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Possible targets of wood modification in brown rot degradation
2014 - IRG/WP 14-40676
Wood modification protects wood from fungal degradation in a non-toxic manner. However, the mechanisms behind the decay resistance in modified wood are currently unknown. The aim of this study was to discuss the i) colonisation, ii) nutrient recognition, iii) transcription, iv) depolymerisation and v) hydrolysis steps in the brown rot degradation progress and explore whether they are inhibited by wood modification, based on new and previously published data from our group. In previous studies, it has been shown that fungi were able to colonise modified wood, to recognise it as a source of nutrients and express genes associated with cellulose degradation while growing on modified wood. In this study, Fenton derived hydroxyl radicals (∙OH) and brown rot cellulases were able to degrade modified wood. We conclude that the five degradation steps discussed in this paper are unlikely targets for wood modification and that wood modification rather inhibits a step that in a schematic overview of the brown rot degradation process is downstream from transcription but upstream of depolymerisation and hydrolysis.
R Ringman, A Pilgård, G Alfredsen, B Goodell, K Richter

The iron reduction by chemical components of wood blocks decayed by wood rotting fungi
2021 - IRG/WP 21-10979
Brown-rot fungi, a group of wood rotting fungi, is well known to be one of major microorganisms that cause the deterioration of wooden buildings in Japan and have been considered to use chelator-mediated Fenton (CMF) reaction in concert with hydrolytic and redox enzymes for degradation of wood cell wall. CMF can be described as a non-enzymatic degradation system that utilizes hydroxyl radicals produced by the reaction of hydrogen peroxide with ferrous iron in the presence of chelator, and therefore, the reduction process of ferric iron present in wood cell walls is one of the key reactions in this process. To date, various candidates such as low-molecular weight aromatic metabolites produced by fungi, proteins, and lignolytic products have been proposed as iron-reducing compounds. In addition, lignin polymer has been also reported to have the ability of iron reduction although detailed mechanism is not still unknown. In the present study, the authors investigated the iron reduction with wood flour, which was expected to contain both a part of soluble compounds and cell wall polymers, using ferrozine assay in order to obtain the knowledge of iron reduction mechanism. We show that iron reduction capacity by decayed wood samples was higher than that by the samples of sound wood.
R R Kondo, Y Horikawa, K Ando, B Goodell, M Yoshida

The effect of a chelator mediated fenton system on activation of TMP fibres and decolorization of synthesized dyes
2004 - IRG/WP 04-50223
The purpose of this work is to improve our current knowledge of the non-enzymatic mechanisms involved in the brown rot degradation of wood, but also to study the potential applications of a chelator-mediated Fenton system in activation of wood fibers and decolorization of synthesized dyes. In this work, Electron Spin Resonance (ESR) spin-trapping techniques were used to study the generation of hydroxyl radicals in a mediated Fenton system. The activation of Thermal Mechanical Pulp (TMP) fibers was also evaluated by ESR measurement of free phenoxy radical generation on solid fibers. The results indicate that low molecular weight chelators can improve Fenton reactions, thus in turn stimulating the free radical activation of TMP fibers. However, the data also show that excessive and prolonged free radical treatment may cause the destruction of fiber phenoxy radicals. A mediated Fenton system was evaluated for decolorization of several types of synthesized dyes as well. The results show that, compared to a neat Fenton process, the mediated Fenton process increased the production of .OH species to increase the decolorization efficiency. The color of a dilute liquid dyes (Carta Yellow RW, Carta Yellow G, or Cartasol Red 2GF) was effectively reduced to a colorless level after 90 minutes of treatment at room temperature by a mediated Fenton process. In conclusion, this study demonstrates the potential for application, but also the complexity of free radical chemistry in biological materials, especially with regard to the chelation of transition metals and the interaction between free radicals. The complexity of the dyes is similar to that of some organic wood preservatives and may provide a means for remediation of preservative contaminants in soils.
Yuhui Qian, B Goodell, J Jellison

