Recent applications of the Dot-ELISA in immunoparasitology

The dot enzyme-linked immunosorbent assay (Dot-ELISA) is a highly versatile solid-phase immunoassay for antibody or antigen detection. The assay uses minute amounts of reagent dotted onto solid surfaces such as nitrocellulose and other paper membranes which avidly bind proteins. After incubation with antigen-specific antibody and enzyme-conjugated anti-antibody, the addition of a precipitable, chromogenic substrate causes the formation of a colored dot on the solid phase which is visually read. The Dot-ELISA has been used extensively in the detection of human and veterinary protozoan and metazoan parasitic diseases, including amebiasis, babesiosis, fascioliasis, cutaneous and visceral leishmaniasis, cysticercosis, echinococcosis, malaria, schistosomiasis, toxocariasis, toxoplasmosis, trichinosis, trypanosomiasis and even ixodid tick infestation. The technique is rapid, easy to perform and interpret, reagent conservative, cost effective and field portable. In addition, the Dot-ELISA may be configured to detect antibodies or parasite antigen in either microtiter plates for large-batch testing or with dipsticks for small numbers of determinations. A slight modification of the Dot-ELISA procedure allows the determination of infection rates of vectors such as ticks and sandflies with parasites.     

Detection and characterization of fungicide resistant phenotypes of Botrytis cinerea in lettuce crops in Greece

The development of resistance to chemical control agents needs continuous monitoring in Botrytis cinerea. 790 isolates from lettuce and other vegetable crops were collected from six widely separated sites in Greece and tested for their sensitivity to 11 fungicides from nine unrelated chemical groups. 44 of the isolates exhibited multiple resistance to fenhexamid (hydroxyanilides), azoxystrobin and pyraclostrobin (QoI’s), boscalid (SDHI’s), cyprodinil and pyrimethanil (anilinopyrimidines), fludioxonil (phenylpyrroles), carbendazim (benzimidazoles) and iprodione (dicarboximides). Thirty per cent of such phenotypes were detected in an experimental glasshouse with lettuce crops, the third year after commencing fungicide applications. The average resistance factor (R_f) for mycelial growth to fenhexamid, pyraclostrobin, boscalid, cyprodinil and fludioxonil, was over 40, 1,000, 100, 700 and 50, respectively. Some strains with high resistance to anilinopyrimidines (14 %) or moderate to fludioxonil (7 %) were detected even in isolates collected from vegetable crops prior to commercial use of these fungicides in Greece. Isolates with fludioxonil moderate resistance and fenhexamid high resistance, were detected for the first time in Greece. The results suggested the high risk in chemical control of grey mould due to development of resistance to most fungicides with site-specific modes of action. Isolates with resistance to fluazinam (phenylpyridinamines) and to chlorothalonil (phthalonitriles) were not found. The inclusion of appropriate multi-site inhibitors like chlorothalonil in fungicide anti-resistance strategies was indispensable.     

Nitrogen uptake in riparian plant communities across a sharp ecological boundary of salmon density

Background

Recent studies of anadromous salmon (Oncorhynchus spp.) on the Pacific Coast of North America indicate an important and previously unrecognized role of salmonid nutrients to terrestrial biota. However, the extent of this uptake by primary producers and consumers and the influences on community structure remain poorly described. We examine here the contribution of salmon nutrients to multiple taxa of riparian vegetation (Blechnum spicant, Menziesii ferruginea, Oplopanax horridus, Rubus spectabilis, Vaccinium alaskaense, V. parvifolium, Tsuga heterophylla) and measure foliar δ¹⁵N, total %N and plant community structure at two geographically separated watersheds in coastal British Columbia. To reduce potentially confounding effects of precipitation, substrate and other abiotic variables, we made comparisons across a sharp ecological boundary of salmon density that resulted from a waterfall barrier to salmon migration.

Results

δ¹⁵N and %N in foliage, and %cover of soil nitrogen indicators differed across the waterfall barrier to salmon at each watershed. δ¹⁵N values were enriched by 1.4‰ to 9.0‰ below the falls depending on species and watershed, providing a relative contribution of marine-derived nitrogen (MDN) to vegetation of 10% to 60%. %N in foliar tissues was slightly higher below the falls, with the majority of variance occurring between vegetation species. Community structure also differed with higher incidence of nitrogen-rich soil indicator species below the waterfalls.

Conclusions

Measures of δ¹⁵N, %N and vegetation cover indicate a consistent difference in the riparian community across a sharp ecological boundary of salmon density. The additional N source that salmon provide to nitrogen-limited habitats appears to have significant impacts on the N budget of riparian vegetation, which may increase primary productivity, and result in community shifts between sites with and without salmon access. This, in turn, may have cascading ecosystem effects in forests adjacent to salmon streams.