Immunological quantification of the nematode parasitic bacterium Pasteuria penetrans in soil

Abstract

Currently, the abundance of Pasteuria penetrans in soils, an unculturable bacterial parasite of root-knot nematodes (Meloidogyne spp,). is estimated by the percentage of nematode juveniles infected with bacteria and the number of spores attached to their cuticle. Indirect immunofluorescence led to detection of free spores directly in soil suspensions using UV light and polyclonal antibodies raised against two P. penetrans populations (ORS-21414-Sen and PPI). Three extraction methods were compared in order to improve spore recovery. A gentle shaking/sieving method recovered more than 90% of the spores inoculated in soils and was more efficient and simple than aqueous two-phase partitioning and polyethylene glycol extractions. All the spores inoculated in sandy or sandy-clay soils were detected with immunofluorescence microscopy. The quantification of the spores was improved using an ELISA technique that showed a good correlation between optical density and spore concentration in inoculated soils. Specific antibodies provide a suitable method to quantify A penetrans and may be used to follow the evolution of the real pool of bacteria either in native suppressive soils or in inoculated ones. (C) 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.