11:30 Coffee break and poster session 11:30-12:15 Giandomenico Spinola, Musei Vaticani, Italy: The

excavation and conservation of the Necropolis of Via Triumphalis, Rome, Italy.

12:15-13:00 Maria Angela Turchetti, Soprintendenza per i Beni

Archeologici della Toscana: Problems of colour preservation in the Etruscan necropolis of Tuscany (Italy): the painted Tombs of Chiusi (Siena) and Sovana, Sorano (Grosseto).

13:00-15:00 Lunch and poster session 15:00-15:45 Laura Bruno, Simona Belleza, Filomena De Leo and

Clara Urzi, Italy, Universitá di Roma2, Italy: A study for monitoring and conservation in the Roman Catacombs of St. Callistus and Domitilla, Rome (Italy).

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15:45-16:30 Angel Fernández Cortes, Museo Nacional de Ciencias Naturales, CSIC, Madrid, Spain: The conservation of the Carmona Necropolis, Spain.

16:30-17:00 Coffee break and poster session 17:00-18:00 Round Table on Conservation of tombs and

catacombs

19:00-20:30 Visit to the Antiquarium (Subterranean display of the

Sevillian settlements from Roman to Moorish times) 19:00-19:05 Arrival and welcome by J.P. González 19:05-19:45 Guided visit to the Archaeological Site. 19.50-20:30 Round table on conservation problems of the site. 20.30-21:30 Informal tapas testing at the Antiquarium terrace Wednesday 26 March 2014

Session 2. Caves

Chairman: Juan Carlos Cañaveras, Universidad de Alicante, Spain.

9:30-10:15 Sergio Sanchez, Museo Nacional de Ciencias

Naturales, CSIC, Madrid, Spain: Two decades of microclimatic monitoring in Altamira Cave: A scientific project for their conservation.

10:15-11:00 José A. Lasheras, Carmen De Las Heras and Alfredo

Prada, Museo Nacional y Centro de Investigación de Altamira, Spain: Altamira and its future.

11:00-11:30 Coffee break and poster session 11:30-12:15 Jean-Michel Geneste. Centre National de Préhistoire,

Périgueux, France and Muriel Mauriac, Conservatrice de la grotte de Lascaux, Bordeaux, France: The conservation of Lascaux Cave, France.

12:15-13:00 Jean-Michel Geneste. Centre National de Préhistoire,

Përigueux, France and Marie Bardisa, Curator of Chauvet Cave, Direction régionale des Affaires culturelles de Rhône-Alpes, France: The conservation of Chauvet Cave, France.

13:00-15:00 Lunch and poster session

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15:00-15:45 Roberto Ontañón. Museo de Prehistoria y Arqueología

de Cantabria. Cuevas Prehistóricas de Cantabria, Spain: The conservation of the prehistoric caves in Cantabria, Spain.

15:45-16:30 Yolanda del Rosal Padial, Cristina Liñan Baena,

Instituto de Investigación Cueva de Nerja, Nerja, Spain, and Mariona Hernández Mariné, Departamento de Botánica, Facultad de Farmacia, Universidad de Barcelona, Spain: The conservation of Nerja Cave, Spain.

16:30-17:00 Coffee break and poster session 17:00-18:30 Round Table on Conservation of Palaeolithic caves 18:30-19:00 Closing session 21:00-24:00 Social dinner Thursday 27 March 2014 9:00-16:00 Guided visit to the Roman Necropolis of Carmona and

the town (upon pre-registration) 17:00-18:00 Return to Seville with stop around 17:30 at the Seville

airport Poster session L. Bruno, I. Ficorella, L. Quici, F. Valentini: Nanographene oxide: a

new material for a non-invasive and non-destructive strategy to remove biofilms from rock surfaces

C. Capel Ferrón, S.E. Jorge Villar, F.J. Medianero Soto, J.T. López

Navarrete, V. Hernández: Raman spectroscopic analysis of prehistoric lithic industry and some fat-burning stone lamps found at the Ardales and Las Palomas de Teba caves in the Guadalteba County (Málaga, Spain).

O.Ya. Chervyatsova, A.S. Pakhunov, L.Yu. Kuzmina, L.V. Leonova,

S.P. Glavatskikh: A study of interaction between Palaeolithic pigments and needle-fiber micro-crystals

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of calcite at Kapova Cave (Shulgan-Tash), Bashkortostan, Russia.

I. Dominguez-Moñino, V. Jurado, M.A. Rogerio-Candelera, C. Saiz-

Jimenez: Human impact on show caves: chewing gums stuck to the walls.

A. Fernandez-Cortes, E. Sanz-Rubio, A. Sanchez-Malo, S. Cuezva, E.

Garcia-Anton, M.J. Afonso, H.I. Chaminé, B. Hermosin, S. Sanchez-Moral: Detection of urban subsurface pollution by rapid multiparametric surveys in 16th century Paranhos spring water tunnel (Porto, Portugal).

A. M. Garcia-Sanchez, A. Z. Miller, V. Jurado, A. Dionísio, V. S. F.

Muralha, M. J. Afonso, H. I. Chaminé: Is the presence of bacterial communities related to the urban contamination sources of the Paranhos spring water galleries?

A. Gómez-Bolea, I. Álvaro, M. Hernández-Mariné, E. Llop, S.

Sammut: Indoors diversity in phototrophic biofilms at St Paul’s Catacombs (Malta).

V. Jurado, A. Nováková, M. Hernandez Marine, C. Saiz-Jimenez:

Cueva del Tesoro, Rincón de la Victoria, Málaga: a treasure of biodiversity.

V. Jurado, L. Laiz, S. Sanchez-Moral, C. Saiz-Jimenez: Pathogenic

microorganisms in Altamira Cave, Spain, related to human activities.

P. M. Martin-Sanchez, C. Saiz-Jimenez: Contribution of culture-

independent methods to cave aerobiology: the case of Lascaux Cave.

A. Z. Miller, A. Dionísio, M. E. Lopes, M. J. Afonso, H. I. Chaminé:

Microbe-mineral interactions in a Portuguese geo-archaeological site.

J. Mulec: Lampenflora as an accompany of mass cave tourism,

problems and solutions: example from Postojna Cave, Slovenia.

A. Nováková, V. Jurado, C. Saiz-Jimenez: Are fungi a real threat for

the conservation of Altamira Cave, Spain?

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A. Novakova, V. Hubka, C. Saiz-Jimenez: Microscopic fungi isolated

from cave air and sediments of the Nerja Cave - preliminary results.

A. Pakhunov, N. Brandt, A. Chikishev: Raman Microscopy and IR

Imaging of the Paleolithic Paintings from Kapova Cave, Southern Ural, Russia.

L. Pašić, J. Mulaomerović: Microorganisms and the natural heritage of

Dinaric karst – a review. G. Piñar, J. Ettenauer, K. Sterflinger: “La vie en rose”: a review of the

rosy discoloration of subsurface monuments. R. Senserrich-Espuñes, B. Mazzei, M.G. Patrizi, S. Bracci, M.

Realini, B. Sacchi & G. Bartolozzi: The mural execution technique of the Cubicle “dei ornai” revealed by laser cleaning.

D. Tapete, Fabio Fratini, B. Mazzei, E. Cantisani, C. Riminesi, R.

Manganelli Del Fà, B. Sacchi, O. Cuzman, M.G. Patrizi, L. Scaletti, P. Tiano: Methodological approach to monitor Roman catacombs: experience in the Catacombs of St Mark, Marcellian and Damasus, Rome, Italy.

D. Tapete, R. Piovesan, E. Cantisani, F. Fratini, C. Mazzoli, L.

Maritan, B. Mazzei: Identification of lime-based mural painting techniques in catacomb environments using well-established criteria of stratigraphic investigation.

C. Urzì, L. Bruno, F. De Leo, L. Krakova, D. Pangallo: New species description, biomineralization processes and biocleaning applications of Roman catacombs-living bacteria.

V.S. Zhitenev: The conservation of Kapova cave, Russia: past and

present.

Plenary Conferences

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THE ROCK-CUT TOMB-CHAPEL OF DJEHUTY (CA. 1470 A. C.) ON THE WEST BANK OF LUXOR, EGYPT José Manuel Galán Centro de Ciencias Humanas y Sociales, CSIC, Madrid, Spain The rock-cut tomb-chapels of Djehuty and Hery (TT 11–12) are located at the northern end of the Theban necropolis, in the central area of the hill of Dra Abu el-Naga. They were hewn at the foothill, and they are interconnected through a third tomb (–399–). The three date to the early XVIIIth Dynasty, in the first half of the 15th century BC. A Spanish-Egyptian archaeological mission has been working in the area since 2002, conducting consecutive annual campaigns in the months of January and February.

The excavations at both sides of the funerary monuments of Djehuty and Hery are bringing back to light part of the ancient necropolis, showing how the tomb-chapels were arranged one next to the other, in a row, as if they were following a street along the foothill. They are separated from one another only by a rock-wall half a meter thick, and for that reason they were easily interconnected in later times. The density of tomb-chapels here is higher than in other areas of the necropolis due to its symbolic location.

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Tomb-chapel of Djehuty (TT 11) Djehuty was a high official under the joint reign of

Hatshepsut–Thutmose III, ca. 1470 BC. He came from Middle Egypt, from the province of Hermopolis (Hemenu), consecrated to the god Thot, the scribe of the gods. Djehuty’s name also related him to the god Thot, since “Djehuty” means “one who belongs to Thot” as Thot is the Greek version of the Egyptian divine name Djehut. This is an important feature in Djehuty’s personality, since he tried to present himself as a cultivated man, well acquainted with the most arcane religious texts, and gifted in the writing and visual composition of inscriptions. His funerary monument was designed as the perfect medium to demonstrate his knowledge and creativity in writing.

As “royal scribe”, he was appointed “overseer of the Treasury” and “overseer of the works (of the craftsmen)”. These two duties were related to each other in as much as they concerned the acquisition, management and redistribution of metals (gold, silver, electrum, bronze, copper), semiprecious stones (turquoise), and exotic timber (cedar wood). As “overseer of the Treasury” he was in charge of collecting taxes from the local governors, and he registered in writing the marvels brought from Punt and directed to the god Amun of Karnak in year 9. As “overseer of works”, he was responsible for “giving instructions and leading the artisans to act regarding the work in” shrines and temples all over Thebes. In Karnak, the noble portal “Presentation of Maat” and two obelisks erected by Hatshepsut between pylons IVth and Vth in year 16 were inlaid with electrum; and the sacred bark of Amun, User-hat-Amun, with gold so that “the Two Lands would be illuminated”. He enhanced with copper and electrum the great doors of Hatshepsut’s Temple of Millions of Years, Djeser-djeseru, and those of the nearby temple of Kha-akhet.

The layout of the inner part of the tomb-chapel has an inverted T-shape, and the walls are fully covered with inscriptions and scenes in relief. Djehuty included in the repertoire three biographical inscriptions, a solar hymn to

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Amun-Ra, two long cryptographic texts, a visual and textual description of the Opening of the Mouth ritual and other funerary ceremonies, together with the standard compositions of the deceased fishing and fowling in the marshes, hunting game in the desert fringe, a pilgrimage boat journey back and forth to Abydos and a couple of banquet scenes.

The inner part of Djehuty’s funerary monument penetrates horizontally 18m inside the rock of the hill, and its excavation was completed during the 9th archaeological campaign. The clearance of the innermost room brought to light the mouth of a funerary shaft (2 x 1m), which was excavated in 2008, and ended being more than 8m deep. At its bottom there was an entrance leading to a big chamber (5.30 x 3.45m, and 1.55m high), excavated in 2009. At the rear end of it there was a second shaft, going down only about 3m. At the bottom there was an entrance leading to second chamber, meant to be Djehuty’s “burial chamber”.

The burial chamber was decorated with passages from the Book of the Dead. Now only two of the original walls and the area of the ceiling corresponding to the first quadrangular design preserve the layer of stucco and the text written on it. Djehuty’s selection of Book of the Dead chapters is particularly relevant because it was under Hatshepsut when the composition reached a considerable length and the sequence of chapters began to be established. Thus, Djehuty’s burial chamber is one of the earliest and longest (34 chapters) Book of the Dead samples that has been preserved. It is worth noticing that it includes one of the earliest versions of chapter BD 125, the so-called ‘negative confession’ during the final judgement.

Djehuty’s burial chamber is in an unstable and uncertain condition. The ceiling has a big hole in the middle and cracks running in all directions, making it advisable not to stay long inside the chamber. We designed and installed a sort of custom-made iron structure to minimize the consequences if one or more blocks happen to fall down by accident. On the other hand, the major problem for the conservation of the painted stucco is the humidity inside the chamber. Due to its

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foothill location and its proximity to the irrigated valley, and the fact that the burial chamber rests 12m below ground level, the underground water table lays very close to it. This circumstance causes that the salts of the limestone bedrock precipitate and migrate to the surface. As the salts appear on the surface, they push the layer of stucco and separate it from the rock. Thus, gypsum and halite salts can easily be seen on the surface, particularly on the lower half of the walls and inside the cracks. Under this environmental condition, we need to avoid drastic and repeated variations in humidity and temperature levels, and for that reason the chamber remains closed as much as possible, so that the natural environmental conditions can be recovered and maintained stabilized. A team of geologists specialized in interior environments (such as prehistoric painted caves) set up autonomous electronic devices inside and outside the burial chamber to monitor all year long the variations in temperature and humidity.

