Six solid substrates were compared for their support of growth of ten isolates of Borrelia burgdorferi, the causative agent of Lyme borreliosis. The following substrates with or without rabbit serum and bovine serum albumin were tested: BSK agar, BHIAM agar, TAROM agar, MEM agar, MKP agar and PMR agar. After incubation in a candle jar and a GasPak for two to four weeks, Borrelia colonies were counted and characterized. Colony morphology was related more to the growth substrate than to the characteristics of the various Borrelia burgdorferi isolates. Culture on PMR agar resulted in the highest recovery rate and the best colony formation, with a size variation of 0.3-3.0 mm. Culture of Borrelia burgdorferi on solid media may facilitate the diagnosis of Lyme borreliosis by enabling easier biological and genetical analysis of isolates and more accurate differentiation of strains.