Visceral leishmaniasis is a systemic infectious disease that can cause to a severe, potentially life-threatening chronic condition in humans. Risk factors for infection in urban areas have been associated with poor living conditions, the presence of sand fly vectors and infected pets. This study aimed to describe sand fly and canine infection in the neighborhoods of human visceral leishmaniasis occurrence in the city of Cuiabá, Mato Grosso State, central-western Brazil, reported between January 2005 and December 2006. A total of 1,909 sand flies were collected. They were predominantly males and the most frequent species were Lutzomyia cruzi (81.25%), Lutzomyia whitmani (13.88%) and Lutzomyia longipalpis (2.62%). The sand fly density was not significantly correlated with the variation of environmental factors. The prevalence of canine visceral leishmaniasis in the neighborhoods studied was 26.82% and it was found that areas with high density of vectors coincided with areas of high prevalence of dogs and those with the highest rates of human cases. The study of vectors and other potential hosts are essential for a good understanding of visceral leishmaniasis and the related public health concerns, aiming at the prevention and control of leishmaniasis in the city of Cuiabá, Mato Grosso State.