Fungi in Fox Spots of a Drawing by Leon Wyczółkowski
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Joanna Karbowska-Berent
Joanna Karbowska-Berent , Ph.D, microbiologist, examines the biodeterioration of historic objects (on paper, parchment, easel and wall paintings, stone and wooden objects) as well as methods for controlling microbial deterioration. Her dissertation (1997) dealt with the role of Streptomycetes in the biodeterioration of historic parchment. She is the co-author and the author of several publications, i.a., of a course book entitled “Microorganisms destroying historic objects and how to fight them” (together with Alicja B. Strzelczyk). At present she works on the disinfection of historic paper items.
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Joanna Jarmiłko
Joanna Jarmiłko , M. Sc., microbiologist, involved in laboratory examination of biodeterioration of historical objects and methods for controlling microbial deterioration.Jolanta Czuczko , Ph.D, graduate of the Nicolaus Copernicus University, is an art conservator. She has been working at the Department of Paper and Leather Conservation in Toruń since 1999 where she is in charge of the Art Conservation Laboratory. She teaches conservation of historic book blocks and co-teaches conservation of paper-based art. Her professional interests lie in the field of conservation of modern works of art. In December 2010, she gained her Ph.D.; her dissertation deals with a typology and conservation issues of paper surfaces used by Leon Wyczółkowski.
Abstract
Many authors dealing with the phenomenon of foxing on paper point out its microbiological origin, but in fact, foxing-causing fungi can rarely be cultured. Leon Wyczółkowski’s drawing “The Market in Gniew” is an exception in this respect: its paper carrier shows foxing as well as numerous hyphae and cleistothecia, visible to the naked eye either as a white and fluffy coating or as a light yellow powder. Our aim was to investigate their role in the formation of foxing. Microscopic observations revealed uncountable amounts of lenticular spores and spherical asci. Incubation of the yellowish powder on microbiological media enabled the isolation of five strains: Eurotium rubrum, E. repens, Aspergillus versicolor, Penicillium solitum, and Penicillium decumbens. Three of them secreted yellow-colored pigments to the culture media. The reproduction of foxing on test papers with the use of these strains was successful. Brown stains resembling foxing appeared on the samples of all four types of paper placed on 1-week-old colonies of Eurotium rubrum. E. repens and Aspergillus versicolor also resulted in the creation of such stains, but only on some types of paper. It was not possible to determine the source of nutrition of the fungi unequivocally.
Zusammenfassung
Schimmelpilze im Bereich von Stockflecken auf einer Zeichnung von Leon Wyczółkowski
Oft wird die Entstehung von Stockflecken auf Papier in Zusammenhang mit mikrobiellem Befall gebracht, aber es gelingt selten, die für die Entstehung von Stockflecken verantwortlichen Schimmelpilze zu züchten. Auf dem Papier der Zeichnung “Markt in Gniew” traten zahlreiche Hyphen und Kleistothezien, die als weiße und flaumige oder hellgelb-braune und pulverige Beläge sichtbar waren, gemeinsam mit Stockflecken auf. Bei Betrachtung unter dem Mikroskop konnten unzählbar viele linsenförmige Sporen und kugelige Sporenkapseln beobachtet werden. Die Inkubation der Proben des hellgelb-braunen Belags auf verschiedenen Nährböden ermöglichte die Kultivierung und Isolierung von fünf Pilzstämmen (Eurotium rubrum, E. repens, Aspergillus versicolor, Penicillium solitum und Penicillium decumbens). Drei von ihnen (Eurotium rubrum, E. repens und Aspergillus versicolor) schieden große Mengen von gelbbraunen Pigmenten in den Nährboden ab. Mit diesen Stämmen konnten Stockflecken auf Testpapieren produziert werden. Die braunen, Stockflecken-ähnlichen Flecken entstanden auf allen vier Probepapieren, die in Kontakt mit einwöchigen Kolonien von Eurotium rubrum gebracht wurden. Die übrigen Pilze (E. repens und Aspergillus versicolor) riefen ebenfalls solche Flecken hervor, allerdings nicht auf allen Testpapieren. Es gelang nicht, die Nahrungsquellen dieser Pilze eindeutig zu bestimmen.
