Mubbashir Hussain1, Shahzad Munir2, Sultan Ayaz3, Bahar Ullah Khattak4, Taj Ali Khan4, Niaz Muhammad4, Muhammad Anees4, Hazir Rahman4, Muhammad Qasim4, Muhammad Ameen Jamal5, Irfan Ahmed6, Kashif Rahim7, Humaira Mazhar4, Noha Watanay8, Mohamed Kasbari9. 1. Vector Borne Diseases Laboratory, Department of Microbiology, Kohat University of Science and Technology, Kohat, KP 26000, Pakistan. Electronic address: mubashirbangash@gmail.com. 2. Department of Plant Pathology, Faculty of Plant Protection, Yunnan Agricultural University, Kunming 650201, China. Electronic address: shazid_10@yahoo.com. 3. College of Animal Husbandry and Veterinary Sciences, Abdul Wali Khan University Mardan, Pakistan. 4. Vector Borne Diseases Laboratory, Department of Microbiology, Kohat University of Science and Technology, Kohat, KP 26000, Pakistan. 5. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China. 6. Yunnan Provincial Key Laboratory of Animal Nutrition and Feed, Yunnan Agricultural University, Kunming 650201, China. 7. Beijing Key Laboratory of Genetic Engineering Drug and Biotechnology, Institute of Biochemistry and Biotechnology, College of Life Sciences, Beijing Normal University, Beijing 100875, China. 8. US Naval Medical Research Unit-3, (NAMRU 3), Cairo, Egypt. 9. French Agency for Health and Safety Animal Health Laboratory, Maisons-Alfort, France.
Abstract
OBJECTIVE: To report presence of Leishmania major in Khyber Pakhtunkhwa of Pakistan, where cutaneous leishmaniasis (CL) is endemic and was thought to be caused by Leishmania tropica only. METHODS: Biopsy samples from 432 CL suspected patients were collected from 3 southern districts of Khyber Pakhtunkhwa during years 2011-2016. Microscopy on Giemsa stained slides were done followed by amplification of the ribosomal internal transcribed spacer 1 gene. RESULTS: Leishmania amastigotes were detected by microscopy in 308 of 432 samples (71.3%) while 374 out of 432 samples (86.6%) were positive by ribosomal internal transcribed spacer 1 PCR. Subsequent restriction fragment length polymorphism confirmed L. tropica in 351 and L. major in 6 biopsy samples. CONCLUSIONS: This study is the first molecular characterization of Leishmania species in southern Khyber Pakhtunkhwa. It confirmed the previous assumptions that anthroponotic CL is the major CL form present in Khyber Pakhtunkhwa province. Furthermore, this is the first report of L. major from a classical anthroponotic CL endemic focus identified in rural areas of Kohat district in southern Khyber Pakhtunkhwa.
OBJECTIVE: To report presence of Leishmania major in Khyber Pakhtunkhwa of Pakistan, where cutaneous leishmaniasis (CL) is endemic and was thought to be caused by Leishmania tropica only. METHODS: Biopsy samples from 432 CL suspected patients were collected from 3 southern districts of Khyber Pakhtunkhwa during years 2011-2016. Microscopy on Giemsa stained slides were done followed by amplification of the ribosomal internal transcribed spacer 1 gene. RESULTS:Leishmania amastigotes were detected by microscopy in 308 of 432 samples (71.3%) while 374 out of 432 samples (86.6%) were positive by ribosomal internal transcribed spacer 1 PCR. Subsequent restriction fragment length polymorphism confirmed L. tropica in 351 and L. major in 6 biopsy samples. CONCLUSIONS: This study is the first molecular characterization of Leishmania species in southern Khyber Pakhtunkhwa. It confirmed the previous assumptions that anthroponotic CL is the major CL form present in Khyber Pakhtunkhwa province. Furthermore, this is the first report of L. major from a classical anthroponotic CL endemic focus identified in rural areas of Kohat district in southern Khyber Pakhtunkhwa.