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Determination of fungal activity in modified wood by means of micro-calorimetry and determination of total esterase activity

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Beech and pine wood blocks were treated with 1,3-dimethylol-4,5-dihydroxyethylen urea (DMDHEU) to increasing weight percent gains (WPG). The resistance of the treated specimens against Trametes versicolor and Coniophora puteana, determined as mass loss, increased with increasing WPG of DMDHEU. Metabolic activity of the fungi in the wood blocks was assessed as total esterase activity (TEA) based on the hydrolysis of fluorescein diacetate and as heat or energy production determined by isothermal micro-calorimetry. Both methods revealed that the fungal activity was related with the WPG and the mass loss caused by the fungi. Still, fungal activity was detected even in wood blocks of the highest WPG and showed that the treatment was not toxic to the fungi. Energy production showed a higher consistency with the mass loss after decay than TEA; higher mass loss was more stringently reflected by higher heat production rate. Heat production did not proceed linearly, possibly due to the inhibition of fungal activity by an excess of carbon dioxide.

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  1. Akitsu H, Norimoto N, Morooka T, Rowell RM (1993) Effect of humidity on vibrational properties of chemically modified wood. Wood Fiber Sci 25:250–260

  2. Belaich JP (1980) Growth and metabolism in bacteria. In: Beezer AE (ed) Biological microcalorimetry. Academic, London, pp 1–42

  3. Bjurman J (1992) ATP assay for the determination of mould activity on wood at different moisture conditions. International Research Group on Wood Protection (IRG/WP 92–2397), Stockholm

  4. Bjurman J (1993) Determination of microbial activity in moulded wood by the use of fluorescein diacetate. Mater Org 28:1–16

  5. Bjurman J (1994) Ergosterol as an indicator of mould growth on wood in relation to culture age, humidity stress and nutrient level. Int Biodeterior Biodegrad 33:355–386

  6. Bjurman J, Wadsö L (2000) Microcalorimetric measurements of metabolic activity of six decay fungi on spruce wood as a function of temperature. Mycologia 92:23–28

  7. Bravery AF (1978) A miniaturized wood block for the rapid evaluation of wood preservative fungicides. International Research Group on Wood Protection (IRG/WP 2113), Stockholm

  8. Criddle RS, Fontana AJ, Rank DR, Paige D, Hansen LD, Breidenbach RW (1991) Simultaneous measurement of metabolic heat rate, CO2 production, and O2 consumption by microcalorimetry. Anal Biochem 194:413–417

  9. Dermoun Z, Belaich JP (1980) Micro-calorimetric studies of Escherichia-coli aerobic growth—theoretical aspects of growth on succinic acid. J Bacteriol 143:742–746

  10. Donath S, Miliz H, Mai C (2004) Wood modification with alkoxysilanes. Wood Sci Technol 38:555–566

  11. Dyckmans J, Flessa H, Lipski A, Potthoff M, Beese F (2006) Microbial biomass and activity under oxic and anoxic conditions as affected by nitrate additions. J. Plant Nutr Soil Sci 169:108–115

  12. Eaton RA, Hale MDC (1993) Wood: decay, pests and protection, 1st edn. Chapman and Hall, London

  13. European Standard (1996) EN 113. Wood preservatives—method of test for determining the protective effectiveness against wood destroying basidiomycetes—determination of the toxic values, CEN, technical committee TC 38

  14. Ginterova A, Lazariva A (1989) Energy transformation of lignocellulosics into fruit bodies of the wood-rotting fungus Pleurotus ostreatus. Folia Microbiol 34:141–145

  15. Goldstein IS, Jeroski EB, Lund AE, Nielson JF, Weaver JW (1961) Acetylation of wood in lumber thickness. For Prod J 11:363–370

  16. Guilbault GG, Kramer DN (1964) Fluorometric determination of lipase, acylase, alpha- and gamma-chymotrypsin and inhibitors of these enzymes. Anal Chem 36:409–412

  17. Hill CAS (2006) Wood modification. Chemical, thermal and other processes. Wiley, Chichester

  18. Hill CAS, Jones D (1996) The dimensional stabilisation of Corsican pine sapwood by reaction with carboxylic acid anhydride. The effect of chain length. Holzforschung 50:457–462

  19. Hill CAS, Jones D (1999) Dimensional changes in Corsican pine sapwood due to chemical modification with linear chain anhydrides. Holzforschung 53:267–271

  20. Hill CAS, Forster SC, Farahani MRM, Hale MDC, Ormondroyd GA, Williams GR (2005) An investigation of cell wall micropore blocking as a possible mechanism for the decay resistance of anhydride modified wood. Int Biodeterior Biodegrad 55:69–76

  21. Kabir FRA, Nicholas DD, Vasishth RC, Barnes HM (1992) Laboratory methods to predict the weathering characteristics of wood. Holzforschung 46:395–401

  22. Kerem ZD, Friesem Y, Hadar Y (1992) Lignocellulose degradation during solid state fermentation: Pleurotus osteratus versus Phanerochaete chrysosporium. Appl Environ Microbiol 58:1121–1127

  23. Kwon JH, Hill CAS, Ormondroyd GA, Karim S (2007) Changes in the cell wall volume of a number of wood species due to reaction with acetic anhydride. Holzforschung 61:138–142

  24. Lande S, Eikenes M, Westin M (2004) Chemistry and ecotoxicology of furfurylated wood. Scand J For Res 19(Suppl. 5):14–21

