Abstract

BACKGROUND--Pseudomonas aeruginosa has been located in the endobronchiolar spaces of patients with cystic fibrosis where nutrients may be limited. In these sites it is thought that adaptation of the pathogen might occur and growth factors, present in relative excess, may thus promote survival of the organism. Auxotrophy of pulmonary isolates of P aeruginosa has previously been shown to be a feature of cystic fibrosis and chronic lung sepsis; auxotrophic isolates have additional nutritional requirements to the prototrophic "wild types" of the species. A study was therefore carried out to determine whether the proportion of auxotrophs differs between stable and acutely ill patients, or correlates with the extent of underlying disease. METHODS--Sputum samples were cultured for P aeruginosa and tested for auxotrophy by spreading serial dilutions of homogenised sputum on to a minimal medium which supports only prototrophs, and a complete medium which supports both nutritional types. The proportion of auxotrophs to prototrophs was determined and growth factors of confirmed auxotrophs were identified. RESULTS--Thirty two (86%) of 37 adults with cystic fibrosis infected with P aeruginosa harboured auxotrophs; methionine dependent mutants were isolated from seven of 16 patients tested (44%). More than 50% of the total number of colonies were auxotrophic in 19 of 26 samples (73%) from patients with acute exacerbations and in only six of 15 samples (40%) from clinically stable patients. In four patients from whom samples in both the acute and stable states were available, the proportion of auxotrophs fell in the sample taken when stable. Auxotrophs predominated in all samples from 11 of those patients with very severe underlying lung disease, in contrast to 13 of 30 samples from patients with less severe disease. There was no association between the percentage of auxotrophs and the presence of other respiratory pathogens. CONCLUSIONS--The majority of adults with cystic fibrosis infected with P aeruginosa harbour auxotrophs in the sputum. A significant proportion of acutely ill patients and those with severe underlying disease have a preponderance of auxotrophs in the sputum compared with stable patients and those with less severe disease.