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The influence of crystalline and amorphous cellulose on extracellular hydrogen peroxide production by brown-rot fungi
1991 - IRG/WP 1482
The production of hydrogen peroxide (H2O2) has been suggested to play a key role in the degradation of wood by wood-rotting fungi. The production of extracellular hydrogen peroxide was studied by a quantitative method which detects the oxidation of the 2,2-azinobis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) by H2O2 and horseradish peroxidase (HRP) in liquid culture medium. The carbon sources used were crystalline and amorphous cellulose. Two brown-rotters, Serpula lacrymans and Poria placenta, were able to produce clearly detectable amounts of extracellular hydrogen peroxide in liquid medium which contained crystalline cellulose as carbon source. No detectable H2O2 was produced in conditions where amorphous medium was used as carbon source. This result suggests that the conformational structure of the substrate may induce H2O2 production by brown-rot fungi.
A-C Ritschkoff, L Viikari

Non-enzymatic Gloeophyllum trabeum decay mechanisms: Further study
2001 - IRG/WP 01-10395
Information will be presented on the mechanisms involved in, and potential application of, non-enzymatic wood decay by brown rot decay fungi. Specifically, the hypothesized role of low molecular weight phenolate derivatives will be discussed in relation to non-enzymatic degradation of wood. The mechanism of binding of iron by cellulose, and binding and reduction of iron by fungal derivatives and model compounds is examined. Positive and negative aspects of potential application of these compounds in the generation of free radicals will be discussed.
B Goodell, J Jellison

Microbial breakdown mechanisms. Mini-Symposium at the 18th IRG meeting, Honey Harbour, Ontario, Canada
1987 - IRG/WP 1327
An understanding of how micro-organisms cause chemical alternation to wood is essential for effective wood preservation. Investigation of the Microbial Breakdown of wood is a very dynamic research field and is increasing in importance as several biocides are being examined as alternativesw to classic wood preservatives. These biocides often do not possess broad spectrum activity. It is esential that the biocides are carefully examined and a good knowledge obtained of how and what they must protect. The aim of this Symposium is to attempt to present an update of research, presented by leaders in the field. The following presentations were held: 1) Intrduction/Overview - L.E. Leightley 2) Morphological observations of brown rot and white rot decay - K. Messner 3) Biochemical aspects or brown and white rot decay - T.L. Highley 4) Influence of variable lignin content on brown rot decay of wood - T. Nilson; G. Daniel 5) Soft rot decay - R.A. Eaton.
L E Leightley

Preliminary studies of the decay mechanism of some brown-rot fungi
1989 - IRG/WP 1402
The importance of the enzymatic degradation of hemicellulose and cellulose by brown-rot fungi is still under discussion. Endo-ß-1,4-xylanase and endoglucanase activities of Coniophora puteana and Poria placenta cultures were measured by the increase in reducing groups. Enzymes were produced in liquid and solid sawdust based culture media. Enzyme activities were measured in two, four and eight week intervals from the beginning of the test. The ability of brown-rot fungi to degrade crystalline and noncrystalline cellulose by an enzymatic pathway was studied by measurement of total cellulase activities from Coniophora puteana and Serpula lacrymans. Low enzyme activities were noted. This result indicates strongly that these fungi possess a complete cellulose-degrading enzymatic pathway.
A-C Ritschkoff, H Viitanen

Biochemical aspects of white-rot and brown-rot decay
1987 - IRG/WP 1319
This paper presents an overview of the decomposition of wood by white- and brown-rot fungi - the most important and potent of known wood-decay fungi. These organisms are unique among cellulose destroyers because of their strong capability to enzymatically degrade lignified material. Special emphasis is given to the following aspects of wood decomposition by white- and brown-rot fungi: (1) effects on the chemical and physical properties of wood, (2) method of invasion and ultrastructural modification of wood, (3) nature and activities of extracellular degrating enzymes, (4) relationship of ultrastructural changes to the degradative enzyme systems, and (5) unique physiological features of the fungi that can be used to control decay.
T L Highley

