In-use testing of the disinfectants; Hibitane (5% w/v Chlorhexidinegluconate), Hibiscrub (4% w/v Chlorhexidinegluconate), Savlon (3% w/v Chlorhexidine/Cetrimide), hydrogen peroxide (6% w/v hydrogen peroxide with stabilizer) and a common household bleach Jik (3.5% w/v sodium hypochlorite), was carried out over a two-month period at a university teaching hospital in Nigeria. Contamination levels were high with 82 (63.1%) of the 130 in-use disinfectants contaminated. However, a few of the stock solutions remained sterile. One hundred and thirty-four isolates were obtained of which 120 (91%) were gram-negative with Pseudomonas species being the commonest, constituting 67.2% of all the isolates. Gram-positive organisms made up the remaining 12 (9.0%) isolates. All the Pseudomonas spp. were resistant to gentamicin, ceftazidime, nalidixic acid and perfloxacin. Contributory factors for the high contamination levels were dilution of disinfectants with tap water, inadequate care of stock solution bottles and long storage of the diluted disinfectants in the wards.