Last season we conducted and completed and thorough photographic documentation of the inner part of Djehuty’s funerary monument, including the walls and ceiling of the burial chamber. A set of ortho-photos was taken, and then joined in the computer to obtain a high resolution single photo of each wall and ceiling, which can then be zoomed in to perceive small details of its decoration and physical condition.

Cleaning, consolidation and restoration of the walls have continued, paying special attention to the inner shrine, corridor and the right side of the transverse hall. Mud was carefully removed from the surface, turning visible parts of inscriptions that were hidden underneath. Some blocks found outside, when digging the courtyard, were relocated in the walls, in their original place, thanks to the collaboration between epigraphers and restorers.

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EXCAVATIONS AND CONSERVATIVE PROBLEMS IN THE ROMAN NECROPOLIS ALONG VIA TRIUMPHALIS IN THE VATICAN Giandomenico Spinola Curator of the Greek and Roman Art Department, Vatican Museums Vatican, in Roman age, was located on the Etruscan bank of the Tiber river and it was very far from the city. Famous for its quarries of clay and the poor quality wine, Vatican had actually housed great parks, both private and imperials: there were located the gardens where emperor Caligula had built the circus and later emperor Nero had executed the Christians who were accused of causing the Great Fire of Rome of 64 a. D. Adjacent to the circus, there were Horti Domitiae, where emperor Hadrian had built his majestic mausoleum, knows as Castel Sant'Angelo. As in all marginal areas, the paths that crossed the Vatican – Via Triumphalis to Veio (now Isola Farnese), Via Cornelia-Aurelia to Caere (now Cerveteri) – were skirted with tombs of all kinds, many of which have been discovered in the current area of the Vatican City.

The Necropolis along Via Triumphalis is of great significance in the context of the rich archaeological heritage of the city of Rome. In fact, another equally vast and varied complex of tombs that represent the lower and middle Roman society doesn't exist; commonly, there are the imperial tombs, the noble tombs of Via Appia but not the tombs of "normal" people. It is possible to observe here a huge variety of burial customs, ranging from the poorest cremation in wood urns to lavish sarcophagi, to the medium-high segment of society with tombs adorned with frescoes and mosaics. Scene of daily life can be observed between a burial and the other: there were Alcimus, slave of emperor Nero assigned to the scenes of the Theatre of Pompeus in Rome; Tiberius Claudius Optatus, archivist of the imperial accounting; Clement, the horseman

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of the “faction of the Azures” who perhaps had competed several time near the circus, and many others. A recent excavation, carried out between 2009 and 2011 in the central area of the Necropolis and currently visible, has linked the sectors of the “Autoparco” (1956-1958 excavations) and Santa Rosa (excavations 2003), until now separated. Among the novelties of the excavation, a designated area for the cremations (ustrino) was discovered; they are rarely preserved in complexes of this type.

This area is a veritable laboratory of archaeological research. Several scientific investigations and assessments are in progress, such as prospecting with a georadar and anthropological analysis. These last analyses were performed by Prof. Henri Duday, University of Bordeaux; cremation was studied in collaboration with the Ecole Française de Rome, and the burials, in collaboration with the service of Anthropology of the Superintendence for Archaeological Heritage of Rome, coordinated by Dr. Paola Catalano.

Restoration and conservation aspects have certainly a primary role in the work that continues in the archaeological site. The monitoring, chemical and optical analysis inside the museum area, allow us to verify climate changes and any damage to graves and preserved artefacts. In parallel, new technologies such as laser cleaning are being used and new methodologies of biorestoration are being tested.

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PROBLEMS OF COLOUR PRESERVATION IN THE ETRUSCAN NECROPOLIS OF TUSCANY (ITALY): THE PAINTED TOMBS OF CHIUSI (SIENA) AND SOVANA, SORANO (GROSSETO) Maria Angela Turchetti Soprintendenza per i Beni Archeologici della Toscana Via della Pergola 65, Firenze, Italy The Tomb of the Monkey is the most important tomb of the Etruscan necropolis of Poggio Renzo, near Chiusi (Siena). Dated around 480-470 BC, it was discovered in 1846 by the Florentine archaeologist Alexander François. The tomb lies 8 meters below the ground level, it is composed of three rooms around a main atrium. The walls of the atrium are decorated with polychrome paintings, showing scenes from funerary games. The tomb itself takes his name from a monkey depicted on a tree. In the front room, two other pictures, apparently monochromatic, probably represent young slaves carrying unknown objects. The state of preservation of the paintings is rather precarious, because of the particular technique used for their realization. In fact, the pigments (hematite, charcoal black and Egyptian blue) were applied on a fragile clay layer on the rock wall. Moreover, a further deterioration of the painting was produced by environmental effects, including the biodegradation.

The funerary hellenistic architecture in tufa of the necropolis of Sovana and the surrounding area has been considered archaeological evidence of great importance ever since the 19th century, when it never failed to arouse the admiring interest of English travelers, who first understood the importance of documenting what appeared before their eyes. Now – thanks to timely and careful analytical work and study – we can imagine with a wealth of color that until recently was almost unknown for these monuments. Though blending today with the natural surroundings thanks to the

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color of the tufa, they once appeared visible at a great distance thanks to their bright decorations. However, at the same time, we recall the verses of Rutilio Namaziano, who summarized incisively the distressing problem of how to ensure for posterity the preservation of a legacy so perishable. Although it is not always possible to find a solution, it is essential to promote campaigns of documentation to enable us – armed with new technologies of analysis and mapping – a thorough anderstanding of these contexts and their criticality.

If it is crucial to combat the relentless deterioration of the material with constant maintenance, emergency relief and monitoring of the deterioration, at the same time we should not forget that only the study of monuments and publishing of results can guarantee these relics of the past another form of conservation and protection, and enhance what it’s possible to call the «formal identity, including the chromatic aspect» of these original examples of Etruscan architecture.

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A STUDY FOR MONITORING AND CONSERVATION IN THE ROMAN CATACOMBS OF ST. CALLISTUS AND DOMITILLA, ROME (ITALY) L. Bruno & S. Bellezza Dept. of Biology, University of Rome ‘Tor Vergata’, Via della Ricerca Scientifica 1, Italy

F. De Leo & C. Urzì Department of Biological and Environmental Sciences, University of Messina, Italy

Roman Catacombs are touristic sites with high religious, archaeological and historical value. They were excavated from the 2nd century onwards, carved through tufa, a soft volcanic rock, outside the walls of the city of Rome. They consist of corridors and cubicula that in some sites are covered by fresco decorations witnessing the Early Christian behavior and art. The peculiar environment in these underground sites along with the artificial lights and the presence of the tourists sustain the growth of biological patina on the exposed rock surfaces. These biofilms are mainly formed by cyanobacteria and chemoorganotrophic bacteria associated to green microalgae and mosses all embedded in a common exopolymeric matrix. The metabolic activity of these biofilms causes aesthetic and structural problems to the substratum by means mineralization processes. Although the sites were annually restored with chemical and mechanical tools the microbial growth occur regularly when the Catacombs are re-opened to the public, underlying the need to employ better and safe intervention strategies.

In the frame of the European project Cats (EVK4 CT-2000-00028) different approaches were undertaken in order to prevent and/or limit the development of this phototrophic biofilms1-3. At first the macro- and microclimatic parameters at selected sites inside the two Catacombs were registered,

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with the characterization of the lithic substrata; moreover, the microbial diversity has been investigated along with the study of the biofilm architecture and the exopolymeric substances produced. In a second step new physical methods were developed and tested to control and prevent biofilm growth using wavelengths in the visible part of the light spectrum that are, at best, poorly used by photosynthesis: a blue monochromatic lamp was installed in the Ocean’s cubiculum and the photosynthetic microorganisms disappeared from the painted surfaces. The CATS data allowed to ascertain the most critical physical, chemical and biological factors that control colonisation of rock surfaces in the two Catacombs suggesting the possibility to employ new non-invasive preventive methods. In the following years a monitoring of the colonization in the site was carried out evidencing the effectiveness of the monochromatic lamp in preventing the phototrophic growth. In order to improve the possibility to use selected lights for the illumination of these sites, able to deter the biofilm development and at the same time to offer a better vision to the tourists, different combinations of monochromatic spectra emissions were tested in laboratory experiments obtaining encouraging results. 1Sanchez-Moral S, Luque L, Cuezva S, Soler V, Benavente D, Laiz L, Gonzalez JM, Saiz-Jimenez C. 2005. Deterioration of building materials in Roman catacombs: the influence of visitors. Sci Total Environ 349: 260–276. 2Bruno L, Billi D, Bellezza S, Albertano P. 2009. Cytomorphological and genetic characterization of troglophilic Leptolyngbya strains isolated from Roman hypogea. Appl Environ Microbiol 75: 608–617. 3Albertano P, Bruno L, Bellezza S. 2005. New strategy for the monitoring and control of cyanobacterial films on valuable lithic faces. Plant Biosyst 139: 311–322.

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THE CONSERVATION OF THE CARMONA NECROPOLIS (SEVILLE, SPAIN) Angel Fernandez-Cortes Geomnia Natural Resources SLNE, Cea Bermudez 14 5-A, 28003 Madrid, Spain and Departamento de Geologia, Museo Nacional de Ciencias Naturales (CSIC), Jose Gutierrez Abascal 2, 28006 Madrid, Spain The necropolis of Carmona is one of the most significant Roman burial sites in southern Spain used during the first and second centuries AD. It comprises about 600 underground tombs excavated in biocalcarenites, most of them are collective mausoleum consisting of an underground chamber accessed by a well-staged and, exceptionally, with some relict stuccos with original roman paintings.

This archaeological site has suffered numerous and intense interventions since its discovery (1868-1869) to present days that in most cases have enhanced the deterioration of tombs. After our preliminary field surveys several causes of deterioration were identified, including: erosion and mechanical and chemical weathering favoured by the petrophysical properties of host rock and the presence of anthropic features from the original excavation of tombs and further conditioning works for the tourism use of this archaeological site.

During the triennium 2007-2009 our research group conducted a comprehensive project to study its conservation status, funded by the Regional Government of Andalusia. This project aimed to characterize the alteration processes in building materials and host-rock of tombs in order to design the near future preventive guidelines and corrective measures to optimize the conservation status of each particular tomb.An integrative assessment of the different geo-environmental factors that interact on a small scale has helped us in the understanding of the past weathering

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evolution of tombs and in foreseeing their possible future deterioration, as well as to define the general and particular safe microclimatic conditions for a suitable conservation.

The scope of this research project was compulsory enlarged due to the multiple factors involved on the weathering processes and the diverse degradation mechanisms. Therefore, it entailed an multidisciplinary approach that required a range of analytical procedures and research tasks grouped on three main work packages: 1) Geomorphological and geological study of the outcrop where the Necropolis is located, with special attention to determine the runoff pathways and the disturbances of the drainage pattern caused by human activities (excavations and tourism adaptation); 2) Monitoring of environmental conditions of tombs (air and surface of rock and building materials); and 3) Petrophysical and mineralogical characterization of host rock, building materials and weathering forms (e.g. salt efflorescences), including water absorption test.

A georreferenced database was created and implemented on interrelated GIS models, including 3D models to assess the seasonal evolution of the intensity and duration of direct isolation at each tomb entrance, which is highly correlated with its inner environmental conditions and some particular decaying mechanisms (i.e. thermal expansion of building materials or condensation/evaporation cycles of porous water at rock surface). A spatial analysis of surface drainage pattern confirms that, in spite the prevailing xeric weather conditions, the random but torrential rains occasionally floods the tombs. The transport of sediments into the tombs is enough to seal the natural drainage and to maintain elevated moisture conditions inside tombs, which favours the cyclic appearance of effective capillary condensation during longer periods. Water condensation and capillary rise are favoured by the high porosity and polymodal pore-size distribution of the calcarenite.

Daily condensation/evaporation cycles involve an intense hydric stress of materials from excavated tombs, with cycles of dissolution/crystallization of salts and the consequent

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granular disintegration of the calcarenite by the crystallization pressure of salts. Additionally, the water availability within surficial porous system of building materials and calcarenite triggers other decay mechanisms, including: relatively rapid appearance of microorganisms causing biodeterioration, clay swelling reducing rock cohesion and calcite dissolution by NaCl- and CO2-rich pore waters provoking a progressive arsenisation of calcarenite.

The integrated analysis of the decay mechanisms and the environmental ranges in which they develop, allowed us to define the safe threshold microclimatic conditions that minimize the main rock-decay processes: air relative humidity below 60% in the natural temperature range in order to inhibit condensation, and low temperature variation inside the tombs and over the ceilings of the more superficial tombs by avoiding high insolation rates.

