Interspecies effects in a ceftazidime-treated mixed culture of Pseudomonas aeruginosa, Burkholderia cepacia and Staphylococcus aureus: analysis at the single-species level

In vitro studies commonly use single bacterial isolates for testing antibiotic susceptibilities. However, interspecies effects that may arise when mixed infections are treated with antibiotics can obviously not be investigated by this approach. In the study presented here, the effect of ceftazidime against a model microbial community consisting of Pseudomonas aeruginosa, Burkholderia cepacia and Staphylococcus aureus was studied in order to reveal effects that only may appear in a ceftazidime-treated mixed culture. Time-kill experiments were conducted with mixed and pure cultures in a defined medium containing 30 mg/L ceftazidime. Interspecies effects were revealed by comparing growth and kill dynamics from time-kill experiments with results from untreated mixed and pure cultures. For species-specific cell enumeration, a quantitative terminal restriction fragment length polymorphism was used. Ceftazidime was measured by HPLC. P. aeruginosa showed only a lytic phase in the ceftazidime-treated mixed culture, but not in the untreated mixed culture nor in the ceftazidime-treated pure culture. On the other hand, S. aureus did not lyse in the ceftazidime-treated mixed culture, while it did in the untreated mixed culture. This finding suggests that the efficacy of ceftazidime against P. aeruginosa was increased by an interspecies effect during co-cultivation with B. cepacia and S. aureus. The latter seemed to be negatively affected by interspecies effects in mixed culture without ceftazidime. The same effect was nullified when ceftazidime was applied to the mixed culture. Further studies are required to reveal the underlying mechanisms.