Chelator-Mediated Fenton Chemistry in Wood Degraded by Fungi
2007 - IRG/WP 07-10618
Wood specimens were colonized by individual isolates of brown rot, white rot, soft rot and blue (sap) stain fungi. Ethyl acetate extracts of the ground wood were analyzed for their iron-reducing capabilities using a ferrozine-based assay. Extracts from wood colonized by brown rot fungi showed a significantly greater iron-reducing capability than extracts from wood colonized by white rot fungi or non-decay fungi. The ability of brown rot fungi to produce compounds, and/or modify the wood components, that reduce iron is of relevance to the “chelator-mediated Fenton mechanism” that has been advanced as a theory in the non-enzymatic degradation of wood by the brown rot fungi.
B Goodell, G Daniel, J Jellison, Yuhui Qian

The Chelator Mediated Fenton System in the Brown Rot Fungi: Details of the Mechanism, and Reasons Why it has Been Ineffective as a Biomimetic Treatment in some Biomass Applications – a Review
2014 - IRG/WP 14-10828
The chelator-mediated Fenton (CMF) reaction requires the action of two types of chelating compounds. The first chelator, oxalate, solubilizes and then sequesters iron, and the second chelator reduces iron. Iron reduction must be controlled near the fungal hyphae to prevent damaging Fenton chemistry from occurring in that location. Similarly, iron reduction must be promoted within the wood/plant cell wall to promote Fenton chemistry in the proximity of the target lignocellulose. The mechanism for that control is reviewed in this paper. Both neat Fenton and the CMF have been examined by researchers seeking to exploit this relatively simple mechanism for biomass conversion and lignocellulose pretreatment systems. This paper reviews why some of that research has not produced useful depolymerization reactions and why excess amounts of reagents have been required. The application of Fenton treatments requires that the reactive oxygen species produced in the reaction be generated within a nanometer of the target substrate (lignocellulose), and for this to occur in biomass treatments using Fenton or CMF systems, iron must first be allowed to bind to the substrate to allow the reactions to proceed within nanoscale proximity to lignocellulose. Further, excess iron in solution and in interstitial space must be removed as this “free” iron will react preferentially with chelators and peroxide preventing appropriate targeted action on lignocellulose.
B S Goodell, M Nakamura, J Jellison

Synergistic effects between 2-HPNO, Irganox 1076 and EDTA on the inhibition of wood degradation by Coriolus versicolor
2004 - IRG/WP 04-30331
The efficiency of 2-hydroxypyridine-N-oxide (2-HPNO) as wood preservative has been investigated. As shown using classical experiments as well as using response surface methodology, the efficiency of 2-HPNO as wood preservative is strongly improved in presence a chelator like EDTA and/or of Irganox 1076 an industrial antioxidant. In these conditions, wood preservative efficiency of the mixture the three previous compounds is quite similar to that of tebuconazole used alone. 2-HPNO exhibits an hydroxamic acid function and is a susbtrate of fungal peroxidase. These properties could explain the observed synergy. The implications of these data for the design of new wood preservation strategies are also discussed.
A Mabicka, S Dumarçay, N Rouhier, M Linder, J P Jacquot, P Gérardin, E Gelhaye

Identification of terminal structures in cellulose degraded by the brown-rot fungus Postia placenta
1989 - IRG/WP 1389
To gain insight into the biochemical mechanism employed by brown-rot fungi to depolymerize cellulose, we identified the end-groups of chemically pure cellulose that had been depolymerized by the brown-rot fungus, Postia placenta. The depolymerized cellulose was acid hydrolyzed and the anion fractions isolated by ion chromatography. Sugar acids were identified by gas chromatographic and mass spectrocopic analysis. Cellulose degraded by Fenton's reagent (H2O2/Fe2+) was also analyzed. The two systems generated the same sugar acids but not in the same quantities. The acids identified include glyceric and erythronic, indicating oxidative cleavage of the vicinal diol carbon-carbon bonds within glucosyl residues in the cellulose polymer. Gluconic and arabonic acids were also identified as major products. No uronic acids were produced in either systems.
T K Kirk, T L Highley, R E Ibach, M D Mozuch

