P54: Genetic assessment of microbial diversity in four geographically separated sterile manufacturing facilities in the United States

Monday, August 12, 2013
Pavilion (Sheraton San Diego)
Bindhu Verghese and Christine Farrance, Accugenix services, Charles River Laboratories, Newark, DE
The current good manufacturing practices require that the quality of finished pharmaceutical products be assured through controlled processes. Environmental monitoring (EM) is, therefore, an important part of the production of aseptic products. Utilizing 16S rDNA and ITS2 gene sequence data, the present study evaluated the bacterial and fungal diversity in EM samples from four geographically separated sterile manufacturing facilities in the United States (Northeast: Plant-A, Southwest: Plant-B, Southeast: Plant-C, Northwest: Plant-D). Over a period of two years, a total of 15410 (A), 6853 (B), 867 (C), and 2155 (D) samples were analyzed from the respective plants. Our analyses have revealed some degree of geographic specificity in the abundance of microbial flora. Plant-A is dominated by Gram-positive bacterial genera (Staphylococcus (29%), Micrococcus (13%), Bacillus (11%)), plant-C by Bacillus (14%), and plant-D by Staphylococcus (23%) and Bacillus (12%). Plant-B, on the other hand, was dominated by Gram-negative bacterial genera (Pseudomonas (20%), Serratia (17%)). Consistently, fungal samples from each plant also showed unique taxonomic profiles. Penicillium, Aspergillus, Cladosporium, Candida and Aureobasidium were the dominant fungal genera with varying species compositions across the plants. Bacterial load was higher during summer. During spring 2012, plant-B showed a clear shift in bacterial diversity; however, the endemic species recolonized the facility by summer 2012. Change in the cleaning regimen may be the likely explanation for the shift. This study highlights the importance of EM in trending and tracking and the awareness that each facility has its own microbial niche that contributes to product and process contamination.