Herpes simplex virus type 1 (HSV-1) infection appears in the form of lesions containing infectious virus that if ruptured, virus can reach wastewater while washing or showering. We aimed to test infectivity of (HSV-1) in wastewater of different chemical composition and if it represents an additional risk factor for infection among wastewater treatment plant workers (WWTPW). Virus with known viral count was inoculated in three autoclaved wastewater (WW) samples collected from Balas, Berka and Arab abou-said WWTPs. Infectious viral particles were counted by Plaque count assay at different time intervals (day 1 to 21). Heavy metals concentrations were detected in WW samples by using atomic absorbtion spectrometer. Finally enzyme linked immunosorbent assay (ELISA) was used to detect anti-HSV-1-antibodies among 52 WWTPW, compared to control group (52 samples). Results showed sharp decrease in viral count starting from day 1 and reached 100 % by day 17. Berka wastewater sample gave the highest % reduction in all tested time points that might be due to the highest Fe and Zn concentrations compared with the other two stations,. Prevalence of anti-HSV-1-IgG among WWTPW (35%) was nearly equal to control group (42%) showing that working in contact with wastewater wasn't an additional risk factor for infection among WWTPW.