The role of oxidation in wood degradation by brown-rot fung
1992 - IRG/WP 92-1562
Brown-rot fungi are suggested to degrade cellulose by oxidation with hydrogen radicalsformed eg. in the conversion of hydrogen peroxide in the Fenton type reactions. The stuctural changes in the holocellulose in wood induced by Fenton's reaction on wood components are very similar to those caused by brown-rot fungi. In this work the effect of the Fenton reaction on wood components was studied. The brown-rot fungi produce extracellular hydrogen peroxide while growing on sawdust medium and on crystalline cellulose. Hydrogen peroxide is apparently formed by the action of an oxidase enzyme. In this work the role of enzymatic oxidation in cellulose degradation by the brown-rot fungus, Poria placenta was studied.
A-C Ritschkoff, J Pere, J Buchert, L Viikari

Investigation of new wood preservation formulations based on synergies between antioxidant, 2-HPNO and Propiconazole
2006 - IRG/WP 06-30401
Wood degradation by fungi is a complex phenomenon taking place in part through oxidative reactions involving many oxidizing agents like free radicals and hydroperoxide. In order to develop novel and more environmentally benign preservative systems, mixtures of Propiconazole and antioxidants with different stabilization mechanisms were investigated to detect possible synergies allowing to reduce the quantity of biocide to prevent wood biodegradation. Results obtained showed that the nature of the antioxidant had an important effect on the potential synergies. The most interesting results were obtained with 2-HPNO and hindered amine and to a weaker extend with H-donors like hindered phenol.
B Bakhsous, S Dumarçay, E Gelhaye, P Gérardin

Gypsum effects on ‘dry rot’ wood degradation as a function of environment
2007 - IRG/WP 07-10624
‘Dry rot’ fungi are a unique group of brown rot fungi that can degrade wood away from ground contact where other fungi fail to colonize. Successfully occupying this niche is partially due to efficient water and nutrient transport, but mobilizing elements, notably calcium (Ca) and iron (Fe), from adjacent building materials has also been implicated in their success. Here we report a series of trials with Serpula himantioides (previously ATCC 36335 S. lacrymans) degrading blocks in the presence or absence of Ca-rich gypsum (aka. drywall, plasterboard), using weight-loss as a measure of decay success. In previous work, pure gypsum did not facilitate dry rot in optimal growth conditions, but it was unclear if moisture or Fe-impurities could alter this dynamic. Commercial drywall was tested in a standard ASTM soil-block trial with spruce blocks and this Serpula wild-type isolate. Gypsum was also made from >99% pure CaSO4 and added in a sand-block trial with variable moisture. Finally, FeSO4 was used to amend gypsum to various Fe contents and the hardened material added to microcosms where spruce was decayed with no external Fe source. Weight-loss in blocks decayed in each experiment was statistically equal between treatments with and without the building material. These results corroborate previous findings, although this dynamic should be tested with other Serpula isolates and in an in-service trial to determine the role of building materials in dry rot.
J Schilling, J Jellison

Degradation of wood veneers by Fenton’s reagents: effects of low molecular weight phenolic compounds on hydrogen peroxide decay and tensile strength loss
2009 - IRG/WP 09-20400
Pine wood (Pinus sylvestris) veneers strips were incubated in acetate buffer containing hydrogen peroxide and iron to mimic mechanisms of brown rot decay and assess the degradation of cellulose through analysis of the tensile properties of the decayed wood. The tensile properties of thin wood strips treated with Fenton system reagents or precursors were determined and correlated to weight loss as reaction conditions were varied. The effect of 2,3‑DHBA concentration on the decay rate of H2O2 in the reaction mixture and the mechanisms of H2O2 decomposition will be discussed. The concentration and ratio of the low molecular weight, phenolic compound 2,3-dihydroxybenzoic acid (2,3‑DHBA) were also studied with regard to the generation of hydroxyl radicals from H2O2 and the resulting weight and strength loss. The pathway and rate of H2O2 decay depended on the pH and the medium (water, buffer) of the reaction mixture. As the concentrations of 2,3-DHBA above an optimal ration with iron, strength loss of the wood strips was reduced. This is likely to have occurred because concentrations of catechols that exceed a 1:1 ratio sequester iron via hexadentate and quadradentate ligand formation as has been previously reported Wood constituents were also shown to reduce Fe(III) to Fe(II) and to drive the Fenton reaction.
Yanjun Xie, R Well, Zefang Xiao, B Goodell, J Jellison, H Militz, C Mai

