This study aimed to evaluate the preservative potential of extracts from teak wood industrial waste against artificial weathering and decay fungi. Teak extracts were obtained in hot water, acetone, ethanol and methanol and tested against brown-rot (Rhodonia placenta, Gloeophyllum trabeum, Neolentinus lepideus) and white-rot fungi (Irpex lacteus). Aging tests included the leaching test and artificial weathering in a QUV accelerated weathering tester. The chemical analysis of the extracts was carried out by gas chromatography-mass spectrometry (GC-MS), and the compounds were compared with the cited literature. The extracts were characterized as quinone derivatives (35 %), mainly tectoquinone (28 %), followed by squalene (8-12 %) and other phenolic compounds (6 %). Lapachol, although found in proportions of 0.7-0.9 %, plays an important role in wood durability and biocidal activity in teak extracts. The results obtained by the chemical analysis indicated that the main compounds identified in this study can act synergistically on the resistance of wood impregnated. Teak extracts ensured high resistance to the decay fungi tested and results showed better performance of acetone extracts. In view of the results obtained, it is recommended to carry out other tests (soil bed and field tests) to verify the long-term effects of the extracts.
Eastern subterranean termites, Reticulitermes flavipes (Kollar), are widely distributed across North America where they are exposed to a broad range of environmental conditions. However, mechanisms for overwintering are not well understood. Wisconsin is a unique location to study mechanisms of cold tolerance as it represents the northern boundary for persistent R. flavipes populations. In this study, we evaluated seasonal shifts in cold tolerance using critical thermal minimum (CTmin) and supercooling point (SCP) and examined how these measurements correlate to changes in the microbial community of the termite gut. Results showed seasonal acclimatization to cold, which is consistent with the use of behavioral freeze-avoidant mechanisms. However, these insects also demonstrated an increased susceptibility to freezing later in the season, which may be tied to changes in gut microbiota. Our results found shifts in the composition of the gut microbiome in R. flavipes between mid- to late summer and early to late fall. These differences may be suggestive of a change in metabolism to adjust to a period of reduced feeding and increased metabolic stress during overwintering. Specifically, results showed an increased abundance of Methanobrevibacter sp. (Euryarchaeota) associated with cold, which may be indicative of a metabolic shift from acetogenesis to methanogenesis associated with overwintering. Further work is needed focusing on specific contributions of certain gut microbes, particularly their role in metabolic adaptability and in providing protection from oxidative stress associated with changes in environmental conditions.
Wood deterioration due to basidiomycetous decay fungi shortens the useful life span of wood and wood-based materials. Prescriptive preservative treatment is the most effective way to reduce the detrimental effects of these microorganisms, particularly in soil contact and areas of critical use (difficult to replace or vital to structure). Current American Wood Protection Association (AWPA) guidelines in the standardized use category system specify 3 zones of severity regarding wood decay fungal hazards but contain very little information on the diversity and abundance of these fungi colonizing soil and wood. In this study, amplicon based sequencing was utilized to compare fungal communities in wood and adjacent soil to provide baseline data on the fungi involved in the process. A thorough understanding of decay hazards is critical for the proper selection and use of wood in soil contact. The goal of this work is to provide baseline data on basidiomycete fungal diversity and species composition in different zones of the existing 3-zone AWPA hazard map as compared to the previous 5-zone hazard map and Scheffer decay indices and discuss the ecological implications for wood decay.
Background Illegal logging is a global crisis with significant environmental, economic, and social consequences. Efforts to combat it call for forensic methods to determine species identity, provenance, and individual identification of wood specimens throughout the forest products supply chain. DNA-based methodologies are the only tools with the potential to answer all three questions and the only ones that can be calibrated “non-destructively” by using leaves or other plant tissue and take advantage of publicly available DNA sequence databases. Despite the potential that DNA-based methods represent for wood forensics, low DNA yield from wood remains a limiting factor because, when compared to other plant tissues, wood has few living DNA-containing cells at functional maturity, it often has PCR-inhibiting extractives, and industrial processing of wood degrades DNA. To overcome these limitations, we developed a technique—organellar microcapture—to mechanically isolate intact nuclei and plastids from wood for subsequent DNA extraction, amplification, and sequencing. Results Here we demonstrate organellar microcapture wherein we remove individual nuclei from parenchyma cells in wood (fresh and aged) and leaves of Carya ovata and Tilia americana, amyloplasts from Carya wood, and chloroplasts from kale ( Brassica sp.) leaf midribs. ITS (773 bp), ITS1 (350 bp), ITS2 (450 bp), and rbcL (620 bp) were amplified via polymerase chain reaction, sequenced, and heuristic searches against the NCBI database were used to confirm that recovered DNA corresponded to each taxon. Conclusion Organellar microcapture, while too labor-intensive for routine extraction of many specimens, successfully recovered intact nuclei from wood samples collected more than sixty-five years ago, plastids from fresh sapwood and leaves, and presents great potential for DNA extraction from recalcitrant plant samples such as tissues rich in secondary metabolites, old specimens (archaeological, herbarium, and xylarium specimens), or trace evidence previously considered too small for analysis.
