Development of Bursaphelenchus xylophilus Populations in Wood Chips with Different Moisture Contents

Bags of Pinus strobus wood chips with moisture contents of 38, 92, 164, and 217% (oven dry weight) were inoculated with Bursaphelenchus xylophilus and incubated at 30 C in order to determine the effect of wood moisture on nematode population development. Nematodes were extracted after 2, 4, 8, and 12 weeks. Population levels were greatest in wood chips with a moisture content of 38% and decreased successively with each higher moisture content. In chips with the three lower moisture contents, populations peaked at 2 weeks, but at 217% moisture, they peaked at 8 weeks. By 12 weeks, nematode populations had declined in wood chips with 92 and 164% moisture contents. The fungi most frequently isolated from the wood chips were Alternaria, Fusarium, Gliocladium, Graphium, Penicillium, Trichoderma, and Mucorales.     

Evaluation of a combined brown rot decay–chemical delignification process as a pretreatment for bioethanol production from Pinus radiata wood chips

Wood chips of Pinus radiata softwood were biotreated with the brown rot fungus (BRF) Gloeophyllum trabeum for periods from 4 and 12 weeks. Biodegradation by BRF leads to an increase in cellulose depolymerization with increasing incubation time. As a result, the intrinsic viscosity of holocellulose decreased from 1,487 cm³/g in control samples to 783 and 600 cm³/g in 4- and 12-week decayed wood chips, respectively. Wood weight and glucan losses varied from 6 to 14% and 9 to 21%, respectively. Undecayed and 4-week decayed wood chips were delignified by alkaline (NaOH solution) or organosolv (ethanol/water) processes to produced cellulosic pulps. For both process, pulp yield was 5–10% lower for decayed samples than for control pulps. However, organosolv bio-pulps presented low residual lignin amount and high glucan retention. Chemical pulps and milled wood from undecayed and 4-week decayed wood chips were pre-saccharified with cellulases for 24 h at 50°C followed by simultaneous saccharification and fermentation (SSF) with the yeast Saccharomyces cerevisiae IR2-9a at 40°C for 96 h for bioethanol production. Considering glucan losses during wood decay and conversion yields from chemical pulping and SSF processes, no gains in ethanol production were obtained from the combination of BRF with alkaline delignification; however, the combination of BRF and organosolv processes resulted in a calculated production of 210 mL ethanol/kg wood or 72% of the maximum theoretically possible from that pretreatment, which was the best result obtained in the present study.     

Delignification of Wood Chips and Pulps by Using Natural and Synthetic Porphyrins: Models of Fungal Decay

Kraft pulps, prepared from softwoods, and small chips of birch wood were treated with heme and tert-butyl hydroperoxide in aqueous solutions at reflux temperature. Analyses of treated pulps showed decreases in kappa number (a measure of lignin content) from about 36 to less than 2, with concomitant increases in brightness (80% increase in the better samples). Analyses of treated wood chips revealed selective delignification and removal of hemicelluloses. After 48 h of treatment, lignin losses from the wood chips approached 40%, and xylose/mannose (hemicellulose) losses approached 70%, while glucose (cellulose) losses were less than 10%. Examination of delignified chips by transmission electron microscopy showed that the removal of lignin occurred in a manner virtually indistinguishable from that seen after decay by white rot fungi. Various metalloporphyrins, which act as biomimetic catalysts, were compared to horseradish peroxidase and fungal manganese peroxidase in their abilities to oxidize syringaldazine in an organic solvent, dioxane. The metalloporphyrins and peroxidases behaved similarly, and it appeared that the activities of the peroxidases resulted from the extraction of heme into the organic phase, rather than from the activities of the enzymes themselves. We concluded that heme-tert-butyl hydroperoxide systems in the absence of a protein carrier mimic the decay of lignified tissues by white rot fungi.     

Influence of substrate wood-chip particle size on shiitake (Lentinula edodes) yield

Wood chips from four commercial hardwood sawmills were screened with 10 US standard sieves (4-0.21 mm) to assess particle size distributions. 96-98% of wood chips were < 4 mm while 95-99% of particles were > 0.21 mm. The majority (mean = 64.5%) of wood chips passed through US standard sieve size 14 (< 1.4 mm). Shiitake (Lentinula edodes) was grown in three crops to determine the effect of four particle size classes (1 = 2.8-4 mm; 2 = 1.7-2.8 mm; 3 = 0.85-1.7 mm; 4 = < 0.85 mm) on mushroom yield. Yields from substrates prepared with wood chips from class 4 (< 0.85 mm) were lower by 27.7%, 12.4% and 2% (mean = 14.9%) for Crops I, II, and III, respectively, when compared to controls. Profiling of wood chips may help growers optimize their production media and reduce production costs.     

