RESUMEN
The active biomonitoring technique has been demonstrated to be an excellent tool for monitoring water quality; however, further improvement of the protocol is urgently needed. The present study was carried out to determine the best options for various methodological aspects of monitoring some metals and metalloids (i.e. Al, As, Cd, Co, Cu, Fe, Hg, Ni, Zn and Pb): i) the type of transplant, ii) pre-exposure washing (with or without cellular extractants), iii) the ratio between moss weight and bag surface area, and iv) the depth at which the bags are exposed. The importance of the different methodological aspects in the outcome of biomonitoring studies was also assessed by considering the results of the present and other previously published studies. Regarding the type of transplant, the traditionally used net bags were the best option for enclosing the moss; in addition, washing the moss with extracellular extractants (i.e. EDTA) prior to exposure increased the sensitivity of the technique and reduced the required exposure time (i.e. one week). For the amount of moss packed in each bag, a ratio of 12.5â¯mgâ¯cm-2 was the best choice. Finally, the depth at which the transplants were exposed did not affect pollutant accumulation (in shallow rivers, reservoirs or dams). Pollutant concentrations were also not affected by the existence of thermocline in deep waters during warmer months. Different methodological aspects involved in applying this technique determine the final concentrations of metals in moss. Although the influence of those was variable, for most elements (i.e. As, Cd, Co, Cu, Hg, Pb, Zn) 80% of the total variance was explained by 3-4 aspects, being species selection, devitalization treatment, duration of exposure, and number of transplants exposed the most important.