Chemical mediated depolymerization of cotton cellulose for the understanding of non-enzymatic fungal decay
2010 - IRG/WP 10-10731
Small, low molecular weight non-enzymatic compounds have been linked to the early stages of brown rot decay as the enzymes involved with holocellulose degradation are found to be too large to penetrate the S3 layer of intact wood cells. The most pronounced of these which were analyzed in this study are hydrogen peroxide, iron, and oxalic. The compounds related to the Fenton reaction: the combination of hydrogen peroxide and iron caused marked lowering of the degree of polymerization in the cotton cellulose after treatment. This was the case for both iron ions; Fe3+ and Fe2+. A 10mM solution of oxalic acid also showed significant depolymerizing effect on cotton cellulose, whereas diluting the oxalic acid with sodium oxalate to obtain a pH gradient, showed that this decreased the effect reducing of oxalic acid. In addition an organic iron chelator, EDTA, was tested but was found to inhibit depolymerization when in solution with chemicals related to Fenton chemistry. Manganese was tested to see if other metals than iron could generate an significant impact on the degree of polymerization of cotton cellulose and the metal showed good depolymerizing properties as a substitute for iron. We conclude that low molecular weight metabolites are capable of effectively depolymerizing cellulose during incipient decay by brown-rot fungi.
A C Steenkjær Hastrup, B Jensen, F Green III

The Use of Micro-Tensile Testing to Assess Weathering Decay and Oxidative Degradation of Wooden Items
2010 - IRG/WP 10-20433
This paper presents a method to study the weathering performance of wooden items and to assess the oxidative degradation of wood via the Fenton reaction. Weathering resistance and photo-stability of wood was tested using pine wood (Pinus sylvestris) veneer strips measuring approximately 60 µm in thickness. The veneer strips were treated using a reactive chemical to impart surface protection. The artificially weathered veneers were characterised regarding tensile strength loss as well as by means of infrared spectroscopy. The chemical treatment reduced the tensile strength of the veneer strips. During artificial weathering, however, the strength loss of the treated veneers was clearly lower than that of the controls. The shape of the veneers was preserved due to the treatment. In a second approach veneers strips were used to study wood degradation by the Fenton’s reagent in order to minic the long-term deterioration of archaeological wood from marine environments. Veneer strips were incubated in acetate buffer (pH 4) containing hydrogen peroxide and Fe ions (Fenton’s reagent) and tensile properties (measured in a zero-span mode) were determined. Varying the type of iron (ferrous or ferric sulphate) with H2O2 did not yield significant differences in the rates of H2O2 and tensile strength reduction. However, increasing the amount of wood material (the number of wood strips) in the reaction mixture increased Fe(III) reduction in solution indicating that wood constituents participated in this reaction. Increasing concentrations of Fe(III) in the reaction mixture resulted in a decrease in H2O2 in solution. Despite an increase in iron concentration and H2O2 decomposition under these conditions, a uniform and consistent strength loss of 30% was observed at all Fe(III) concentrations tested. At fixed Fe(III) concentrations, increasing the H2O2 concentration linearly increased the strength loss of the veneers.
C Mai, Yanjun Xie, Zefang Xiao, P D Evans, H Militz

Microbiological degradation of wooden piles in building foundations
1988 - IRG/WP 1370
White rot, soft rot and bacterial attack have been detected in softwood piles under buildings. In some cases bacteria were found to be the main degradation organisms in the studied piles. The water content of degraded piles was very high. The compression strength was quite low also in the piles deteriorated by bacteria. The density of wood was very variable, and the degree of degradation could not be evaluated according to density analyses.
L Paajanen, H Viitanen

