Application of Cytosense flow cytometer for the analysis of airborne bacteria collected by a high volume impingement sampler

Abstract

Characterization of airborne bacterial cells requires efficient collection, concentration, and analysis techniques, particularly to overcome the challenge of their dilute nature in outdoor environments. This study aims to establish a rapid and reliable approach for quantification of bacteria in air samples. To do this, a high volume impingement sampler was applied to collect airborne bacteria from a wastewater treatment plant (WWTP). The bacterial cell density was estimated by a Cytosense flow cytometer (Cytobouy) and compared to quantitative PCR (qPCR) data based on 16S rRNA genes. The average bacterial cell density measured by Cytosense ranged from 1.1 to 2.5 × 10⁴ cells m^-3 of air and that estimated by qPCR ranged from 0.08 to 3.8 × 10⁴ cells m^-3 of air. Regression analysis showed no systematic difference in bacterial cell densities between two methods applied when the cells were analyzed in vivo, and statistical tests confirmed that Cytosense counts of unfixed samples provided realistic values. Bacterial cell densities and the amount of DNA extracted from the sample were significantly correlated with relative humidity on a sampling day. The results showed that the present method was reliable to estimate bacteria densities from the outdoor environment, and the analysis given by Cytosense was faster and more sensitive than qPCR method. In addition, the Cytosense gave valuable information about cell characteristics at different sampling conditions.

Keywords: Airborne bacteria; Flow cytometry; Impingement sampler; Wastewater treatment plant; qPCR.