Massive sequencing of the V3–V4 hypervariable region of bronchoalveolar lavage from patients with COVID-19 and VAP reveals the collapse of the pulmonary microbiota


 Background. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic is a predisposing factor for the development of healthcare-associated infections, of which ventilator-associated pneumonia (VAP) is one.
 
 Hypothesis. VAP is caused by ESKAPE bacteria and other pathogens not detected by microbiological culture.
 
 Aim. To elucidate the bacterial pathogens of severe coronavirus disease 2019 (COVID-19) and VAP patients by massive sequencing and to predict their degree of relationship with the age and sex of the patients.
 
 Methods. Analysis of ribosomal libraries of the V3–V4 hypervariable region obtained by Illumina sequencing of bronchoalveolar lavages from COVID-19 and VAP (first wave) patients from Hospital Juárez de México.
 
 Results. 
 
 Acinetobacter
 
 and 
 
 Pseudomonas
 
 were the main bacterial genera in the bronchoalveolar lavages (BALs) analysed. Other members of the ESKAPE group, such as 
 
 Enterococcus
 
 and 
 
 Klebsiella
 
 , were also identified. Taxonomic composition per patient showed that non-ESKAPE genera were present with significant relative abundances, such as 
 
 Prevotella
 
 , Stenotrophomas, 
 
 Enterococcus
 
 , 
 
 Mycoplasma
 
 , 
 
 Serratia
 
 and 
 
 Corynebacterium
 
 . Kruskal–Wallis analysis proved that VAP acquisition is an adverse event that is not influenced by the sex and age of COVID-19 patients.
 
 Discussion. Metagenomic findings in COVID-19/VAP patients highlight the importance of implementing comprehensive microbiological diagnostics by including alternative tools for the detection of the causal agents of healthcare-associated infections (HAIs).
 
 Conclusions. Timely identification of bacteria ‘not sought’ in diagnostic bacteriology laboratories will allow specific and targeted treatments. Implications for the restricted diagnosis of VAP causative agents in COVID-19 patients and the presence of pathogens not detected by classical microbiology are analysed and discussed.