Résumé
Microorganismes dans les taches de foxing des dessins de Leon Wyczółkowski
De nombreux auteurs traitant des taches de foxing sur papier mentionnent leur possible origine microbiologique, mais ces micro-organismes peuvent rarement être cultivés. Le support papier du dessin de Leon Wyczółkowski’s “The Market in Gniew” fait exception. En effet, des hyphae et cleistothecia étaient présents sur la surface du papier à l’emplacement de taches de foxing. Ils étaient visibles à l’œil nu, soit sous la forme d’un revêtement blanc et duveteux, soit sous la forme de poudre jaune-marron pale. L’objectif de cette recherche est l’explication du rôle de ceux-ci dans la formation de taches de foxing. L’observation microscopique a révélé une très grande quantité de spores lenticulaires et d’asci sphériques. L’incubation des éléments poudreux jaune-marron pale sur support microbiologique a permis l’isolation de cinq souches : : Eurotium rubrum, E. repens, Aspergillus versicolor, Penicillium solitum and Penicillium decumbens. Trois d’entre eux secrétaient des pigments jaune-marron pale dans le milieu de culture. Les essais de reproduction de taches de foxing sur des papiers tests avec ces souches se sont révélés positifs. Des taches brunes, ressemblant à des taches de foxing, sont apparues sur les quatre types de papiers placés au contact de cultures âgées d’une semaine de Eurotium rubrum et E. repens; Aspergillus versicolor a aussi induit la formation de telles taches, mais seulement sur les échantillons de certains types de papiers. Il n’était pas possible de déterminer de façon explicite quelle était la source de nutrition de ces micro-organismes. Une question émerge alors : ce type de taches devraient elles être désignées par “taches de foxing”, “taches ressemblant a du foxing”, “foxing biologique” ou bien classifiés comme un type de bio détérioration causée par des micro-organismes/moisissures?
About the authors
Joanna Karbowska-Berent, Ph.D, microbiologist, examines the biodeterioration of historic objects (on paper, parchment, easel and wall paintings, stone and wooden objects) as well as methods for controlling microbial deterioration. Her dissertation (1997) dealt with the role of Streptomycetes in the biodeterioration of historic parchment. She is the co-author and the author of several publications, i.a., of a course book entitled “Microorganisms destroying historic objects and how to fight them” (together with Alicja B. Strzelczyk). At present she works on the disinfection of historic paper items.
Joanna Jarmiłko, M. Sc., microbiologist, involved in laboratory examination of biodeterioration of historical objects and methods for controlling microbial deterioration.
Jolanta Czuczko, Ph.D, graduate of the Nicolaus Copernicus University, is an art conservator. She has been working at the Department of Paper and Leather Conservation in Toruń since 1999 where she is in charge of the Art Conservation Laboratory. She teaches conservation of historic book blocks and co-teaches conservation of paper-based art. Her professional interests lie in the field of conservation of modern works of art. In December 2010, she gained her Ph.D.; her dissertation deals with a typology and conservation issues of paper surfaces used by Leon Wyczółkowski.
Appendix 1
ATP measurements
The ATP is the primary energy unit of all living cells. ATP is restricted to cell activity not present to a notable amount in resting cells, e.g. spores and conidia, but its great advantage is the estimation of the activity or vitality of the microorganisms, because if microorganisms are dead, the object does not need any disinfection, but only cleaning. It can be quantitatively determined by measuring the amount of light generated with luciferase-luciferin reagent in a bioluminometer (Rakotonirainy et al. 1995; Karbowska-Berent and Kotala 2006). The oxidation of luciferin is catalyzed by luciferase in the presence of Mg2+ ions and ATP molecules. During the reaction, ATP releases energy which is turned into light photons. The intensity of the yellow-green light is measured at the wavelength of 562 nm and the result is expressed in Relative Light Units (RLU).
The samples of the deposit to be tested for ATP were collected with a sterile and chemically clean cotton swab from the surface of a 3 × 3 cm square. The reaction was carried out using commercially available tests (Merck) and the ATP amount was measured with a HY-LiTE 2 bioluminometer (Merck). On the basis of our earlier observations, an ATP level not exceeding 200–300 RLU is usually considered to be low and caused by contamination from the air and dust, but we also take into consideration the ATP level of the unaffected areas on the same object.
Appendix 2
Culture media: dichloran agar DG18 (Merck) for xerophiles, malt extract agar (MEA) from Merck, SNA (Synthetic Nutrient Agar: KH2PO4 1 g, KNO3 1 g, MgSO4 × 7H2O 0.5 g, KCl 0.5 g, glucose 0.2 g, sucrose 0.2 g, agar 20 g, distilled water 1 L) and the medium for cellulolytic fungi according to Aschan and Norkrans (1953).
