In the Old World, cutaneous leishmaniasis (CL) is zoonoses and natural vertebrate hosts of CL parasites are mammals. This study was carried out on natural infection rates of Leishmania parasites in reservoir hosts in one new focus of zoonotic cutaneous leishmaniasis (ZCL) and in suspected reservoir in an old focus of ACL in Iran. The sampling of rodents using Sherman traps was carried out and PCR technique was used for detection and identification of Leishmania species in Bahreman district, Kerman province, southeast of Iran. In addition, the smears were taken from suspicious lesions in stray dogs in the city of Kerman, the center of Kerman province. Simultaneously, pieces of lesion (1 9 191 cm) were taken for further histopathological examination. Overall, 25 rodents were collected and identified, including Meriones libycus and Rhombomys opimus. Amastigotes were observed in 33 % of the R. opimus by microscopic examination and indentified as Leishmania major by PCR technique. Four suspicious dogs out of 391 stray dogs showed no Leishmania species. To the best of our knowledge, this is the first isolation and identification of L. major from R. opimus in Kerman province, where ZCL has been present in recent years. Therefore, R. opimus is considered as the main animal reservoir host in Bahreman ZCL focus. In ACL focus such as the city of Kerman, dogs had no role in CL infection as reservoir host.

Benzoxonium chloride is an anti-infective agent that is used as anti-septic drugs for disinfection of the mucus membrane, skin surface and anti-bacterial, and it is also found to be effective against cutaneous leishmaniasis. The present study aims to evaluate the leishmanicidal activity of benzoxonium chloride and niosomal forms against Leishmania tropica stages. Benzoxonium chloride niosomes were prepared by the thin film hydration method and evaluated for morphology, particle size and release study and encapsulation efficiency. This study measured the cytotoxicity, leishmanicidal activity against promastigote and intra macrophage amastigote, apoptosis, and mRNA transcripts by quantitative real time PCR (qPCR) of free solution and niosomal-encapsulated benzoxonium chloride. Span/Tween 60 niosomal formulation of benzoxonium chloride showed superior physical stability and high encapsulation efficiency (96%) than the other forms. Release from the formulations showed that the Span/Tween 60 containing drug had a milder gradient so that 10% of the drug was not released after 4 h. The benzoxonium chloride and niosomal forms inhibited the in vitro growth of promastigote and amastigote forms of L. tropica after 48 h of incubation and represented IC 50 values of 90.7 ± 2.7 and 25.4 ± 0.6 μg/ mL, respectively. The rate of apoptosis in niosomal formulations was approximately equal to the positive control (meglumine antimoniate) at the same concentration. Also, an increase in the concentration of this drug reduced the expression of IL-10, but increased the expression of IL-12. The niosomal formulations provided improved anti-leishmanial activities of benzoxonium chloride and played an immunomodulatory role as the mode of action in the treatment of anthroponotic CL.

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