Fenton's reagent as a modification tool in brown-rot decay
1996 - IRG/WP 96-10155
A biomimetic approach was used to clarify the role and importance of the Fenton-type reaction in the carbohydrate degradation by brown-rot fungi. Spruce sawdust and microcrystalline cellulose were modified in the H2O2/Fe(II) treatment. The degree of hydrolysis of the pretreated spruce sawdust was clearly increased with the complete cellulase (Econase), purified endoglucanase from Trichoderma reesei and endoglucanase of Poria placenta. The oxidative pretreatment of microcrystalline cellulose decreased the hydrolyzability of pure cellulose with the complete cellulase, but the hydrolyzability with both purified endoglucanase of Trichoderma reesei and endoglucanase from Poria placenta was increased. Thus, after oxidative treatment with Fenton's reagent the hydrolysis of both pure cellulose and wood was substantially increased.
M Rättö, A-C Ritschkoff, J Buchert, L Viikari

Mechanisms of Protection by NHA Against Fungal Decay
2002 - IRG/WP 02-10429
Treating wood with the water-borne sodium salt of N'-N-naphthaloylhydroxylamine (Na-NHA) protects wood against decay and termite damage. Initial testing indicated little or no inhibition of sapstain fungi, molds, or soft-rot fungi by Na-NHA, suggesting that the mechanism by which this compound protected wood was complex and not that of a broad-spectrum biocide. Previously, we (Green et al, 1997) suggested that the protective mechanism was due to Na-NHA complexing with calcium ions to disrupt fungal metabolism, and/or binding of Na-NHA to the calcium in pit membranes, which acts as a physical barrier to fungal colonization. More recent experiments suggest an alternate mechanism. Specifically, pKa measurements of Na-NHA show that the sodium salt will be essentially fully protonated when impregnated into SYP which is naturally acidic (pH at or below 5.5). Furthermore, the protonated form (H-NHA) was more fungicidal than Na-NHA against two white- and two brown-rot fungi, with the bioactivity approaching that of several commercial organic fungicides. The moderate fungicidal activity of H-NHA is not surprising; the compound is a polyaromatic hydrocarbon much like many of the compounds in creosote. By precipitating mainly at the pit membranes, H-NHA or Ca-NHA serves as a fungicidal barrier to inhibit fungal hyphae from spreading to adjacent cells through the pits. Studies of H-NHA combined with three commercial organic biocides showed that only one combination was synergistic, which further suggests that NHA protects wood by a biocidal mechanism rather than being simply a compound is extremely water insoluble and will thus not leach from wood in outdoor exposure. NHA may also have metal chelating and/or antioxidant properties which further assists in protecting wood. non-biocidal additive.
F Green III, W Henry, T Schultz

Hydrolysis of bordered pits during colonization of conifers by brown-rot fungi
1995 - IRG/WP 95-10103
Brown-rot decay results in rapid reduction in degree of polymerization (DP) of holocellulose with concomitant strength loss (MOR) without removing lignin. Development of new methods of wood protection will require focusing on early events in the sequence of depolymerization. Bordered pit membranes (sapwood) represent a readily available source of non-lignified carbohydrate, ie. pectin and cellulose. Commercial pectinases (Pectinol) and Trichoderma sp. have been shown to degrade pit membranes and increase penetration of preservatives. Brown-rot fungi have previouely been shown to produce oxalic acid (OA) during the decay process. Plant pathogens have been shown to degrade pectin by the synergistic action of OA and polygalacturonase (PG). The OA solubilizes the pectin by chelating the Ca++ and the PG hydrolyses the a-1,4 linkages. We have demonstrated the ability of Postia placenta, Gloeophyllum trabeum and Serpula incrassata to use pectin as a sole carbon sourse and to produce OA and PG on both liquid media and wood. Aspergillus niger and Trichoderma sp. also produce PG on wood but no OA or weight loss. The optimal pH of brown-rot polygalacturonase activity is circa 4.0. As the pH of the wood drope below pH 4, due to acid production during decay, there is a progressive decrease in PG activity and the possibility of acid catalyzed hydrolysis of pit membranes is suggested by increased permeability. We hypothesize that pectin utilization is an essential step during incipient brown-rot decay which helps to initiate fungal metabolism and promote the spread of fungal hyphae between tracheids.
F Green III, J L Tschernitz, T A Kuster, T L Highley

The identification of the carbohydrate degrading enzymes from the crude extract of brown-rot fungus Gloeophyllum trabeum
1991 - IRG/WP 1483
The brown-rot fungus, Gloeophyllum trabeum, produces a pattern of carbohydrate degrading enzymes during the wood decay. In liquid sawdust media the activities of endo-b-1,4-gluganase and endo-b-1,4-xylanase were at the maximum after 5-6 weeks cultivation. The production of enzymes started immediately after inoculation suggesting that the degradation of hemicellulose and easily degradable parts of cellulose is an initial step of wood decomposition.
A-C Ritschkoff, J Buchert, L Viikari

