A rapid staining-assisted wood sampling method for PCR-based detection of pine wood nematode Bursaphelenchus xylophilus in Pinus massoniana wood tissue

For reasons of unequal distribution of more than one nematode species in wood, and limited availability of wood samples required for the PCR-based method for detecting pinewood nematodes in wood tissue of Pinus massoniana, a rapid staining-assisted wood sampling method aiding PCR-based detection of the pine wood nematode Bursaphelenchus xylophilus (Bx) in small wood samples of P. massoniana was developed in this study. This comprised a series of new techniques: sampling, mass estimations of nematodes using staining techniques, and lowest limit Bx nematode mass determination for PCR detection. The procedure was undertaken on three adjoining 5-mg wood cross-sections, of 0.5 · 0.5 · 0.015 cm dimension, that were cut from a wood sample of 0.5 · 0.5 · 0.5 cm initially, then the larger wood sample was stained by acid fuchsin, from which two 5-mg wood cross-sections (that adjoined the three 5-mg wood cross-sections, mentioned above) were cut. Nematode-staining-spots (NSSs) in each of the two stained sections were counted under a microscope at 100· magnification. If there were eight or more NSSs present, the adjoining three sections were used for PCR assays. The B. xylophilus – specific amplicon of 403 bp (DQ855275) was generated by PCR assay from 100.00% of 5-mg wood cross-sections that contained more than eight Bx NSSs by the PCR assay. The entire sampling procedure took only 10 min indicating that it is suitable for the fast estimation of nematode numbers in the wood of P. massonina as the prelimary sample selections for other more expensive Bx-detection methods such as PCR assay.

Porosity in ion-exchanged and acid activated clays evaluated using n-nonane pre-adsorption

The applicability of the n-nonane pre-adsorption method for characterising the porosity in clays is presented. Na-SD, a Na+-exchanged purified bentonite, and materials obtained by Al3+-exchange and acid treatments of Na-SD and SAz-1 were used. Nitrogen adsorption isotherms, at -196 ºC, were determined before and after n-nonane pre-adsorption on each of the samples. In all materials, n-nonane remained adsorbed in ultramicropores after outgassing at 25 ºC. Outgassing at higher temperatures (50, 75 and 200 ºC) removed nonane and ultramicropores became available for nitrogen adsorption. All treatments on Na-SD led to increase in micropore volume. Larger ultramicropore and supermicropore volumes were obtained for Na-SD acid activated with HCl at 95 ºC than for treatments at 25 ºC with HCl or following Al3+-exchange (Al-SD), and increased with increasing acid concentration to 3 M. Activation with 4 M HCl led to the largest pore volume with contribution from mesopores. However, the specific external surface area was the same as that obtained for Na-SD, Al-SD and for most of the other acid activated samples. Treatments at 95 ºC with 1 M and 6 M HCl promoted increase in specific external surface area. The micropore volumes and specific external surface area for SAz-1 treated with 1 M HCl at 95 ºC were larger than those of Al-SAz-1, but lower than those obtained for corresponding materials derived from Na-SD. The n-nonane pre-adsorption method enabled micropore volumes and specific external surface areas to be obtained for all samples.

Dynamical modeling of biogas production supported by an experimental anaerobic digestion process

This paper presents the development of a combined experimental and numerical approach to study the anaerobic digestion of both the wastes produced in a biorefinery using yeast for biodiesel production and the wastes generated in the preceding microbial biomass production. The experimental results show that it is possible to valorise through anaerobic digestion all the tested residues. In the implementation of the numerical model for anaerobic digestion, a procedure for the identification of its parameters needs to be developed. A hybrid search Genetic Algorithm was used, followed by a direct search method. In order to test the procedure for estimation of parameters, first noise-free data was considered and a critical analysis of the results obtain so far was undertaken. As a demonstration of its application, the procedure was applied to experimental data.