The aim of this study was to evaluate the efficacy of biological and chemical remediation of chromated copper arsenate (CCA) treated Corymbia citriodora poles, removed from service after 30 years. The presence of arsenic (As), chromium (Cr) and copper (Cu) was quantified by inductively coupled plasma optical emission spectrometry (ICP-OES). Twelve species of decay fungi were used for the biological remediation assay. For chemical remediation oxalic, citric, maleic and ethylenediamine tetraacetic (EDTA) acids were used for 24 and 48 h. In biological remediation, copper-tolerant brown-rot fungi, Wolfiporia cocos, Antrodia xantha and Fibroporia radiculosa, performed the best results, with the highest removals for As (59-85 %) and Cr (38-61 %). Cu was the most easily extracted, with removals above 60 % among the tested fungi, with the best results (90-98 %) for F. radiculosa, Coniophora puteana, Antrodia vaillantii and Postia placenta. In chemical remediation, the extraction time of 48 h was the most effective, and oxalic acid generally reached the highest removals. The EDTA + oxalic acid combination reached the highest value for Cu extraction (98 %).
Shou sugi ban, also known as yakisugi, or just sugi ban, is an aesthetic wood surface treatment that involves charring the surface of dimensional lumber, such as exterior cladding. The goal of this research is to examine the effect of shou sugi ban on the flammability and decay resistance of wood. Several species and variants of commercially available sugi ban were tested. The flammability was examined from the heat release rate curves using the oxygen consumption method and cone calorimeter. Durability was examined with a soil block assay for one white-rot fungus and one brown-rot fungus. The testing showed that the shou sugi ban process did not systematically improve the flammability or durability of the siding.
Brown rot fungi utilize iron as part of a chelator-mediated Fenton (CMF) reaction during wood biodegradation. Research suggests these fungi reduce Fe3+ to promote oxygen radical generation resulting in depolymerization of the wood cell wall. High levels of Mn are also found in wood decayed by brown rot fungi. However, little is known about the oxidation states of Fe and Mn during the decay process. X-ray absorption near edge spectroscopy (XANES) can be used to examine metal oxidation states and coordination chemistry. XANES experiments were conducted on wood decayed by Gloeophyllum trabeum over 2–8 weeks with results showing that Mn2+ and Fe3+ predominated for metal oxidation states. However, Fe2+ was present at sites of greater fungal growth In certain cases, the μXANES measurements showed that the fraction of Fe2+ in the wood samples was as high as 50%. Localized areas of reduced iron corresponded with areas of greater fungal hyphal mass which is in agreement with how brown rot fungi decay wood via the CMF reaction. The limited change in oxidation state of Mn observed in wood with active fungal activity suggests that the role of manganese in CMF biodegradation chemistry should be further explored.
Copper tolerance of brown-rot basidiomycete decay fungi can lessen the efficacy of copper-containing wood preservatives for wood products in-service. The purpose of this study was to evaluate wood mass loss and differential expression of three genes that have putative annotations for copper-transporting ATPase pumps (FIBRA_00974, FIBRA_04716, and FIBRA_01430). Untreated southern pine (SP) and SP treated with three concentrations of ammoniacal copper citrate (CC, 0.6, 1.2, and 2.4%) were exposed to two copper-tolerant Fibroporia radiculosa isolates (FP-90848-T and L-9414-SP) and copper-sensitive Gloeophyllum trabeum isolate (MAD 617) in a 4-week-long standard decay test (AWPA E10-19). Decay of copper-treated wood was inhibited by G. trabeum (p = 0.001); however, there was no inhibition of decay with increasing copper concentrations by both F. radiculosa isolates. Initially, G. trabeum and one F. radiculosa isolate (L-9414-SP) highly upregulated FIBRA_00974 and FIBRA_04716 on copper-treated wood at week 1 (p = 0.005), but subsequent expression was either not detected or was similar to expression on untreated wood (p = 0.471). The other F. radiculosa isolate (FP-90848-T) downregulated FIBRA_00974 (p = 0.301) and FIBRA_04716 (p = 0.004) on copper-treated wood. FIBRA_01430 expression by G. trabeum was not detected, but was upregulated by both F. radiculosa FP-90848-T (p = 0.481) and L-9414-SP (p = 0.392). Results from this study suggest that all three test fungi utilized different mechanisms when decaying copper-treated wood. Additionally, results from this study do not provide support for the involvement of these putative gene annotations for copper-transporting ATPase pumps in the mechanism of copper-tolerance.
