Essential genes forHaemophilus parainfluenzaesurvival and biofilm growth

Abstract

AbstractHaemophilus parainfluenzae(Hp) is a Gram-negative, pleomorphic rod, highly prevalent and abundant as a commensal in the human oral cavity, and an infrequent extraoral opportunistic pathogen.Hpoccupies multiple niches in the oral cavity, including the tongue dorsum, keratinized gingiva, and the supragingival plaque biofilm. As a member of the HACEK group,Hpis also known to cause infective endocarditis. Additionally, case reports have identifiedHpas the causative agent of meningitis, septic arthritis, chronic osteomyelitis, septicemia, and a variety of other infectious diseases. Little is known about howHpinteracts with its neighbors in the healthy biofilm nor about its mechanisms of pathogenesis as an extraoral opportunistic pathogen. To address these unknowns, we identified the essential genomes of twoHpstrains and the conditionally essential genes for their growth inin vitrobiofilms aerobically and anaerobically. Using transposon insertion sequencing (TnSeq) with a highly saturatedmarinertransposon library in two strains, the ATCC33392 type-strain (Hp392) and a commensal oral isolate EL1 (HpEL1), we show that the essential genome ofHp392 andHpEL1 is composed of 395 and 384 genes, respectively. The core essential genome, consisting of 341 essential genes conserved between both strains, was composed of genes associated with genetic information processing, carbohydrate, protein, and energy metabolism. We also identified conditionally essential genes for aerobic and anaerobic biofilm growth, which were associated with carbohydrate and energy metabolism in both strains ofHp. Additionally, RNAseq analysis determined that most genes upregulated during anaerobic growth are not essential forHp392 anaerobic biofilm survival. The completion of this library and analysis under these conditions gives us a foundational insight into the basic biology ofH. parainfluenzaein differing oxygen conditions, similar to itsin vivooral habitat. Further, the creation of this library presents a valuable tool for further investigation into conditionally essential genes for an organism that lives in close contact with many microbial species in the human oral habitat.