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TWO DECADES OF MICROCLIMATIC MONITORING IN ALTAMIRA CAVE: A SCIENTIFIC PROJECT FOR THEIR CONSERVATION Sergio Sanchez-Moral Museo Nacional de Ciencias Naturales, MNCN-CSIC, José Gutiérrez Abascal 2, 28006 Madrid, Spain Management and conservation of tourist caves must involve a complete knowledge of physicochemical parameters of both subterranean and external surrounding environments. The factors affecting preservation of valuable cultural heritage in underground environments, especially those with archaeological features such as prehistoric rock paintings and engravings, are mainly: exchanges of energy and matter between cave and external atmosphere, physicochemical properties of the karst water and presence of visitors inside cave.

Rock art in caves is located within a dynamic natural environment whose intrinsic characteristics differ substantially from those found in a museum hall. These unique characteristics of the karst environments make them particularly vulnerable to human activities. In these environments, paintings made 15 or 20 thousand years ago have been preserved until today because the environmental conditions experienced during millennia were optimal for the rock art conservation, and have prevented the damage and the disappearance of the paintings. Therefore, the first step aimed at conserving the rock art should be focused on the study of the environmental characteristics of the karst system, including the inner and outer conditions.

In the case of Altamira Cave, and over a period of fifteen years (1997-2012), a microclimatic recording of their environmental conditions was carried out in four different stages by the research team of National Museum of Natural Sciences (Spanish National Research Council, MNCN-CSIC):

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In the first period (1996-1999), in the framework of an EU-project (ENV4-CT95-0104), the studies were focused on quantifying the impacts on the physicochemical conditions caused by the entrance of visitors in the Polychrome Hall. During this period of micro-environmental monitoring, Altamira Cave was exposed to a restricted visitation scheme, effective from 1982 to September 2002, based on previous studies conducted by the University of Cantabria in the 1980s. Increases in average temperature and CO2 content were quantified after a detailed assessment of the impact induced on the microclimate at the Polychrome Hall by a total of 900 visiting groups. Continual and almost daily opening of the access door as well as the entry of consecutive groups in the Polychrome Hall caused cumulative disturbances in the microenvironment, originating a general increase in the average interior temperature.

Each daily series of visiting groups generated an average impact of 0.22ºC on the air temperature, and 440 ppm on the CO2 content. The average value of thermal impact caused by each series of daily visits was of the same order of magnitude as the monthly thermal oscillation range under natural conditions. Moreover, increases in both parameters after the visit often were cumulative throughout the day and did not recover between consecutive groups.

The average time needed to restore the previous microclimatic conditions after a series of daily visits was quantified in 17 hours for air temperature. The recovery time was even greater for CO2 levels in the air. The anthropogenic CO2 assimilated by the interior atmosphere would begin to causing the transformation of gas phase CO2 into CO2 dissolved in liquid phase (dripping and condensed waters). The kinetics of this chemical reaction requires about five minutes, and once begun, the water acquires the characteristics of a dissolvent and begin a corrosion processes on the carbonated surfaces. The pitting generated by these corrosion processes are preference niches for the establishment and development of microbial colonies.

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One of the most immediate data reached in this period was to note that documentation works developed within the cavity to construct the painting replica led to a strong disturbance of the cave microclimate and a significant increase in artificial lighting time. During 1998 the maximum temperature increased strongly, reaching 16.15ºC, which means an increase of 1.3ºC above the 1997 level. On 29 June 1998, the daily increase in temperature was close to 2ºC. The major and more continuous disturbances were detected during January and February 1999, with daily accumulated increments between 0.5 and 1.7ºC.

The development of phototrophic microbial communities in the ceiling of the Polychrome Hall, detected in April 2002, was directly related to all those environmental disturbances previously described. Once the cave was closed in September 2002, the

Ministry of Culture appointed again the MNCN-CSIC team to survey the cave from 2003 to 2005. Discrimination between natural environmental fluctuations and disturbances caused by human activities is essential in the study of decay processes taking place in caves with rock art. This was the main goal of the project during this second phase of studies, which included a continuous monitoring of the microclimate of the cave over two annual cycles. Researches were focused on the cave as an ecosystem depending on both microclimate and geochemical characteristics of the underground environment and energy-matter exchange with the external atmosphere.

The 2002 closing represented a clear benefit for conservation of the paintings: The green phototrophic colonizations did not continue to progress, a progressive thermal stabilization was observed and the corrosion rate of the paintings’ host rock decreased. The final report arising from this research recommended the renewal of the entrance door to the cave and the installation of a second door in order to reduce the rates of exchange of matter and energy between the cave and the outer atmosphere. Under natural conditions, Altamira Cave shows high microclimatic stability

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throughout the annual cycle, with low energy exchange with the outside. The temperature inside the cavity varies annually according to a cyclical-seasonal pattern which is reflected in the sinusoidal tendency of the evolution curve. Despite the prevailing high thermo-hygrometric stability (about 1.5ºC annual thermal amplitude and indoor relative humidity values near saturation), Altamira Cave shows an outgassing stage during summer season and a gas recharge phase during wet and rainy periods. In summer, CO2 concentrations are relatively steady near 500 ppm. Winter concentrations are much higher and more erratic, sometimes exceeding 5000 ppm, with weekly-monthly isolated degassing events. The two main trace gases (CO2 and 222Rn) show significant similarities in their seasonal evolution, with the lowest values during summer and peaks in winter and early spring. This indicates that the annual-scale variations in CO2 are due in large part to variable ventilation of the cave, which is most efficient in summer. In the third phase of the study (2007-2009), the main

conclusions coincided with those obtained from the previous stages. However, new data indicated that the data obtained in 1997-1999 had considerably underestimated the real impact generated by the visitor’s entry into the area which house the paintings (ceiling). To sum up, the conclusions obtained in the 2007-2009 study were the following:

- The entry of visitors caused a substantial increase in the thermo-hygrometric oscillation and in the CO2

concentration in the interior air as well as changes in the temperature relation between different areas (Crossing and Polychrome Hall). This occasionally triggered important changes in the local air circulation.

- The air warming in the Polychrome Hall as a consequence of visitors stay was especially sharp near the ceiling in the entryway and inside the Hall. The effects recently detected indicated that the real thermal increase on the ceiling has at least doubled regarding the data obtained from the previous monitoring system. Also the thermal

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increase generated by the visits to the Polychrome Hall was linked to the decrease in density of the hall air. This allows the mobilization of the air from the interior of the Polychrome Hall to mix with that of the nearby corridors.

- The experimental visit which took place in March 2007 demonstrated the effect of the mobilization of the air mass on the ceiling of the Polychrome Hall generated by visitor circulation. The air displacement happens throughout the visitor route and contributed to the progress of the air mass with water micro-particles toward the interior of the cave and thereby enhancing the phenomena of condensation and microorganism dispersion to the interior of the cave.

- Before the installation of the new doors, the entrance area of Altamira Cave promoted the entry of warm, wet air into the cave during the summer season (June-September) resulting in the formation of water microparticle clouds (hydroaerosols) moving toward the interior of the cave. Each hydroaerosol acted as potential adherence nucleus for dust particles (solid aerosols) and spores in the air and increased liquid water and organic matter availability favoring the development and dissemination of microbial colonies.

- The installation of a new access door in 2007, equipped with a thermal insulation system, followed 20 m after by a second door, reduced the entry of airborne particles, the condensation rate in the entrance area, and the metabolic activity of the main visible microbial colonies. In the last phase of our studies (2010-2012) the data

confirmed the tendency to thermal stabilization observed since the closing of the cave and especially since the installation of the second gate. During the 1997-1998 period thermal oscillation in Polychrome Hall was 1.66ºC. Closing the cave favored a first thermal cycle stabilization during 2004-2005 (1.54ºC) and after the installation and operation of the second gate led to a new reduction of thermal oscillation to a value of 1.28ºC in the last annual cycle monitored.

However, during this last period new archaeological excavations were conducted in an external area immediate to

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the cave entrance. During and after the archaeological excavations of October 2010 there was a sudden drop in air temperature in the Entrance Hall. The temperature decrease was transmitted directly to the area of the Crossing causing short-period oscillations in the Polychrome ceiling from the end of November 2010. These temperature fluctuations in Polychrome were not detected in any previous cycle in winter season. Also in the Entrance Hall could be seen, from the autumn of 2010, an increase of short-period thermal fluctuations during winter as a result of the reduction of the insulating effect. Thermographic studies showed how this phenomenon causes the intense cold air intake during winter and favors the condensation processes in the Entrance Hall.

As an overall conclusion, our data showed that human activity inside the cave or in its surroundings (e.g., recent archeological excavations) caused microenvironmental disturbances in the cave which would be detrimental to the conservation of rock art. These disturbances have greatly impacted the production and dispersion of bacterial and fungal spores in the cave air.

The extensive data compiled over the past 15 years do not recommend a cave reopening. In the case of a reopening, the entry of visitors could reactivate the deterioration mechanisms previously described. This would particularly affect the Polychrome Hall because is the most fragile area of the cave due to its special morphogeometric characteristics (small dimensions, low level of ventilation) and hydrokarstic dynamic (very slow infiltration levels).

Therefore, the conservation of rock art should consider the maintenance or restoration of the natural conditions in a strictest possible way. The corrective actions as far as possible should be limited to those parameters involved in a clear and rapid disappearance of the cave paintings, although this phenomenon is due to natural causes.

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ALTAMIRA AND ITS FUTURE José A. Lasheras, Carmen De Las Heras y Alfredo Prada Museo Nacional y Centro de Investigacion de Altamira, Santillana del Mar, Spain When Marcelino Sanz de Sautuola discovered Altamira and became the first to publish Magdalenian paintings, the oldest and only known art of its kind at the time, he understood their value and fragility, foresaw their future and protected them by placing a gate at the cave mouth before making his find public. If they had come down to us more or less well-preserved, it was only because of the relatively favourable and sufficiently stable geological and environmental conditions inside the cave that housed them. Ever since, people have always taken an interest in Altamira's past and worried about its future. Nevertheless, in the 1960s and 70s the pressure of the present began to threaten its future: the constant flow of tourists in those years—more than 160,000 people each year from 1960 to 1977—was a serious conservation risk that endangered the paintings' very survival.

The situation changed when the Spanish government took ownership of the cave in 1978 and assigned it to the Museum of Altamira. Like any museum, by definition it was entrusted with researching and preserving its collections and making the public aware of their intrinsic value.

A team at the University of Cantabria was assigned the project of researching and evaluating the condition of the paintings. Their principal conclusion was that the number of visitors needed to be drastically reduced, and to this end they created a mathematical physics formula for calculating ideal environmental parameters which was implemented after 1982. However, twenty years later the appearance of green algae on the ceiling made it necessary to close the cave again.

A team from the Spanish National Research Council (Consejo Superior de Investigaciones Cientificas - CSIC) was

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brought in to conduct a new study of the environmental conditions affecting the current state and future preservation of the paintings, which was concluded in 2009. Among their conclusions, the researchers suggested that the best conservation strategy was to keep the cave closed to the public indefinitely.

Shortly afterwards, in 2012, the Ministry of Education, Culture and Sport launched a conservation programme, spearheaded by the ministry's own Spanish Cultural Heritage Institute (IPCE) and due to conclude in August, at which time the institute is expected to propose, if possible, a plan for reopening Altamira to visitors while also guaranteeing its proper conservation.

New knowledge obtained from research into aspects that affect the paintings and current technical conservation criteria will be taken into account in the upcoming decision-making process and the future management of this World Heritage Site.

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THE CONSERVATION OF LASCAUX CAVE, FRANCE

Jean Michel Geneste Centre National de Préhistoire, Sous-Direction de l’Archéologie, Périgueux, France Muriel Mauriac Conservatrice de la grotte de Lascaux, Direction régionale des Affaires culturelles d’Aquitaine, Bordeaux, France

The Lascaux cave has been discovered in September 1940, protected and classified as Historic Monument in the same year. After extensive modifications of its entrance that caused profound destabilization of its natural bioclimatic system, the cave has been opened to the public in 1948.

But after important microbiological and climatic diseases Lascaux has been closed to visits in 1963. After returning to apparently normal equilibrium and the cave remaining closed to the public, a new bioclimatic crisis suddenly arose in 2001 with an apogee in 2007-2008.

At each stage in the history of the conservation of the cave the French government has implemented all the scientific resources necessary to ensure the preservation of the painted and engraved walls. Several scientific committees have been constituted. Their archives are important and provide numerous data on the multidisciplinary scientific history and on all the activities developed for the preservation of the cave.

All the committees have coordinated the scientific research necessary to understand the functioning of the cave and to create or adapt the tools and equipment required to ensure its continued health. Several research programs have been and are actually developed in several directions: geology, hydrogeology, origin of the carbon dioxide, and microbiology, at national and international levels.

