Lippia javanica is widely distributed throughout South Africa where it is used extensively in traditional herbal preparations. An infusion of the leaves is commonly used as a decongestant for colds and coughs. A preliminary study indicated that the essential oil chemistry varies dramatically both within and between natural plant populations. As the antimicrobial activity may be directly related to the specific composition of the oil, the activity may also fluctuate. The aerial parts of Lippia javanica were collected from various localities in southern Africa to study the essential oil composition and the antimicrobial activity thereof. The hydrodistilled essential oils were analysed by GC/MS and a cluster analysis was performed on the essential oil dataset. From 16 samples (representing five natural populations), 5 chemotypes were identified; a myrcenone rich-type (36-62%), a carvone rich-type (61-73%), a piperitenone rich-type (32-48%), an ipsenone rich-type (42-61%) and a linalool rich-type (>65%). The myrcenone and linalool chemotypes have been mentioned in the literature but the carvone, ipsenone and piperitenone chemotypes have not previously been reported for Lippia javanica. Time kill studies were performed on three microbial respiratory isolates to document the scientific rationale of using Lippia to treat respiratory complaints in traditional herbal medicine. Klebsiella pneumoniae, Cryptococcus neoformans and Bacillus cereus showed reduction in microbial populations with the strongest bacteriostatic effect observed for Klebsiella pneumoniae.