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Detection of Wood Destroying Fungi Using DNA Microarray Technology
2010 - IRG/WP 10-20435
Wood decay fungi of the phylum Basidiomycota cause serious damage to wooden constructions and building elements. The elimination and the appraisement of fungal decay require an assured species identification. Conventional fungal diagnostics are mainly based on morphological characteristics by macro- and microscopy analysis. For some years, standardised and highly sensitive molecular methods focussing on DNA analysis are increasingly used. However, these methods still have some disadvantages in routine diagnostics, like long operating time and infeasibility of analysing mixed samples, where several fungi occur simultaneously in one sample. To overcome these drawbacks, a DNA microarray for the confident identification of the 27 most important wood destroying basidiomycetes was developed that is already used in the laboratory routine. First of all, species-specific DNA probes were generated on the basis of validated rDNA ITS sequences from 121 fungal isolates. The specificity of these DNA probes was assured using the polymerase chain reaction (PCR) and microarray detection technology. Afterwards, the microarray detection procedure that is based on the arrayed ligation reaction (ALR) was optimized and validated. For a comprehensive field testing and validation of the newly developed detection technique, 36 samples from decayed timber have been collected. The test specimens were analysed by microarray analysis as well as by conventional methods like microscopy analysis and ITS sequencing determination in parallel. Thereby, at 33 samples the fungus species could be addressed directly by microarray analysis. In the case of three samples, the concentration and the quality of the DNA preparations were insufficient for a distinct diagnosis. In these samples the conventional morphological identification as well as the sequencing determination failed, too. The comparison methods provided a higher level of failure, in which 22 % (ITS sequencing), respectively 33 % (conventional diagnostics) of the samples could not be identified to the species level. The field test showed that the microarray based diagnostic system ensures a fast and reliable identification of wood decay fungi. Since January 2010, the Mycotype® BasidioQS Microarray Detection Kit is available as the first commercial product. This DNA microarray was developed in a research cooperation between the Institute of Wood Technology, Dresden (IHD) and the Biotype Diagnostic GmbH, Dresden.
K Jacobs, N Rangno, W Scheiding, B Weiss, D Müller, C Hiller, W Brabetz
Questionnaire - Fungal decay types
1985 - IRG/WP 1265
Improvements of monitoring the effects of soil organisms on wood in fungal cellar tests
1996 - IRG/WP 96-20093
Accelerated testing the durability of preservative treated timber in a so called "fungal cellar" or "soil-bed" to evaluate its performance in ground contact is widespread practice. In order to obtain a more accurate and reproducible estimate of preservative performance, several institutes, among them the BAM in Berlin, have routinely carried out static bending tests in addition to visual examination. These tests were usually performed with a defined maximum load or deflection path regardless of the remaining degree of elasticity of the test specimens. Recent studies at the BAM revealed that by modifying the method, i.e. by restricting the applied load to the non-destructive interval for each individual test specimen, the calculated modulus of elasticity (MOE) reflect the changing strength properties caused by biological deterioration and allow within a relatively short time valuable predictions on the service life of the treated timber in soil contact.
I Stephan, S Göller, D Rudolph
The restricted distribution of Serpula lacrymans in Australian buildings
1989 - IRG/WP 1382
Temperature data has been gathered over a number of years, not only for flooring regions of various buildings in Melbourne, but also within roof spaces and external to the buildings. Findings are discussed in relation to the distribution of Serpula lacrymans within Australia, its restriction to certain types of building construction and its restriction to flooring regions. The subfloor spaces of badly-ventilated, masonry buildings are highlighted as being better suited than are the subfloor spaces of, for example, Japanese buildings for the activity of this fungus. Hence Serpula lacrymans is very restricted in its distribution in Australia, yet where it is active it does grow rapidly and causes rapid flooring failures.
J D Thornton
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
The accelerated field simulator (= fungal cellar)
1982 - IRG/WP 2170
G C Johnson, J D Thornton, H Greaves
An attempt to evaluate wood resistance against fungal decay in non-sterile conditions by measuring the variation of resistance to bending test
1988 - IRG/WP 2308
The main object of this work was to determine the variation of strength on large test specimens of wood (800 x 45 x 45 mm³) when exposed to accelerated fungal attacks close to natural conditions, out of test vessels. The modulus of elasticity (MOE) and the modulus of rupture (MOR) have been assessed. Thereby, the natural resistance of the wood species to fungal decay, the efficiency of preservative as well as the treatment applied are discussed. The wood tested is a guianese secondary species (Couma guianensis). The fungi tested are two guianese strains of brown and white rot. The exposure time is 12 weeks. No mould contamination has been recorded by use of a selective fungicide. The results obtained show that it is possible to infest in nonsterile conditions large wood specimens. Furthermore, modulus of rupture appears to be the most reliable criterion. The investigation, that requires limited equipment and staff could be performed in any tropical research station as it has been done at CTFT, French Guiana center.
