title: Molecular and microscopical investigation of the microflora inhabiting a deteriorated Italian manuscript dated from the thirteenth century.
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. 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.
authors.affiliation: Department of Microbial Ecology, University of Vienna, Althanstrasse 14, Vienna, Austria.
authors.forename: Astrid
authors.initials: A
authors.lastname: Michaelsen
authors.forename: Guadalupe
authors.initials: G
authors.lastname: Piñar
authors.forename: Flavia
authors.initials: F
authors.lastname: Pinzari
mesh_terms: D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D044822:Biodiversity; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004783:Environmental Microbiology; D005658:Fungi / Q000235:genetics / Q000302:isolation & purification*; D008373:Manuscripts as Topic; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial; D004271:DNA, Fungal
references.citation: Microbiol Res. 2008;163(2):121-31
references.pmid: 17686620
references.citation: J Microbiol Methods. 2001 Jun;45(2):77-87
references.pmid: 11311392
references.citation: Appl Environ Microbiol. 2002 Mar;68(3):1047-54
references.pmid: 11872449
references.citation: Mycopathol Mycol Appl. 1959 Dec 30;12:1-45
references.pmid: 13811731
references.citation: J Bacteriol. 2000 Apr;182(7):1910-5
references.pmid: 10714996
references.citation: FEMS Microbiol Ecol. 2001 Jan;34(3):229-241
references.pmid: 11137603
references.citation: Appl Environ Microbiol. 1993 Mar;59(3):695-700
references.pmid: 7683183
references.citation: Appl Environ Microbiol. 2000 Apr;66(4):1444-52
references.pmid: 10742225
references.citation: Lett Appl Microbiol. 2007 Jul;45(1):87-94
references.pmid: 17594466
references.citation: Food Microbiol. 2008 Apr;25(2):413-21
references.pmid: 18206785
references.citation: Appl Environ Microbiol. 1996 Apr;62(4):1405-15
references.pmid: 8919802
references.citation: J Appl Microbiol. 2003;94(1):25-34
references.pmid: 12492920
references.citation: Nucleic Acids Res. 1990 Apr 25;18 Suppl:2237-317
references.pmid: 1692117
references.citation: Food Microbiol. 2008 Sep;25(6):831-6
references.pmid: 18620976
references.citation: J Appl Microbiol. 2006 Sep;101(3):620-7
references.pmid: 16907812
references.citation: Antonie Van Leeuwenhoek. 2001 Dec;80(3-4):275-86
references.pmid: 11827213
references.citation: J Antibiot (Tokyo). 1996 Nov;49(11):1101-4
references.pmid: 8982338
references.citation: Biodegradation. 2001;12(4):259-63
references.pmid: 11826908
references.citation: FEMS Microbiol Ecol. 2002 Dec 1;42(3):327-37
references.pmid: 19709292
references.citation: J Gen Appl Microbiol. 2008 Dec;54(6):399-407
references.pmid: 19164883
references.citation: Curr Microbiol. 1995 May;30(5):255-8
references.pmid: 7766152
references.citation: Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
references.pmid: 9254694
references.citation: Phytopathology. 2000 Apr;90(4):384-9
references.pmid: 18944588
references.citation: J Appl Microbiol. 2004;97(6):1228-35
references.pmid: 15546414
references.citation: J Appl Microbiol. 1998 Jun;84(6):1069-84
references.pmid: 9717292
references.citation: Microb Ecol. 2002 Apr;43(3):329-40
references.pmid: 12037611
references.citation: Appl Environ Microbiol. 2003 Sep;69(9):5389-97
references.pmid: 12957927
references.citation: J Gen Appl Microbiol. 1999 Feb;45(1):29-37
references.pmid: 12501399
references.citation: Mycopathologia. 1978 Sep 1;64(1):13-6
references.pmid: 714145