Performance of oak,beech and spruce beams after more than 100 years in service

The aim of this study was to investigate differences in the mechanical and fungicidal properties of three different wood species (English oak (Quercus sp.), common beech (Fagus sylvatica) and Norway spruce (Picea abies)) that had been in indoor use for several decades, compared to control specimens of freshly cut timber. The collected material was cut into smaller samples prior to further analysis. Extractive content, mechanical, fungicidal and sorption properties were determined according to standard procedures. The obtained results showed that the mechanical properties of oak wood do not deteriorate over the investigated time frame. On the other hand, the resistance of oak wood against fungi decreases over time. The reason for this is yet to be confirmed; it may be due to degradation of secondary metabolites. Similar results have been reported for spruce wood. There were no statistically significant differences in the mechanical properties of old and new spruce wood. In contrast to oak wood, there were also no significant differences in fungicidal properties, bearing in mind that spruce wood has lower durability than oak wood. Aging of beech wood resulted in a considerable decrease in the tested mechanical properties but showed no significant differences in fungicidal properties. Old beech wood specimens were moderately deteriorated by insects and fungi, which was the reason for the loss of bending and compressive strength. Our results confirm that most of the relevant properties do not deteriorate with time and that wood can be reused for a variety of other applications even after decades in service.     

Distribution of glucomannans and xylans in poplar xylem and their changes under tension stress

Present work investigated glucomannan (GM) and xylan distribution in poplar xylem cells of normal- (NW), opposite- (OW) and tension wood (TW) with immunolocalization methods. GM labeling was mostly detected in the middle- and inner S(2) (+S(3)) layer of NW and OW fibers, while xylan labeling was observed in the whole secondary cell wall. GM labeling in vessels of NW and OW was much weaker than in fibers and mostly detected in the S(2) layer, whereas slightly stronger xylan labeling than fibers was detected in the whole secondary cell wall of vessels. Ray cells in NW and OW showed no GM labeling, but strong xylan labeling. These results indicate that GMs and xylans are spatially distributed in poplar xylem cells with different concentrations present in different cell types. Surprisingly, TW showed significant decrease of GM labeling in the normal secondary cell wall of gelatinous (G) fibers compared to NW and OW, while xylan labeling was almost identical indicating that the GM and xylan synthetic pathways in fibers have different reaction mechanisms against tension stress. Unlike fibers, no notable changes in GM labeling were detected in vessels of TW, suggesting that GM synthesis in vessels may not be affected by tension stress. GM and xylan was also detected in the G-layer with slightly stronger and much weaker labeling than the normal secondary cell wall of G-fibers. Differences in GM and xylan distribution are also discussed for the same functional cells found in hardwoods and softwoods.     

Fungal wood decay in the presence of fly ash as indicated by gravimetrics and by extractability of low molecular weight volatile organic acids

A mycoremediation system of wood substrate and fungal inoculum was established on an organically poor fly ash deposit at the ?o?tanj Thermal Power Plant, Slovenia. A mini-block method with beech wood samples was used to select a suitable fungal isolate for inoculation into the mycoremediation system. Pleurotus ostreatus ZIM 76 proved to be the most appropriate for further tests. Compared to the mass loss in a non-contaminated medium, wood degradation with the selected fungal isolate was reduced by 51% on wood substrate contaminated with 17% (w/w) fly ash. The deterioration processes of wood chips on inoculated and non-inoculated experimental plots were studied by ionic chromatographic analyses of volatile organic acids in water extracts. The presence of P. ostreatus did not influence the concentrations of acetic and formic acids in water extracts from wood chips collected at experimental plots in different intervals from May 2006 to November 2007 comparing inoculated vs. non-inoculated plots.     

