“…Several approaches have been used to define reference conditions, among them the use of physical and chemical water variables (Detenbeck and Cincotta, 2008;Zhu and Chang, 2008;Casatti et al, 2009), water quality and habitat data (Astin, 2007), historical data on environmental conditions or on the aquatic fauna (Hu et al, 2007;Qadir and Malik, 2009), maximum scores of the IBI for less altered sites (Bozzetti and Schulz, 2004), gradients of human interference and occupation (Kanno et al, 2010), or protocols for environmental assessment (Oliveira et al, 2011). Some studies did not clearly state if or how reference conditions were defined (Van Liefferinge et al, 2010). The WFD associates reference conditions with stream typology; the biological attributes of the reference site should represent the range of conditions that are expected to occur naturally within each type of stream (Stoddard et al, 2006).…”