ABSTRACT
Microbiological monitoring of pharmaceutical industry aseptic areas is an important procedure to assess the efficiencyof contamination control measures in these areas. Once the permitted microbiological level has been exceeded, themicrobial contaminant should be identified, and the actions to eliminate this agent should be adopted. The objective ofthis study was to identify the filamentous fungi and yeasts isolated from the pharmaceutical industry environment by thematrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) proteomic approachas well as to analyze fungal universal region polymerase chain reaction- restriction fragment length polymorphism(PCR-RFLP) capacity in discriminating inter- and intraspecies differences in these isolates. The MALDI-TOF methodwas able to identify 100% of the samples to the genus level; however, only 68.42% of the samples were identifiedto the species level. Candida guilliermondii, Penicillium spp., Rhodosporidium toruloides, and Aspergillus specieswere the most prevalent microorganisms among the samples. The PCR-RFLP analysis of the fungal universal regionrevealed a heterogeneous profile in the analyzed samples. The isolates of the same species, collected from differentsampling points, presented the same restriction profile. MALDI-TOF and PCR-RFLP methods of the fungal universalregion were able to, respectively, identify and characterize the genetic diversity and differentiate interspecifically theyeast and fungi samples analyzed in this study. The genetic profiles established by the PCR-RFLP methods of thisstudy can be used to create a database for in-house identification of yeasts and fungi in the laboratory, where this studywas carried out.