Effect of acetylation on decay resistance of wood against brown-rot, white-rot and soft-rot fungi
1989 - IRG/WP 3540
Effect of acetylation on decay resistance of wood was investigated using wood blocks of Cryptomeria japonica, Pinus densiflora, Albizia falcata and Fagus crenata. Blocks were treated with uncatalyzed acetic anhydride for different lengths of time and exposed to Tyromyces palustris, Serpula lacrymans, Coriolus versicolor and unsterilized soil. The action of OH-radical on acetylated wood was also examined using Fenton's reagent. The enhancement of decay resistance by acetylation was revealed clearly for all cases of exposures but varying with fungal and wood species used. For a brown-rot fungus Tyromyces palustris, the weight loss reached almost nil in all woods at 20 WPG (weight percent gain) of acetylation, after the striking decrease from 10 to 15 WPG. For a white-rot fungus Coriolus versicolor, it was counted until 12-15 WPG in the perishable hardwoods used, but not in a softwood Cryptomeria japonica, even at 6 WPG. In cases of another brown-rotter Serpula lacrymans and soil burial, effect of acetylation was intermediate between Tyromyces palustris and Coriolus versicolor. Anti-degradation mechanism by acetylation was discussed, from these weight loss - weight gain relationships, and the IR-and 13C-NMR spectral analyses of fungus-exposed wood.
M Takahashi, Y Imamura, M Tanahashi

Fungal and bacterial attack of CCA-treated Pinus radiata timbers from a water-cooling tower
1991 - IRG/WP 1488
Transmission electron microscopy of decaying CCA-treated Pinus radiata timbers from an industrial water cooling tower showed presence of a thick biofilm covering some areas of the wood. The biofilm contained various morphologically distinct forms of microorganisms embedded in a slime. The study provided evidence of the activity of soft rot fungi and tunnelling and erosion bacteria in wood cells. The extent of damage to wood cells due to microbial activity varied, combined fungal and bacterial attack having the most damaging impact.
A P Singh, M E Hedley, D R Page, C S Han, K Atisongkroh

Ultra-structural observations on the degradation of wood surfaces during weathering
1987 - IRG/WP 2280
Radiata pine (Pinus radiata D. Don) sapwood was converted into blocks with a transverse face about 5 mm square and measuring 8 mm longitudinally. Transverse (T.S.), Radial (R.L.S.) and Tangential (T.L.S.) surfaces were prepared and specimens exposed to the weather inclined at 45° facing equatorially for periods of between 20-60 days. After 30 days exposure erosion of the middle lamella was observed followed after 40 days exposure by extensive separation of individual fibres at the interface of the middle lamella and secondary wall. Degradation of the S2 layer of the cell wall revealed corrugations orientated parallel to the fibre axis suggesting preferential removal of cell wall components. Further degradation proceeded by progressive delamination and checking of the S2 and erosion of the S3 cell wall layer. In addition to the above changes preferential degradation of the rays was observed in radial (R.L.S.) and tangential (T.L.S.) longitudinal surfaces.
P D Evans, S Thein

Resistance of Alstonia scholaris vestures to degradation by tunnelling bacteria
1992 - IRG/WP 92-1547
Electron microscopic examination of vessels and fibre-tracheids in the wood of Alstonia scholaris exposed to tunnelling bacteria (TB) in a liquid culture showed degradation of all areas of the secondary wall. The highly lignified middle lamella was also degraded in advanced stages of TB attack. However, vestured pit membranes and vestures appeared to be resistant to degradation by TB even when other wall areas in Alstonia scholaris wood cells were severely degraded. The size comparison indicated vestures to be considerably smaller than TB, and we suspect that this may primarily be the reason why vestures in Alstonia scholaris wood were found to be resistant to degradation by TB.
A P Singh, T Nilsson, G F Daniel

