Endotracheal tube biofilm inoculation of oral flora and subsequent colonization of opportunistic pathogens

Abstract Endotracheal (ET) tubes accumulate a biofilm during use, which can harbor potentially pathogenic microorganisms. The enrichment of pathogenic strains in the biofilm may lead to ventilator-associated pneumonia (VAP) with an increased morbidity rate in intensive care units. We used quantitative PCR (qPCR) and gene surveys targeting 16S rRNA genes to quantify and identify the bacterial community to detect fastidious/nonculturable organisms present among extubated ET tubes. We collected eight ET tubes with intubation periods between 12 h and 23 d from different patients in a surgical and a medical intensive care unit. Our qPCR data showed that ET tubes were colonized within 24 h. However, the variation between patients was too high to find a positive correlation between the bacterial load and intubation period. We obtained 1263 near full-length 16S rRNA gene sequences from the diverse bacterial communities. Over 70% of these sequences were associated with genera of typical oral flora, while only 6% were associated with gastrointestinal flora. The most common genus identified was Streptococcus (348/1263), followed by Prevotella (179/1263), and Neisseria (143/1263) with the highest relative concentrations for ET tubes with short intubation periods, indicating oral inoculation of the ET tubes. Our study also shows that even though potentially pathogenic bacteria existed in ET tube biofilms within 24 h of intubation, a longer intubation period increases the opportunity for these organisms to proliferate. In the ET tube that was in place for 23 d, 95% of the sequences belonged to Pseudomonas aeruginosa , which is a bacterial pathogen that is known to out compete commensal bacteria in biofilms, especially during periods of antibiotic treatment. Harboring such pathogens in ET biofilms may increase the chance of VAP, and should be aggressively monitored and prevented.