Appendix 3
Selected characteristics of the test paper
| Symbol of the paper | General characteristic | Identification of fibers | Sizing |
| X | Laid paper, Chinese | Long-fibrous pulp – paper-mulberry fiber, substantial addition of cellulose pulps from straws, most probably rice straws | Lack of starch and gelatin |
| Y | Handmade laid Japanese Atsu-Shi paper (Japico) | Long-fibrous pulp – kozo nonwoven fabric, addition of cellulose pulp from conifers | Lack of starch and gelatin |
| Z | European, woven water-color paper, textured | Long-fibrous pulp – cotton | Gelatin, lack of starch |
| V | European, laid (Hahnemühle) | Cellulose pulps from conifers and deciduous trees | Gelatin, lack of starch |
Appendix 4
The ability to acidify the culture medium was tested, having the following composition: glucose 10 g, lactose 10 g, yeast extract 5 g, CaCO3 5 g, peptone 1 g, agar 20 g, bromocresol purple 20 mg, distilled water 1 L. The change in the color of bromocresol purple from purple to yellow during the growth of fungi in the zone around the colony proved that the colony was secreting acids to the medium. The estimation of the acidified zones was performed after a week of incubation at 27°C.
Amylolytic activity was tested on the medium containing: starch 30 g, NaNO3 3 g, KH2PO4 1 g, MgSO4 0,5 g, KCl 0,5 g, FeSO4 traces, agar 20 g, distilled water 1 L. The zones of starch decomposition were developed after 2 weeks of incubation with the use of Lugol’s solution (Walczak et al. 2013).
Proteolytic activity was determined on Frazier’s medium containing: gelatin 40 g, agar 20 g, and distilled water 1 L. The zones of gelatin’s decomposition were revealed after a week of incubation after treatment with Frazier’s reagent (Burbianka et al. 1983).
The culture media were poured into Petri dishes, 10 mL per plate, in order for the decomposition zones to be comparable. All tests were carried out in duplicates. The fungi were inoculated to the center of the Petri dishes with the appropriate media. The diameters of the colonies as well as the acidification or decomposition zones were measured each time, where the coefficient of activity W was calculated according to the following formula:
where
R = diameter of the decomposition zone
K = diameter of the colonies
The ability of fungi to acidify and decompose starch and gelatin was estimated according to the following scale:
0–lack (W = 0)
1–low (W = 1)
2–medium (2 > W > 1)
3–strong (4 > W > 2)
4–very strong (W > 4).
Cellulolytic activity was measured as by Nol et al. (1983). Strips (15 × 1 cm) of sterile Whatman paper were hung within test tubes, fixed by Kapsenberg’s caps. The strips were in contact with 5 mL of liquid Czapek medium lacking sucrose, to which a suspension of 0.2 mL conidia of the test fungus had been added. Test tubes were held for as long as 8 or 24 weeks at 27°C and tested frequently by a strong wrist-flicking movement for weakening of the strength of paper.
Capability for growth in conditions of low water activity was tested on MEA with an addition of 8% NaCl and 20% NaCl as well as for comparison on MEA without the addition of NaCl. The fungi were applied to the center of the Petri dishes with appropriate media and incubated at 27°C. After 4 weeks the intensity of growth of the colonies with the following scale was assumed as the result:
0–lack of growth
1–weak growth or growth of mycelium only
2–medium growth, but clearly weaker than in the control
3–growth as in the control
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©2014 by Walter de Gruyter Berlin / Boston
Articles in the same Issue
- Frontmatter
- The “Schinkel’s Legacy” Project at the Kupferstichkabinett Berlin
- ATWISE: An Innovative Apparatus for Efficiently Documenting Watermarks in Manuscripts
- Methods of Creating, Testing and Identifying Traditional Black Persian Inks
- Fungi in Fox Spots of a Drawing by Leon Wyczółkowski
- Fungal Biodeterioration of Paper: How are Paper and Book Conservators Dealing with it? An International Survey
Articles in the same Issue
- Frontmatter
- The “Schinkel’s Legacy” Project at the Kupferstichkabinett Berlin
- ATWISE: An Innovative Apparatus for Efficiently Documenting Watermarks in Manuscripts
- Methods of Creating, Testing and Identifying Traditional Black Persian Inks
- Fungi in Fox Spots of a Drawing by Leon Wyczółkowski
- Fungal Biodeterioration of Paper: How are Paper and Book Conservators Dealing with it? An International Survey