Protection mechanisms of modified wood against decay by white and brown rot fungi
2010 - IRG/WP 10-10713
The resistance of beech and pine wood blocks treated with 1,3-dimethylol-4,5-dihydroxyethylene urea (DMDHEU) against T. versicolor and C. puteana increased with increasing WPG. Full protection (mass loss below 3%) was reached at WPGs of approximately 15% (beech) and 10% (pine). Metabolic activity of the fungi in the wood blocks was assessed as heat or energy production determined by isothermal micro-calorimetry. Fungal activity in the wood decreased with increasing WPG. Still, activity was detected even in wood blocks of highest WPG and showed that the treatment was not toxic to the fungi. The infiltration of untreated and DMDHEU-treated wood blocks with nutrients and thiamine prior to fungal incubation did not result in an increased mass loss caused by the fungi. This shows that the destruction or removal of nutrients and vitamins during the modification process has no influence on fungal decay. In order to study the effect of cell wall bulking and increased surface area, the cell wall integrity was partly destroyed by milling and the decay of the fine wood flour was compared to that of wood mini-blocks. The mass losses caused by the fungi, however, also decreased with increasing WPG and showed comparable patterns like in the case of mini-blocks.To study the effect of the chemical change of cell wall polymers, cellulose was treated with DMDHEU and the product was subjected to hydrolysis by a cellulase preparation. The release of sugar during the incubation was clearly reduced as compared to untreated cellulose. Pre-treatment of modified cellulose with Fenton’s reagent increased the amount of released sugar due to the cellulase activity. Pine micro-veneers were subjected to Fenton’s reagents in acetate buffer over 48h. While untreated specimens and veneers treated with low DMDHEU concentration displayed strong and steady tensile strength loss, veneers treated to a higher WPG did hardly show tensile strength loss.
C Mai, P Verma, Yanjun Xie, J Dyckmans, H Militz

Fungal Attack on Lignin and Cellulose: Elucidation of Brown- and White-Rot Mechanisms Comparing Biomimetic and In-Vivo Degradation Patterns
2010 - IRG/WP 10-10714
This paper examines research and hypotheses that have been developed over several years on wood degradation mechanisms. This information is combined with new data and analyses to explain why wood decay patterns caused by brown-rot fungi and specific types of white-rot fungi are different. New data, including work with both biomimetic studies on low molecular weight compounds, degradative enzymes, and wood decayed by brown- and white-rot fungi support a role for these compounds, which results in different types of fungal decay. Specifically the presence or absence of low molecular weight phenolate compounds that bind and reduce iron to generate oxygen radicals is related to brown-rot, as well as “selective white-rot” decay of wood. Free radicals generated by the low molecular weight systems are important in opening up the structure of wood in advance of, or concurrent with, enzymatic attack in both brown-rot and selective white-rot decay. “Simulataneous white-rot” fungi do not typically posses a highly expressed low molecular weight phenolate system and this may help to explain the erosion pattern of decay observed in decay by this type of white-rot. New analytical techniques including Pyrolysis-molecular beam mass spectrometry and 13C-labeled tetramethylammonium hydroxide thermochemolysis are used to provide new information, particularly regarding how lignin is attacked, and either repolymerized or solubilized depending on the type of fungal attack. Discussion of the mechanisms involved, and how new wood protection schemes may be developed to exploit these mechanisms is reviewed.
V Arantes, B Goodell, A M F Milagres, Yuhui Qian, T Filley, J Jellison, S Kelley

A survey of factors affecting decay resistance of wood modified with acetic anhydride
2012 - IRG/WP 12-40594
From the range of information published, acetylation appears well suited to provide adequate protection against biological attack for materials derived from typically non-durable wood species. Acetylated wood is now commercially available both in Europe and in the USA. But still there are a lot of unanswered questions related to fungal decay mechanisms in acetylated wood. The paper summarize existing knowledge and highlight future research opportunities related to fungal deterioration of acetic anhydride modified wood. In addition statistical analyses based on previously published data were performed to quantify what factors contribute most to the performance (calculated as test sample/control). The results showed that WPG can explain approximately 50 % of the performance, measured as test sample/control (T/C), for acetic anhydride treated wood. Other of the applied variables, like wood species or type of fungus can reduce the variance in T/C by additional 15 %.
G Alfredsen, P O Flæte, H Militz

