The essential oils from two native species from Guatemala were studied for their chemical composition and the dichloromethane and methanol extracts for their biological activity. A GC-MS analysis of the essential oil from Piper jacquemontianum Kunth, Piperaceae, showed 34 constituents, consisting mainly of linalool (69.4%), while Piper variabile C. DC. essential oil had 36 constituents, camphor (28.4%), camphene (16.6%) and limonene (13.9%) being the major components. Dichloromethane extracts of both species were cytotoxic against MCF-7, H-460 and SF-268 cell lines (<7 μg/mL). Dichloromethane extract of P. jacquemontianum was slightly active against bacteria (0.5 mg/mL), was active against promastigotes of Leishmania (20.4-61.0 μg/mL), and epimastigotes of Trypanosoma cruzi (51.9 μg/mL). The methanol extract of P. variabile showed antimalarial activity against Plasmodium falciparum F32 (4.5 μg/mL), and the dichloromethane extract against Leishmania (55.8-76.3 μg/mL) and T. cruzi (45.8 μg/mL). None of the extracts from the two species was active against Aedes aegypti larvae and Artemia salina nauplii.