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Moore, D.M. (1992). Lichen Flora of Within the frame of EC project ENVA- Great Britain and Ireland. 710 pp. Natural CT95-0104 microbial colonization in both History Museum, London. caves was studied by conventional isolation and cultivation techniques. Wirth, V. (1995). Die Flechten Baden- Sampling was done by touching the rock Württembergs Teil 1 & 2. Ulmer, between the paintings with sterile cotton Stuttgart. [Superb colour photographs, swabs and suspending the adherent keys, notes on ecology, etc.; in German.] in sterile 0.15 sodium phosphate buffer solution. Additionally contact Macrophotographs on line plates with different culture media and http://dbiodbs.univ.trieste.it/web/lich/arch_icon pieces of rock or soil material were used http://www.lias.net/index.cfm for isolation of microorganisms from different sites in the caves. Ecology Nimis, P.L. (1993). The Lichens of Italy. Pure cultures of actinomycete isolates An annotated catalogue. Museo were classified by morphological, Regionale di Scienze Naturali. selected physiological and Monographie XII. Torino. pp. 897. chemotaxonomic methods. On the basis of a polyphasic taxonomic approach in Purvis, O.W., Coppins, B.J., most of the cases a tentative genus Hawksworth, D.L., James, P.W. and affiliation was possible. Moore, D.M. (1992) Lichen Flora of Great Britain and Ireland. 710 pp. Natural As the result of our study it was stated History Museum, London. that actinomycetes were the most abundant microorganisms in the two Glossary caves (Groth et al. 1999, 1999a, 2001). Hawksworth, D.L., Kirk, P.M., Sutton, The term actinomycetes refers to all B.C. and Pegler, D.N. (1995) Ainsworth Gram-positive bacteria with a high G+C & Bisby's Dictionary of the Fungi. Eighth content (> 55 mol %) in their DNA. Edition, XII + 616 pp. International Actinomycetes are morphologically Mycological Institute. CAB International. diverse and comprise different mophological types; irregular rods, cocci and filamentous or mycelium forming ACTINOMYCETES IN CAVES organisms.

Ingrid Groth1, Cesareo Saiz- The cave isolates could be affiliated to 2 11 different families of the order Jimenez Actinomycetales, class . 1 Hans-Knöll-Institute for Natural Products This fact indicates a high phylogenetic Research, Jena, Germany. diversity which has been developed 2 Instituto de Recursos Naturales y within these special biotopes (Table 1). Agrobiologia, Sevilla, Spain. Furthermore it was demonstrated by fatty acid profiles (MIDI system), Parietal paintings are valuable works of utilization patterns of 95 different carbon art giving evidence of prehistoric life. In sources (BIOLOG-System) combined with many cases these paintings were morphological studies that there was an detected by chance after thousands of equally high diversity at the or years as it happened upon the Altamira strain levels within the isolates being Cave in Spain and the Grotta dei Cervi in affilated to genera. Italy. While the Altamira Cave (discovered in 1879) had been opened to The predominant actinomycete isolates tourists reaching a daily flow of up to from both caves belong to the genus 3000 visitors in the 1970's, the access Streptomyces. Members of this genus to the Grotta dei Cervi (discovered in are known for their versatile metabolic 1970) has been restricted only to activities. In dependence on scientists. environmental or cultural conditions in the laboratory actinomycetes are able to

