The possibility of vertical transmission of Borrelia burgdorferi sensu lato in Ixodes persulcatus Schulze, 1930 ticks was studied in the progeny of 20 females collected from the vegetation in an active focus of ixodid tick-borne borrelioses (ITBB) located in the Perm oblast, Russia, where Borrelia garinii and B. afzelii are circulating. The presence of Borrelia DNA was detected by the PCR method after feeding and egg laying in 16 engorged females (80.0%), as well as in 36.5 ± 7.2% samples containing 20 eggs each and in 21.4 ± 4.2% samples containing 10 eggs each. The respective rates of individual egg infection were 0.4-8.0% and 0.5-23.0%. PCR analysis of 370 eggs (one egg per sample) and 781 unfed larvae hatched from the same egg masses (1, 10, 20, 40, and 50 larvae per sample) failed to reveal the presence of Borrelia DNA. Negative results were also obtained in experiments on inoculating the BSK II medium with the egg and larval materials. Microscopic analysis of 1,683 smear preparations of eggs and 1,416 preparations of unfed daughter larvae revealed spirochete-like cells in 7 (0.4 ± 0.3%) and 13 (0.9 ± 0.5%) preparations, respectively; typical Borrelia cells were found in seven preparations of larvae (0.5 ± 0.4%). Only 1 out of 16 infected females transmitted Borrelia vertically, through the eggs to the larval progeny. The infection rate in this progeny was about 7%, and the prevalence of Borrelia in individual larvae was 0.4-0.8 cells per 100 microscopic fields. These data do not allow the conclusion that transovarial transmission of B. burgdorferi sensu lato in the I. persulcatus tick is an established fact. However, they show that, even if such transmission is possible, its probability is very low.