Fifty samples of water were taken from 10 wells in the southern Iraqi city of Muthanna to investigate microbiological contamination in the wells, specifically Pseudomonas aeruginosa, coliform, and fecal coliform bacteria. Toxic ions (Cd⁺², B⁺³, pb⁺² and No^-3) were examined as part of the chemical contamination in the wells. Some phenotypic diagnostic tests, namely Gram stain, were performed to identify bacteria that were Stain-negative or positive. The presumptive, confirmatory tests demonstrated the presence of coliform bacteria and fecal coliform contamination in the well water. The bacteria P. aeruginosa were also found and separated using Cetrimed Agar media via biochemical and cultural analyses. The average most probable number (MPN) of bacteria in the well water was found to be varied. The findings demonstrated that all the hazardous ions under investigation were present at low amounts in comparison to the standard limits established by the World Health Organisation (WHO).