Effects of doxycycline on the endosymbiont Wolbachia in Dirofilaria immitis (Leidy, 1856)--naturally infected dogs.

Dirofilaria immitis carries intracellular endosymbiotic bacteria of the genus Wolbachia, known to be vital for the worms and sensitive to tetracycline antibiotics. With the purpose of studying the interaction between D. immitis and the endosymbiont Wolbachia sp., heartworm naturally infected microfilaremic or antigenemic dogs were treated with doxycycline (10mg/kg/day of the drug in three cycles of 21 days each, with 6-month intervals). Blood samples were collected on days 0, 7 and 21 of each treatment as well as on day 111 after the beginning of each cycle. A final sample was collected on day 723 from the beginning of the first treatment. The samples were examined for the presence and number of microfilariae and the presence of antigen as well as the presences of D. immitis and Wolbachia sp. DNA using PCR (polymerase chain reaction). With this approach, an evaluation of the effect of doxycycline on antigenemia and on the presence of Wolbachia sp. DNA in dogs with heartworm infection was possible. Doxycycline treatment did not alter the detection of adult parasite antigens with the exception of two animals, though the number of animals carrying Wolbachia sp. DNA decreased, despite the presence of the microfilariae. The effect of the antibiotic therapy on the worms may have interfered with the transmission of heartworm disease because the population of microfilariae and the number of microfilaremic dogs were reduced and the microfilariae positive samples that were found did not test positive for Wolbachia sp. in many cases. These findings suggest that in areas were doxycycline is extensively used D. immitis transmission may be impaired by the reduction on the number of microfilariae and on the endosymbiotic bacteria in the larvae turning them incapable of completing development once they infected a new host.