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Lascaux cave is very fragile due to its shallow underground depth, its small dimensions and High CO2 levels. More recently a climatic simulator has allowed researchers to model the climatic functioning inside the cave since its discovery. Using this preventive conservation tool and the very precise 3D model of the entire cave, it is possible:

- to estimate the role played by natural and anthropogenic parameters in the changes of the internal climate

- to test the possibilities of improvement of the climatic assistance system, before operating them.

- the impact of human presence on the climate, and the feasibility of certain modifications to the cave structure.

Microbiology is the second main approach implicated in the conservation of the cave. In the long term the fundamental research aims to understand the microbial ecology of the cave and in the short term the survey is increased, especially in the sensitive areas of the cave and of the painted panels, with different types of monitoring such as comparative photographic control.

The main methodological aspect of this research and health control is to link the techniques used to identify and follow the evolution of the microorganisms with their function in the natural equilibrium or in other words the role of biological diversity.

The internal work and studies of the actual Lascaux Scientific committee are now followed and culminated in the “sanctuarisation” of the environmental landscape on which Lascaux is located. This natural environment with its actual transformations is a major element of the global conservation plan.

The fact that it is definitely not any more possible to open Lascaux cave to the public has led to the decision to build a new complete replica which will be located in Montignac, for preservation purpose, at the bottom of the hill, away from the original.

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THE CONSERVATION OF CHAUVET CAVE, FRANCE Jean Michel Geneste Centre National de Préhistoire, Sous-direction de l’Archéologie, Périgueux, France Marie Bardisa Curator of Chauvet Cave, Direction régionale des Affaires culturelles de Rhône-Alpes, France The decorated cave of Vallon-Pont-d’Arc, in Ardèche, known as Chauvet Cave, discovered on 18 December 1994 by Jean-Marie Chauvet and his companions in the Gorges de l’Ardèche, has been under study since 1988. The professional team was first directed by Jean Clottes, followed by Jean-Michel Geneste since 2002.

From the beginning of this research, in response to a call for proposals by the Ministry of Culture and Communication for the scientific study of the Chauvet-Pont d’Arc Cave, certain principles were defined, inspiring the research and conservation actions that would be undertaken. This is the first time in the world that a multidisciplinary research team as diverse as this was conceived and composed for the study of a major rock art site. This would not have been possible without the motivation and financial support of the Ministry of Culture and Communication. Since then, several teams following a similar model have been formed elsewhere, particularly in the United States, South Africa and Australia. Diverse conditions must be meet for this type of teamwork to function.

This vast cavern is a still active karstic system located under the high part of the cliffs of the Ardeche Gorges. The entrance was abruptly closed off from the outside by a rock fall around 22,000 years ago, tightly sealing and protecting the archaeological contents inside. From the moment of its discovery, this site was thus recognized for the exceptionally good preservation of the drawings and engravings on its walls,

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as well as of the paleontological remains and traces of human and animal presence on the floors and walls. This cave, occupied between 36,000 and 29,000 years ago was immediately protected and the decision was made not to open it to the public due to the fragility of its art works, its confined space, and its inaccessibility at the time of the discovery.

Much thought was given to determining how the entrance would be modified and what types of equipment would be allowed inside. A raised metallic path was installed in the areas where the discoverers had walked on the ground.

The research itself was first organized in function of these paths and when possible was extended into areas that are accessible without risk; these are zones that are covered with a thick layer of hard calcite on which a secondary system of removable paths can be installed.

In parallel with these first modifications, climatic and biological studies of the cavity were undertaken. This research was realized by the Laboratoire de Recherches des Monuments Historiques (Laboratory of Research on Historic Monuments), and the Laboratoire Souterrain (Underground Laboratory) of the CNRS of Moulis on the climatic evolution of the cave in order to ensure that the time spent inside would cause as little harm as possible. These studies led to the definition of a set of rules that has since been applied to all visits, including those associated with research activities. In addition to the climatic regulation of this cave system, variations in C02 and Radon levels are today the most restricting parameters determining the conditions for human presence in the cave.

Due to the remarkable preservation of the ground surfaces – on which we find many traces and remains of human and animal presence –our foremost priority must be to ensure the preservation of this cave, including its walls, climate and floors. The ultimate goal is to enable to pass on a cavern that is fully intact, where all types of future research will still be possible.

The question of how to transmit and share knowledge is fundamental at this site, unique in the history of the

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development of modern human thought. Some answers have been found in the creation of an ambitious cave replica and learning center, as well as in a strict policy permitting only exceptional visits in the original cavern.

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THE CONSERVATION OF THE PREHISTORIC CAVES IN CANTABRIA, SPAIN Roberto Ontañón Museo de Prehistoria y Arqueología de Cantabria, Cuevas Prehistóricas de Cantabria, Spain The subterranean domain houses one of the distinguishing elements of the cultural heritage of Cantabria. The thousands of limestone cavities that burrow its subsoil endow this region of great natural wealth and provide for an important touristic resource. The relevance of this heritage is reflected in the large number of visitors every year and has been endorsed by the inclusion of ten caves of Cantabria in the UNESCO’s World Heritage List.

However, different risk factors, natural and anthropogenic, exogenous and endogenous, threaten the preservation of this extraordinary heritage. This requires the adoption of precautionary measures that eliminate or at least minimize those risks. These measures range from the legal and administrative instruments of protection up to the conservation studies and actions, always guided by criteria of preventive conservation.

The conservation of this fragile heritage thus becomes an interdisciplinary activity, aimed at protecting these unique ecosystems and parietal manifestations they have preserved for millennia. In Cantabria, recent years have seen great advances in the knowledge of the caves and the state of conservation of rock art. The task has been directed mainly to detect existing problems and the implementation of means to solve them, using traditional techniques (topographic and geodetic, physical-chemical, biological) and investigating the potential of new technologies (computer modelling, imagery).

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As an always provisional conclusion, today we can say that the state of conservation of decorated caves in Cantabria is satisfactory and that the risk factors are under strict control.

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THE CONSERVATION OF NERJA CAVE (SOUTHEASTERN SPAIN) Yolanda del Rosal Padial & Cristina Liñán Baena Fundación Cueva de Nerja. Instituto de Investigación. Ctra. de Maro, s/n, 29787- Nerja Málaga, Spain Mariona Hernández Mariné Departamento de Productos Naturales, Biologi ́a Vegetal y Edafologi ́a, Facultad de Farmacia, Universidad de Barcelona, 08028 Barcelona, Spain Nerja Cave is developed within dolomitic marbles belonging to Sierra Almijara, some four kilometres East of Nerja in the province of Malaga (Spain). The cave, with a nearly horizontal development, has a total surface of 35,000 m2. It was discovered in 1959 and prepared for visitors a year later, after its touristic opening works were carried out. With a current average of about 350,000 visitors per year, Nerja Cave represents a very important natural and cultural resource for the tourism industry of the region where it is located. The important archaeological site of the cavity, with a chrono-cultural and paleoenvironmental sequence between, at least, 25,000 and 3,600 years before present, and more than three hundred catalogued rupestrian paintings, belonging to the Upper Paleolithic and Late Prehistory, motivated its declaration as Good of Cultural Interest with the category of Archaeological Zone. In addition, Nerja Cave is catalogued as Geological Interest Place with International Relevancy. Moreover, exclusive animal species together with singular speleothems raise Nerja Cave as highlight of the Natural Heritage of Andalusia. Almost since its discovery, many research projects have been carried out in the cave, studying its geology, hydrogeology, microclimate and the microbiota. The results have enabled the identification of natural or anthropogenic factors that represent a risk to the conservation

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of the cultural and natural heritage of Nerja Cave and the design and application of the most appropriate preservation measures. Among these, the delimitation of the area of protection of the Good of Cultural Interest, the determination of the carrying capacity of the cavity and the gradual replacement of unsuitable materials previously used in the tourist track Additionally, specific protocols are being developed to implement the change of light quality and intensity, to reduce the development of photosynthetic organisms which cause biodeterioration and/or aesthetic damage to surfaces. Moreover, the use of non-destructive techniques to monitor lit areas has been implemented.

Poster Communications

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NANOGRAPHENE OXIDE: A NEW MATERIAL FOR A NON-INVASIVE AND NON-DESTRUCTIVE STRATEGY TO REMOVE BIOFILMS FROM ROCK SURFACES L. Bruno* Dept. of Biology, University of Rome ‘Tor Vergata’, Via della Ricerca Scientifica 1, Rome, Italy I. Ficorella Dep. of Chemistry, University of Rome ‘Tor Vergata’, Via della Ricerca Scientifica 1, Rome, Italy

L. Quici University of Perugia, PhD student in Biochemical Science and Biotechnology course, Perugia, Italy F. Valentini Dep. of Chemistry, University of Rome ‘Tor Vergata’, Via della Ricerca Scientifica 1, Rome, Italy Biodeterioration of rocks is a phenomenon caused by chemical and physical reactions due to the growth of microorganisms on the lithic faces. This causes structural and aesthetic damages to the substrata resulting hazardous for the cultural heritage1. Outdoor monuments and stone work of art surfaces are worldwide extensively covered by biological patina, named biofilms, mainly constituted by cyanobacteria and associated microorganisms2. Conventional restoration methods are based on harmful mechanical and chemical treatments that often induce damages to the substrata and are unsafe for the human health.

Our purposes are to provide non-invasive and non-destructive tools having better performances than that obtained using traditional procedures. In this work the antimicrobial activity of nano-grafene oxide (GO) on some

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strains of cyanobacteria has been investigated. The eco-sustainable GO, obtained by using a Green-Chemistry approach, represents a functionalized multilayer carbon based nanomaterial, exhibiting excellent chemical-physical and mechanical properties3. The antimicrobial effects of GO has been already studied in nano-medicine field applications, also combined with its biocompatibility effects. For the first time the GO’s antibacterial properties were tested on cyanobacteria, using GO’s nanodispersions at different concentrations, from 85 μg\ml to 1mg\ml. Among of all the most representative cyanobacterial strains, collected in Roman hypogea and isolated in culture, Oculatella subterranean and Scytonema julianum were selected for this study. Cyanobacteria treated with GO showed a reduction of the photosynthetic activity at high GO concentrations, while low concentrations didn’t sort significant effects. These preliminary results are particularly promising in the design of innovative nano-based technologies for the conservation of Cultural Heritage, in indoor and outdoor archeological places. 1Urzì C, De Leo F, Bruno L, Albertano P. 2010. Microbial

Ecology, 60: 116-129. 2Rossi F, Micheletti E, Bruno L, Adhikary SP, Albertano P, De

Philippis R. 2012. Biofouling 28:215–224. 3Xu Y., Shi, G. 2011. Journal of Materials Chemistry 21: 3311-

3323.

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RAMAN SPECTROSCOPIC ANALYSIS OF PREHISTORIC LITHIC INDUSTRY AND SOME FAT-BURNING STONE LAMPS FOUND AT THE ARDALES AND LAS PALOMAS DE TEBA CAVES IN THE GUADALTEBA COUNTY (MÁLAGA, SPAIN) C. Capel Ferrón SCAI, Universidad de Málaga, Spain S.E. Jorge Villar Área de Geodinámica Interna, University of Burgos, Spain and Centro Nacional de Investigación sobre la Evolución Humana (CENIEH), Burgos, Spain F.J. Medianero Soto Escuela Taller Parque Guadalteba, Guadalteba County, Málaga, Spain J.T. López Navarrete Departamento de Química Física, Universidad de Málaga, Spain V. Hernández Departamento de Química Física, Universidad de Málaga, Spain

We report on the Raman spectroscopic analysis of a series of prehistoric lithic tools found in different archaeological sites at the Guadalteba County (Málaga, Spain), which were further investigated by X-ray diffraction followed by Rietveld refinement. The bulk and patinas of the different specimens were firstly analyzed in the own archaeological sites by means of a handheld Raman spectrometer. Then, selected lithic tools were studied in more detail in the laboratory, by means of high-quality bench-top FT-Raman and micro-Raman spectrometers. Raman spectroscopy was found to be a useful technique to characterize, in a quick, easy, low-cost and non-

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destructive way, the mineral composition of the bulks and patinas of all the lithic tools.

The relative intensity of the Raman band at 501 cm-1 due to moganite, a naturally abundant silica polymorph, was found to significantly vary with respect to the strongest Raman-active symmetric stretching vibration of α-quartz at 465 cm-1, among the different lithic tools under analysis. The precise bulk wt% of moganite was then accurately quantified by means of high-quality XRD, followed by Rietveld refinement, and found to range between 2% and 15%.

We also analysed by means of Raman spectroscopy a series of fat-burning stone lamps as well as other likely remains of fire-linked activities of firing processes documented in different places inside the Ardales Cave. Calcite was found to be the main mineral component, what is logic since all portable closed-circuit bowl lamps, having shallow, circular or oval depressions designed to retain the melted fuel, were built from apical fragments of stalagmites. Other minor mineral components were α-quartz, dolomite, hematite and limonite. Phosphate was also evidenced in one of the portable stone lamps. Whose uncertain origin could result either from bone remains used as fuel, or from bat excrements. Black ashes were identified as being due to carbon, with different graphitization degrees.

