The study involved 120 samples collected from wards of the Al-Diwaniya Teaching Hospital, Iraq during the period from 5 April to 5 June 2016. The results showed that the percentage of the fungal contamination in the wards was 77/120 (64.1%), highest of the fungal contamination was recorded in the floors and walls and in the air 18 (90%), followed by patients (Nose, Ear and Hands) 14 (70%) and lowest contamination was in the instruments 3 (15%) when compared. A total of 125 fungal isolates were identified and the isolates belong to 9 genera comprising of Aspergillus spp with 52 isolates (mainly A.  niger= 24, A. flavus= 19, A. fumigates= 5 and A. ochraceus= 4). Penicillium spp with 17 isolates, Alternaria alternata with 14 isolates, Candida albicans with 11 isolates, Fusarium  spp 9 isolates, Mucor spp 7 isolates, Rhizopus stolonifer 7 isolates, Cryptococcus spp 5 isolates and Rhodotorula spp 3 isolates. The highest occurrence and frequency percentages of fungi recorded for A.  niger reaching (18.947%) and (19.2%) respectively, followed by A. flavus reaching (16.842%) and (15.2%) respectively with significant differences when compared with other fungi isolated in this study, while the lowest occurrence and frequency percentages recorded for Rhodotorula spp reaching (2.105%) and (2.4%) respectively. The antiseptics (Dettol, Hibitane and Formalin) have high inhibitory activity against the growth of all fungi studied when used at original concentration which was 10% compared with other concentrations 7.5, 5 and 2.5%. Formalin showed highly significant effect in inhibitory activity for the growth of fungi from Dettol and Hibitane, the percentage of inhibition when using Formalin at concentration 10% ranged between (85.2-95.4%) while for Hibitane between (68.1-79.7%) and for Dettol between (40.6-65.8%) at the same concentration.