Respiratory infections are common causes of morbidity and mortality worldwide. We sought to assess the multiple antibiotic resistance (MAR) index, fitness and virulence potential in Pseudomonas aeruginosa from patients with lower respiratory tract infections. Isolates were assessed for antimicrobial susceptibility, in vitro competitive fitness, and pigment, elastase and rhamnolipid production. Oxidative stress tolerance was determined on both planktonic and biofilm cells, and virulence potential was tested in a plant model. Mean MAR index for isolates was 0.34 (range 0.17-0.50). Whilst isolates exhibited good biofilm formation in the presence of ciprofloxacin, there was no significant difference in biofilm production over the concentration range assessed. Several drug-resistant strains were out-competed by a susceptible strain even in the presence of antibiotic. H 2 O 2 exerted a greater oxidative stress than tert-butylhydroperoxide and, as expected, biofilms were more resistant than planktonic cells. Whilst most (81 %) isolates were pigmented there was no significant difference between pigmented and non-pigmented isolates when elastolytic activity was compared (P.0.05). More than half of the isolates produced the quorum sensing mediator rhamnolipid and infection of the plant model by bacteria occurred whether elastase or rhamnolipid was present or absent. These data suggest that non-pigmented strains of P. aeruginosa might pose an equally significant microbiological threat as pigmented strains even though pigment production appeared to be strongly associated with elastase expression. Whilst dual expression of elastase and rhamnolipid by these bacteria would cause severe tissue damage (as seen in the plant model), non-production of either does not prevent bacteria from causing serious infection.

Klebsiella pneumoniae is an important opportunistic pathogen with significant potential for virulence and multidrug resistance. Treatment failure often occurs because the pathogen may couple virulence and drug resistance with the stringent response. This study assessed the role of the spoT gene in environmental and nutritional stress tolerance, exopolysaccharide capsule production and biofilm formation. spoT mutants were constructed using the lambda red recombinase technique, and mutant and wild‐type (WT) strains were exposed to limiting concentrations of carbon (glucose), phosphate and aminoacid, and environmental stresses of ethanol, salt and heat. Cell viability, capsule production and cell length were assessed as well as the ability to grow biofilm under antibiotic pressure using gentamicin and ceftazidime. spoT mutants were more susceptible to stresses versus WT; the reverse was true for survival during biofilm susceptibility assay (p < 0.05), especially when carbon and phosphate were present. spoT mutants were elongated and lacked a capsule versus WT and non‐starved strains. The inability to produce capsule in mutants before and after starvation was likely a general effect of spoT mutation. These data suggest that the spoT‐mediated stringent response is important for K. pneumoniae in conditions of nutrient limitation, environmental stress and antimicrobial pressure.

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