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The use of immunofluorescence labelling for detecting Ophiostoma piceae in radiata pine
1998 - IRG/WP 98-10268
The primary objective of the current investigation is to understand fungal interactions of dominant sapstaining fungi in radiata pine using confocal laser scanning microscopy (CLSM). In the current paper, an immunofluorescence labelling technique was developed for detection of Ophiostoma piceae using a monoclonal antibody. The primary antibody was labelled with Oregon green 514 Goat anti-mouse IgG fluorescent dye before examining with CLSM. The study showed that immunofluoresence labelling was achieved satisfactorily for detecting fungal hyphae of Ophiostoma piceae but not for some other important sapstaining fungi that occur in radiata pine.
Ying Xiao, B Kreber, C Breuil
Use of the enzyme-linked immunoassay (ELISA) and immunocytochemistry to detect soft rot in preservative treated and untreated wood
1990 - IRG/WP 2347
A polyclonal antibody was raised against mycelial extracts from the soft rot fungus Phialophora mutabilis. In ELISA assays, the antibody showed strong reactions with its own antigen and strong to moderate cross reactions with 6 other Phialophora soft rot species. With the exception of Ceratocystis albida, the antibody gave only weak or negative reactions with 11 other mold, blue stain and rot fungi chosen for their frequent occurrence on treated wood.The antibody also showed a high cross reactivity with extracellular filtrates from Phialophora mutabilis and was able to detect filtrates containing ca 40 ng/ml-l of protein. TEM-immunogold labelling of wood degraded by Phialophora mutabilis showed specific localization of the antibody on the fungal cell wall and certain intracellular structures. Extracellular labelling within soft rot cavities and sites of erosion decay were also noted. The antibody was assessed in ELISA assays for detecting the presence of the fungus and soft rot in untreated and preservative treated wood blocks of pine and birch degraded for periods ranging from 1-12 months. Phialophora mutabilis was detected in samples from all wood blocks at low or high weight losses. Highest ELISA readings were recorded for wood blocks with highest substrate losses.
G F Daniel, T Nilsson
Development of a new characterization method to analyse Cypermethrin penetration in wood material by immunolabelling
2015 - IRG/WP 15-30674
The preservative efficacy of organic biocides is strongly related to their capacity of penetration and retention within wood tissues. The specific detection of the pyrethroid insecticide cypermethrin is currently obtained after extraction followed by chemical analysis by chromatography techniques. However visualizing the insecticide molecule within the wood structure requires specific probes together with microscopy techniques. Therefore, the aim of the present work was to apply a new methodology based on antibody-antigen recognition and electronic microscopy to visualize directly cypermethrin in wood material. A polyclonal antibody directed against cypermethrin was developed and implement it on Pinus sylvestris wood samples coated with technical cypermethrin. The antibody was tested on cypermethrin-impregnated wood and the specific recognition of the insecticide was visualized in transmission electron microscopy (TEM). The immunogold-TEM assay evidenced the capacity of the synthetic biocide to penetrate in the wood. The depth of penetration was measured on sections taken at increasing distances from the coated surface of the wood. In the present conditions of application, cypermethrin was shown to penetrate up to the 6-9 mm zone below the surface. Such results correlated with chemical analyses carried out by GC-ECD after extraction. In addition, the immuno-TEM investigation allowed visualizing, for the first time at the ultrastructure scale of resolution, that cypermethrin was able to diffuse within the secondary wood cell walls. The scarce labeling of the compound middle lamella shows that the chemical does not diffuse freely in this part of the cell walls. The results suggest that the adsorption within the cell walls is an essential factor of the retention of cypermethrin and of its permanence in wood-treated materials and products.
S Tapin-Lingua, K Ruel, J-P Joseleau, D Messaoudi, O Fahy, M Jequel, M Petit-Conil