Enhancing Our Understanding of Brown Rot Mechanisms through Catalytic Pretreatment and Cellulase Cocktail
2018 - IRG/WP 18-10909
A catalytic mechanism, described as the “chelator-mediated Fenton” (CMF) mechanism, is proposed to mimic the non-enzymatic action of brown rot fungi. A CMF treatment was used together with an enzymatic cocktail to study how wood was deconstructed and solubilized. This was done in-part to determine if the treatment mimicked the action of brown rot fungi, but also to explore improved treatment processes for bioprocessing of woody biomass. Our data suggest that the CMF mechanism is highly efficient in overcoming the lignin recalcitrance barrier to solubilize wood. Multiple pulses (up to 4 pulses) of CMF treatment were able to solubilize a majority of both the lignin and cellulose of wood at room temperature, using a hydrogen peroxide concentration of only 1%. Using a single pulse of the CMF system as a pretreatment allowed more wood residue to be retained, and enzymatic action on this pretreated wood was enhanced compared to control wood. In separate experiments, significantly greater solubilization of both sugars and lignin occurred when a single-pulse CMF pretreatment was used prior to enzymatic action than by enzymatic action alone on unmodified wood. This work suggests a key reason why the brown rot fungi have abandoned many of their extracellular enzymes to produce only a select suite of cellulases, which the fungus employs after prior modification of the cell wall using a CMF mechanism. This research further suggests that the CMF mechanism may have potential to be adapted for bioprocessing of woody biomass to produce sustainable fuels and bioproducts in the future.
S Tabor, L Orjuela, D Contreras, G Alfredsen, J Jellison, S Renneckar, B Goodell

Using X-ray scattering to elucidate the mechanisms behind the moisture and fungal decay resistance of epoxybutene modified wood
2019 - IRG/WP 19-40854
Chemical modification of the hydroxyl groups of wood can improve the properties of wood by providing moisture and biological resistance, as well as dimensional stability. Southern pine solid wood was chemically modified to various weight percentage gains (WPG) with epoxybutene (EpB, 8%-38% WPG). After modification, specimens were extracted with a toluene: ethanol (2:1) solution for 2 hours or water leached for 2 weeks. The equilibrium moisture content (EMC) at 30%, 65% and 90% relative humidity (RH) and 27 C was determined on all specimens. Laboratory soil block decay evaluations against the brown-rot fungus Gloeophyllum trabeum was performed and weight loss calculated by mass loss. Biological efficacy was found, and the biological resistance correlated with the lowering of the equilibrium moisture content, suggesting that the mechanism of efficacy was due to moisture exclusion. To assist in understanding the mechanism of effectiveness, small angle x-ray scattering (SAXS) and wide-angle x-ray scattering (WAXS) were both performed. Preliminary WAXS results showed that the modification did not significantly change the cellulose crystalline lattice parameters. Preliminary SAXS results showed that epoxide addition led to an increased polydispersity in the microfibril alignment and broader microfibril angle distributions, thus, suggesting that modification may target regions outside the microfibrils. Further experimentation is underway to confirm these results.
R E Ibach, N Plaza

Confocal laser scanning microscopy of a novel decay in preservative treated radiata pine in wet acidic soils
1997 - IRG/WP 97-10215
Light microscopy of radiata pine (Pinus radiata D. Don) field test stakes (20x20x500mm3) exposed in wet acidic (pH 3-4) soil for 12 - 24 months showed predominance of an unusual type of decay characte-rised by tunnelling attack of wood cell walls. After two years decay was moderate to severe in wood treated to ground contact CCA specifications and also equivalent retentions of creosote, and a number of new generation preservatives. Relative to other New Zealand temperate test sites and also an Australian tropical site, the New Zealand acidic soil test site was very aggressive. Correlative scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to elucidate the micromorphology of this attack. Tunnels of diameter 0.2-5 µm were present throughout all layers of the cell wall, and their orientation was not related to cellulose microfibril orientation. They also showed no preference for particular cell wall layers, indicating a lignin degrading capability. CLSM images showed that living, connecting fungal hyphae were present in the cell lumina and tunnels. This type of attack was predominant in wood that was highly saturated with water whereas wood that was less moist was predominantly attacked by classical white rot. Ongoing isolation and incubation studies in conjunction with further microscopy should enable identification of the fungal species involved.
R N Wakeling, Ying Xiao, A P Singh

