From 2012 to 2014, tick collections for tick and tick-borne pathogen surveillance were carried out in two areas of Nakai District, Khammouane Province, Laos: the Watershed Management and Protection Authority (WMPA) area and Phou Hin Poun National Protected Area (PHP NPA). Throughout Laos, ticks and tick-associated pathogens are poorly known. Fifteen thousand and seventy-three ticks representing larval (60.72%), nymphal (37.86%) and adult (1.42%) life stages were collected. Five genera comprising at least 11 species, including three suspected species that could not be readily determined, were identified from 215 adult specimens: Amblyomma testudinarium Koch (10; 4.65%), Dermacentor auratus Supino (17; 7.91%), D. steini (Schulze) (7; 3.26%), Haemaphysalis colasbelcouri (Santos Dias) (1; 0.47%), H. hystricis Supino (59; 27.44%), H. sp. near aborensis Warburton (91; 42.33%), H. sp. near darjeeling Hoogstraal and Dhanda (5; 2.33%), H. sp. near lagrangei Larrousse (3; 1.4%), H. spp. (16; 7.45%), Rhipicephalus haemaphysaloides (Supino) (5; 2.33%), and R. (Boophilus) microplus (Canestrini) (1; 0.47%). These collections, together with the literature to date, provide evidence for the occurrence of at least 22 ixodid tick species, representing six genera, in Laos. Here we present new records for at least four tick species from WMPA area, Nakai District, Khammouane Province, where tick-borne pathogens may circulate. These preliminary results should serve as a framework for further molecular investigations of putative tick vectors and their pathogens in Laos.