Identification and phylogenetic characterization of Alkalihalophilus strain isolated in a pharmaceutical industry using polyphasic techniques

Introduction:The production of medicines and vaccines must comply with strict Good Manufacturing Practice (GMP) standards to ensure the safety of the manufactured products.Therefore, the identification of microorganisms is essential for environmental monitoring and quality control of these processes.Proper identification of microorganisms in production areas is critical to ensure safety standards are maintained and that the final product is free from contamination. Objectives:The aim of this study was to identify, at species level, the strain B1521/24, isolated from bioburden analysis of a viral vaccine production area at an immunobiological facility. Methodology:The strain was initially analyzed by MALDI Biotyper (Bruker), but it could not be identified.The 16S rRNA full gene of the B1521/24 strain was then sequenced, and the data obtained was analyzed using the EzBioCloud platform, considering the threshold 98.7% for classification at species level.For species delimitation, a maximum likelihood phylogenetic tree was constructed using MAFFT online, MEGA XI, jModelTest, IQ-TREE, and FigTree software. Results:The analysis revealed two possible identities for strain B1521/24: Alkalihalophilus pseudofirmus (99.66%) and Alkalihalophilus lindianensis (98.71%).After phylogenetic analysis, the B1521/24 strain was identified as Alkalihalophilus pseudofirmus, a Gram-positive, rod-shaped, motile, spore-forming, mesophilic species that is obligately aerobic and commonly found at lake shores. Conclusion:The confirmation of the strain's identification as Alkalihalophilus pseudofirmus allowed its inclusion in the MALDI Biotyper database, which will facilitate its identification the next time it is isolated.Furthermore, expanding the MALDI Biotyper database with in-house strains helps improve the laboratory's identification capacity and consequently process deviation investigations and risk analysis, highlighting the significant technological and scientific importance of this study for the pharmaceutical industry.