Little is known regarding the internal dissemination of initial cutaneous lesions and tissue tropism of Leishmania (Viannia) braziliensis populations in naturally infected dogs. The aim of this study was to investigate genetic polymorphisms of L. (V.) braziliensis populations in different anatomic sites of naturally infected dogs by using polymerase chain reaction (PCR) and low-stringency single specific primer-PCR (LSSP-PCR) techniques. The amplified products were analyzed by LSSP-PCR to investigate the genetic variability of the parasite populations present in different anatomical sites. Twenty-three out of the 52 samples gave PCR-positive results. The existence of L. (V.) braziliensis strains that remained restricted to cutaneous lesions and others showing characteristics of dissemination to internal organs and healthy skin was observed. LSSP-PCR and numerical analyses revealed that parasite populations that do not disseminate were genetically similar and belonged to a separate phenetic cluster. In contrast, populations that showed spreading to internal organs displayed a more polymorphic genetic profile. Despite the heterogeneity, L. (V.) braziliensis populations with identical genetic profiles were observed in popliteal and cervical lymph nodes of the same animal. Our results indicate that infection in dogs can be manifested by dissemination and tissue tropism of genetically distinct populations of L. (V.) braziliensis. 1. Introduction Leishmaniasis is a protozoal disease caused by different species of the genus Leishmania, and it is transmitted by the bite of female phlebotomine insects of the genus Lutzomyia in the New World. Cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) are endemic in large areas of the tropics, subtropics, and the Mediterranean basin. Around 350 million people in 88 countries are at risk of contracting leishmaniasis, and this number is believed to be inaccurate due to the flaws in the case detection data used to estimate the disease prevalence in many endemic countries [1]. In the state of Rio de Janeiro, Leishmania (Viannia) braziliensis is the most prevalent species that is responsible for CL. Transmission mainly occurs in periurban areas, where the primitive rain forest vegetation is being replaced by disorganized human occupation. Adaptation of the vector Lutzomyia intermedia to the domiciliary and peridomiciliary environments, as well as the presence of infected humans, dogs, and horses, has been observed [2–4]. In the canine population, L. (V.) braziliensis infection is manifested by cutaneous lesions,
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