Biodeterioration of Paper (Fungi)
Biodeterioration of Paper (Fungi)
Biodeterioration of Paper (Fungi)
DOI: 10.1002/masy.200650609
Summary: Biodeterioration phenomena represent a complex of physical and chemical alteration processes in various materials, such as those constituting the objects
that represent our cultural heritage. The biodegradation of paper is conditioned by
several variables such as the materials from which cellulose is obtained, the
manufacturing processes employed, the occurrence of other affecting substances
such as lignin or metallic compounds, and by the environmental conditions in which
papers are conserved. In this study, biodeterioration of paper was artificially induced
in order to evaluate the role of a range of chemical and physical variables on damage
caused by cellulolytic fungi. A variable pressure SEM instrument was used to
characterise paper samples with different fibre origins, and alterations obtained
in vitro. Two fungal strains, Aspergillus terreus Thom and Chaetomium globosum
Kunze, which are cellulolytic species frequently associated with paper spoilage, were
used to produce stains with characteristics close to those observable on art objects
made from paper. The stains obtained on the different samples of paper were
compared at both low and high magnification, in order to visualize the macro- and
microscopic characteristics of paper fibres, inorganic constituents, impurities, and
the deteriorating agents related to the spoiled areas. During this survey it was
observed that single paper characteristics can strongly influence the intensity and the
results of the fungal action. For example, the activity of a fungal strain on paper
grades containing fibres of the same origin, but with different sizing, led to the
formation of profoundly different stains and alterations. Moreover fungal structures,
analysed by low vacuum SEM, in areas on paper corresponding to the stains appeared
in different physiological states suggesting an important effect of paper constituents
on fungal growth and their sporulating ability.
Keywords: cellulose; degradation; fungi; paper; SEM
Introduction
Biodeterioration phenomena represent a
complex of natural physical and chemical
spoilage processes in various materials,
such as those composing most of the objects
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In this study paper samples with artificially induced damage were studied using a
variable pressure SEM instrument. The
variable pressure SEM technique proved to
be particularly useful because it allowed for
the direct observation of a non-conductive
material such as paper, and its chemical
characterisation without the need of surface metallization. With this equipment the
spots and stains occurring on naturally
aged and artificially biodeteriorated papers,
could be observed at the right magnification
in order to study, in the spoiled areas of the
samples, the microscopic characteristics of
cellulose fibres, inorganic constituents,
impurities, and deteriorating agents. A set
of samples with experimentally induced
alterations was obtained as reported in the
materials and methods section. The set up
of the experimental protocol was governed
by the aim of studying paper biodeterioration phenomena in simplified models in
which some of the variables could be
controlled. Paper samples with artificially
induced stains and spots were then analysed
macro- and microscopically and the behaviours of the different components in the
built-up model-systems were evaluated.
The results of this paper were obtained in
the framework of the CNR Project Progetto Finalizzato Beni Culturali, Sottoprogetto 3 of the Consiglio Nazionale delle
Ricerche (CNR- Rome) with the cooperation of ICB-CNR Rome, with the cooperation of the ICPL, and ING - Ministero per i
Beni e le Attivita` Culturali.
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Table 1.
Caption of paper samples and their characteristics
Sample N8
1
2
3
4
5
6
0
Filling material
Sizing
Pulp pH
rosin
alkyl ketene dimer
rosin
alkyl ketene dimer
rosin
alkyl ketene dimer
none
4.55.0
7.58.0
4.55.0
7.58.0
4.55.0
7.58.0
6.57.0
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SEM Analysis
The analysis of paper samples before and
after fungal growth was conducted with a
variable pressure SEM instrument (LEO
1450 VP, Carl-Zeiss Electron Microscopy
Group) that allows the direct observation
of non-conductive materials. This SEM
technique permitted the description of
paper surfaces without preparation by
means of metallization. The possibility of
observing uncoated specimens, with a not
conductive surface, allowed appreciating
the signals given by backscattered electrons
that are the result of electron-nucleus
interaction. The characteristics of a backscattered electron image are dependent
primarily upon the nature of the specimen.
The intensity of elastically scattered electrons in the signal significantly influences the image contrast, and this intensity,
in turn, depends upon the average atomic
number of the specimen and the incident
angle of the primary beam on the specimen.[14] Differences in atomic number of
the specimen give rise to appreciable contrast in the image. The differences in chemical composition, and therefore in average
atomic number of the organic (vegetable
fibres and fungal mycelium) and inorganic
materials allowed for well contrasted observation of filling materials and ions distribution on paper surfaces, and the fungal
structures growing on them.[14]
Figure 1.
A conidiophore of Aspergillus terreus with chains of
conidia. Picture obtained with an Olympus AX60
optical microscope.
Figure 2.
An open perithecia (fruit-body) of Chaetomium globosum; meiotic spores can be seen among the perithecial hairs (sterile hyphae). Picture obtained with an
Olympus AX60 optical microscope.
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Figure 3.
Paper samples obtained inoculating fungal spores of Chaetomium globosum (samples on the left) and Aspergillus
terreus (samples on the right) on the paper grades 1 to 6, as listed in Table 1. Picture obtained with a Photo Stylus
Epson Scanner.
Figure 4.
SEM picture at 775. Paper sample n. 5 made of softwood and sized with rosin, kaolin and alum. Arrows indicate
the not germinated spores of C. globosum deposited on paper samples and incubated at 75% of RH.
2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Figure 5.
SEM picture at 1.800. Paper sample n. 1 made of hemp and sized with rosin, kaolin and alum. Arrows indicate a
conidiophora of A. terreus with short chains of conidia, developed at 75% RH.
2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Figure 6.
SEM picture at 499. Paper sample n. 1 made of hemp and sized with rosin, kaolin and alum. Arrows indicate
conidiophores of A. terreus with conidia, developed at 100% RH.
2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Figure 7.
SEM picture at 600. Paper sample n. 2 made of hemp and sized with AKD and calcium carbonate. Arrows
indicate the walls (w) of a broken perithecia of C. globosum with spores (s) developed at 100% RH.
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Figure 8.
SEM picture at 406x. Paper sample n. 5 made of chemical pulp of softwood, sized with rosin, kaolin and alum and
incubated at 100% RH. Arrows indicate perithecia of C. globosum with well developed sterile hyphae, but no
visible spores.
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Conclusions
In this study we have observed that not only,
and not always, fibres in paper influence the
intensity and the results of the biodeteriorating action of fungi, but other factors also
profoundly and intrinsically act to define the
event of degradation. For example, fungal
activity on paper containing different types of
sizing led to very different stains and damage.
Moreover, SEM observation of fungal structures in corresponding stained areas showed
that despite an apparent homogeneous
growth of the fungal strain, different physiological situations could be found, suggesting a
sharp and varied effect of paper constituents
on the different phases of the fungal life cycle.
Further studies are needed in order to explain
some of the observed phenomena, but there
is plenty of evidence to suggest that natural
alterations occurring in paper should be
studied, considering the large number of
variables that interact in causing these
processes.
[1] D. Allsopp, K.J. Seal, Introduction to Biodeterioration. Edward Arnold, London 1986.
[2] R. Kowalik, Restaurator, 1980, 4, 99.
[3] F. Gallo, Biological Factors in the Deterioration of
Books. ICCROM, Rome, 1985.
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