Gry Alfredsen

Comparison of quantitative real-time PCR, chitin and ergosterol assays for monitoring colonization of Trametes versicolor in birch wood

Abstract This paper describes the use of quantitative real-time PCR for monitoring colonization of birch wood (Betula pubescens) by the white-rot fungus Trametes versicolor in an EN113 decay experiment. The wood samples were harvested after 4, 8, 12, 16 and 20 weeks of incubation. The mass loss was in the range of 4–40%. Chitin and ergosterol assays were conducted for comparison. Second-order polynomial fits of the mass loss of decayed wood versus chitin, ergosterol and DNA gave correlations (r2) of 0.87, 0.61 and 0.84, respectively. Compared to the other two assays employed, real-time PCR data correlated best with the relative mass loss of decayed samples 4–8 weeks after inoculation, while the saturation and decline of DNA-based estimates for fungal colonization 16–20 weeks after inoculation indicated that the DNA assay is not suited for quantification purposes in the late stages of decay. The impact of conversion factors, extraction efficiency, inhibitory compounds and background levels in relation to the three detection assays used is discussed.

Comparison of chitosans with different molecular weights as possible wood preservatives

Fungi cause serious problems in wood utilization, and environmentally benign wood protection is required as an alternative to traditional chemicals. Chitosan has shown promising antimicrobial properties against several microorganisms. In this study, we present the characterization of and antifungal properties of a commercial chitosan formulation developed for impregnation of wood. A broad range of chemical and mycological methods were used to evaluate the uptake, fixation, and antifungal properties of chitosan for wood preservation. The results show that the higher the uptake of chitosan the lower the relative recovery of chitosan in wood after leaching, and the higher the molecular weight of chitosan the higher the recovery. Chitosan with high molecular weight proved to be more efficient against decay fungi than chitosan with low molecular weight. The fungi tested on chitosan-amended nutrient agar medium were totally inhibited at 1% (w/v) concentration. In decay studies using small wood blocks, 4.8% (w/v) chitosan concentration gave the best protection against brown rot fungi.

Screening of chitosan against wood-deteriorating fungi

Chitosan, a derivative of the natural amino polysaccharide chitin, has proven effective as a potential environmentally benign antimicrobial component. Few studies have focused on chitosan applied to wood against wood-inhabiting and decaying fungi. In these screening studies several mycological experiments were performed to screen chitosan as a potential wood-protecting agent. Growth studies on chitosan-amended media showed total inhibition of Poria placenta, Coriolus versicolor and Aspergillus niger using 1% w/v concentration. Chitosan with high average molecular weight (MW) was more efficient against mould and staining fungi than chitosan with low MW. Agar plate leaching tests showed only a small leaching effect using a 5% concentration on A. niger and P. placenta. Decay testing with P. placenta demonstrated efficacy using 5% and 2.5% concentrations in unleached samples. Leaching decreased the efficacy of chitosan and further investigations are needed to improve the fixation in wood.

Antifungal effect of bark extracts from some European tree species

The inhibitory effect of methanol bark extracts from six deciduous and three coniferous European tree species were bioassayed against eight fungi from the different damage categories, brown rot, white rot, canker and blue-stain. This is the first report providing data on the antifungal activity of several Europaen tree species against fungi within these damage categories. Generally the decay fungi were more inhibited by the bark extracts than the blue-stain fungi, while the lowest inhibition was found among the cancer fungi. The main pattern found between the fungal groups in relation to the bark extracts in this study is believed to be caused by the route of ingress. Acer platanoides bark extract proved to be the most efficient bark extract tested, significantly reducing the growth rate of all tested fungi. Betula pubescens bark extract generally gave the weakest reduction in growth rate. In this study, the conifer bark extracts were in general more active against the canker and blue stain ascomycete fungi than the deciduous trees extracts.

qPCR as a tool to study basidiomycete colonization in wooden field stakes

Abstract Molecular methods are emerging also as useful tools for wood protection studies. The aim of the present study was to evaluate quantitative real-time polymerase chain reaction (qPCR) as a tool for investigating details of the colonization pattern of basidiomycete decay fungi in wood samples after 6 years of soil exposure. Samples of Pinus sylvestris L. (heartwood without treatment), furfurylated P. sylvestris sapwood and Cu-HDO treated P. sylvestris sapwood was in focus. The qPCR method based on basidiomycete DNA content in the wood had the highest sensitivity, while the ergosterol assay was more sensitive than the chitin assay. Visual rating was compared with laboratory analyses and was found to be correlating well with qPCR. This study demonstrates that qPCR in combination with microscopy provides relevant data about basidiomycete colonization in wooden material.

The combined effect of wetting ability and durability on outdoor performance of wood: development and verification of a new prediction approach

Comprehensive approaches to predict performance of wood products are requested by international standards, and the first attempts have been made in the frame of European research projects. However, there is still an imminent need for a methodology to implement the durability and moisture performance of wood in an engineering design method and performance classification system. The aim of this study was therefore to establish an approach to predict service life of wood above ground taking into account the combined effect of wetting ability and durability data. A comprehensive data set was obtained from laboratory durability tests and still ongoing field trials in Norway, Germany and Sweden. In addition, four different wetting ability tests were performed with the same material. Based on a dose–response concept, decay rates for specimens exposed above ground were predicted implementing various indicating factors. A model was developed and optimised taking into account the resistance of wood against soft, white and brown rot as well as relevant types of water uptake and release. Decay rates from above-ground field tests at different test sites in Norway were predicted with the model. In a second step, the model was validated using data from laboratory and field tests performed in Germany and Sweden. The model was found to be fairly reliable, and it has the advantage to get implemented into existing engineering design guidelines. The approach at hand might furthermore be used for implementing wetting ability data into performance classification as requested by European standardisation bodies.

