Determination of the species of the sand flies the vector of cutaneous leishmaniasis that is endemic in

M. Emin LİMONCU a, İ. Cüneyt BALCIOĞLU b, Seray ÖZENSOY TÖZ c, Samiye DEMİR d, Hakan KAVUR e, V. Özge ERMİŞ f, Yusuf ÖZBEL c

aCelal Bayar University, Vocational School of Health Services, , bCelal Bayar University, Medical Faculty, Department of Parasitology, Manisa, Turkey cEge University, Medical Faculty, Department of Parasitology, Izmir, Turkey dEge University Science Faculty, Department of Zoology, Izmir, Turkey ͤ Cukurova University Faculty of Medicine, Department of Microbiology fCelal Bayar University Science Faculty, Department of Zoology, Manisa, Turkey

INTRODUCTION AND PURPOSE Molecular Studies :

Different clinical manifestations of leishmaniasis infections which caused by leishmania parasites in different species are transmitted by phlebotomine Of 428 female sand flies samples 326 of them diagnosed and classified according to blood-fed/eggs sand flies. Two clinical form of leishmaniasis; visceral leishmaniasis(VL) and cutaneous leishmaniasis have been reported in Turkey. presence properties and 29 pools were prepared by putting 3 to 19 members in each of them. Thirteen of these pools were prepared from the samples that have been collected from Mut, 13 of Visceral leishmaniasis (VL), caused by L. infantum , is endemic in the Aegean and Mediterranean region and occurs sporadically in other regions. them from and 3 of them from . Leishmania DNA was investigated in the DNA Cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum , is endemic in west and southeast . samples found in these pools by real time PCR method using ITS1 gene region. DNA isolation was performed by using ZR insect/Tissue DNA kit-5 TM (cat. No. D 6015). İn 2009, promastigotes forms of L. İnfantum leading to CL, were found in P. tobbi , as a proven vector, at province which close to Mersin. The aim of this study was to determine the sand fly fauna in various areas of the Mersin province where cutaneous leishmaniasis caused by L. tropica are seen and to determine the potential of Phlebotomus and other species for vectoring besides showing the presence of this parasite species directly and with molecular biological methods.

MATHER İALS AND METHODS

Fıgure 5. The real time PCR result of pool numbers 2 for ITS1 gene region Study Area

Mersin province where is located in the south east of the Mediterranean region has spread of 15853 km 2 land. Mean temperature of days is 36.6 ºC, mean temperature of the nights is 16.3 ºC in Mersin during the summer. During the winter season it has 24.0 ºC and 3.6 ºC, respectively.

Sandfly Collection

The sand flies were collected between on the 10-17 July 2010. The study was carried out with the data of the 8 different locations of Anamur, Mut, and Silifke disticts in Mersin where KL patients living area. A total of 52 CDC light traps were settled down into the patient ´s houses and their animal shelters. After the sand flies collected, the gut dissections and determination of species has been studied. The identification of sand flies species were done by several keys and previous drawing.

Figure 6. The real time PCR result of pool numbers 6 for ITS1 gene region

According to real time PCR results : Figure 1. Placing the CDC light traps indoor and outdoor Figure 3. DNA isolation steps ‹ Pool number 2 that contains female P. Sergenti samples from Anamur ‹ Pool number 6 that contains female P. Sergenti samples from Mut Identification of Sand Flies RESULTS are found positive in terms of ITS 1 gene region and type of parasite has been detected as L.Tropica.

P. sergenti

P. alexandri

0% P. neglectus/syriacus 0% 0% Conclusions 1% 1% P. tobbi 0% In the previous faunistic studies that were carried out In Medditerean region of Turkey, neighbourhood of Mersin, In that have found 6 11% P. mascitti 1% Phlebotomus species (P . tobbi, P. syriacus, P. perfiliewi, P. papatasi, P. sergenti ve P. simici ), and also another study that performed in Hatay has 3% 30% P. papatasi found 3 Phlebotomus species (P. sergenti, P. papatasi ve P. syriacus ). In our studies we have detected 9 species belonging to genus phlebotomus ; P. papatasi, P. sergenti, P. alexandri, P. tobbi, P. syriacus/P. neglectus, P. simici, P. halepensis, P. brevis ve P. mascittii and 4 species belonging to 10% P. simici sergentomiya ; S. minuta , S. dentata, S. fallax, S. pawlowski.

P. halepensis A study that carried out in Iran showed that the L. Tropica is the cause of CL cases. The study was based on the determination of kDNA and ITS1 gene regions on female P.sergenti species and the determination were done by using the nested PCR and RFLP methods. 12% P. brevis 1% 18% Leishmania (L.) tropica and L. infantum caused by CL in Turkey. L. tropica in the western part of Turkey transmits by P. similis , and in the eastern 12% S. minuta part of Turkey, especially in Şanliurfa transmits by P. sergenti .

S. dentata In this study, the sand fly fauna in Mersin where CL is an endemic disease was determined. P. S. fallax sergenti was identified as the dominant species and it was concluded that P. sergenti might be S. pawlowski the probable vector of L. tropica .

Sergentomyia sp.

?

Fıgure 2. Sand fly identification steps Figure 4 Sand fly specieses and their density in study area