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Banknotes - the most adored, cherished, and highly respected item in human life - are one of the most vastly circulating and traded items between hands of citizens. As a result of this fast and frequency in circulation of banknotes between handlers, who are of different personal hygiene, medical status, saving of banknotes habits, geographical habitats and business, banknotes are more borne to contamination with different types of pollutants. A number of studies have found multidrug-resistant and virulent strains on banknotes. These have the potential to cause serious infections that are hard to treat, to disseminate in health care and community settings, and to spread antimicrobial resistant determinants to other bacteria. This study was designed to assess the banknotes handlers hygiene and microbial (Bacteria, Yeast and Moulds) contamination of Sudanese banknotes.