Operators are the primary source of cleanroom contamination, with the majority of their detritus identified as skin squames and their associated microorganisms. To reduce contamination, operatives are required to wear a specific arrangement of specialist garments. However, bacteria can evade this clothing and tarnish outer surfaces whilst operators work, with adverse implication for cleanroom environment and product sterility. Gender plays a significant role in bacterial dispersion, with male rates being in excess of female counterparts. Currently there is a lack of published literature evaluating the effect of gender on contamination of cleanroom garments. Such information would assist cleanroom facilities to more robustly assess and mitigate operator-associated contamination risks.

Background Contamination of sterile pharmaceutical products can have serious consequences, in worst case scenario resulting in patient death. Cleanroom operators are the primary source of microbial contamination, where the surface of their specialist sterile clothing garments is subject to such contamination during wear. In turn these garments become a transmission vector for microorganisms within the cleanroom environment. Insight into identification of predominant bacterial isolates from garment surfaces would help to establish their original source and probable contamination route. This should assist possible intervention strategies to mitigate against this contamination. Aim The research aimed to determine identity of representative bacterial isolates recovered from the surface of cleanroom operators’ garments following wear within a cleanroom. Methods Following isolation and purification of bacterial isolates, 16S rRNA gene sequencing was used to establish species identity for isolates recovered from the surface of male and female operators’ garments following wear within the cleanroom environment. Results Of the 47 isolates recovered from the surface of garments, 16S rRNA gene sequencing successfully identified 94 % to genus level and 77 % to species level. Most were confirmed as Gram - positive bacteria; predominantly species of Staphylococcus, Micrococcus and Bacillus. The isolates recovered from the surface of female operatives’ garments were more diverse than those retrieved from male counterparts. Conclusion Most isolates recovered from garments were found to be skin commensals, with nearly 70% attributed to the operators within the environment. The remainder were credited to contamination of garments with species of environmental origin. Whilst most bacteria identified present minimal threat to healthy individuals, certain of these are opportunistic pathogens, presenting a hazard for immunocompromised and/or those with underlying health conditions.