12 COALITION No. 4, March 2002 produce a wealth of structural diverse Our studies in the two different caves chemical compounds including pigments revealed that mass growth of and organic acids which may contribute actinomycetes seemed to be controlled in to biodeterioration of works of art. an undisturbed environment, like Grotta dei Cervi, by natural factors (low temperature, high humidity and limited Table 1: Taxonomic diversity of actinomycetes from the Altamira Cave and Grotta dei Cervi amount of nutrients). While microbial Family Genus colonization in the Grotta dei Cervi did Brevibacteriaceae Brevibacterium not result in visible damages of the Dermabacteraceae Brachybacterium paintings over thousands of years, the Gordoniaceae Gordonia deteriorations observed in the caves of Microbacteriaceae Agromyces Altamira and Lascaux after their opening Microbacterium Micrococcaceae Arthrobacter clearly demonstrate the negative effects Micrococcus of mass tourism within a relative short Rothia period. Micromonosporaceae Micromonospora Nocardiaceae Nocardia Rhodococcus To combine both access to works of art Nocardioidaceae Nocardioides in underground environments and to Nocardiopsaceae Nocardiopsis preserve these early manifestations of Pseudonocardiaceae Amycolatopsis prehistoric life, basic multidisciplinary Saccharothrix studies are necessary. Concerning Streptomycetaceae Streptomyces microbial colonization both noncultural (Rölleke et al. 1996) and cultural techniques should be combined. While Furthermore Cañaveras et al. (1999) molecular approaches based on detection suggested that actinomycetes may play a and identification of DNA sequences role in the formation of moonmilk encoding for 16S rRNA or 23S rRNA deposits in hypersaline environments of equally include culturable and some caves as actinomycetes (mainly nonculturable bacteria, classical isolation streptomycetes) were isolated from and cultivation techniques selectively hydromagnesite and needle fiber support the growth of only a minor aragonite deposits in the Altamira Cave. fraction of the really existing bacteria in a This assumption was supported by the special biotope. However pure cultures of fact that numerous actinomycete isolates selected bacteria obtained by isolation were able to produce either calcite or procedures enable scientists to study the both calcite and vaterite crystals under metabolic activities of these organisms special laboratory conditions (Groth et al. and contribute to a better understanding 2001). of their role in biodecay processes. Furthermore living cultures are a It is now generally accepted that prerequisite for the discovery and biodeterioration of outdoor monuments is description of novel taxa from these not only caused by phototrophic interesting but up to now less studied microorganisms and fungi covering the environments. The occurrence of novel surfaces with green or dark biofilms but bacteria was confirmed by recent is rather a concerted action of diverse publications of two novel genera isolated procaryotic and eucaryotic from a Chinese cave (Groth et al. 1999, microorganisms. Among the 2002). heterotrophic bacteria actinomycetes predominate in hypogean environments. References They are able to colonize nearly all Cañaveras J.C., Hoyos M., Sanchez-Moral S., Sanz-Rubio habitats and can adapt to very poor and E., Bedoya J., Soler V., Groth I., Schumann P., Laiz L., Gonzalez I., Saiz-Jimenez C. (1999). Microbial highly saline environments, e.g. communities associated with hydromagnesite and stalactites. By formation of dormant cells needle-fiber aragonite deposits in a karstic cave actinomycetes can survive unfavourable (Altamira, Northern Spain). Geomicrobiol. J. 16, 9-25. Groth I., Saiz-Jimenez J. (1999). Actinomycetes in conditions. hypogean environments. Geomicrobiol. J. 16, 1-8. Groth I., Vettermann R., Schuetze B., Schumann P., Saiz- Jimenez C. (1999a). Actinomycetes in karstic caves of

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Northern Spain (Altamira and Tito Bustillo). J. Preliminary programme Microbiol. Meth. 36, 115-122. Groth I., Schumann P., Schuetze B., Augsten K., Kramer I., Stackebrandt E. (1999b). Beutenbergia cavernae gen. November 8th nov., sp. nov., an L-lysine-containing actinomycete isolated from a cave. Int. J. Syst. Bacteriol. 49, 1733- Session 1. Biodeterioration Processes: 1740. organic/inorganic substrates Groth I., Schumann P., Laiz L., Sanchez-Moral S., Session 2. Diagnostic techniques Cañaveras J. C. (2001). Geomicrobiological study of the Grotta dei Cervi, Porto Badisco, Italy. (traditional and innovative) Geomicrobiol. J. 18, 241-258. Session 3. Intervention procedures: Groth I., Schumann P., Schuetze B., Augsten K., Stackebrandt E. (2002). sinensis gen. nov., chemical (biocides) - Pysical/mechanical sp. nov. and sp. nov., two novel (laser, UV) actinobacteria isolated from a cave. Int. J. Syst. Evol. Microbiol. 52, 77-84. Session 4. Bioremediation Rölleke S., Muyzer G., Wawer C., Wanner G., Lubitz W. (1996). Identification of bacteria in a biodegraded wall November 9th painting by denaturing gradient gel electrophoresis of PCR-amplified gene fragments coding for 16S rRNA. Session 5. Biohazard in Restoration: Appl. Environ. Microbiol. 62, 2059-2065. Allergenic and pathogenic, toxicity. Session 6. Health protection guidelines Session 7. COALITION questionnaire. Forthcoming activities Send your preliminary inscription form (to be distributed) to [email protected] QUESTIONNAIRE ON THE APPLICATION OF MOLECULAR BIOLOGICAL TECHNIQUES IN CONSERVATION INTERNATIONAL CONFERENCE ON A questionnaire is being prepared by MOLECULAR BIOLOGY AND CULTURAL Caroline Kyi on behalf of COALITION to HERITAGE know the real impact on molecular tools into the conservation field and its An International Conference on Molecular feasibility in microbiological assessment. Biology and Cultural Heritage will be This will be distributed in April to the organized about 2-8 March 2003 in COALITION list and it is expected that Seville (Spain). we will obtain a high number of respondents. We ask you to collaborate This International Conference is open to with us in this task. The results will be researchers, conservators, restorers and presented in the Florence Advanced Cultural Heritage authorities. The data Course (8-9 November, 2002) and collected and reviewed by COALITION published in the Newsletter. partners, during the project, will be disseminated in the Conference, and the invited lectures and contributions will be published in a monographic book. ADVANCED COURSE ON BIODETERIORATION OF THE CULTURAL Preliminary registration forms (to be HERITAGE distributed) should be sent to [email protected] Organized by COALITION in collaboration with the Opificio delle Pietre Dure, Florence, Italy. Call for papers Florence, 8-9 November 2002

Addressed to biologists, conservators This newsletter is open to external and restorers under 35 years old. contributions. These include short communications and notes (maximum 2 Cost of participation: 100 €, includes pages), or critical comments (1 page) on registration fee, printed material and the topics covered by COALITION. coffe-lunch breaks. Send your contributions by e-mail to: [email protected]

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