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A STUDY OF INTERACTION BETWEEN PALAEOLITHIC PIGMENTS AND NEEDLE-FIBER MICRO-CRYSTALS OF CALCITE AT KAPOVA CAVE (SHULGAN-TASH), BASHKORTOSTAN, RUSSIA

O.Ya. Chervyatsova State Nature Reserve «Shulgan-Tash», Bashkortostan, Burzyan area, Russia A.S. Pakhunov Russian State University for the Humanities, Moscow, Russia L. Yu. Kuzmina Institute Biology, Ufa Scientific Center RAS, Ufa, Russia L.V. Leonova & S.P. Glavatskikh Institute of Geology and Geochemistry, Ural Scientific Center RAS, Yekaterinburg, Russia

Kapova (Shulgan-Tash) cave is located on the western slope of the Southern Urals in the Paleozoic framing of the Bashkir Anticlinorium in Carboniferous Visean-Serpukhovian chemogenic limestone. The cave entrance is situated at the mouth of the karst canyon on the right side of the Belaya River Valley. The length of the investigated part is 3050 m, the amplitude is 165 m, and the volume is 180,510 m3. Paleolithic paintings in the cave were discovered by Dr. Alexander Ryumin in 1959. Rock paintings were found in four halls. Up to now, over 50 images are known; they are mainly intricate geometric symbols and images of Pleistocene animals (Mammuthus primigenius, Coelodonta antiquitatis, Bison priscus, Equus sp.) Radiocarbon dating of cultural deposits showed that the cave paintings belonged to the Upper Paleolithic Period (14,000-17,000 years ago).

The speleothems, composed of fiber micro-crystals of calcite (NFC), have been investigated in the Kapova Cave. In

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2013, as a result of our study, a wide dissemination of fiber micro-crystals of calcite (NFC) in the cave was revealed. This type of deposits covered approximately 40% of the cave walls. Dripping water, which produced crystallization NFC, was characterized by weak supersaturation relative to calcium carbonate (computing potential deposition CaCO3 0,4-11 mg/l). The minimum air temperature of NFC growth was +4.0ºC. Data on the distribution, morphological characteristics, mineral (XRD analysis) and trace element composition (EDX and ICP-MS analysis), microcrystalline structure of the fiber micro-crystals (SEM observations) are presented. It is noted that in certain microclimatic conditions, coralloids and thick micrite crusts, except well-known moonmilk, can be formed out of the fiber micro-crystals. Growth of NFC plays a crucial role in the processes of mineral formation on paintings at the «first floor» of the cave. NFC-mineralization influenced the state of conservation of the paintings at the western wall of the hall of Znakov («Signs») by the redistribution of pigment particles in the mass of newly formed sediments. Some pictures halls Kupol'nyj ("Dome") and Znakov ("Signs") were partially covered of micrite crusts, presumably formed by recrystallization of fiber micro-crystals of calcite.

This report discusses the hypothesis of abiotic and biogenic genesis for fiber micro-crystals of calcite.

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HUMAN IMPACT ON SHOW CAVES: CHEWING GUMS STUCK TO THE WALLS I. Dominguez-Moñino, V. Jurado, M.A. Rogerio-Candelera & C. Saiz-Jimenez Instituto de Recursos Naturales y Agrobiología, IRNAS-CSIC, Avda. Reina Mercedes 10, 41012 Sevilla, Spain Cave tourism represents the main threat for a cave and for their conservation. Visitors cause important changes on the microclimate and introduce organic carbon and microorganisms into caves. The visitors leave behind skin cells, hair, soil trapped on the shoe sole and occasionally vomits, feces and urine. Recently we faced a new human impact on show caves: chewing gums stuck to the walls. We have found chewing gums on the walls of Cueva del Tesoro, Rincón de la Victoria, Málaga, Spain. The bacteria colonizing the chewing gums were represented by members of Actinobacteria, Alphaproteobacteria and Firmicutes while the fungi were members of the Ascomycota, order Eurotiales and Hypocreales. From the bacteria were noteworthy the isolation of Brachybacterium fresconis, Microbacterium aerolatum, Sphingopyxis italica and Bacillus litoralis and from the fungi the presence of Aspergillus, Fusarium and Penicillium species.

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DETECTION OF URBAN SUBSURFACE POLLUTION BY RAPID MULTIPARAMETRIC SURVEYS IN 16TH CENTURY PARANHOS SPRING WATER TUNNEL (PORTO, PORTUGAL) Angel Fernandez-Cortes, Enrique Sanz-Rubio & Angel Sanchez-Malo Geomnia Natural Resources SLNE, Cea Bermudez 14 5-A. 28003. Madrid, Spain Soledad Cuezva

Laboratorio de Petrologia Aplicada, Universidad de Alicante, Spain Elena Garcia-Anton Museo Nacional de Ciencias Naturales, MNCN-CSIC, José Gutiérrez Abascal 2, 28006 Madrid, Spain Maria Jose Afonso & Helder I. Chamine

Laboratório de Cartografia e Geologia Aplicada, Instituto Superior de Engenharia do Porto (ISEP), Instituto Politécnico do Porto, Rua Dr. A. Bernardino Almeida, 431, 4200-072 Porto; and Centro GeoBioTec-UA, Portugal Bernardo Hermosin

Instituto de Recursos Naturales y Agrobiologia de Sevilla, IRNAS-CSIC, Spain Sergio Sanchez-Moral

Museo Nacional de Ciencias Naturales, MNCN-CSIC, José Gutiérrez Abascal 2, 28006 Madrid, Spain The Subterranean Cultural Heritage is often located in urban areas with significant influence of human activity. Atmospheric and water pollution on these subterranean sites can contribute to damage artistic and cultural elements which

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houses or, even, it imposes restrictions on the time of its opening to tourism. Therefore, a detailed knowledge of the environmental processes underlying the pollutants’ storage on subsoil environments is vital for understanding and managing the Subterranean Cultural Heritage.

Here, we show the main results of the environmental monitoring programme conducted in the 16th century Paranhos spring water tunnel excavated in granites under the city of Porto (NW Portugal). The main objective is to detect and identify potential contaminated areas using innovative monitoring procedures including: 1) gas tracing providing detailed snapshots of CO2 (and its isotopic signal 13CO2), CH4 contents in the subsurface atmosphere, by using wavelength scanned cavity ring-down spectroscopy; 2) continuous monitoring of main microclimatic parameters of tunnel air (temperature, relative humidity and CO2 and 222Rn levels) at three locations and external weather conditions, and 3) periodical analysis of inorganic and organic tracer of groundwater contamination.

The rapid (few-hours long) multi-parametric surveys, encompassing simultaneous and spatial-distributed measurements of tracer gases and microenvironment parameters along 900 m of these galleries, have enabled to detect polluted areas and pollution sources of hazardous substances as well as the influence of spatiotemporal changes of tunnel atmosphere under different weather conditions. The deepest areas of Paranhos tunnel (-18 m below surface, on average) displays a high stability of microenvironmental conditions, e.g. air temperature annually oscillates below 1.5 ºC, with a higher inertia of temperature compared to seasonal changes at exterior. Conversely, the annual amplitude of air temperature at most superficial areas of tunnel (< -5 m below surface) reaches up to 3.3 ºC. Two opposite processes in function of air temperature trend are clearly noted in these shallow areas: 1) strong air exchange with the atmosphere during the colder months; and 2) thermal stratification of air when the temperature at exterior is well above the air

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temperature of tunnel. These areas nearby to the surface are topographically higher than the rest of tunnel. This geomorphologic feature of tunnel favours a motionless trap of warm and less dense air in these shallow areas and, consequently, it contributes to the gas entrapment process: CO2 content rises from 700 ppm to values above 1000 ppm, on average, with sharp increments above 2000 ppm when mean daily outside temperature exceeds 20 ºC. Radon content of air presents high levels along all tunnel locations, ranging 7-34 kBq/m3 with an annual average around 16 kBq/m3. The high 222Rn levels make it necessary to adopt preventive measurements for the health of visitors to tunnel against the danger arising from ionizing radiations, in accordance to the guidelines from International Commission on Radiological Protection and the EU recommendations.

A Keeling model tuned to the carbon isotope signal data (13CO2), with three-component mixture, corroborates the edaphic origin of CO2 found in tunnel air. CO2-soil source is more obvious in those shallow areas located at higher elevation but closer to soil, where a trap air parcel is often found due to hidden air exchange with outer atmosphere. In contrast, the areas of tunnel at lower altitudes display a heavier 13CO2 signal due to a higher influence of air renewal with exterior, even though the thickness of rock is greater in these areas. Methane content of air corroborates this general spatial pattern of air parcels, so the most isolated areas displays lower CH4-concentrations relative to those contents close to atmospheric background (~1.8-1.9 ppm) registered in the rest of areas. Anomalous CH4-contents of air above atmospheric background (ranging 3.4-22.8 ppm) were registered in some tunnel locations revealing the possible presence of natural gas leaks or leakage of gases from the wastewater pipelines of adjacent houses. Analysis of groundwater in those areas of tunnel with seepage points also revealed two prevailing areas with high potassium concentrations and nitrates content above acceptable limits for the drinking water. The presence of other indicators as

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calcium nitrate also confirmed the presence of wastewater pipeline leakage.

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IS THE PRESENCE OF BACTERIAL COMMUNITIES RELATED TO THE URBAN CONTAMINATION SOURCES OF THE PARANHOS SPRING WATER GALLERIES? A. M. Garcia-Sanchez, A. Z. Miller & V. Jurado Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, Sevilla, Spain A. Dionísio CERENA/CEPGIST, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal V. S. F. Muralha Vidro e Cerâmica para as Artes (VICARTE), Faculdade de Ciências e Tecnologia, UNL, Portugal M. J. Afonso & H. I. Chaminé Laboratório de Cartografia e Geologia Aplicada, Instituto Superior de Engenharia do Porto (DEG|ISEP), Porto; Centro GeoBioTec|UA, Portugal Subsurface engineering environments, such as mines, tunnels and water galleries, may have cultural significance, in addition to their inherent geological characteristics. The 16th century Paranhos spring water galleries located in Porto city (NW Portugal), were excavated throughout the granitic bedrock to supply water for the public fountains for more than five centuries. Nowadays, due to groundwater quality degradation and the high deterioration level of the granite galleries, the water is no longer used for public purposes. This subsurface granite heritage represents a favourable habitat for a wide variety of microorganisms, which participate in the formation processes of secondary minerals and dissolution of rock components. Due to the large number of anthropogenic contamination sources and urban surface activities along the course of the Paranhos spring water galleries, such as garages, petrol stations and in situ sanitation and sewer network, this subterranean environment shows high vulnerability to

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contamination. In fact, previous hydrogeochemical studies showed a nitrate- and sulfate-enriched composition for the groundwater probably resulting from urban drainage, sewer leakage and agricultural activities. In this study, we identify and characterize the bacterial communities in biogenic ferromanganese deposits coating the weathered subsurface granite in order to understand the relationship between their presence and the surface potential contamination sources. Molecular biology techniques, enrichment and isolation cultures, field emission scanning electron microscopy, transmission electron microscopy and micro-Raman spectroscopy were conducted. Thirty five different bacterial strains were isolated, most of them belonging to Bacillus, Pseudomonas and Ensifer genera. Many of these were represented by the species B. vietnamensis, B. thuringiensis, P. anguilliseptica, P. umsongensis, P. otitidis, P. linyingensis, P. alcaligenes, P. resinovorans and E. adhaerens. The presence of Pseudomonas spp., are usually associated with contaminated environments, which is consistent with the presence of contamination sources. Moreover, micro-Raman analysis pointed out the presence of copper phthalocyanine, a blue pigment chiefly used in the manufacture of enamels, printing inks, and automotive finishes. Its presence can be related with the garages and petrol stations on the surface of the Paranhos spring water galleries. Electron microscopy observations of isolates and enrichment cultures for Mn- and Fe-oxidizing bacteria revealed the presence of Mn- and Fe-rich precipitates upon bacterial cells, which suggests that these bacteria induce the precipitation of manganese oxides. We were thus able to isolate bacteria probably involved in the formation of the black ferromanganese oxides coating the Paranhos granite surfaces. This clearly demonstrates that the interest in the geomicrobiology of man-made subterranean environments lies not only in the fact that some of them can provide useful information for general and applied microbiology but also for representing geomining sites of particular geological and cultural relevance that needs conservation and maintenance interventions.

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INDOORS DIVERSITY IN PHOTOTROPHIC BIOFILMS AT ST PAUL’S CATACOMBS (MALTA) Antonio Gómez-Bolea, Isabel Álvaro, Mariona Hernández-Mariné & Esteve Llop Universitat de Barcelona, 08028 Barcelona, Spain Stephanie Sammut Malta Heritage, Malta St Paul’s Catacombs are an archaeological site dating from the 4th century, located within the urban area of Rabat (Malta), but in a semiurban environment. The catacombs were used during different periods, and after the 15th century were abandoned. They were rediscovered, cleared and examined during the 19th century.