Leaching and decay resistance of alder and pine wood treated with copper based wood preservatives

The objective of this study was to determine the leaching and decay resistance of Alder (Alnus glutinosa subsp. barbata) and Scots pine (Pinus sylvestris) wood samples treated with copper based preservatives. Samples were treated with CCA, ACQ, Tanalith E and Wolmanit with different concentrations. Scots pine samples were exposed the mini-block test against brown rot fungi (Poria placenta) and Coniophora puteana while alder wood samples were tested against brown rot fungi (Coniophora puteana) and white rot fungi (Coriolus versicolor). Regarding to leaching test, treated samples were impregnated with 300 ml of distilled water and after 6, 24, 48 and thereafter at 48-hour intervals, the leachate was removed and replaced with fresh distilled water according to AWPA E11. Samples of each leachate were collected and retained for copper analysis. Amount of copper released from treated wood during the leaching test was chemically analyzed with Atomic Absorption spectroscopy. Perchloric acid procedure for the digestion of wood was used according to AWPA A7-97. The amount of copper component (Qd), the cumulative quantities leached (Qc) and the average daily fluxes (FLUX) were calculated. Results shows that CCA treated samples release less copper compared to other copper based preservatives used in this study. Highest mass losses were obtained from the leached samples treated with 1% of ACQ-2200 against decay fungi.

Carbon footprint including effect of carbon storage for selected wooden facade materials

Abstract The outer facade constitutes a substantial volume of the total consumption of materials used in a building and the need for maintenance of the facade makes it especially interesting from a life cycle perspective. The range of wooden materials and products used for facades has different impacts over the life cycle, but the analyses so far have not included a time-adjusted global warming potential (GWP). Wooden facade materials were assessed with respect to their life cycle carbon footprint based on the environmental product declarations (EPD) and adjusted according to EN15804. The results showed low impacts of untreated Scots pine heartwood (≈1 kg CO2-eq. per square meter [m2]), medium for thermally modified Scots pine, coated Norway spruce and oil/copper–organic preservative-treated Scots pine (1–5 kg CO2-eq. per m2) and high for furfurylated Scots pine and acetylated Radiata pine (5–10 kg CO2-eq. per m2). The results with time-adjustment showed that these methods have a potential large effect on the carbon footprint of wooden claddings. The inclusion of biogenic carbon flows and timing seems to be more important than the difference between the product when biogenic carbon is not included. This fact highlights the importance time-adjusted GWP would have for wood products EPDs.

Quantification of fungal colonization in modified wood: quantitative real-time PCR as a tool for studies on Trametes versicolor.

Abstract Traditional wood preservatives based on biocides are effective against wood-deteriorating organisms because of their toxicity. By contrast, modified woods are non-toxic by definition. To investigate the efficiency of various wood modifications, quantitative real-time polymerase chain reaction (qPCR) was used to profile the DNA amounts of the white-rot fungus Trametes versicolor (L.) [Lloyd strain CTB 863 A] during an 8-week-long growth period in treated Pinus sylvestris (L.) sapwood. The studied wood was modified by acetylation, furfurylation, and thermal treatment. The traditional wood preservatives bis-(N-cyclohexyldiazeniumdioxy)-copper (Cu-HDO) and chromated copper arsenate (CCA) were used as references, whereas untreated P. sylvestris (L.) sapwood served as a control. The maximum levels of fungal DNA in native wood occurred at the end of the experiment. For all wood treatments, the maximum fungal DNA level was recorded after an incubation period of 2 weeks, followed by a decline until the end of the trial. For the preservative-treated woods, Cu-HDO showed the lowest level of fungal DNA throughout the experiment, indicating that exploratory hyphal growth is limited owing to the phytotoxicity of the treatment. The other treatments did not inhibit the exploratory hyphal growth phase. We conclude that qPCR studies of hyphal growth patterns within wood should provide a powerful tool for evaluating and further optimizing new wood protection systems.

The activity of rot fungi (Postia placenta) during drying and rewetting cycles measured by isothermal calorimetry

Rot fungi are a major problem in the construction sector, and method to study under which moisture and temperature coefficients they grow are therefore of significant interest. Measurements of heat production rate have been made on wood samples with the brown rot fungus Postia placenta at different moisture contents (MCs). The results clearly show the heat production rate (a measure of respiration rate and fungal activity) is moisture‐dependent. For most cases, less heat was produced when the MC was decreased, and more heat was produced when the MC was increased. It was also found that when the MC increased after a dry period, the increase in activity was significantly delayed. However, if the moisture state was then kept constant at a high level, the activity slowly increased, showing that the fungi need time to recover back to the original activity level after drying. Isothermal calorimetry is a measurement technique well suited for the study of the activity of wood‐decaying fungi as a function of temperature and moisture content.