Multiwall Carbon Nanotubes Induce More Pronounced Transcriptomic Responses in Pseudomonas aeruginosa PG201 than Graphene, Exfoliated Boron Nitride, or Carbon Black

Carbonaceous and boron nitride (BN) nanomaterials have similar applications and hydrophobic properties suggesting common release pathways and exposure to bacteria. While high nanomaterial concentrations can be bactericidal or growth-inhibitory, little is known regarding bacterial transcriptional responses to non-growth-inhibitory nanomaterial concentrations. Here, using one strain of Pseudomonas aeruginosa—a clinically and environmentally important bacterial taxon—we analyzed the comparative transcriptomic response to carbonaceous or BN nanomaterials. We show that, at non-growth-inhibitory, equal mass concentrations (10 mg/L), multiwall carbon nanotubes (MWCNTs) induced differential regulation of 111 genes in P. aeruginosa, while graphene, BN, and carbon black caused differential regulation of 44, 26, and 25 genes, respectively. MWCNTs caused the upregulation of genes encoding general stress response (9 genes), sulfur metabolism (15), and transport of small molecules (7) and downregulation of genes encoding flagellar basal-body rod proteins and other virulence-related factors (6), nitrogen metabolism (7), and membrane proteins (12), including a two-component regulatory system CzcS/R. Because two-component systems are associated with antibiotic resistance, the antibiotic susceptibility of P. aeruginosa was tested following MWCNT exposure. In MWCNT-treated cultures, the minimal inhibitory concentrations (MICs) of meropenem and imipenem decreased from 0.06 to 0.03 μg/mL and from 0.25 to 0.125 μg/mL, respectively. Taken together, whole genome analysis indicated that, in the absence of growth inhibition, nanomaterials can alter bacterial physiology and metabolism. For MWCNTs, such alterations may include downregulation of antibiotic resistance pathways, suggesting that pre-exposure to MWCNTs could potentially render bacteria more susceptible to carbapenems which are often the last resort for the globally concerning, highly antibiotic resistant P. aeruginosa.