L N Trong
A short note on fungal decay in K33-treated poles
1982 - IRG/WP 1169
Soft rot cavities and erosion of the lumen have been found in K33-treated Pinus sylvestris poles from the years 1956-66 by microscopic studies. Poor treatment quality has been proved for some of these poles. The microscopy showed an unusual pattern of attack, and pre-treatment decay is suspected but not yet proved. Sounding the poles and using the Pilodyn indicated decay, but poking did not. Quantification of the attacks was possible only by microscopic studies.
Biological and chemical observation on the early fungal colonization of TBTO treated Swedish redwood stakes
1984 - IRG/WP 3311
Data on the early fungal colonization of Swedish redwood stakes, impregnated with 1% TBT0 / 0.5% dieldrin solution, both by double vacuum impregnation and immersion processes are presented. Results of chemical analyses of wood samples from the outer 1 mm of separate painted and unpainted stakes, exposed over the same twelve month period, are also discussed.
R Hill, A H Chapman, A Samuel, K Manners, G Morton
Evaluation of new creosote formulations after extended exposures in fungal cellar tests and field plot tests
2000 - IRG/WP 00-30228
Although creosote, or coal tar creosote, has been the choice of preservative treatment for the railroad industry since the 1920s, exuding or "bleeding" on the surface of creosote-treated products has been one incentive for further enhancements in creosote production and utility (Crawford et al., 2000). To minimize this exuding problem, laboratories such as Koppers Industries Inc., USA, and Commonwealth Scientific and Industrial Research Organization (CSIRO), Division of Chemical and Wood Technology, Melbourne, Australia, have developed changes in processing of coal tar that produce distillates with fewer contaminants. This "clean distillate" is then used to formulate "clean creosote" as a preservative. These new, unique creosote formulations are being investigated as part of a program to enhance the use of regionally important wood species in the United States. Four retention levels of each of two new creosote formulations creosote, one pigment-emulsified creosote (PEC) and one creosote formulation that meets the AWPA Standard C2-95 for P1/P13 creosote (AWPA, 1995a), were applied to two softwood species and two hardwood species. Two laboratory procedures, the soil-block and fungal cellar tests (accelerated field simulator), were used to evaluate the four creosote formulations. These procedures characterized the effectiveness of the wood preservatives. The soil-block tests were used to determine the minimum threshold level of the preservative necessary to inhibit decay by pure cultures of decay fungi. In general, the soil block tests showed there was little difference in the ability of the four creosote formulations to prevent decay at the three highest retention levels as summarized in a previous report by Crawford and DeGroot (1996). The soil-block tests will not be discussed in this report. Fungal cellar tests expose treated wood to mixtures of soil-borne fungi that promote accelerated attack. Crawford and DeGroot (1996) discussed the evaluation of the creosote formulations after 17 months of exposure in the USDA Forest Service, Forest Products Laboratory (FPL), fungal cellar. At that point in time data from the fungal cellar tests showed that softwoods are protected better than hardwoods for all four formulations of creosote tested. This report will discuss exposure of the fungal cellar stakes upto 36 months. In addition, field stake tests are being used to verify service life of the new creosote formulations in vivo. Results from accelerated tests are indicative of field performance, but the correlation between laboratory and field results is still being investigated. Field stake tests are regarded as critical, long-term evaluations that provide results most directly related to the performance of treated products in service. In this study, we report on the performance of the creosote formulations after five years of exposure in field tests.
D M Crawford, P K Lebow, R C De Groot
A direct method for testing plywood and particle boards against fungal decay
1984 - IRG/WP 2214
A method directly inspired from the French standard testing method of the resistance of particle boards against fungal decay (AFNOR N° 51.295 May 1980) is described. But in that experimentation, the infestation is localized and realized in non sterile conditions. Small blocks of Fagus sylvatica (60 x 20 x 10 mm³) used as " inoculates " are infested with basidiomycetes, in Kolle flask for 4 to 6 weeks, then tightly pressed against the middle part of the test specimens (190 x 15 x 15 mm³). The lower part of the inoculates is plunged in vermiculite kept constantly humid by water containing a selective fungicide. After twelve weeks of exposure in non sterile conditions, in a green house with constant temperature around 20°C, the test specimens are then submitted to a static bending test until fracture. The comparison of the fracture-stress between control test specimens and the specimens exposed to wood rotting basidiomycetes permits to evaluate the resistance of the studied materials against fungal decay.
