An investigation and examination of the levels and types of bacterial contamination on the surface of clean room operators' garments.

Abstract

The contamination of sterile pharmaceutical products is a serious event which has in the worst case scenario led to patient death. Operators are the primary source of clean room contamination, with the majority of their detritus being identified as skin squames and their related microorganisms. The ability of operator associated bacterial contamination to disseminate through specialist garments worn in the clean room environment is apparent in the literature. However, despite the fibres of such garments being identified as a suitable substrate for bacteria to adhere to and grow upon, the bacterial bioburden of the surface of clean room operators garments is an area which severely lacks in published research. Reported here is the recovery, enumeration and comparison of the levels of bacteria on the surface of reusable antistatic carbon filament polyester clean room garments, using the direct agar contact method, following their laundering with and without terminal gamma sterilisation, immediately following their donning with operators dressing wearing either no gloves, non “ sterile gloves or sterile clean room gloves, and following their wear within the clean room environment, with respect to gender. The aforementioned method, with its recovery efficiency shown to be unaffected by agar composition (NA or TSA), recovered bacteria from the surface of garments laundered with and without gamma sterilisation. Such terminal decontamination was shown to reduce the surface bacterial bioburden of the garments, especially at the chest and umbilicus regions, which were shown to harbour higher levels of bacteria than the other sites tested. The direct agar contact method, showing an increase in recovery efficiency following a 48 hour agar incubation period as opposed to a 24 hour period, also recovered bacteria from the surface of clean room garments donned by operators dressing wearing either no gloves, non “ sterile gloves and sterile clean room gloves. Bacteria were transferred onto the surface of these garments via the hand borne route, with the chest and oral cavity regions being found to harbour more bacteria than the other sites tested. Overall, glove type was shown to have no effect upon the resultant bacterial bioburden of the surface of the garments, suggesting expensive clean room gloves could be substituted for their cheaper non “ sterile equivalents or no gloves during the donning process without subsequently increasing the surface bacterial bioburden of the garment. The direct agar contact method also recovered bacteria from the surface of clean room garments worn by male and female operators, following their working period within a clean room environment. Gender was found to significantly affect the surface bacterial bioburden of the garments, with the surface of those garments worn by male operators being more contaminated than the surface of those worn by their female counterparts. In addition, the donning of a clean room hood was shown to reduce the levels of bacteria at the chest and posterior cervicis regions of suits worn by both genders. Overall, the direct agar contact method was identified as a successful tool to recover, enumerate and estimate the surface bacterial bioburden of reusable antistatic polyester carbon filament clean room garments. Finally, using 16S rRNA gene sequencing, found to be more reliable and accurate at identifying unknown isolates than traditional phenotypic first - stage tests, which were subsequently found to misidentify > 85 % of the isolates tested, a self - selected representative number of isolates recovered from the surface of garments during the laundering and gender comparison studies were predominantly identified as skin commensal species of Staphylococcus and Micrococcus, as well as environmental species of Bacillus. The knowledge contained within this thesis, with respect to clean room operators and their specialist garments, contributes towards improving contamination control standards within clean room facilities.