Comprehensive Oropharyngeal Microbiota Screening

Material from tonsillar crypts were taken, placed, and transported in AMIES transport medium at room temperature within 24 h and cultivated on two Columbia blood agar plates with and without optochin disk, Brucella blood agar, Chocolate agar with oleandomycin disc, Mannitol salt agar, MacConkey agar, and Sabouraud dextrose agar plates. Columbia blood agar, Mannitol salt agar, MacConkey agar, and Sabouraud dextrose agar plates were incubated at 36 ± 1 °C for 24–48 h aerobically. A Brucella blood agar plate was incubated in a BD GasPak™EZ pouch system at 36 ± 1 °C for up to five days. A Columbia blood agar plate with an optochin disc incubated in a CO₂ incubator at 36 ± 1 °C for 24–48 h was used for the cultivation of Streptococcus pneumoniae. A Chocolate agar plate with an oleandomycin disc incubated in a CO₂ incubator at 36 ± 1 °C for 24–48 h was used for the cultivation of Haemophylus spp. We took note of the common oropharyngeal microbiota as described by the European Society of Clinical Microbiology and Infectious Diseases [16 ]. Microorganisms that are not part of the common oropharyngeal microbiota were considered as potential pathogens. The identification of the considered pathogens was performed using a Microflex LT (Bruker Daltonics flex Analysis version 3.4, Bruker Daltonics GmbH & Co. KG, Bremen, Germany) matrix-assisted laser desorption ionization–time-of-flight mass spectrometer (MALDI–TOF MS) system.