Effect of medium-term degradation of beech wood by erosive (Phanerochaete chrysosporium) and lignin-selective (Ceriporiopsis subvermispora) strains of white rot fungi on its selected physical properties
2004 - IRG/WP 04-40292
At the Faculty of Wood Sciences and Technology a fungal delignification of normal and tension beech wood by erosive and lignin-selective strains white-rot fungi has been studied. The pre-treatment of both kind of wood samples was accompanied by partial delignification and apparent changes of their physical properties influencing the polar liquids penetration.
R Solár, S Kurjatko, M Mamonová, J Hudec

Detoxification of preservatives: Tri-n-butyltin oxide as a biocide
1982 - IRG/WP 1156
T E Dudley-Brendell, D J Dickinson

The present classification of wood degradation factors
1994 - IRG/WP 94-10071
The revised version of classification of wood degradation factors is presented after discussion and remarks sended by IRG-Members. The classification contains the biotic and abiotic factors in two aspects: etiological and symptomatical.
J Wazny

Options for accelerated boron treatment: A practical review of alternatives
1985 - IRG/WP 3329
Boron wood preservatives are almost exclusively applied by momentary immersion and block diffusion storage. Alternative techniques are described which can be used to accelerate boron treatment. Diffusion coefficients have been derived to define the acceleration of diffusion with increasing temperature. Schedules are described for pressure impregnation of green timber, involving steam conditioning, evacuation and alternating pressure method treatment. Timber Preservation Authority penetration and retention requirements can be met in approximately 4-5 h. The optimum schedule, however, included a 12 hour holding period between steaming and preservative treatment. A method of applying boron preservatives as a vapour is described, Trimethyl borate vapour reacts with wood moisture to form boric acid. The kinetics of this reaction, however, are very fast. This limits treatment to timber dried to very low wood moisture contents.
P Vinden, T Fenton, K Nasheri

Studies of the distribution and degradation of tributyltin naphthenate in double-vacuum treated wood
1983 - IRG/WP 3230
The effects of forced solvent evaporation by kilning redwood (Pinus sylvestris) that has been double-vacuum treated with tributyltin naphthenate (TBTN) have been investigated. Contrary to previous studies reported, it has been shown that forced evaporation can have a considerable influence on the losses of the fungicide. It has been found that, whether the solvent is allowed to evaporate slowly or the evaporation is forced by kilning, the TBTN breaks down considerably in freshly treated wood. In view of the implications of this work for the long-term effectiveness of TBTN further studies are called for.
J Jermer, M-L Edlund, W Hintze, S V Ohlsson

Accelerated ageing of preservatives in treated wood
1988 - IRG/WP 3476
New preservatives are tested in the laboratory and often in field tests before they are used commercially. Some preservatives, however, tested in the laboratory do not show the expected stability when used in service. The differences between laboratory tests and practical use can never be completely eliminated but must be minimized as far as possible by relevant testing methods. Studies of the effect of different accelerated ageing procedures on the chemical degradation and the wood preserving capacity of six different fungicides or combinations thereof have been carried out. Chemicals tested were tributyltinoxide (TBTO), tributyltin naphthenate (TBTN), furmecyclox, benzalkoniumchloride (AAC) + guazatin and pentachlorophenol. The ageing procedures included exposure of test specimens in a wind tunnel (according to EN 73), in an oven at 40°C, 60°C and 70°C, leaching (according to EN 84) and combinations of these procedures. The influence of the different accelerated ageing procedures on the chemical degradation and toxic effect of different fungicides was obvious and, for some procedures and chemicals, comparable with experiences from practice.
M-L Edlund, B Henningsson, B Jensen, C-E Sundman

Contribution to study of the degradation caused in Pinus spp. poles used in field test
1989 - IRG/WP 1417
The study of the degradation produced by soil natural microflora on wood in contact with it in the field, has been going on for several years now. Our contribution to this aim in the present work has dealt with the possible relationship of the microorganisms in the soil. The microscopic visualization of wood colonization by the microorganisms, and the chemical analysis of the degraded wood compared with the undergraded.
M T De Troya, A Garcia, M J Pozuelo, A M Navarrete, A Cabanas

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