L N Trong
Are fungal cellar tests really necessary?
1989 - IRG/WP 2333
During the past decade the range of methodology used to evaluate wood preservative potential has significantly expanded. At the forefront of these new tools available to the scientist·is the fungal cellar. This technique, as currently applied, involves the exposure of treated and untreated samples to conditions of moisture and temperature which ensure optimum fungal attack. By comparison data with that obtained on similar preservative systems in conventional stake tests, acceleration factors for predicting performance have been developed. This discussion paper examines the philosophy influencing the development of fungal cellar testing, and provides an alternative viewpoint.
J N R Ruddick
Isolation and identification of the fungal flora in treated wood
1976 - IRG/WP 144
J F Levy
Preliminary note on the fungal problem of rubber wood
1983 - IRG/WP 3246
Susceptibility of rubber wood to fungal attack limits its wider utilisation. Fungal problems encountered in treating rubber wood with boron compounds by diffusion process have been discussed. Sodium pentachlorophenoxide and 2-thiocyanomethylthio benzothioazole (TCMTB) were investigated for possible control of fungal growth during diffusion storage and their performance has been reported.
Defining fungal decay types - Final proposal
1988 - IRG/WP 1355
The term soft rot is proposed for all forms of decay caused by Ascomycetes and non-basidiomyceteous Fungi imperfecti. The terms brown rot and white rot should be used only for decay caused by Basidiomycetes. Brown rot is characterized by extensive depolymerization of the cellulose and limited lignin degradation. White rot is characterised by significant degradation of the lignin component in wood.
Targeting fungal proteinases to prevent sapstain on wood
1995 - IRG/WP 95-10097
Discoloration of wood, caused by a variety of sapstaining fungi, leads to periodic losses in the Canadian lumber export industry. Proteolytic enzymes are thought to be necessary for retrieval of nitrogen during fungal growth on wood. The major extracellular proteinase of Ophiostoma piceae, a representative sapstaining fungus, was purified to homogeneity and its inhibition pattern characterised. Classic serine proteinase inhibitors inhibited the activity of the purified proteinase in a dose-dependent manner. Several chelating agents, detergents, heavy metals and PQ-8, a currently used commercial antisapstain formulation, also caused a reduction in proteolytic activity in vitro. Selected inhibitory compounds were then tested for their effect on growth of Ophiostoma piceae in artificial media and on wood. Heavy metals and several chelators inhibited growth on media containing protein or inorganic nitrogen, suggesting that they were toxic to the fungus rather than specific to the proteinase. However, chymostatin and PQ-8 did appear to be specific proteinase inhibitors. These products caused a decrease in growth on a protein-supplemented medium which induces proteinase production, but had little effect on growth on medium containing inorganic nitrogen. Visual assessment of lodgepole pine sapwood samples inoculated with Ophiostoma piceae also identified PQ-8 as an effective inhibitor of fungal growth. Other pure chelators and serine proteinase inhibitors did not perform well on wood. While some compounds showed promise in these tests, definitive testing and commercial application are constrained by the current lack of stable, cheap, non-toxic, specific serine proteinase inhibitors.
L D Abraham, D E Bradshaw, A Byrne, P I Morris, C Breuil
Fungal defacement of water-stored softwoods
1993 - IRG/WP 93-10009
Sapstain, mould and basidiomycete defacement of untreated sawn boards of Scots and Corsican pine was recorded during a 14 week field trial. Boards were cut from freshly felled logs and from logs previously maintained under water sprinklers for 6 months and ca. 4 years. The incidence of sapstain defacement or the freshly felled wood was very servere aftcr 2 weeks and remained so throughout the board trial. Fungal defacement of wood previously stored under sprinklers for 6 months was progressive and was severe by 14 weeks. In contrast, the level of defacement of boards from long lerm water-stored logs remained very low after 14 weeks exposure. The marked differences in susceptibility to fungal infestation of boards from freshly felled and water-stored logs are discussed in terms of wood sugar levels and the bacterial populations present in the wood.
