Survival of Beauveria brongniartii in the soil after application as a biocontrol agent against the European cockchafer Melolontha melolontha

The survival of the entomopathogenic fungusBeauveria brongniartii and its efficacyagainst the European cockchafer, Melolontha melolontha, was examined over 16months after application as a biologicalcontrol agent in different soil types inSwitzerland. In the absence of M. melolontha,the reduction of the number of the CFU in thesoil was nearly 90%. In soils with highorganic content and increased catalase activitythe decline in the CFU density of B.brongniartii was more pronounced. At siteswhere M. melolontha was present, thesurvival was significantly longer. Theapplication of B. brongniartii resultedin a maximal infection rate of 75% in the M. melolontha population. The number of CFU ofB. brongniartii in the soil started todecrease only when the M. melolonthapopulation was reduced during the epizootic(reduction of 46% on average). The rapiddecrease of the fungus in the absence of thehost gives further evidence of the highspecificity of the fungus and that asaprophytic multiplication without the host isunlikely. The fungus was isolated on aselective medium as well as by baiting withGalleria mellonella. The latter methoddid not reveal the significant differencesmentioned above, which leads to the assumptionthat the two methods may select for differentfungal strains.     

Schistosoma mansoni: a diamidinophenylindole probe for in vitro death of schistosomula

The following fluorochromes were studied as probes for discrimination between living and dead Schistosoma mansoni schistosomula: ethidium bromide (EB), propidium iodide (PI), diamidinophenylindole (DAPI), and carboxyfluoresceine diacetate (C-FDA). While schistosomula stained with EB, PI, or C-FDA showed leakage of fluorochrome into the medium, this was not the case with DAPI. Dead schistosomula, which were stained with DAPI, showed an intense blue fluorescence, while living schistosomula were not stained even after prolonged incubation. In addition, the low DAPI concentration (1 microgram/ml) in the medium proved not to be toxic to the schistosomula, nor did it cause any background fluorescence. These properties make DAPI an ideal probe: the viability of S. mansoni schistosomula in cytotoxicity tests can be continuously monitored in tissue culture trays, using an inverted microscope with simultaneous transmitted light and incident fluorescent light illumination.     

Effect of Host Plant Age on Population Development and Pathogenicity of Ditylenchus destructor on Peanut

The effect of inoculating peanut, Arachis hypogaea cv. Sellie, with Ditylenchus destructor at timed intervals after planting and with different initial nematode population densities (Pi) was tested in greenhouse experiments. Final nematode population densities (Pf) in hulls and seeds were greater (Pf < 0.001) in plants inoculated at or before 9 weeks after planting. Pod disease symptoms correlated positively with the Pf in the pods. The seedgrade of peanuts inoculated at or before 9 weeks after planting was reduced, whereas grade of peanuts from plants inoculated at 15 weeks or later was not reduced. Peanut plants inoculated 12 weeks after planting with a Pi of 10-100 had a lower Pf (P < 0.05) than plants with a Pi of 250 to 8,000. Seed of plants with a Pi of 250 or less could be marketed as choice edible seed, whereas those with a Pi of 500 or more were of reduced seedgrade. These results suggest that as few as 500 nematodes per plant at 12 weeks after planting can build up to injurious levels before harvest. A nematicide should therefore be active for longer than 12 weeks after planting to sufficiently suppress the population.     

Using direct amplification and next-generation sequencing technology to explore foliar endophyte communities in experimentally inoculated western white pines

Fungal endophytes can influence survivability and disease severity of trees. Here we characterized the endophyte community in Pinus monticola (western white pine), an important species in the northwest USA, largely decimated by pathogenic fungi. We also assessed the ability to successfully inoculate seedlings with desirable endophytes, with the long-term goal of providing a protective microbiome and added defense from pathogens. P. monticola seedlings were inoculated in the field with potential pathogen antagonists and fungi isolated from healthy mature trees. Following inoculations direct amplification and next generation sequencing were used to characterize fungal endophyte communities, and explore interspecific competition, diversity, and co-occurrence patterns in needle tissues. Negative co-occurrence patterns between inoculated fungi and potential pathogens, as well as many other species, suggest early competitive interactions. Our study explores early endophyte community assemblage and shows that fungal inoculations may influence tree growth.