Biopulping process design and kinetics

Biopulping is the solid-state fermentation of wood chips as a pretreatment for mechanical pulping processes. The two organisms that are currently of the greatest interest for biopulping are the white-rot fungi, Phanerochaete chrysosporium and Ceriporiopsis subvermispora. P. chrysosporium has been shown to successfully biopulp wood (33% energy savings; 39% improvement in tear index) without the need for sterilization of the wood or nutrient supplementation. Demonstrating the practical and economical feasibility of the biopulping process requires process modeling based on accurate kinetic data. Techniques to monitor dry weight loss and growth rate as functions of time using carbon dioxide production data have been developed. Growth was shown to be linear with time on unsupplemented chips and exponential with time on supplemented chips.     

Influence of Pleurotus ostreatus inoculation on wood degradation and fungal colonization

The influence of Pleurotus ostreatus inoculation on wood degradation and on fungal community structure was studied. The experiments were performed on an organically poor fly ash deposit covered with a 10 cm layer of beech wood chips inoculated with P. ostreatus isolate ZIM76. Compared to non-inoculated wood chips, inoculation increased the temperatures and relative humidities and, in the first 6 months, accelerated Klason lignin degradation by 9% and also, after 17 months, increased iron translocation into wood chips by 30%. After 6 months, PCR-DGGE showed 22-28 and 13-21 fungal taxa in non-inoculated and P. ostreatus-inoculated beech chips, respectively. The differences in number of taxa and in the fungal community structure (based on Dice coefficient) between non-inoculated and inoculated wood chips diminished with time. The results indicate that the naturally occurring processes of wood degradation are as efficient as those occurring in sites inoculated with P. ostreatus.     

Evaluating mulches together with Heterorhabditis zealandica (Rhabditida: Heterorhabditidae) for the control of diapausing codling moth larvae,Cydia pomonella (L.) (Lepidoptera: Tortricidae)

The potential of using an entomopathogenic nematode, Heterorhabditis zealandica Poinar, together with different test mulches (pine chips, wheat straw, pine wood shavings, blackwood and apple wood chips) to control diapausing codling moth, Cydia pomonella (L.) larvae was evaluated. Mesh cages were identified as a suitable larval-containment method. High levels of codling moth mortality were obtained when using pine wood shavings as mulch (88%) compared to pine chips, wheat straw, blackwood and apple wood chips (41–88%). Humidity (>95% RH) has to be maintained for at least 3 days to ensure nematode survival. It was also proven that nematodes had the ability to move out of infected soil into moist mulch, to infect the codling moth larvae residing at heights of up to 10 cm. Field experiments showed the importance of climatic conditions on nematode performance. Low temperatures (<15°C) recorded during the first trial resulted in low levels of control (48%), as opposed to the 67% mortality recorded during the second trial (temperatures ranged between 20 and 25°C). Low levels of persistence (<10%) were recorded in the mulches post-application. The study conclusively illustrated some of the baseline requirements fundamental to the success of entomopathogenic nematodes together with mulches for the control of codling moth.     

Biodegradation of Abies bornmülleriana (Mattf.) and Fagus orientalis (L.) by the white rot fungus Phanerochaete chrysosporium

The effects of Phanerochaete chrysosporium, a white rot fungus, on the chemical composition of Abies bornmülleriana and Fagus orientalis wood chips were investigated. After the chips were inoculated with the fungus, 20-, 40- and 60-day samples were analysed in order to determine the influence of fungal treatment on the chemical components of the cell walls, and the fibre properties of both species were measured. As a result of P. chrysosporium growth, both types of wood had slight relative increases in percentage cellulose. Percentage holocellulose showed statistically significant decreases and solubility values increased considerably. The lignin ratio for F. orientalis decreased significantly in relation to zero-time control samples.     

Steam pretreatment and enzymatic production of d-glucose from Eucaluptus globulus

Batch steam pretreatment followed by enzymatic hydrolysis of Eucalyptus globulus chips of two ages (5 and 12 years) was assessed. After 24 h of hydrolysis, young wood samples yielded cellulose conversions close to 80%, while almost complete conversion was obtained at 48 h. Increasing wood concentration from 7% to 10% (by weight) caused cellulase end-product inhibition. In mature wood samples, where the rate of glucose accumulation was slower, cellulase inhibition was not observed. Both samples showed higher conversion values when the cellulase loading was increased from 20 to 30 FP IU dry g⁻¹.     

