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We collected 50 subsamples of the moss Scleropodium purum from each of three sampling sites and determined the concentrations of As, Cd, Cu, K, Hg, Ni, Pb, Se, and Zn in each subsample. We then calculated the number of subsamples required to determine significant differences in the mean concentrations of two sampling sites. We found that to differentiate between an uncontaminated sampling site and another, slightly contaminated site, 30 subsamples are required from each. On the basis of these results and because, to date, there are no studies that justify the application of the previously proposed recommendations, studies of local variability in other areas, and under different conditions, with other contaminants and moss species must be undertaken. For posterior comparison of data on variability as well as application of the recommendations associated with such results, the general use of a single definition of a subsample is required. We propose the following definition of a subsample: "1 g dry weight (approx.), collected within a 25-cm radius of a node, selected at random from a 1 x 1-m sampling grid placed in a sampling site."

In this study, we investigated if the different sampling designs normally used in extensive bryomonitoring surveys of air pollution allow compliance with the objectives proposed in the 'Heavy Metals in European Moss' project. For this, we have used data on concentrations of As, Cu, Se and Zn from five moss sampling surveys carried out in Galicia (NW Spain), using three types of sampling design: regular grid, sampling sites inside squares and the strategy recommended in the guidelines for European surveys. To study the effect of the interaction between the sampling design and the spatial pattern of the presence of contaminants, we have used a simulated and a real pattern onto which we superimposed the three sampling designs considered. We found that only random sampling and grid sampling allow compliance with the proposed objectives, and we discuss the advantages of the latter over the former. The data corresponding to studies using different sampling designs are not comparable, and in future large-scale surveys, the same design must be used if the results are to be compared and if valid conclusions are to be reached. The scale of the grid determines the scale to which the objectives are complied with, so that for correct standardization of the technique, and taking into account the most commonly used grid sizes, we recommend a grid size of 30 x 30 or 32 x 32 km. When additional samplings are required, the size of the grid size should be 30/2(n) or 32/2(n) km.

Moss (Scleropodium purum) transplants were used to study bioconcentration originating from dry and bulk deposition, by measuring the tissue contents of Al, As, Ca, Cu, Fe, Hg, Mn, Ni, Pb, Se and Zn. Furthermore, a laboratory experiment was carried out to determine the sequence of maximum concentration and affinity of Al, Cu, Fe, Hg, Ni, Pb and Zn, in S. purum. We found that in many of the transplants, for the same period of exposure, higher levels of metals were accumulated via dry deposition than via bulk deposition. This result may be explained by the simple washing action of the rain on the surface of the moss, and by the existence of processes that provoke the loss of some of the accumulated elements: intercationic displacement and leaching caused by acid precipitation. Modelling of the final bioconcentration observed, as a balance of inputs and outputs of elements, revealed that this terrestrial moss does not integrate, but rather concentrates atmospheric deposition, and there exists a state of unstable equilibrium between inputs and outputs of elements, a state that is determined by the characteristics of the surrounding environment. On the basis of the results obtained in this study, we can conclude that at present, it is not possible to extrapolate the calibrations between the concentrations of elements accumulated in a certain species of moss to values of atmospheric deposition (bulk deposition) from one place to another with different environmental conditions.