title: Leishmaniasis in Turkey: first clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regions.
abstract: To report isolation of Leishmania major strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L. major between 2011 and 2014.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.affiliation: Department of Parasitology, Faculty of Medicine, Celal Bayar University, Manisa, Turkey.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Özbilgin
authors.affiliation: Department of Parasitology, Faculty of Medicine, Mustafa Kemal University, Hatay, Turkey.
authors.forename: Gülnaz
authors.initials: G
authors.lastname: Çulha
authors.affiliation: Department of Dermatology, Faculty of Medicine, Akdeniz University, Antalya, Turkey.
authors.forename: Soner
authors.initials: S
authors.lastname: Uzun
authors.affiliation: Department of Dermatology, Faculty of Medicine, Dicle University, Diyarbakɩr, Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Harman
authors.affiliation: Department of Dermatology, Faculty of Medicine, Çukurova University, Adana, Turkey.
authors.forename: Suhan Günaştı
authors.initials: SG
authors.lastname: Topal
authors.affiliation: Clinic of Dermatology, Atatürk State Hospital, Antalya, Turkey.
authors.forename: Fulya
authors.initials: F
authors.lastname: Okudan
authors.affiliation: Department of Microbiology, Faculty of Medicine, Harran University, Şanlɩurfa, Turkey.
authors.forename: Fadile
authors.initials: F
authors.lastname: Zeyrek
authors.affiliation: Department of Medical Biology, Faculty of Medicine, Ege University, İzmir, Turkey.
authors.forename: Cumhur
authors.initials: C
authors.lastname: Gündüz
authors.affiliation: Vocational School of Health Sciences, Celal Bayar University, Manisa, Turkey.
authors.forename: İpek
authors.initials: İ
authors.lastname: Östan
authors.affiliation: Department of Parasitology, Faculty of Medicine, Ege University, İzmir, Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Karakuş
authors.affiliation: Department of Parasitology, Faculty of Medicine, Ege University, İzmir, Turkey.
authors.forename: Seray
authors.initials: S
authors.lastname: Töz
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, Acɩbadem University, İstanbul, Turkey.
authors.forename: Özgür
authors.initials: Ö
authors.lastname: Kurt
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, Acɩbadem University, İstanbul, Turkey.
authors.forename: Işın
authors.initials: I
authors.lastname: Akyar
authors.affiliation: Department of Dermatology, Faculty of Medicine, Akdeniz University, Antalya, Turkey.
authors.forename: Ayşegül
authors.initials: A
authors.lastname: Erat
authors.affiliation: Department of Dermatology, Faculty of Medicine, Çukurova University, Adana, Turkey.
authors.forename: Dilek
authors.initials: D
authors.lastname: Güngör
authors.affiliation: Department of Medical Biology, Faculty of Medicine, Ege University, İzmir, Turkey.
authors.forename: Çağla
authors.initials: Ç
authors.lastname: Kayabaşı
authors.affiliation: Department of Parasitology, Faculty of Medicine, Celal Bayar University, Manisa, Turkey.
authors.forename: İbrahim
authors.initials: İ
authors.lastname: Çavuş
authors.affiliation: National Reference Center for Leishmaniases, Montpellier University Hospital, Montpellier, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Bastien
authors.affiliation: National Reference Center for Leishmaniases, Montpellier University Hospital, Montpellier, France.
authors.forename: Francine
authors.initials: F
authors.lastname: Pratlong
authors.affiliation: Department of Medical Microbiology, Faculty of Medicine, Acɩbadem University, İstanbul, Turkey.
authors.forename: Tanıl
authors.initials: T
authors.lastname: Kocagöz
authors.affiliation: Department of Parasitology, Faculty of Medicine, Ege University, İzmir, Turkey.
authors.forename: Yusuf
authors.initials: Y
authors.lastname: Özbel
mesh_terms: D000818:Animals; D002417:Cattle; D004199:Disease Vectors; D005260:Female; D007527:Isoenzymes / Q000032:analysis; D018320:Leishmania major / Q000235:genetics / Q000302:isolation & purification*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis / Q000469:parasitology* / Q000473:pathology; D008297:Male; D008322:Mammals / Q000469:parasitology; D008807:Mice, Inbred BALB C; D060888:Real-Time Polymerase Chain Reaction; D012720:Severity of Illness Index; D012867:Skin / Q000469:parasitology* / Q000473:pathology; D014421:Turkey
chemical_list: D007527:Isoenzymes
keywords: ITS1; ITS1 RT-PCR; Leishmania major; Leishmaniasis; Leishmaniose; RT-PCR; Turkey; Turquie; Turquía; enriched NNN medium; leishmaniasis; medio NNN enriquecido; milieu NNN enrichi; modelo murino; modèle murin; mouse model