Biofilms colonizing illuminated stone surfaces of these catacombs are mainly composed by subaerial phototrophic species. The composition of these biofilms depends mainly on water availability and light. Light comes from two main sources, artificial and natural. Artificial light represents a more constant source; it comes from bulbs, which are switched on eight hours daily. Natural light is projected from doors and external vents, and is more variable, depending on several factors such as orientation of entrance, season of the year, or cloudiness.

Forty five taxa are reported from 21 catacombs. Cyanobacteria are the most diversified with 24 taxa; among them, the most frequent are Chroococcidiopsis sp. and Leptolyngbya nostocorum. Chlorophyta are represented by 13 taxa, with Desmococcus olivaceus, Chlorella minutissima and Muriella sp. as the most frequent. Bacillariophyta has five representatives, with Diadesmis contenta and Hantzschia amphyoxis being the most common. Rhodophyta are represented by two species, growing only in areas with natural

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light. There are also one lichenized Ascomycete and protonemata of mosses.

Cyanobacteria seem to prevail in the photoautotrophic community from surfaces with natural illumination. On the other side, chlorophytes dominate in artificially illuminated surfaces.

Light is the main but not the only factor to drive phototrophic biofilms; other factors, such water availability or substrate characteristics, determine changes in diversity and abundance of species.

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CUEVA DEL TESORO, RINCÓN DE LA VICTORIA, MÁLAGA: A TREASURE OF BIODIVERSITY Valme Jurado Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, Avenida Reina Mercedes, 10, 41012 Sevilla, Spain Alena Nováková Institute of Soil Biology, Biology Centre AS CR v.v.i., Na Sádkách 7, 370 05 České Budějovice, Czech Republic Mariona Hernandez Marine Departamento de Productos Naturales, Biología Vegetal y Edafología, Facultad de Farmacia, Universidad de Barcelona, 08028 Barcelona, Spain Cesareo Saiz-Jimenez Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, Avenida Reina Mercedes, 10, 41012 Sevilla, Spain All niches of the planet Earth are colonized by microorganisms and caves are not an exception. The great adaptability of microorganisms permits to establish themselves in caves where develop different types of microbial communities. La Cueva del Tesoro harbours a great biodiversity barely explored. To the light of the ongoing projects and studies, Cueva del Tesoro revealed to harbour a rich bacterial and fungal diversity. In the last few years two new species of Aspergillus and three species of bacteria were isolated and described. In addition, the phototrophic communities showed to be of relevance regarding the study of parasitized microorganisms. If the removal and cleaning of phototrophic communities is carried out, this could endanger the conservation of the cave diversity due to the relationships between all cave inhabitants and will prevent further studies that could be carried out. We propose to declare Cueva del

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Tesoro a reservoir of biodiversity and encourage scientists to carry out their taxonomic studies in this interesting cave. We consider that the rock art present in the cave, mostly in non visited and impacted galleries, can be well preserved in this largely dry cave and can be easily controlled. In this cave, biological diversity should be considered worthy of preservation.

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PATHOGENIC MICROORGANISMS IN ALTAMIRA CAVE, SPAIN, RELATED TO HUMAN ACTIVITIES Valme Jurado, Leonila Laiz, Instituto de Recursos Naturales y Agrobiologia, CSIC, Avenida Reina Mercedes 10, 41012 Sevilla, Spain Sergio Sanchez-Moral Museo Nacional de Ciencias Naturales, MNCN-CSIC, José Gutiérrez Abascal 2, 28006 Madrid, Spain Cesareo Saiz-Jimenez Instituto de Recursos Naturales y Agrobiologia, CSIC, Avenida Reina Mercedes 10, 41012 Sevilla, Spain With today’s leisure tourism, the frequency of visits to many caves makes it necessary to know about possible potentially pathogenic microorganisms in caves, determine their reservoirs, and inform on the consequences of such visits for people who are immunosuppressed, undergoing chemotherapy, or have lowered defences. Unfortunately, information on this topic is not well known, and visitors remain unaware.

Determining the extent of the potential danger of cave visits is of great interest ― not only scientific, but also medical and social. Our data reveal that Altamira Cave visits could represent a potential danger to visitors. In addition to the presence of bacteria related to human activities (e.g. Staphylococcus epidermidis, Ochrobactrum anthropi), other opportunist microorganisms, whose existence and possible development in humans is currently unknown, were isolated.

The most-frequent diseases produced by the microorganisms identified in the cave are located in the respiratory system and skin, therefore it is recommended that in the case of a visit, the visitors should wear protective masks and clothings. Visits to any subterranean environment should

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possibly no longer be looked upon as a simple, riskless tourist activity, rather as one with potential risks for human health, particularly for the ill and elderly.

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CONTRIBUTION OF CULTURE-INDEPENDENT METHODS TO CAVE AEROBIOLOGY: THE CASE OF LASCAUX CAVE Pedro M. Martin-Sanchez

Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, Avda. Reina Mercedes 10, 41012 Sevilla, Spain and BAM Federal Institute for Materials Research and Testing, Division 4.1 Biodeterioration and Reference Organisms, Fabeckstrasse 60, 14195 Berlin, Germany Cesareo Saiz-Jimenez

Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, Avda. Reina Mercedes 10, 41012 Sevilla, Spain Lascaux Cave in France contains one of the most notorious examples of rock art in the world made up of numerous Palaeolithic paintings and engravings. Twenty years after its discovery, in the 60’s of the past century, several crises threatened the natural bioclimatic equilibrium of the cavity forcing its permanent closure to the public visits. Three main microbial outbreaks sequentially disturbed the cave environment: green colonizations of the algae Bracteacoccus minor (1960-1970), white mycelial masses of Fusarium solani (2001-2006), and black stains caused by the fungi Ochroconis lascauxensis and Acremonium nepalense (2001-present).

Except a few previous studies, the majority of papers available on the microbiology of Lascaux Cave were published from 2009 and contributed to the understanding of the complex processes of microbial colonization and biodeterioration of the cave and its paintings. They covered a variety of topics such as the composition of the microbial communities (bacteria, fungi and amoebae) dwelling on different substrata, with special attention to the black stains, the relationships between the microorganisms and arthropods, the management, such as for example, the intense biocide treatments, etc.

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An aerobiological study focused on the cultivable bacteria and fungi of Lascaux Cave was also carried out in 2010. These data offered a complete inventory of airborne microorganisms isolated from diverse halls in two different seasons.

In order to evaluate new aerobiological methods, the aim of the present study was to characterize the airborne bacterial and fungal communities from the Passageway, a key area connecting the main axes of Lascaux Cave, using culture-independent methods such as the construction of clone libraries based on ribosomal genes.

The results were compared with those previously achieved by culturing in the same area, and discussed according to the knowledge acquired in last years on the microbial ecology of the cave. Based on these data, we can conclude that culture-independent methods provided valuable information identifying the microorganisms more precisely and rapidly, avoiding the limitations imposed by culturing. In addition, these molecular techniques showed a significant higher sensitivity. Therefore, such methods should be included in future aerobiological studies in caves or any other environment to complement the traditional culture methods.

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MICROBE-MINERAL INTERACTIONS IN A PORTUGUESE GEO-ARCHAEOLOGICAL SITE A. Z. Miller Instituto de Recursos Naturales y Agrobiologia de Sevilla (IRNAS-CSIC), Sevilla, Spain; CERENA/CEPGIST, Instituto Superior Técnico, U Lisboa, Lisboa, Portugal A. Dionísio CERENA/CEPGIST, Instituto Superior Técnico, U Lisboa, Lisboa, Portugal M. E. Lopes, M. J. Afonso & H. I. Chaminé Laboratório de Cartografia e Geologia Aplicada, Instituto Superior de Engenharia do Porto (DEG|ISEP), Porto; Centro GeoBioTec|UA, Portugal This study concerns the secondary mineral deposits found in an abandoned mining heritage of particular geological and cultural significance. The Aveleiras mine is an old water mine, belonging to St. Martin of Tibães Monastery, near Braga (NW Portugal), which through the last three centuries supplied the monastery until it was converted in a wolfram mine in 1940-62. By initiative of the Tibães Monastery, it will be partially rehabilitated into a geosite and, in the near future, into a geo-mining museum. Secondary mineral deposits were observed in this metasedimentary subsurface environment, including black ferromanganese deposits, mucolite-like stalactites and blue-green deposits, which cover the walls and ceiling of the Aveleiras mine. The formation of these speleothem features are potentially induced by bacterial communities. In order to evaluate their biogenicity, morphological and geochemical characterization were conducted. Scanning electron microscopy examinations revealed the presence of microorganisms and microbial mats concealed within these mineral deposits, particularly within

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the black ferromanganese deposits. Mineral encrustations were found on biogenic filament-like forms consisting of tubular mineralized filaments. The typical morphology of biogenic manganese oxides consisting of Mn-rich fibril aggregates embedded in extracellular polymeric substances (EPS) was also found. Elemental analyses performed by energy dispersive X-ray analysis (EDS) showed enrichment in manganese and iron, which evidences a relation of microorganisms to the formation of these secondary deposits. In addition, actinobacteria-like filaments, coccoid cells with echinulate ornamentation and additional rod-shaped bacteria could be observed in all samples, which form biofilms and/or microbial mats. The mucolite-like stalactites consisted of gelatinous substance of beige color rich in S and Fe. The blue-green color deposits were characterized as copper-arsenic deposits due to its enrichment in Cu and As, and depletion in Al, Si and Fe.

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LAMPENFLORA AS AN ACCOMPANY OF MASS CAVE TOURISM, PROBLEMS AND SOLUTIONS: EXAMPLE FROM POSTOJNA CAVE, SLOVENIA Janez Mulec Karst Research Institute, Research Centre of the Slovenian Academy of Sciences and Arts, Titov trg 2, SI-6230 Postojna, Slovenia Postojna Cave is one of the most known cave equipped for tourists display in Europe. Artificial lighting in the cave is responsible for light eutrophication and development of lampenflora, a community of mainly phototrophic organisms that is involved in biodeterioration of substrata. High number of visitors in Postojna Cave, exceeding 500.000 annually, and a relatively long lighting period with an average of 1000 hrs/sector/year in the tourist part of the cave, gives a boost for lampenflora which is a serious problem for preservation of speleothems and other objects of cultural value. The first publication that reported on the presence of mosses around lamps in Postojna Cave dated in 1941 what gives one of the oldest record on the presence of green plants deep in the underground. Totally 34 different species of mosses and ferns were identified in lampenflora community in Postojna Cave. Eucladioliths were found around some lamps in the cave. The majority of microscopic phototrophs in lampenflora belonged to Cyanobacteria (59%), followed by Chrysophyta (25%) and Chlorophyta (16%). In Postojna Cave till 2010, the procedure to remove lampenflora was based on active chlorine (sodium dichloroisocyanurate dihydrate, C3H4Cl2N3NaO5) in working concentration of 100 mg/l. In poor ventilated sections of the cave, chlorine odour remained for a month after application what was probably related with slow release of chlorine. In 2011 a new procedure was adopted to remove lampenflora which is based on 15% v/v solution of hydrogen peroxide (pH 7.0-7.5). This solution is suitable only for treatment of

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insensitive calcite surfaces without the presence of cave animals. Based on laboratory data and field experiences, it is recommended to apply solution with hydrogen peroxide within 20 minutes after the solution is prepared. Once hydrogen peroxide is buffered, it becomes unstable and losses its biocidal activity. In Postojna Cave, one set of application was consisted of three consecutive applications with one week between each application. Usually two sets of applications were applied in a year. When application was carried out during the wet period, the effectiveness of lampenflora removal was lower. In a six-month experiment in Postojna Cave with different LED lamps that emitted white light of different quality, the growth of Chlorella vulgaris, a frequent microscopic algae in lampenflora, was rather undisturbed. Unfavourable light quality for photosynthesis reflected in this organism in elevated biosynthesis of accessory photosynthetic pigments. Selection of emission spectra that resembles natural light is from the aspect of lampenflora growth on a long term less important, and more attention should be given to lighting regime and constant removal of re-emerging lampenflora patches around lamps.

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MICROSCOPIC FUNGI ISOLATED FROM CAVE AIR AND SEDIMENTS OF THE NERJA CAVE: PRELIMINARY RESULTS Alena Novakova Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology AS CR, v.v.i, Vídeňská 1083, 142 20 Praha 4 and Institute of Soil Biology, Biology Centre AS CR, v.v.i., Na Sádkách 7, České Budějovice, Czech Republic Vit Hubka Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology AS CR, v.v.i, Vídeňská 1083, 142 20 Praha 4 and Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01, Praha 2, Czech Republic Cesareo Saiz-Jimenez Instituto de Recursos Naturales y Agrobiologia de Sevilla, Avenida Reina Mercedes, 10, 41012 Sevilla, Spain Previous studies on fungal community in the Nerja Cave (Malaga, Andalusia) were focused only on a capture of fungal spores from cave atmosphere and outdoor air by the Hirst-type volumetric pollen traps, it means without cultivation methods. Results of these studies showed the presence of different spore types belong to Oomycota, Myxomycota, but mainly to Ascomycota and Basidiomycota including their anamorph states, where spores of Penicillium/Aspergillus were dominated. Our study was targeted on cultivable microscopic fungi. Several sites inside the cave, mainly in the Touristic Gallery (2011 and 2012) and in the Upper Gallery (2012) were selected and a few soil samples above the cave were collected. The isolation from outside air was also used to compare with cave environment. Two isolation methods were used for this study – the gravity settling technique for the isolation of air-borne microfungi inside and outside the cave and the dilution plate method for the isolation of microfungi from cave sediment and soil above the cave. The CFU counts

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of airborne cultivable microfungi were found much lower in comparison with previous published data from volumetric traps. Nevertheless a rich microfungal spectrum was isolated from cave air as well as from cave sediments. Up to now, sixteen and six species of the genus Aspergillus were identified from cave sediments and cave air, respectively, including one novel species. The study points to a rich diversity of Aspergillus species in Nerja Cave and suggest the need to investigate the reasons for such a predominance of this fungal genus.