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

Field trial with poles of Scots pine treated with six different creosotes
1996 - IRG/WP 96-30115
In the middle of the 50's field trials with creosote-treated poles were started in France, Germany and Sweden. The trials were initiated by WEI (Western-European Institute for Wood Preservation). Six different creosotes were used and 40 poles per creosote were installed at each test field. Results after 39 years of exposure in Simlangsdalen, Sweden are reported. Poles treated with a heavy creosote were less decayed than poles treated with medium-heavy creosotes. Poles treated with a light creosote were most decayed.
Ö Bergman

Questionnaire - Fungal decay types
1985 - IRG/WP 1265
T Nilsson

JWPA method for testing effectiveness of surface coatings with preservatives against decay fungi
1981 - IRG/WP 2164
In 1979 JWPA established a new method for testing effectiveness of surface coatings in accordance with practical use of preservative-treated lumber. Comparing the new testing method with JIS A 9302, a few new trials - size of wood specimen, weathering procedure, and decay-test procedure - are incorporated.
K Tsunoda

Utilization of curcumin for detection of presence of boron in wood
1982 - IRG/WP 3191
It has been shown that curcumin is not a reliable reagent for detecting boron in wood that has been attacked by fungi
M-L Edlund

Co-operative studies on determining toxic values against wood-destroying Basidiomycetes: Progress report to May 1989
1989 - IRG/WP 2339
This document reports progress on the co-operative study between nine laboratories set up following the proposals contained in Document IRG/WP/2316. Results have been received from two laboratories. Toxic values data have been established successfully using the test fungus Coniophora puteana but problems have been encountered with the other test fungi.
A F Bravery, J K Carey

Monographic cards for wood-destroying fungi. [Fiches monographiques pour les champignons lignivores]
1970 - IRG/WP I 5B
C Jacquiot

On Donkioporia expansa (Desm.) Kotl. & Pouzar
1986 - IRG/WP 1285
Donkioporia expansa is found more often in houses than realised until now. Virulence tests according to EN 113 show not only an attack of oak, but also of other hardwoods and even soft-woods.
G Buchwald

Nondestructive Evaluation of Oriented Strand Board Exposed to Decay Fungi
2002 - IRG/WP 02-20243
Stress wave nondestructive evaluation (NDE) technologies are being used in our laboratory to evaluate the performance properties of engineered wood. These techniques have proven useful in the inspection of timber structures to locate internal voids and decayed or deteriorated areas in large timbers. But no information exists concerning NDE and important properties of wood composites exposed to decay fungi. For our pilot study on several types of wood composites, we examined the relationship between nondestructive stress wave transmission, decay rate and the bending properties of OSB exposed to the brown-rot fungus, Gloeophyllum trabeum (MAD-617). The following measurements were taken: stress wave transmission time (pulse echo test method), static bending test (ASTM D3043-95), and decay (expressed as percent weight). Stress wave measurements correlated with strength loss and with increasing rate of fungal decay. Stress wave NDE has great potential as a method for inspection of wood composite load-bearing (in-service) structures, detection of decay in laboratory tests, assessment of chemical additives to improve wood composite durability, and prediction of long term composite performance.
B Illman, V W Yang, R J Ross, W J Nelson

Moisture content levels and decay of hemlock
1986 - IRG/WP 1287
As a model of decay conditions of wooden members in wooden houses, a decay test was set up in which samples of western hemlock (Tsuga heterophylla) under 4 moisture levels were examined. Each week the samples were weighed and if the weights indicated that their moisture contents were lower than the expected levels, distilled water was added. Every 8 weeks 3 samples from each condition were oven dried at 60°C for 48 hours, up to 48 weeks. After 48 weeks, 3 samples from each condition were oven dried every 16 weeks. The results obtained were as follows: After examining the samples for 96 weeks at 27°C, the mean weight loss of the hemlock samples kept at about 50-100% moisture content level was larger than those of the other levels. If the samples were dried every 8 weeks, the amount of decay in them was not significant. Decay was also not significant in the samples kept at approximately 20-30% moisture content level.
K Suzuki

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