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    Leishmaniasis in Turkey: Determination of Leishmania Species by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS)
    (Iranian Scientific Society Medical Entomology, 2014) Culha, Gulnaz; Akyar, Isin; Zeyrek, Fadile Yildiz; Kurt, Ozgur; Gunduz, Cumhur; Toz, Seray Ozensoy; Ostan, Ipek
    Background: Cutaneous leishmaniasis (CL) is endemic in Southeastern Anatolia, mainly in Sanliurfa and Hatay provinces, and the causative agents are mostly Leishmania tropica and less frequently L. infantum. Here, we report the first MALDI-TOF analyses of Leishmania promastigotes obtained from the cultures of two CL cases from Osmaniye and Hatay provinces who were initially diagnosed by microscopy, culture and identified as L. infantum with Real-Time PCR (RT-PCR). Methods: Samples obtained from the skin lesions of patients were initially stained with Giemsa and cultivated in NNN medium. Examination of the smears and cultures revealed Leishmania amastigotes and promastigotes, respectively. The promastigotes (MHOM/TR/2012/CBU15 and MHOM/TR/2012/MK05) obtained from the cultures of both patients were used for RT-PCR targeting the ITS-1 region in the SSU of rRNA. The reference strains of four Leishmania species (L. infantum, L. donovani, L. tropica and L. major) were initially assessed with MALDI-TOF and their data were added to MALDI-TOF Biotyper Library. Results: Both RT-PCR and MALDI-TOF analyses indicated that the causative agent in both patient samples was L. infantum. Conclusion: Despite disadvantages such as requirement of culture fluid with nothing but promastigotes and high cost, MALDI-TOF analysis may be a fast, sensitive and specific diagnostic tool in especially large-scale research studies, where the cost declines, relatively.
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    Leishmaniasis in Turkey: molecular characterization of Leishmania from human and canine clinical samples
    (Wiley-Blackwell Publishing, Inc, 2009) Toz, Seray Ozensoy; Nasereddin, Abedelmajeed; Ozbel, Yusuf; Ertabaklar, Hatice; Culha, Gulnaz; Sevil, Naser; Alkan, M. Ziya
    Human leishmaniasis, both visceral and cutaneous, and canine leishmaniasis have been reported in Turkey for centuries. However, the advent of new diagnostic tools during the last 30 years has led to the recognition that leishmaniasis is an important public health problem throughout the country. In most disease foci both canine and human leishmaniases exist together and identification of parasite species causing these diseases is a pre-requisite for understanding disease epidemiology. A total of 109 samples obtained from human and canine leishmaniasis cases were examined using internal transcribed spacer 1 PCR followed by restriction fragment length polymorphism analysis. Our results indicate that two species, Leishmania tropica and Leishmania infantum, are primarily responsible for cutaneous and visceral leishmaniasis, respectively, in Turkey. However, a new focus of human cutaneous leishmaniasis caused by L. infantum in Hatay region is described. This finding further stresses the importance of Leishmania species molecular characterization in prescribing appropriate therapy and understanding the disease's transmission in different endemic foci.
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    A New Approach for Determining the Spatial Risk Levels for Visceral and Cutaneous Leishmaniasis related with the Distribution of Vector Species in Western Part of Turkey using Geographical Information Systems and Remote Sensing
    (Kafkas Univ, Veteriner Fakultesi Dergisi, 2012) Olgen, M. Kirami; Ozbel, Yusuf; Balcioglu, I. Cuneyt; Demir, Samiye; Simsek, Fatih; Toz, Seray Ozensoy; Ertabaklar, Hatice
    Leishmaniases are present in two clinical forms, as visceral and cutaneous, in Turkey showing a tendency of spreading throughout the country. The aim of the present study was to produce a new model for determining the spatial risk levels using the data in a selected study site in the western part of Turkey. The results of entomological studies in this leishmaniasis focus indicated the presence of suspected vector species Phlebotomus (Larroussius) tobbi and P (Larroussius) neglectus for the visceral, P (Paraphlebotomus) similis for cutaneous forms of the disease. The new risk model was developed based on univariate and multivariate binary logistic regression analyses of geographical variables as altitude, aspect, Normalized Difference Vegetation Index (NDVI), Enhanced Vegetation Index (EVI) and Land Surface Temperature (LST) values related to the distribution of these three species. The results of the new model were used to produce the risk maps and the potential distribution areas of the incriminated vector species with the use of geographical technologies which allowed the identification of the leishmaniasis risk levels that may provide useful information to guide the control program interventions.
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    A Real-Time ITS1-PCR Based Method in the Diagnosis and Species Identification of Leishmania Parasite from Human and Dog Clinical Samples in Turkey
    (Public Library Science, 2013) Toz, Seray Ozensoy; Culha, Gulnaz; Zeyrek, Fadile Yildiz; Ertabaklar, Hatice; Alkan, M. Ziya; Vardarli, Asli Tetik; Gunduz, Cumhur
    Human visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter-and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L. infantum and 6.52% as L. tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. Hovewer, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey.
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    Spatial distribution of phlebotomine sand flies in the Aydin Mountains and surroundings: the main focus of cutaneous leishmaniasis in western Turkey
    (Soc Vector Ecology, 2011) Ozbel, Yusuf; Balcioglu, I. Cuneyt; Olgen, M. Kirami; Simsek, Fatih M.; Toz, Seray Ozensoy; Ertabaklar, Hatice; Demir, Samiye
    An entomological survey was conducted to determine the spatial distribution of phlebotomine fauna and understand the effect of environmental factors. The entomological survey was carried out during 2006-2007 in a study area in the rural area of Aydin province, near the Kusadasi town where VL, CL, and canine leishmaniasis (CanL) are endemic. In 2006 and 2007, 132 locations were sampled using sticky traps mainly on embankments. Detailed environmental and meteorological information was also collected for each location. The results of entomological studies indicated that the probable vectors are Phlebotomus tobbi and P. neglectus for VL and CanL, and P. similis for CL in this western leishmaniasis focus. The data revealed a correlation between their presence and spatial variables such as altitude, sampling site location, and humidity. The distribution areas of probable vector species in this study area allowed the identification of risk levels, which may provide useful information to guide the leishmaniasis research in endemic regions.