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ARE FUNGI A REAL THREAT FOR THE CONSERVATION OF ALTAMIRA CAVE? Alena Nováková Institute of Soil Biology, Ceské Budejovice, Czech Republic Valme Jurado & Cesareo Saiz-Jimenez Instituto de Recursos Naturales y Agrobiologia, IRNAS-CSIC, Sevilla, Spain Altamira Cave is classified as a cave threatened by fungi. An inventory of the fungi present inside the cave was carried out between 2007 and 2009 in order to assess the potential threat for the cave conservation. The fungi can roughly be grouped according to their origin as plant saprophytes introduced in the cave through airborne spores, entomopathogenic fungi introduced by arthropods, coprophilous fungi growing on rodent feces and soil-borne fungi. The role of fungi as responsible of outbreaks and the measures needed for their control are discussed.

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RAMAN MICROSCOPY AND IR IMAGING OF THE PALAEOLITHIC PAINTINGS FROM KAPOVA CAVE, SOUTHERN URAL, RUSSIA Alexander Pakhunov Russian State University for the Humanities, Moscow, Russia Nikolay Brandt & Andrey Chikishev Physics Department and International Laser Center, Moscow State University, Moscow, Russia Twenty two samples from fifteen paintings from Kapova Cave (Southern Urals, Russia) were studied with the help of Raman microscopy and scanning electron microscopy. In addition, 12 paintings were imaged in the visible and infrared light. The images were used in development of the sampling strategy and characterization of the paintings.

The main component of the paintings is hematite which consists of crystallized hematite particles with a size of up to 10 m and disordered hematite particles with a size of less then 1 m. Goethite is found in a few samples. Carbon particles are identified in 10 samples from 10 paintings. All the paintings are clearly seen in IR light. Carbon deposits are also detected in some hematite grains. Anatase and rutile TiO2 are the trace white components. The bulk composition of the substrate from the first and the second floor samples is calcite and aluminosilicates, respectively.

IR photography is a powerful method for studying altered paintings as well as pattern decoding. This method provides additional information on the composition of paintings.

The results obtained demonstrate the applicability of micro-analytical methods and IR photography to the studies of Palaeolithic pigments and benefitial role of IR imaging in determining the composition of the paintings from the Kapova Cave.

This work is partially supported by RFBR grant 13-06-00277.

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MICROORGANISMS AND THE NATURAL HERITAGE OF DINARIC KARST – A REVIEW L. Pašić Department of Biology, Biotechnical Faculty, University of Ljubljana, Slovenia J. Mulaomerović Centre for Karst and Speleology, Sarajevo, Bosnia and Herzegovina The Dinaric karst is the largest continuous karst landscape in Europe. It extends from north-eastern Italy through Slovenia, Croatia, Bosnia and Herzegovina, Montenegro to Albania. Its extreme northwest part is the cradle of the world's study of karst phenomena with recorded show cave visits and scientific studies dating back to the 17th century. Its Škocjan Caves are recognized as a UNESCO site of World Natural Heritage, while its karst landscape, Vjetrenica cave and unique medieval monolithic tombstones are placed on its Tentative List. The subterranean fauna of the region is amongst the World's best studied and the region is acknowledged as one of the richest in the terms of biodiversity. Growing concern for the preservation of underground natural and cultural heritage has led to increased interest in its microbiota. Accordingly, in the last decade, the underground microbiota of Dinaric karst received substantial academic interest. It yielded a number of studies that focused on the diversity and the structure of autochthonous and allochthonous microbial communities found in the region. Microbes that inhabit underground habitats are now being recognized as central for key ecosystem functions but also as main culprits in biodeterioration of underground natural heritage. Hence, here presented study aims not only to introduce the natural and cultural heritage of Dinaric karst, but also to summarize current knowledge on microbial communities that inhabit

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these fragile ecosystems as well as to present the proposed guidelines for the preservation of heritage sites in this region.

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“LA VIE EN ROSE”: A REVIEW OF THE ROSY DISCOLORATION OF SUBSURFACE MONUMENTS G. Piñar, J. Ettenauer & K. Sterflinger University of Natural Resources and Life Science, Vienna Institute of Biotechnology (VIBT), Vienna, Austria The variety of biodeterioration phenomena affecting our cultural heritage is determined by several factors, such as the chemical composition and nature of the material itself, the climate and the exposure of the object. A well-known biodeterioration phenomenon, widely observed on monuments located in central and south Europe, is the rosy discoloration suffered by different materials, mainly stone, building materials and wall paintings but also burial-related materials, located in subterranean and non-subterranean environments. The affected monuments are under different climatic conditions, but as common denominator, all of them have a relatively low UV irradiations and constructional problems that enable water infiltrations. The migration of water, mainly through the walls, produces further crystallisation of salts on the surfaces leading to the formation of saline environments that provide optimal growth conditions for halophilic microorganisms. This study offers a review of the microbiota associated with well-investigated monuments affected by the rosy discoloration phenomenon and reveals intriguing similarities among the microbiota detected on the different locations studied.

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THE MURAL EXECUTION TECHNIQUE OF THE CUBICLE “DEI FORNAI” REVEALED BY LASER CLEANING R. Senserrich-Espuñes Facultad de Bellas Artes, Universidad de Barcelona, Barcelona, Spain B. Mazzei Pontificia Commissione di Archeologia Sacra, Roma, Italy M.G. Patrizi Impresa individuale, Roma, Italy S. Bracci, M. Realini & B. Sacchi Istituto per la Conservazione e la Valorizzazione dei Beni Culturali, ICVBC-CNR, Sesto Fiorentino, Italy G. Bartolozzi Istituto Fisica Applicata “Nello Carrara”, IFAC-CNR, Sesto Fiorentino, Italy During the year 2010 an intervention of conservation-restoration was carried-out in the cubicle "dei Fornai", in Domitilla’s Catacombs, in Rome. The cubicle is located in a section of the first floor of the catacomb, topographically assignable to an advanced Constantinian age. It presents a particularly monumental spaces and an extremely unique pictorial decoration because the subjects and topics represented.

One of the most common problems of wall paintings found in Rome’s catacombs is a dark film based on calcium carbonate which is mainly located on the vaults and upper parts of the walls of the hypogeal environments. His tonality varies from light to dark gray -even black- and has the consistency of a thin veil that covers the wall surface, becoming extremely compact and tenacious on numerous occasions.

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Being these overlying layers not removable using the traditional techniques of cleaning, since in most cases and due to the particular conditions of underground environments, the conservation of the wall paintings would be compromised, it was decided to apply a cleaning protocol using laser technology, with the scientific support of the Progetto Temart [1, 2].

Due to the unexpected cleaning level achieved, some decorative details relative to the technical execution of the paintings emerged, which at first were not discernible, such as the subtle marks of some instruments used during the creative process, whose imprints remained caught in the pictorial matter.

From the point of view of the execution technique it has been found that, compared with other works not clearly defined or of hasty implementation, the cubicle "dei Fornai" presents iconographic motifs and chromatic ranges perfectly consistent with the type of hypothesized owner, a personage who, by rank and social position, would be within the Annona directive summit, the organization that was managing the food supplies in Rome.

To deepen and try to clarify this dichotomy, we proceeded to making a diagnosis campaign characterized by the exclusive use of non-invasive techniques: Infrared Spectroscopy in the total reflection (TR FT-IR), X-ray Fluorescence Spectrometry (XRF), Colorimetric Measures, Digital Microscopy (DM) Visible and Induced Luminescence (VIL). The analyses carried out on pigments also show one particularly meaningful information that can be associated with the art historical and iconographic studies concerning the cubicle "dei Fornai". The presence of mixed colors or with a percentage of calcite denotes amplitude in the color range, absolutely unusual regarding to the standards registered on catacomb painting. This information confirms, therefore, advanced hypotheses about the allocation of the cubicle to a socially high customer with considerable economic capacity.

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[1] Patrizi M. G., Mazzei B., Mascalchi M., Siano S.; “Rimozione laser di pellicole scure su dipinti murali in ambiente ipogeo: il caso di studio del cubicolo ‘dei fornai’ nelle catacombe di Domitilla a Roma” in “Atti del convegno APLAR 3, Bari 2010,” Padova, 2010, pp. 73-83.

[2] Patrizi M. G., Senserrich R., Mazzei B., Mascalchi M., Agresti J., Osticioli I., Siano S.; “Il cubicolo ‘dei fornai’ nelle Catacombe di Domitilla a Roma, considerazioni a conclusione della pulitura laser”, in “Atti del convegno APLAR 4, Roma 2012”, Saonara (PD) 2013, 143-154.

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METHODOLOGICAL APPROACH TO MONITOR ROMAN CATACOMBS: EXPERIENCE IN THE CATACOMBS OF ST MARK, MARCELLIAN AND DAMASUS, ROME, ITALY Deodato Tapete CNR-ICVBC, Via Madonna del Piano 10, 50136, Florence, Italy, and Department of Geography, Durham University, Durham, UK Fabio Fratini CNR-ICVBC, Via Madonna del Piano 10, 50136, Florence, Italy Barbara Mazzei Pontificia Commissione di Archeologia Sacra, Via Napoleone III 1, 00185, Rome, Italy Emma Cantisani, Cristiano Riminesi, Rachele Manganelli Del Fà, Barbara Sacchi & Oana Cuzman CNR-ICVBC, Via Madonna del Piano 10, 50136, Florence, Italy Maria Gigliola Patrizi Individual enterprise, Via Cornelio Celso 18, 00161, Rome, Italy Luciano Scaletti & Piero Tiano CNR-ICVBC, Via Madonna del Piano 10, 50136, Florence, Italy

Roman Catacombs running underneath the ground level in Rome (Italy) are world-renowned for their extraordinary heritage of mural paintings produced by Christian communities since late 2nd - early 3rd centuries A.D. Since 1852 the Pontifical Commission for Sacred Archaeology (PCAS), Vatican, supervise the protection and conservation of these subterranean networks and, in the last years, is seeking to develop feasible and sustainable solutions to prevent

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alteration of the painted surfaces. As part of this strategy, microclimate monitoring is currently exploited by PCAS to understand better the physic-chemical interactions between the indoor environment and the exposed surfaces.

In this regard, as of 2012 PCAS and the Institute for the Conservation and Valorization of Cultural Heritage (ICVBC), belonging to the National Research Council of Italy (CNR), have initiated the scientific research project HYPOGEA aimed to: 1) develop a methodological approach of monitoring suitable to this particular type of subterranean context; and 2) investigate the conditions triggering one of the most common alteration processes observable in different hypogeum systems (e.g. North Korean tombs and Jewish catacombs), i.e. the abiotic and biotic dynamics of calcium carbonate crystallizations.

This conservation topic strongly relates to a various range of issues, including but not limited to: (i) aesthetic and conservation impacts due to a progressive coverage of the painted and decorative surfaces under thick and opaque incrustations, up to totally hidden pictorial details; (ii) selection of appropriate methods for cleaning which can effectively couple the operability under hypogean microclimate conditions with the selectivity of the removal activity; (iii) identification of restoration materials assuring a compatible application and temporal stability within hypogean environments.

This paper illustrates the rationale of the methodological approach of monitoring and diagnostics currently tested in the Cubicle of the Twelve Apostles, within the Catacombs of St Mark, Marcellian and Damasus. Critical discussion is also conducted with regard to the operational issues associated to monitoring of Roman Catacombs (e.g., types of sensors to install and their location), as well as about the solutions chosen in this pilot study to balance conservation needs, costs constraints and technical requirements to achieve sound microclimate measurements.

Outcomes of the first year of monitoring are discussed by analysing the trends of the microclimate parameters of T, RH,

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air CO2 content and air mass movement, jointly with the surface and stratigraphic evidences retrieved from micro-photogrammetric, colorimetric and petrographic investigations of mural painting samples purposely applied on the naked rock walls and monitored simultaneously with the acquisition of the microclimate measurements.

The picture obtained so far is allowing us to assess the feasibility, sustainability and effectiveness of our methodological approach, in light of its potential exploitation also in other cubicles across the huge networks of catacombs managed by PCAS.

