Do heavy metals affect bacterial communities more in small repeated applications or in a single large application?

Heavy metals from anthropogenic sources accumulate slowly but steadily, leading to high metal concentration levels in soil. However, the effect of each heavy metal on soil bacterial communities is usually assessed in lab-oratories by a single application of individually spiked metals. We evaluated the differences between single individual application and repeated individual applications of Cr, Cu, Ni, Pb, and Zn on bacterial communities, through pollution-induced community tolerance (PICT), using bacterial growth as the endpoint (3H-leucine incorporation method). We found that PICT development was higher when soil was spiked in individual single application than individual repeated applications for Cu, Ni and Zn. In contrast, bacterial communities did not show different tolerance between singly or repeatedly when soil was spiked with Cr. In the case of Pb any in-crease of bacterial community tolerance to this metal was found despite high doses applied (up to 2000 mg kg-1). These results are relevant for the interpretation of the effects of heavy metals on soil microbes in order to avoid laboratory overestimations of the real effects of heavy metals on soil microbes.