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Microb Ecol. 2010 July; 60(1): 69–80.
Published online 2010 May 7. doi: 10.1007/s00248-010-9667-9.
PMCID: PMC2917558
Copyright © The Author(s) 2010
Molecular and Microscopical Investigation of the Microflora Inhabiting a Deteriorated Italian Manuscript Dated from the Thirteenth Century (((gets the popcorn)))
Astrid Michaelsen,1 Guadalupe Piñar,2 and Flavia Pinzari3,4
1Department of Microbial Ecology, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria
2Institute of Applied Microbiology, Department of Biotechnology, University of Natural Resources and Applied Life Sciences, Muthgasse 18, 1190 Vienna, Austria
3Laboratorio di Biologia, Ministero per i Beni e le Attività Culturali, ICPAL - Istituto Centrale per il Restauro e la Conservazione del Patrimonio Archivistico e Librario, Via Milano, 76, 00184 Rome, Italy
4Dept. of Plant Biology, School in Ecological Sciences, Sapienza University of Rome, Rome, Italy
Flavia Pinzari, Phone: +39-06-48291215, Fax: +39-06-4814968, Email: [email protected].
Corresponding author.
Received March 1, 2010; Accepted March 12, 2010.
Abstract
This case study shows the application of nontraditional diagnostic methods to investigate the microbial consortia inhabiting an ancient manuscript. The manuscript was suspected to be biologically deteriorated and SEM observations showed the presence of fungal spores attached to fibers, but classic culturing methods did not succeed in isolating microbial contaminants.
Therefore, molecular methods, including PCR, denaturing gradient gel electrophoresis (DGGE), and clone libraries, were used as a sensitive alternative to conventional cultivation techniques. (((The heavy guns of media rot analysis.))) DGGE fingerprints revealed a high biodiversity of both bacteria and fungi inhabiting the manuscript. DNA sequence analysis confirmed the existence of fungi and bacteria in manuscript samples.
A number of fungal clones identified on the manuscript showed similarity to fungal species inhabiting dry or saline environments, suggesting that the manuscript environment selects for osmophilic or xerophilic fungal species. Most of the bacterial sequences retrieved from the manuscript belong to phylotypes with cellulolytic activities.
Introduction
Biodeterioration of paper and parchment in ancient books and documents represents a cause of great concern for libraries and archives all over the world. The study of the mechanisms underlying the microbiological attack of historical materials has been widely practiced and still represents one of the main focuses of those institutions and laboratories that are involved in cultural heritage conservation.
Microbial investigations based on cultivation strategies are not reliable because they yield only a limited fraction of the present microbial diversity [26]. The application of molecular biology techniques on cultural heritage environments has shown that new spoiling taxa and unsuspected microbial consortia ((("unsuspected microbial consortia," ow, my stomach hurts))) are involved in the discoloration and biodeterioration of paintings and monuments [44]. Restricted sampling from art and documental objects results in additional problems for representative floristic analyses.
Filamentous fungi colonize different organic and inorganic materials and play an important role in biodeterioration processes [30, 35, 45, 57]. They can tolerate desiccation, high salt concentrations, and heavy metal compounds that are present in inks and pigments and are thus frequent inhabitants on paper-supported objects [14, 57, 58].
The fungal and bacterial communities that can develop on a book are similar to the communities of decomposers that, in natural environments, transform nutrients bound in dead organic matter into low molecular or inorganic forms, making them available to plants. (((Sort of like literary critics, then. In fact they're a lot like literary critics.)))
The development and maintenance of a fungal community on a shelf of a library or in a single book depends on the spores that reach the material’s surface, on the microenvironment (temperature, relative humidity, light), on the water activity of the substrate, and on the casual events that help colonization of materials (insect dispersion, human contamination, external sources of fungal diversity) [14]. (((I was wrong – they're more like the reader fan-base.))
A library or a single book can be compared to a virgin land that can be reached by some colonizing organisms that behave like pioneer species on a nude soil. (...)
Materials and Methods
History of the Object
A manuscript dated back to 1293 from Italy...
