In this research we evaluate the effects of the method used for estimating the potential surface available for benthic macroinvertebrates in macrophyte and unvegetated habitats on several metrics and habitat preference of aquatic macroinvertebrates in the upper catchment of the Henares River (Guadalajara, Central Spain). Three sampling sites were selected: a well-preserved stream (site A), a stream with no wood riparian vegetation (site B), and a straightened and deforested reach (site Q. Two habitats were selected in each site: unvegetated habitat (i.e., substrata without macrophytes) and macrophyte habitat (i.e., substrata covered by macrophytes). In each habitat, six macroinvertebrate samples (including all macrophytes or mineral particles) were collected using a Hess sampler. Diversity and density of major families were referred to the surface of the Hess sampler ( = Hess surface method) and to the actual surface of either mineral particles or macrophytes actual surface method). In general, for the actual Surface method, biomass, richness, dominance, and diversity metrics were higher in the mineral habitat than in the macrophyte habitat. This trend was different for the Hess surface method. In general, densities turned out to be higher in the unvegetated habitat than in the macrophyte habitat when using the actual Surface method, but the reverse occurred when using the Hess Surface method. This fact is relevant for river biomonitoring, especially when reaches with different dominant Substrates (macrophytes vs mineral) are compared using just one of the methods. It is concluded that the macrobenthic metrics and density values are influenced by the method used to estimate the potential available surface for aquatic macroinvertebrates.
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