The still running HYPOGEA project is also providing a springboard for the design of management policies enabling, in near future, public opening in safety conditions for the conservation of mural paintings. In this regard, one of the main objectives of PCAS is to ensure accessibility to this subterranean heritage for its valorization.

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IDENTIFICATION OF LIME-BASED MURAL PAINTING TECHNIQUES IN CATACOMB ENVIRONMENTS USING WELL-ESTABLISHED CRITERIA OF STRATIGRAPHIC INVESTIGATION

Deodato Tapete CNR-ICVBC, Via Madonna del Piano 10, 50136, Florence, Italy, and Department of Geography, Durham University, Durham, UK Rebecca Piovesan Departimento di Geoscienze, Universita di Padova, Italy Emma Cantisani & Fabio Fratini CNR-ICVBC, Via Madonna del Piano 10, 50136, Florence, Italy Claudio Mazzoli & Lara Maritan Departimento di Geoscienze, Universita di Padova, Italy Barbara Mazzei Pontificia Commissione di Archeologia Sacra, Via Napoleone III 1, 00185, Rome, Italy Inner surfaces of Roman catacombs preserve an extraordinary heritage of mural paintings and plaster decorations dating back 2nd – 5th century AD produced by ancient fossori, whose artistic and technical skills built upon the Roman tradition of lime-based mortars, with capabilities of developing practical solutions that demonstrate adaption to the hypogean environment (Mazzei, 2005; Tapete et al., 2013).

What painting technique was employed is frequently debated and it is not always obvious whether the painted surfaces under examination can be classifiable as true frescos or other type of technique. This is also due to the effect exerted by the microclimate conditions characterizing the indoor environment at the time of execution and/or in the subsequent period during lime hardening. It is well known,

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indeed, that hypogean microclimate with RH persistently close to saturation and high rate of CO2 can lead to longer times for carbonation in depth. Furthermore, calcium carbonate crystallisation processes frequently occurring on the surface and across the stratigraphy in hypogean paintings can alter the overall stratigraphic structure and add layers that hide the former stratigraphic features.

In this regard, the recent laboratory-based research by Piovesan et al. (2012) has demonstrated that well-established criteria of stratigraphic analysis based on evidences retrieved from Optical Microscopy (OM) and Scanning Electron Microscopy (SEM) on cross-sections of mural paintings can effectively help to identify clearly the pictorial technique of aerial paintings, thereby allowing fresco to be distinguished from lime-painting.

This paper aims to present the results obtained by applying the methods by Piovesan et al. (2012), together with that by Tapete et al. (2013) for petrographic study of surface crystallisations in hypogean paitings, on a set of historical samples from Roman catacombs in Rome (St. Tecla, Domitilla, and Priscilla)

The microstratigraphy was examined to detect analogies and differences with typical stratrigraphy of fresco and lime-paint painting techniques, using the following objective criteria: (i) the thickness of the pigment-bearing layers (thinner in fresco), (ii) the outline of the painting surface (rough for the fresco, smooth for the lime paint), and (iii) the number of Ca-rich carbonation layers (single for the fresco, one exterior and one interior for the lime paint). All the samples were analysed, in both thin and thick transversal polished section, by OM and SEM.

The considered criteria demonstrated very reliable on these materials, although with some interesting variations in the catacomb environment. In particular, the thickness of the Ca-rich layers, especially in the fresco paintings, resulted always higher than the corresponding layer produced in aerial conditions, with a reasonable explanation with regard to H2O-CO2 exchange between air and painted surface.

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Most of the samples showed surface crystallisation processes due to dissolution-precipitation reactions triggered by surface condensation of CO2-rich water. The penetration of surface incrustations into the paint layer can mask the presence of the exterior Ca-rich layer. Combined SEM and OM analyses at high magnification led us to recognize the textural remnant (like a ‘ghost’) within the incrustation itself due to alteration of the exterior Ca-rich layer. In such cases, the mural painting can be reasonably classified as a lime-paint technique, because both the Ca-rich layers are detected.

Piovesan, R., Mazzoli, C., Maritan, L., Cornale, P. 2012. Archaeometry 54: 723-736.

Mazzei, B. 2005. In: Climatic Stabilization and Building Physics. Sustainable Approaches to Safeguarding the World Cultural Heritage”, München-Berlin: 65-78.

Tapete, D., Fratini, F., Mazzei, B., Cantisani, E., Pecchioni, E. 2013. Periodico di Mineralogia 82: 503-527.

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NEW SPECIES DESCRIPTION, BIOMINERALIZATION PROCESSES AND BIOCLEANING APPLICATIONS OF ROMAN CATACOMBS-LIVING BACTERIA Clara Urzì

Dept. of Biological and Environmental Sciences, University of Messina, Viale F. Stagno d’Alcontres, 31, 98166 Messina, Italy

Laura Bruno Dept. of Biology, University of Rome ‘Tor Vergata’, Via della Ricerca Scientifica snc, Italy Filomena De Leo Dept. of Biological and Environmental Sciences, University of Messina, Viale F. Stagno d’Alcontres, 31, 98166 Messina, Italy Lucia Krakova & Domenico Pangallo Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska cesta 21, 84551 Bratislava, Slovakia In the frame of the project CATS (2000-2003) and the following updates (2009-2011), many informations were achieved in the study of microflora colonizing the rocky surfaces of the Roman Catacombs of St Callistus and Domitilla.

The characterization and description of microorganisms present on the deteriorated surfaces was not only a mere list of species present (some of them even described as new species, such as Kribbella sancticallisti, K. catacumbae, K. albertanoniae, Oculatella subterranea), but all together gave useful information on the events that occurred on those delicate environments and allowed to better plan the strategies to be used for each site.

In particular, during the CATS project samples were taken in the site CSC13 called Ocean’s cubiculum, and after it a blue lamp were used in order to control (successfully) the phototrophic microflora. Before the treatment, the site could

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have been considered an undisturbed environment; the microbial population associated to the specific pattern of colonization showed the predominance of Actinobacteria up to 70% of the total microflora isolated in correspondence of white patinas as well as associated to the phototrophic microflora. After 10 years, the majority of phototrophic species disappeared from the site while the percentage of Actinobacteria was still high along with an increasing number of Gram negative and spore forming bacteria that were isolated from the site. These data showed that a stress factor was introduced in the site, but still the microclimatic conditions favored the heterotrophic microbial growth. Therefore these results showed the necessity to associate a biocide treatment to the blue lamp applications.

Gram negative strains belonging to the genera Stenotrophomonas, Chitinophaga, Lysobacter were isolated in correspondence of the former phototrophic based biofilm and degradative tests showed a very wide panel of hydrolytic activity (e.g. proteolytic, pectinolytic, etc) that were connected to the organic debris released by the dead cells.

Due to this enzymatic capacity, some selected strains were tested with good results for bioremoval of organic products. It was clear that the screening of their degradative ability and the improvement of protocols for their use were mandatory for any intervention of bioremediation.

In the site CSC13, cotton like, white alteration were often associated specifically to the ability of great variety of bacterial species to precipitate CaCO3 causing esthetical and mechanical (bio)-deterioration. Very often the cause was due to water content and to the presence of wide variety of Actinobacteria, Gram negative bacteria as well as spore forming microorganisms. In particular, Bacillus strains were associated to the cotton-like structures. From applicative point of view, this “natural” inclination of these bacteria to produce solid phase crystals once controlled and driven to conservative purposes could be used for the consolidation and protection of limestone and marble items and surfaces.

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Last but not less important, other aspects that still need to be better studied concern the evaluation of hazardous risk of the microorganisms due to volatile microbial metabolites to human health through the production of allergens, toxins or the direct contact with pathogens. This is particularly true for these special indoor habitats such as catacombs due to the increasing mass tourism. In fact, it is well known that several actinobacteria produce volatile compounds and that some of Gram negative strains commonly isolated from these sites, like Stenotrophomonas maltophilia have pathogen relatives in other environments. Studies should not be only limited to the recognition of microorganisms/biodeteriogens present, but also to their impact on the human health.

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THE CONSERVATION OF KAPOVA CAVE, RUSSIA: PAST AND PRESENT

V.S. Zhitenev

Moscow State University, Russia

Kapova Cave is located in the southern Urals, Russia. It is located on the Belaya river, in the State Nature Reserve "Shulgan-Tash". The cave is a system of halls, galleries and corridors, distributed at three hypsometric levels, the lower of which is occupied by the river. In 1959 a zoologist, A.V. Ryumin, discovered cave paintings on the middle and upper levels of the cave. Archaeological researches were carried out by O.N. Bader (1960-1978), V.E. Scelinsky (1982-1991), T.I. Scerbakova and V.G. Kotov (2000s) (Bader, 1965; Scelinsky V.E,, Sirokov V.N., 1998). From 2008, researches in the Kapova cave are carried out by Southern Urals Archaeological expedition of the Moscow State University (Zhitenev, 2011, 2012). Kapova cave is one of the largest caverns in the region. It was an important touristic and speleological object before the discovery of wall paintings. The flow of tourists increased after the discovery of wall paintings. A myth about healing power of clay/calcite in the cave and the popularity of speleotherapy promoted the rise of visitor’s stream. For a long time, pass into the chambers with wall paintings was accessible to all tourists accompanied by guides. Chambers with paintings were completely closed for the mass flow of tourists in the middle of 2000s. During Bader's works, the main problem was the protection of caves by ordering sightseeing activities and protecting the wall paintings from barbaric visitor’s activity. In the same period, the special staff of the expedition under the leadership of O.N. Bader revealed some paintings under calcite layers on the middle level. On the upper level, wall paintings generally were not covered by calcite. Shchelinsky's

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team found an overgrowing already open paintings. Many of these paintings were damaged by tourists. Only geologists worked in the cave from 1992 to 2004 (without archaeological observation). Intensive scientific researches began after closing of tourist’s activities in the cave. The State Nature Reserve "Shulgan-Tash" staff and the invited experts conduct researches (speleoclimate, microbiology) in order to preserve the Palaeolithic paintings. A special program is devoted to the study of wall paintings pigments and their comparison with the pigments of the cultural layers. Works on the cave for the preservation at modern scientific level are at an initial stage. The research was supported by RFBR grant 13-06-00277.

AUTHORS LIST Afonso, M.J. 50, 54, 64 Álvaro, I. 56 Bardisa, M. 34 Bartolozzi, G. 75 Belleza, S. 19 Bracci, S. 75 Brandt, N. 71 Bruno, L. 19, 43, 84 Cantisani, E. 78, 81 Capel Ferrón, C. 45 Chamine, H.I. 50, 54, 64 Chervyatsova, O.Ya. 47 Chikishev, A. 71 Cuezva, S. 50 Cuzman, O. 78 De las Heras, C. 30 De Leo, F. 19, 84 Del Rosal Padial, Y. 39 Dionísio, A. 54, 64 Dominguez-Moñino, I. 49 Ettenauer, J. 74 Fernández-Cortés, A. 21, 50 Ficorella, I. 43 Fratini, F. 78, 81 Galán, J.M. 11 Garcia-Anton, E. 50 Garcia-Sanchez, A.M. 54 Geneste, J.M. 32, 34 Glavatskikh, S.P. 47 Gómez-Bolea, A. 56 Hermosin, B. 50 Hernandez Mariné, M. 39, 56, 58 Hernández, V. 45 Hubka, V. 68 Jorge Villar, S.E. 45 Jurado, V. 49, 54, 58,

60, 70 Krakova, L. 84 Kuzmina, L.Yu. 47 Laiz, L. 60 Lasheras, J.A. 30 Leonova, L.V. 47 Liñán Baena, C. 39 Llop, E. 56

Lopes, M.E. 64 López Navarrete, J.T. 45 Manganelli del Fà, R. 78 Maritan, L. 81 Martin-Sanchez, P.M. 62 Mauriac, M. 32 Mazzei, B. 75, 78, 81 Mazzoli, C. 81 Medianero Soto, F.J. 45 Miller, A.Z. 54, 64 Mulaomerović, J. 72 Mulec, J. 66 Muralha, V.S.F. 54 Nováková, A. 58, 68, 70 Ontañón, R. 37 Pakhunov, A.S. 47, 71 Pangallo, D. 84 Pašić, L. 72 Patrizi, M.G. 75, 78 Piñar, G. 74 Piovesan, R. 81 Prada, A. 30 Quici, L. 43 Realini, M. 75 Riminesi, C. 78 Rogerio-Candelera, M.A. 49 Sacchi, B. 75, 78 Saiz-Jimenez, C. 49, 58, 60,

62, 68, 70 Sammut, S. 56 Sanchez-Malo, A. 50 Sanchez-Moral, S. 24, 50, 60 Sanz-Rubio, E. 50 Scaletti, L. 78 Senserrich-Espuñes, R. 75 Spinola, G. 15 Sterflinger, K. 74 Tapete, D. 78, 81 Tiano, P. 78 Turchetti, M.A. 17 Urzì, C. 19, 84 Valentini, F. 43 Zhitenev, V.S. 87