Sulfite pretreatment (SPORL) for robust enzymatic saccharification of spruce and red pine

This study established a novel process using sulfite pretreatment to overcome recalcitrance of lignocellulose (SPORL) for robust and efficient bioconversion of softwoods. The process consists of sulfite treatment of wood chips under acidic conditions followed by mechanical size reduction using disk refining. The results indicated that after the SPORL pretreatment of spruce chips with 8–10% bisulfite and 1.8–3.7% sulfuric acid on oven dry (od) wood at 180 °C for 30 min, more than 90% cellulose conversion of substrate was achieved with enzyme loading of about 14.6 FPU cellulase plus 22.5 CBU β-glucosidase per gram of od substrate after 48 h hydrolysis. Glucose yield from enzymatic hydrolysis of the substrate per 100 g of untreated od spruce wood (glucan content 43%) was about 37 g (excluding the dissolved glucose during pretreatment). Hemicellulose removal was found to be as critical as lignin sulfonation for cellulose conversion in the SPORL process. Pretreatment altered the wood chips, which reduced electric energy consumption for size reduction to about 19 Wh/kg od untreated wood, or about 19 g glucose/Wh electricity. Furthermore, the SPORL produced low amounts of fermentation inhibitors, hydroxymethyl furfural (HMF) and furfural, of about 5 and 1 mg/g of untreated od wood, respectively. In addition, similar results were achieved when the SPORL was applied to red pine. By building on the mature sulfite pulping and disk refining technologies already practiced in the pulp and paper industry, the SPORL has very few technological barriers and risks for commercialization.     

Effects of bacterial treatments on wood extractives

Bacterial strains were isolated from spruce wood chips and their ability to reduce the content of wood extractives was studied. Strains were screened by cultivation on liquid media containing wood extractives as the major nutrient. Some bacterial species could decrease remarkably the amount of extractives in the liquid media and reduced the amount of triglycerides, steryl esters and total extractives by 100, 20 and 39%, respectively. Spruce wood chips were treated in controlled conditions with selected bacteria to test their effects on the chips. All the bacteria grew well on wood chips. The effect of bacterial metabolism on wood extractives was significant. Bacterial treatments reduced the amount of lipophilic extractives by 16-38% in 1 week of treatment and up to 67% in 2 weeks. The most efficient strain removed 90, 66 and 50% of triglycerides, steryl esters and resin acids, respectively, in 2 weeks. These results indicate that bacteria may be promising agents for the removal of extractives for improved pulping and papermaking processes.     

Novolak PF resins prepared from phenol liquefied Cryptomeria japonica and used in manufacturing moldings

The wood of Japanese cedar (Cryptomeria japonica) was liquefied in phenol with H2SO4 and HCl as catalysts. The liquefied wood was reacted with formalin to prepare the novolak PF resin. The results showed that the reaction of liquefied Japanese cedar with formalin was an exothermic reaction, and formed a solid-like resin without extra heating. Two novolak PF resins were prepared from the liquefied wood which were identified as SF and CF that using the liquefied wood with H2SO4 and HCl as catalyst respectively. The novolak PF powder displayed thermo-melting characteristic. The resins of SF and CF had weight average molecular weight of 3638 and 3941 respectively and melting temperature of 149.4 degrees C and 127.5 degrees C respectively. Both of the novolak resins could be used to make moldings with good performance by mixing the novolak resin with wood powder, hardener and zinc stearate at the weight ratio of 60:30:10:1 and hot-pressed under 200 degrees C for 10min.     

Biopulping of wood chips with Phlebia brevispora BAFC 633 reduces lignin content and improves pulp quality

The white-rot fungus Phlebia brevispora BAFC 633 produces laccases in large proportions. In this work P. brevispora BAFC 633 was grown on Pinus taeda wood chips in 10-L bioreactors. To select the biopulping experimental conditions, we analyzed the variables affecting enzymatic laccase activity in the culture supernatants, indicating that the suitable incubation temperature was 30 °C in order to promote enzyme stability. Phlebia brevispora BAFC 633 secreted 744 U/g of laccase, selectively removing lignin during biotreatment of wood chips, causing a reduction in Kappa number and 10% weight loss, and creating a more open structure and better access to the pulping liquor, which would require less chemical consumption, thus diminishing the environmental impact of the chemical pulping process.These results support the biotechnological potential of P. brevispora BAFC 633 for biopulping processes and enhance the potential for bioprospecting native isolates of the microflora of our country's natural environment.     

