## 2 Materials and Methods
### Sample Preparation and Analyses for Biomass and HMs  
In the laboratory, the invertebrate samples for biological analysis were dried in an incubator at 40 \({}^{\circ}\)C and then weighed to obtain the total dry mass. The animals for HM analysis were identified to the lowest taxonomic level and stored according to species. The soil and biological (invertebrates and plant leaves) samples for HM analysis were dried in an incubator at 40 \({}^{\circ}\)C. The samples were ground into powder by a mortar and pestle and digested on a hot plate using aqua regia solution (conc. HCl and conc. HNO\({}_{3}\) in ratio of 3:1) for the soil samples and conc. HNO\({}_{3}\) and H\({}_{2}\)O\({}_{2}\) at a ratio of 3:2 for the biological samples. The digested samples were analyzed for some HMs (As, Cu, Zn, Pb, and Mo for the soil samples and Cu, Zn, and Pb for the biological samples) using an atomic absorption spectrometer (AAS) according to the guidelines of the Federal State Institution  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Sampling Site Code** & **N/Lat** & **E/Long** & **Sampling Site Location** \\ \hline S-50 & 41\({}^{\circ}\)06\({}^{\prime}\)13.7\({}^{\prime\prime}\) & 44\({}^{\circ}\)39\({}^{\prime}\)35.4\({}^{\prime\prime}\) & Investigation site located about 50 m north of the Alaverdi \\  & & & copper smelter \\ \hline S-100 & 41\({}^{\circ}\)06\({}^{\prime}\)16.5\({}^{\prime\prime}\) & 44\({}^{\circ}\)39\({}^{\prime}\)36.6\({}^{\prime\prime}\) & Investigation site located about 100 m north of the Alaverdi \\  & & & copper smelter \\ \hline S-200 & 41\({}^{\circ}\)06\({}^{\prime}\)18.4\({}^{\prime\prime}\) & 44\({}^{\circ}\)39\({}^{\prime}\)38.2\({}^{\prime\prime}\) & Investigation site located about 200 m north of the Alaverdi \\  & & & copper smelter \\ \hline S-300 & 41\({}^{\circ}\)06\({}^{\prime}\)21.2\({}^{\prime\prime}\) & 44\({}^{\circ}\)39\({}^{\prime}\)41.0\({}^{\prime\prime}\) & Investigation site located about 300 m north of the Alaverdi \\  & & & copper smelter \\ \hline S-700 & 41\({}^{\circ}\)06\({}^{\prime}\)32.8\({}^{\prime\prime}\) & 44\({}^{\circ}\)39\({}^{\prime}\)47.2\({}^{\prime\prime}\) & Investigation site located about 700 m north of the Alaverdi \\  & & & copper smelter \\ \hline S-800 & 41\({}^{\circ}\)05\({}^{\prime}\)57.7\({}^{\prime\prime}\) & 44\({}^{\circ}\)39\({}^{\prime}\)07.9\({}^{\prime\prime}\) & Investigation site located about 800 m southwest of the Alaverdi copper smelter \\  & & & Alaverdi copper smelter \\ \hline S-R & 40\({}^{\circ}\)52\({}^{\prime}\)53.2\({}^{\prime\prime}\) & 44\({}^{\circ}\)34\({}^{\prime}\)54.6\({}^{\prime\prime}\) & Reference site located about 25 km southwest of the Alaverdi copper smelter \\ \hline \hline \end{tabular}
\end{table}
Table 1: Coordinates of sampling sites located at different distances from the Alaverdi copper smelter.  
Figure 1: Location of the study area in Armenia and investigation sites as coded in Table 1.  
Federal Center for Analysis and Assessment of Technogenic Impact (FSI FCAATI-2011) [14]. All chemicals used were of certified analytical grade. Deionized water was used for the preparation of calibration standards and in the analyses. All glassware used was pre-washed with 10% HNO\({}_{3}\), followed by rinsing with distilled water prior to use. To ensure that the AAS remained calibrated during the experiments, certified reference materials were analyzed for soil and biological samples [14].