  25. Larsson C, Liden G, Niklasson C, Gustafsson L (1991) Calorimetric control of fed-batch cultures of Saccharomyces cerevisiae. Bioprocess Eng 7:151–155

  26. Ljungholm K, Noren B, Skold R, Wadsö I (1979) Use of micro-calorimetry for the characterization of microbial activity in soil. Oikos 33:15–23

  27. Militz H (1991) Die Verbesserung des Schwind- und Quellverhaltens und der Dauerhaftigkeit von Holz mittels Behandlung mit unkatalysiertem Essigsäureanhydrid. Holz Roh Werkst 49:147–152

  28. Militz H (1993) Treatment of timber with water-soluble dimethylol resin to improve their dimensional stability and durability. Wood Sci Technol 27:347–357

  29. Mohebby B, Militz H (2002) Soft rot decay in acetylated wood. Chemical and anatomical changes in decayed wood. International Research Group on Wood Protection (IRG/WP 02-40231), Stockholm

  30. Norimoto M (2001) Chemical modification of wood. In: Hon DNS, Shiraishi N (eds) Wood and cellulosic chemistry. 2nd edn. Dekker, New York, pp 573–598

  31. OECD SIDS (2000) SIDS Initial Assessment Report for 10th SIAM, UNEP Publications, (Tokyo, 15–17 March 2000), http://www.inchem.org/documents/sids/sids/1854268.pdf

  32. Ohkoshi M, Kato Suzuki A, Hayashi K, Ishihara N (1999) Characterization of acetylated wood decayed by brown rot and white rot fungi. J Wood Sci 45:69–75

  33. Ölz R, Larsson K, Adler L, Gustafsson L (1993) Energy flux and osmoregulation of Saccharomyces cerevisiae grown in chemostats under NaCl stress. J Bacteriol 175:2205–2213

  34. Papadopoulos AN, Hill CAS (2002) The biological effectiveness of wood modified with linear chain carboxylic acid anhydrides against Coniophora puteana. Holz Roh Werkst 60:329–332

  35. Ritschkoff AC, Rättö M, Nurmi A, Kokko H, Rapp A, Militz H (1999) Effect of resin treatment on fungal degradation reactions. International Research Group on Wood Protection (IRG/WP 99-10318), Stockholm

  36. Rowell RM (1983) Chemical modification of wood. For Prod Abstr 6:366–382

  37. Rowell RM (2006) Acetylation of wood—journey from analytical technique to commercial reality. For Prod J 56(9):4–12

  38. Saxena G, Lysek G (1993) Observation of nematophagous fungi in natural soils by fluorescence microscopy and their correlation with isolation. Mycol Res 97:1005–1011, Part 8

  39. Scheffer TC (1986) O2 requirements for growth and survival of wood-decaying and sapwood-staining fungi. Can J Bot 64:1957–1963

  40. Schnürer J, Rosswall T (1982) Fluorescein diacetate hydrolysis as a measure of total microbial activity in soil and litter. Appl Environ Microbiol 43:1256–1261

  41. Sparling GP (1983) Estimation of microbial biomass and activity in soil using microcalorimetry. J Soil Sci 34:381–390

  42. Swisher R, Carroll GC (1980) Fluorescein diacetate as an estimator of microbial biomass on coniferous needle surface. Microb Ecol 6:217–226

  43. Verma P, Mai C, Krause A, Militz H (2005) Studies on the resistance of DMDHEU treated wood against white-rot and brown-rot fungi. International Research Group on Wood Protection (IRG/WP 05-10566), Stockholm

  44. Vor T, Dyckmans J, Flessa H, Beese F (2002) Use of microcalorimetry to study microbial activity during the transition from oxic to anoxic conditions. Biol Fert Soils 36(1):66–71

  45. Weigenand O, Humar M, Daniel G, Militz H, Mai C (2008) Decay resistance of wood treated with amino-silicone compounds. Holzforschung 62:112–118

  46. Wieser W (1986) Bioenergetik. Thieme, Stuttgart, pp 35–46

  47. Xie Y, Bjurman J, Wadsö L (1997) Microcalorimetric characterization of the recovery of a brown rot fungus after exposures to high and low temperature, oxygen depletion and drying. Holzforschung 51:201–206

  48. Yano H, Minato K (1993) Controlling the timber of wooden musical instruments by chemical modification. Wood Sci Technol 27:287–293

  49. Yasuda R, Minato K (1994) Chemical modification of wood by non-formaldehyde cross-linking reagents—Part 1. Improvement of dimensional stability and acoustic properties. Wood Sci Technol 28:101–110

  50. Yusuf S (1996) Properties enhancement of wood by cross-linking formation and its application to the reconstituted wood products. Wood Res 83:140–209

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The authors are grateful to the German Science Foundation (DFG) for supporting this project. They would also like to thank Prof. Jody Jellison and Prof. Barry Goodell for fruitful discussions and for their sound advice.

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Correspondence to Carsten Mai.

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Open Access This is an open access article distributed under the terms of the Creative Commons Attribution Noncommercial License (https://creativecommons.org/licenses/by-nc/2.0), which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

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Verma, P., Dyckmans, J., Militz, H. et al. Determination of fungal activity in modified wood by means of micro-calorimetry and determination of total esterase activity. Appl Microbiol Biotechnol 80, 125 (2008). https://doi.org/10.1007/s00253-008-1525-z

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  • Brown rot
  • Fungal activity
  • Micro-calorimetry
  • Total esterase activity
  • White rot
  • Wood modification