The European hedgehog, Erinaceus europaeus Linnaeus, 1758, is a common host of Ixodes ricinus L. and I. hexago-nus Leach, vectors of the Lyme disease spirochaete, Borrelia burgdorferi sensu iato. To investigate whether hedgehogs are reservoirs for li- burgdorferi, hedgehogs were captured in a suburban area suitable for both tick species and in an urban area where /. ricinus is absent. The infection status of the hedgehogs was determined by xenodiagnosis using I. ricinus and I. hexagonus larvae. /. hexagonus and/or I. ricinus were found on all hedgehogs (n = 8) from the suburban area. In contrast, only I. hexagonus was infesting animals (n = 5) from the urban area. A total of 12/13 hedgehogs harboured B. burgdorferi infected ticks. Xeno-diagnostic I. ricinus and I. hexagonus larvae that fed on hedgehogs became infected. The results clearly show that European hedgehogs are reservoir hosts of the Lyme disease spirochetes. DNA of B. burgdorferi sensu stricto, В. garinii and В. afzelii was detected in culture from ear biopsy and needle aspiration material and characterized by using a genospecies-specific PCR assay. One hedgehog presented a mixed infection of the skin with fi. burgdorferi sensu stricto and fi. garinii. This study also identifies an enzootic transmission cycle in an urban area involving E europaeus and /. hexagonus. The close association of /. hexagonus with the burrows of its hosts mean that the risks of contact between /. hexagonus and humans may be low.