The Recovery and Transfer of Aerosolized Listeria Innocua

Abstract

Airborne pathogenic bacteria can present a significant public health risk. Pathogenic Listeria monocytogenes can colonize numerous surfaces as well, through direct and indirect cross contamination. The physical environment can also affect the transmission and viability of Listeria (distance from the source, temperature, humidity, air flow). The purpose of this work was to explore the ability of Listeria innocua (a surrogate for L. monocytogenes) to contaminate a surface after it has become aerosolized in a bioaerosol chamber and a walk-in cooler.

L. innocua was nebulized into a 154 L biosafety chamber (~5 log CFU in 1 mL) at two relative humidity (RH) levels (83% and 65%). Oxford Listeria agar plates, stainless steel coupons and polyethylene (HDPE) coupons in the chamber were exposed to the aerosolized bacteria for 5, 10, 20 or 40 minutes. Also, at these times, air samples (100 L) were collected on to gelatin filters which were transferred to Oxford agar plates. In the second part of the research, L. innocua was nebulized into an 11 m3 walk-in cooler where RH ranged from ~29-37%. Aerosolized bacteria were collected on to Oxford agar plates for 10 min intervals and with 50 or 100 L air samples.

Recovery of L. innocua from steel, plastic and agar was significantly higher at 83% RH (2.7 cells/cm2) compared to 65% RH (0.45 cells/cm2). Mean cell recovery from air samples (gelatin filters) was significantly higher (p<0.05) when collected 5 or 10 minutes after nebulization at 83% humidity (mean 2.2 CFU/L) compared to collection after 20 or 40 minutes or compared to all times under 65% humidity (mean 0.4 CFU/L). Recovery from HDPE coupons (1.21 CFU/cm2) was 2.5 X recovery from Oxford agar (0.49 CFU/cm2). In the walk-in cooler, total estimated mean recovery from Oxford media at 10 min after nebulizing was 0.48%, but only 0.04% for samples collected after 60 minutes. The recovery of L. innocua from air samples after 60 min was one-fourth of the number recovered 5 min after nebulizing. No significant difference in recovery was found between plates at different distances (2 – 2.5 m) from the nebulizer in the walk-in cooler. Understanding the survival of aerosolized Listeria and how it can colonize over time on a food contact surface will enhance our efforts to prevent transmission on a small and large scale. The food industry will be able to implement better safety measures to prevent contamination by Listeria species.