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Since its high toxicity and mobility, hexavalent chromium is considered to be a high priority pollutant. This study was performed to carry out a remediation test along with toxicity assessment in a water sample collected from the saturated zone of a historically Cr (VI)-contaminated site known as a Puliyanthangal lake (Tamilnadu, India) using nanoscale zero-valent iron (nZVI). The water samples were examined before and after the nZVI application by means of microbial cultivation tests, phospholipid fatty acid analysis (PLFA) and toxicological tests. The present experimental results revealed that Cr (VI) is considerably adsorbed on nZVI nanoparticles and it could be a cost-effective method for the in situ remediations of Cr (VI). In addition, standard plate count assay showed the dose-dependent decrease in the bacterial cell viability in lake water sample after the addition of nZVI. Keywords: Iron nanoparticles, Green synthesis, Heavy metal degradation, E. coli, Groundwater remediation, Toxicity