Ahmet Özbilgin1, Gülnaz Çulha2, Soner Uzun3, Mehmet Harman4, Suhan Günaştı Topal5, Fulya Okudan6, Fadile Zeyrek7, Cumhur Gündüz8, İpek Östan9, Mehmet Karakuş10, Seray Töz10, Özgür Kurt11, Işın Akyar11, Ayşegül Erat3, Dilek Güngör5, Çağla Kayabaşı8, İbrahim Çavuş1, Patrick Bastien12, Francine Pratlong12, Tanıl Kocagöz11, Yusuf Özbel10. 1. Department of Parasitology, Faculty of Medicine, Celal Bayar University, Manisa, Turkey. 2. Department of Parasitology, Faculty of Medicine, Mustafa Kemal University, Hatay, Turkey. 3. Department of Dermatology, Faculty of Medicine, Akdeniz University, Antalya, Turkey. 4. Department of Dermatology, Faculty of Medicine, Dicle University, Diyarbakɩr, Turkey. 5. Department of Dermatology, Faculty of Medicine, Çukurova University, Adana, Turkey. 6. Clinic of Dermatology, Atatürk State Hospital, Antalya, Turkey. 7. Department of Microbiology, Faculty of Medicine, Harran University, Şanlɩurfa, Turkey. 8. Department of Medical Biology, Faculty of Medicine, Ege University, İzmir, Turkey. 9. Vocational School of Health Sciences, Celal Bayar University, Manisa, Turkey. 10. Department of Parasitology, Faculty of Medicine, Ege University, İzmir, Turkey. 11. Department of Medical Microbiology, Faculty of Medicine, Acɩbadem University, İstanbul, Turkey. 12. National Reference Center for Leishmaniases, Montpellier University Hospital, Montpellier, France.
Abstract
OBJECTIVE: To report isolation of Leishmania major strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L. major between 2011 and 2014. METHODS: Initial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L. major. Developing lesions were observed, and the thickening of footpads was measured weekly. RESULTS: Melting curve analyses of 18 isolates showed a peak concordant with L. major, and two of them were confirmed by isoenzyme analyses as L. major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3 weeks, which even progressed to extremity amputation. CONCLUSION: Cutaneous leishmaniasis-causing L. major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L. major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L. major infections in Turkey, contrary to the infections in neighbouring countries.
OBJECTIVE: To report isolation of Leishmania major strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L. major between 2011 and 2014. METHODS: Initial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L. major. Developing lesions were observed, and the thickening of footpads was measured weekly. RESULTS: Melting curve analyses of 18 isolates showed a peak concordant with L. major, and two of them were confirmed by isoenzyme analyses as L. major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3 weeks, which even progressed to extremity amputation. CONCLUSION:Cutaneous leishmaniasis-causing L. major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L. major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L. major infections in Turkey, contrary to the infections in neighbouring countries.
Authors: Mehmet Karakuş; Abed Nasereddin; Hüseyin Onay; Emin Karaca; Ahmet Özkeklikçi; Charles L Jaffe; Katrin Kuhls; Ahmet Özbilgin; Hatice Ertabaklar; Samiye Demir; Yusuf Özbel; Seray Töz Journal: PLoS Negl Trop Dis Date: 2017-04-12
Authors: Yusuf Özbel; Seray Töz; Clara Muñoz; Maria Ortuño; Zarima Jumakanova; Pedro Pérez-Cutillas; Carla Maia; Cláudia Conceição; Gad Baneth; André Pereira; Yves Van der Stede; Céline M Gossner; Eduardo Berriatua Journal: Zoonoses Public Health Date: 2022-05-26 Impact factor: 2.954