The effects of leaf litter on moisture content and fungal decay development in above-ground wood specimens were assessed. Untreated southern pine specimens were exposed with or without leaf litter contact. Two types of leaf litter were evaluated; aged (decomposed) and young (early stages of decomposition). The moisture content of specimens was monitored, and specimens were periodically removed for visual evaluation of decay development. In addition, amplicon-based sequencing analysis of specimens and associated leaf litter was conducted at two time points. Contact with either type of leaf litter resulted in consistently higher moisture contents than those not in contact with leaf litter. Visually, evident decay developed most rapidly in specimens in contact with the aged leaf litter. Analysis of amplicon-based sequencing revealed that leaf litter contributes a significant amount of the available wood decay fungal community with similar communities found in the litter exposed wood and litter itself, but dissimilar community profiles from unexposed wood. Dominant species and guild composition shifted over time, beginning initially with more leaf saprophytes (ascomycetes) and over time shifting to more wood rotting fungi (basidiomycetes). These results highlight the importance of the contributions of leaf litter to fungal colonization and subsequent decay hazard for above-ground wood.
Living organisms require copper for several cellular processes. Yet intracellular concentrations of copper must be regulated to avoid toxicity. Not much is known about mechanisms of copper regulation in wood decay fungi. However, one putative annotation for a copper homeostasis CutC gene (FIBRA_00129), found in other brown-rot wood decay fungi, has been annotated in Fibroporia radiculosa. The aim of this study was to evaluate wood mass loss and differential expression of FIBRA_00129 during initial decay of untreated and copper-treated wood by two copper-tolerant F. radiculosa isolates (FP-90848-T and L-9414-SP) compared with copper-sensitive Gloeophyllum trabeum. Untreated southern pine (Pinus spp.) and ammoniacal copper citrate treated southern pine at three concentrations (0.6%, 1.2%, and 2.4%) were used in a 4-week-long standard decay test. Results showed G. trabeum was unable to decay copper-treated wood while both F. radiculosa isolates successfully decayed southern pine at all copper concentrations. G. trabeum and F. radiculosa L-9414-SP showed no detectable FIBRA_00129 expression over the course of this study. F. radiculosa FP-90848-T showed greater FIBRA_00129 downregulation on copper-treated wood than on untreated wood (P = 0.003). Additionally, there was greater FIBRA_00129 downregulation in F. radiculosa FP-90848-T at week 3 compared with other weeks (P = 0.015). Future studies are needed to further evaluate FIBRA_00129 during the decay process to determine its potential role in copper-tolerance.
Acetylation is one of the most common types of wood modification and is commercially available throughout the world. Many studies have shown that acetylated wood is decay resistant at high levels of acetylation. Despite its widespread use, the mechanism by which acetylation prevents decay is still not fully understood. It is well known that at a given water activity, acetylation reduces the equilibrium moisture content of the wood cell wall. Furthermore, linear relationships have been found between the acetylation weight percent gain (WPG), wood moisture content, and the amount of mass loss in decay tests. This paper examines the relationships between wood moisture content and fungal growth in wood, with various levels of acetylation, by modifying the soil moisture content of standard soil block tests. The goal of the research is to determine if the reduction in fungal decay of acetylated wood is solely due to the reduction in moisture content or if there are additional antifungal effects of this chemical treatment. While a linear trend was observed between moisture content and mass loss caused by decay, it was not possible to separate out the effect of acetylation from fungal moisture generation. The data show significant deviations from previously proposed models for fungal moisture generation and suggest that these models cannot account for active moisture transport by the fungus. The study helps to advance our understanding of the role of moisture in the brown rot decay of modified wood.