M A Powell, R A Eaton
Efficacy of some extractives from Pinus heartwood for protection of Pinus radiata sapwood against biodeterioration. Part 1: Fungal decay
1995 - IRG/WP 95-30072
Chemical compounds thought to contribute to the natural durability of heartwood of Pinus spp. were either chemically synthesised in the laboratory or extracted from the heartwood of Pinus elliottii or Pinus caribaea. These compounds included the stilbenes, pinosylvin and its mono- and di-methyl ethers, and the flavonoids, pinobanksin and pinocembrin. Small blocks of Pinus radiata sapwood were impregnated with methanolic solutions of pure compounds or heartwood extracts, to a range of retentions extending above and below the concentration of each compound known to occur in the heartwood of Pinus spp.. Fungicidal efficacy of these compounds has been evaluated by exposure of treated blocks to pure cultures of a white and a brown rot, in addition to an unsterile soil test.
M J Kennedy, J A Drysdale, J Brown
Susceptibility of angiosperm sapwood to white-rot fungal colonization and subsequent degradation: a hypothesis
1997 - IRG/WP 97-10211
It has long been recognized that angiosperm sapwood in nature is relatively easily and preferentially degraded by white-rot fungi. This susceptibility to white-rot fungi is generally believed to be mainly caused by the structure and concentration of angiosperm lignin. However, an explicit explanation as to why lignin structure makes a particular wood vulnerable to white-rot colonisation and subsequent degradation has apparently never been given. We propose that free phenolic groups in wood, such as those present in the lignin or heartwood extractives, can act as free radical scavengers (antioxidants) which disrupt the various white-rot free radical degradative mechanisms. Consequently the presence of a relatively high free phenolic "density", such as that present in gymnosperm sapwood or angiosperm heartwood, may inhibit white-rot degradation. Conversely, white-rot fungi may find wood with a relatively low free phenolic content, such as angiosperm sapwood, easy to colonize. The complex structure of angiosperm wood, in which different cell types have different amounts and types of lignin -- and consequently different levels of free phenolic "densities" -- influences the susceptibility of angiosperm wood to initial white-rot colonisation and, perhaps, also the subsequent decay rate. In addition to the free phenolic ``density" other factors, some as yet unknown, undoubtedly also affect the decay resistance of a particular wood.
T Schultz, D D Nicholas
Preventing fungal attack of freshly sawn lumber using cinnamon extracts
2007 - IRG/WP 07-30432
The potential for using cinnamon oil as an anti-mold and stain compound was investigated on ponderosa pine sapwood. Cinnamon oil was highly effective when used in ethanol, but its activity declined when it was mixed with only water. Attempts to enhance water solubility with surfactants improved solution stability, but had no apparent effect on biological activity. Further studies with other co-solvents are planned
Shujun Li, C Freitag, J J Morrell
Flow charts for termite and decay tests to determine the natural durability of Japanese cedar (Cryptomeria japonica D. Don)
2008 - IRG/WP 08-20385
This paper deals with the experimental flow charts that were used for determination the effects of fungal decay and termite attack on Sugi heartwood during the course of the study of “Comparative studies of natural durability of Japanese cedar (Cryptomeria japonica D. Don) among the geographic cultivate”, which was carried out by Usta et al (2006).
I Usta, S Doi
Evaluation of the decay caused by Chaetomium globosum Kunze, in the course of time
1987 - IRG/WP 2288
The main research done on soft rot has been directed to determining, by microscopic study, the different stages of penetration into the wood of the fungi that cause it. On the basis of the information furnished by this research, in this work we have tried to quantify its effect, by evaluating the weight loss caused by Chaetomium globosum Kunce in wood of Pinus sylvestris L. and Fagus sylvatica L. in the course of time. The results obtained corroborate the existence of succesive stages of decay which is shown by large differences in the weight loss.
M T De Troya, A M Navarrete
Termiticidal chemicals derived from tropical tree resins
1991 - IRG/WP 1477
To test the hypothesis that defensive chemicals protect tropical primary forest trees against biological attack, a bioassay and fractionation program was conducted in Indonesia. Fresh dipterocarp resins were fed in no-choice tests to Neotermes dalbergiae termites on 4.5 cm filter papers, or tested for inhibition of fungal growth. Fractionation of biologically active resins via flash column chromatography, followed by subsequent bioassay and analytical chemical studies, revealed that several sesquiterpene compounds inhibited fungal growth and killed 50% of test termites in 3-7 days. Toxic fractions contained caryophyllene, caryophyllene oxide, alloaromadendrene, and other compounds. From the relatively non-toxic a-gurjunene, novel termiticidal compounds were synthesized, indicating the potential for manufacture of insecticides from natural products.
A Messer, K McCormick, D Richardson, Sunjaya, H Hagedorn, J Meinwald