Background & Objectives: Leishmania infantum is the causative agent of visceral leishmaniasis (VL). Based on isoenzyme typing of a few isolates from patients and domestic dogs, this parasite was considered to predominate in the Kaleybar focus of VL in northwest Iran. There is no report of sandfly infections in this region so this study was aimed to investigate the infection of the sandflies in the field.

  Methods: Sandflies were sampled using sticky paper and CDC traps. Morphological identifications were carried out based on characters of the head and abdominal terminalia. DNA extracted from sandflies abdomens and thoraxes. ITS-rDNA gene of parasite was detected and identified as Leishmania after sequencing.

  Results: Out of 146 sandflies 9 were found to be infected with Leishmania. For first time, three Leishmania species (L. infantum, L. tropica and L. major) were identified in sandflies simultaneously in the region. Among the all sandflies only one Phlebotomus perfiliewi (vector of VL) was found to be infected with L. infantum. All Isolates were confirmed by sequencing of ITS-rDNA gene.

  Conclusion: However, Leishmania tropica and L. major were found more than L. infantum in sandflies in Kaleybar but it could not conclude that these two species of Leishmania are causative agents of VL. Because many criteria should be considered to incriminate an agent or vector of the disease.

  Background & objectives: The leishmaniases are considered among the major infectious diseases affecting public health in several regions. There are many chemical agents which are effective in treatment of visceral leishmaniasis. But, overall treatment of visceral leishmaniasis is often difficult. Thus, identification of new chemotherapeutic agents is important for treatment of disease. Since targeting of the ergosterol synthesis pathway of Leishmania may be useful therapeutically, the aim of this study was to investigate the effect of alone or in combination of amiodarone and ketoconazole on Leishmania infantum.

  Methods : To obtain logarithmic promastigotes of L. infantum, the parasites were cultured in BHI medium with FCS 10% together with antibiotics of penicillin and streptomycin and incubated at 24° C. Amastigote forms were obtained in BHI medium supplemented with 20% FCS at pH of 5.5 which incubated in 37° C. L.infantum susceptibility to amiodarone and ketoconazole was evaluated by proliferation of parasites in the absence or presence of these drugs with MTT assay. For evaluation of antiproliferative synergism against promastigotes and axenic amastigotes, fractional inhibitory concentrations (FIC) were calculated. An isobologram curve was constructed too.

  Results: Amiodarone produced a marked reduction in the viability of L.infantum promastigotes and axenic amastigotes. On the other hand ketoconazole induced a dose dependent effect on the parasites proliferation for promastigotes and axenic amastigotes. When the drugs were used in combination, the results indicated clear synergistic as shown by a concave isobologram and FIC value.

  Conclusion: The present study represents the evidence that the combination of amiodarone plus ketoconazole acts synergistically in controlling L. infantume in vitro. It is possible that amiodarone could be used in combination with ketoconazole to combat infection at low doses, thus reducing its side effects such as cardiotoxicity, thyroid dysfunction and pulmonary fibrosis.