To avoid the use of toxic synthetic chemicals due to their potential environmental impacts, the feasibility of using heartwood extracts of Tectona grandis and Cedrus deodara as wood preservatives against the subterranean termite, Reticulitermes flavipes and two basidiomycete decay fungi, Trametes versicolor and Rhodonia placenta, was investigated in laboratory experiments. There were no significant differences in feeding for R. flavipes fed solvent-extracted and non-extracted T. grandis in choice and no-choice tests with 100% mortality. Reticulitermes flavipes ignored non-extracted C. deodara (mass loss 1.93%) in choice tests and consumed significantly more solvent extracted C. deodara (mass loss 33.4%) with 64.9% mortality. Complete termite mortality (100%) was observed after exposure to non-extracted C. deodara versus 53% mortality when fed on extracted C. deodara in a no-choice test. When extracted and non-extracted blocks of each wood species were exposed to decay fungi, durability of both heartwood species was reduced post extraction. Extracts removed from wood shavings via Soxhlet extraction were used to treat non-durable southern pine and cottonwood. Both extracts imparted termite resistance to the non-durable species. Weight losses of both non-durable species were reduced at the highest extract concentration tested (10 mg ml−1), and were inversely related to extract concentrations and retentions. Significantly higher termite mortality was observed at the maximum extract concentration tested for either extract. Water leaching of non-durable wood species treated with T. grandis extract did not reduce termite resistance, and no significant difference between mortality of termites on leached versus non-leached samples was observed. Conversely, the weight loss of wood treated with C. deodara extract was significantly greater post leaching. T. grandis and C. deodara extracts showed no protective effects at tested concentrations against decay fungi when applied to non-durable southern pine or cottonwood.
The goal of this study was to demonstrate how synchrotron-based X-ray fluorescence microscopy (XFM) can be used to better understand the mechanisms of copper tolerance in wood decay fungi. Copper is a major component in commercial wood preservatives as it is toxic to many wood decay fungi. However, certain fungi are copper tolerant and can attack preservative-treated wood, resulting in structural damage to treated wood members. Here we used large-field XFM to visualize six different elements (K, Ca, Mn, Fe, Cu, and Zn) in the mycelia and wood inoculated with four different species of brown rot wood decay fungi. Wood blocks were partially dipped into a solution of copper sulfate, exposed to fungi in malt extract agar petri dish assays for nine weeks, and then imaged and compared to blocks that were partially dipped in water. The blocks were imaged immediately adjacent to an end-matched control that was placed in malt extract agar petri dish assays for 9 weeks, but not exposed to the fungi so that the differences in the elemental distributions could be directly compared. The colonized wood and mycelia were rich in K, Ca, Mn, and Fe; however, the elements and the spatial distribution in the mycelia and wood differed across fungal species. The most interesting results were the maps showing the copper distribution. While three of the four fungi grew on the copper-rich region of the wood, only one species, Fibroporia radiculosa, dramatically reduced the copper concentration in the region of fungal growth.
Soil samples were collected from field sites in two AWPA (American Wood Protection Association) wood decay hazard zones in North America. Two field plots at each site were exposed to differing preservative chemistries via in-ground installations of treated wood stakes for approximately 50 years. The purpose of this study is to characterize soil fungal species and to determine if long term exposure to various wood preservatives impacts soil fungal community composition. Soil fungal communities were compared using amplicon-based DNA sequencing of the internal transcribed spacer 1 (ITS1) region of the rDNA array. Data show that soil fungal community composition differs significantly between the two sites and that long-term exposure to different preservative chemistries is correlated with different species composition of soil fungi. However, chemical analyses using ICP-OES found levels of select residual preservative actives (copper, chromium and arsenic) to be similar to naturally occurring levels in unexposed areas. A list of indicator species was compiled for each treatment-site combination; functional guild analyses indicate that long-term exposure to wood preservatives may have both detrimental and stimulatory effects on soil fungal species composition. Fungi with demonstrated capacity to degrade industrial pollutants were found to be highly correlated with areas that experienced long-term exposure to preservative testing.
For decades, wood scientists and preservative formulators have employed the monocultured soil bottle assay to test efficacy of wood treatment in the laboratory as a rapid predictor of field performance. This study examines the effects of bicultured soil bottle assays on the decay by common wood decay fungi. Mycelial interactions were noted in early stages of colonization. With only two exceptions, a single fungus was apparent in each soil bottle, indicating dominance. The dominant fungi were not always the most efficient wood rots, and the rot type, white or brown, did not affect the dominance outcome on the preferred wood type.
This paper examines how wood decay fungi affect the electrical resistance (resistivity) of wood to determine whether an electrical resistance probe could be deployed as a remote sensor in a wall to detect wood decay. Electrical resistance measurements were taken on wood blocks exposed to Gloeophyllum trabeum in a standard soil bottle tests (AWPA E10) as a function of time. To understand how and why resistance changed with time, results were compared against resistance measurements taken in sterilized, un-inoculated wood at high moisture contents; concentrations of certain mineral ions in the wood were also measured. Results indicate that most of the changes in electrical resistance in the soil bottle test could be described by changes in moisture content alone. When the moisture effects were removed, decay fungi decreased the resistance at small amounts of weight loss but increased the resistance at high weight loss. The results presented herein help to explain the mechanism by which empirical, electrical resistance nondestructive testing devices work and also the limitations on using resistance measurements to detect wood decay.