Transmission of canine leishmaniosis (CanL) is described in three consecutive generations of female Boxers living in a non-endemic environment in the Czech Republic. Infection of the first generation female likely occurred during a breeding visit to Italy and the dog died with typical clinical signs of the disease but without definitive laboratory diagnosis. The second and third generation offsprings never left the Czech Republic, suffered from clinical CanL confirmed by polymerase chain reaction and serology, and were apparently infected by transplacental transmission. Persistence of CanL in the Czech Republic over 7 years with a suspected origin in an endemic region and progression of infection through subsequent generations in a non-endemic country exemplifies that this disease may establish itself also in areas where no obvious vectors are present. (C) 2017 Published by Elsevier B.V.