Exploring Muscle Activation during Nordic Walking: A Comparison between Conventional and Uphill Walking

Nordic Walking (NW) owes much of its popularity to the benefits of greater energy expenditure and upper body engagement than found in conventional walking (W). Muscle activation during NW is still understudied, however. The aim of the present study was to assess differences in muscle activation and physiological responses between NW and W in level and uphill walking conditions. Nine expert Nordic Walkers (mean age 36.8±11.9 years; BMI 24.2±1.8 kg/m²) performed 5-minute treadmill trials of W and NW at 4 km/h on inclines of 0% and 15%. The electromyographic activity of seven upper body and five leg muscles and oxygen consumption (VO₂) were recorded and pole force during NW was measured. VO₂ during NW was 22.3% higher at 0% and only 6.9% higher at 15% than during W, while upper body muscle activation was 2- to 15-fold higher under both conditions. Lower body muscle activation was similarly increased during NW and W in the uphill condition, whereas the increase in erector spinae muscle activity was lower during NW than W. The lack of a significant increase in pole force during uphill walking may explain the lower extra energy expenditure of NW, indicating less upper body muscle activation to lift the body against gravity. NW seemed to reduce lower back muscle contraction in the uphill condition, suggesting that walking with poles may reduce effort to control trunk oscillations and could contribute to work production during NW. Although the difference in extra energy expenditure between NW and W was smaller in the uphill walking condition, the increased upper body muscle involvement during exercising with NW may confer additional benefit compared to conventional walking also on uphill terrains. Furthermore, people with low back pain may gain benefit from pole use when walking uphill.     

Conversion of Japanese red pine wood (Pinus densiflora) into valuable chemicals under subcritical water conditions

A comparative study on the decomposition of Japanese red pine wood under subcritical water conditions in the presence and absence of phosphate buffer was investigated in a batch-type reaction vessel. Since cellulose makes up more than 40-45% of the components found in most wood species, a series of experiments were also carried out using pure cellulose as a model for woody biomass. Several parameters such as temperature and residence time, as well as pH effects, were investigated in detail. The best temperature for decomposition and hydrolysis of pure cellulose was found around 270 °C. The effects of the initial pH of the solution which ranged from 1.5 to 6.5 were studied. It was found that the pH has a considerable effect on the hydrolysis and decomposition of the cellulose. Several products in the aqueous phase were identified and quantified. The conditions obtained from the subcritical water treatment of pure cellulose were applied for the Japanese red pine wood chips. As a result, even in the absence of acid catalyst, a large amount of wood sample was hydrolyzed in water; however, by using phosphate buffer at pH 2, there was an increase in the hydrolysis and dissolution of the wood chips. In addition to the water-soluble phase, acetone-soluble and water-acetone-insoluble phases were also isolated after subcritical water treatment (which can be attributed mainly to the degraded lignin, tar, and unreacted wood chips, respectively). The initial wood:acid ratio in the case of reactions catalyzed by phosphate buffer was also investigated. The results showed that this weight ratio can be as high as 3:1 without changing the catalytic activity. The size of the wood chips as one of the most important experimental parameters was also investigated.     

Two-stage fungal biopulping for improved enzymatic hydrolysis of wood

A novel two-stage, whole organism fungal biopulping method was examined for increasing the yield of enzymatic hydrolysis of wood into soluble glucose. Liriodendron tulipifera wood chips (1 g) were exposed to liquid culture suspensions of white rot (Ceriporiopsis subvermispora) or brown rot (Postia placenta) fungi and incubated at 28 °C, either alone in single-stage 30 day (one fungal species applied) or two-stage 60 day (both fungal species applied in alternative succession) treatments. Fungi grew in all treatments, but did not significantly decrease the percent carbohydrate content of the wood. Two-stage treatments differed significantly in mass loss depending on order of exposure, suggesting additive or inhibitory fungal interactions occurred. Treatments consisting of C. subvermispora followed by P. placenta exhibited 6 ± 0.5% mass loss and increased the yield of enzymatic hydrolysis by 67-119%. This significant hydrolysis improvement suggests that fungal biopulping technologies could support commercial lignocellulosic ethanol production efforts if further developed.