Browsing by Author "Töz S."
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Item Evaluation of conjunctival swab sampling in the diagnosis of canine leishmaniasis: A two-year follow-up study in Çukurova Plain, Turkey(Elsevier B.V., 2015) Karakuş M.; Töz S.; Ertabaklar H.; Paşa S.; Atasoy A.; Arserim S.K.; Ölgen M.K.; Ziya Alkan M.; Durrant C.; Özbel Y.The diagnosis of canine leishmaniasis (CanL) in symptomatic and asymptomatic dogs is a very important and problematic public health issue in Turkey. A longitudinal study was carried out on dogs in selected villages in the Çukurova Plain in Turkey, from July 2011 to June 2013, where cutaneous (CL) and visceral (VL) leishmaniasis is endemic. The study aimed to determine the prevalence of CanL and to evaluate the early diagnostic performance of the non-invasive conjunctival swab nested PCR (CS n-PCR) test in comparison with the Indirect Fluorescent Antibody Test (IFAT). The consecutive blood and CS samples from a representative number of dogs (80-100 dogs/each survey) were collected in a cohort of 6 villages located in the area. Clinical symptoms, demographic and physical features about each dog were noted and lymph node aspiration samples were obtained from selected dogs with lymphadenopathy. In four surveys during the period, a total of 338 sets (blood and CS) of samples from 206 dogs were obtained, such that 83 dogs were sampled more than once. In the cross-sectional analysis, the CanL prevalence was found to be 27.18% (between 7.14% and 39.13%) by IFAT and 41.74% (between 29.03% and 46.66%) by CS n-PCR. The isolated strains were identified as Leishmania infantum MON-1 (n = 9) and MON-98 (n = 2) by MLEE analysis. Genetic studies targeting the Hsp70 and ITS1 regions performed on 11 dog isolates also showed two clear separate groups. According to IFAT results, 24 of the 83 dogs sampled more than once showed seroconversion (n = 19) or a four-fold increase in Ab titers (n = 5), while 17 were positive in the initial screening. Forty-two dogs stayed negative during the whole period. The natural Leishmania exposure rate was detected as 31.14% in the study area. CS n-PCR only detected Leishmania infection earlier than IFAT in 8 dogs. No statistical difference was found after the analysis of demographical and physical data. The results indicated that (i) circulation of the dog population is very common in settlements in the Çukurova Plain, but the disease prevalence is high and stable, (ii) the performance of CS n-PCR for detecting Leishmania-dog contact is higher than IFAT, (iii) and some of the parasites isolated from dogs have different zymodemes and/or genotypes from previous human and sand fly isolates; suggesting the probability of two different cycles of leishmaniasis in this particular area. This hypothesis should be supported by future studies targeting vectors and reservoirs. © 2015 Elsevier B.V.Item Comparing the Efficacy of Commercially Available Insecticide and Dimeticone based Solutions on Head Lice, Pediculus capitis: in vitro Trials(2015) Balcıoğlu I.C.; Karakuş M.; Arserim S.K.; Limoncu M.E.; Töz S.; Baştemur S.; Öncel K.; Özbel Y.OBJECTIVE: Head lice infestation is a public health and social problem for almost all countries worldwide. For its treatment, insecticide and dimeticone-based solutions are currently available in the markets in many countries. We aimed to compare the efficacy of commercially available anti-head lice shampoos containing insecticide and physically effective products with different percentages of dimeticone using an in vitro technique.; METHODS: Head lice specimens were collected from primary school children using special plastic and metal combs. Anti-head lice products were commercially purchased and used directly. The specimens were placed one by one in 5-cm Petri dishes containing a slightly wet filter paper and were kept in a plastic cage at 28±2°C and 50%±20% relative humidity. A standardized protocol was used for testing all the products, and mortality data were obtained after 24 h. Two control tests were performed with each batch of trials. For each product and control, 10-20 head lice specimens were used, and the results were statistically analyzed.; RESULTS: Our study demonstrated that among all the tested products, two products containing mineral oils [5.5% dimeticone & silicone (patented product) and dimeticone (no percentage mentioned in the prospectus) & cyclopentasiloxane] were found to be more effective for killing head lice in vitro.; CONCLUSION: Physically effective products can be repetitively used because they are non-toxic and resistance to them is not expected. To control the infestation at a public level, the use of these products needs to be encouraged with respect to their cost price.Item Detection of Leishmania major and Leishmania tropica in domestic cats in the Ege Region of Turkey(Elsevier B.V., 2015) Paşa S.; Tetik Vardarli A.; Erol N.; Karakuş M.; Töz S.; Atasoy A.; Balcioğlu T.C.; Emek Tuna G.; Ermiş T.V.; Ertabaklar H.; Özbel Y.Leishmaniosis is a group of diseases caused by different species of Leishmania parasites in mammalian species. The aim of the present study was to investigate the presence of Leishmania spp. DNA in cats using real time polymerase chain reaction (RT-PCR) assays targeting internal transcribed spacer (ITS1) and heat-shock protein 70 gene (Hsp70) regions with Leishmania species-specific primers and probes. Blood samples were collected from 147 cats (73 female; 74 male) in the endemic regions for zoonotic visceral leishmaniasis in the western provinces of Turkey and analyzed using two RT-PCR assays. Additionally, Hsp70 RT-PCR products were sequenced. ELISA assays for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) were also carried out for 145 of the 147 samples.Overall, 13/147 (8.84%) cats were positive for Leishmania by RT-PCR (4 L. major and 9 L. tropica). FIV and FeLV antibody and/or antigen was detected in 4 and 5 cats among Leishmania DNA positives, respectively. To the best of our knowledge, this study is the first to investigate and report the presence of L. major and L. tropica infections in a large group of domestic cats in Turkey. The results obtained indicate that species identification of Leishmania is essential for epidemiological understanding and that clinical signs alone are not indicative for leishmaniosis in cats, as it is in dogs. This study suggests that extensive research should be carried out in cat populations in order to fully understand the role of cats in the epidemiology of the disease. © 2015 Elsevier B.V.Item sandflyDST: a dynamic web-based decision support tool for the morphological identification of sandflies present in Anatolia and mainland Europe, and user study(Blackwell Publishing Ltd, 2016) Karakülah G.; Karakuş M.; Suner A.; Demir S.; Arserim S.K.; Töz S.; Özbel Y.Species identification of sandflies is mainly performed according to morphological characters using classical written identification keys. This study introduces a new web-based decision support tool (sandflyDST) for guiding the morphological identification of sandfly species present in Anatolia and mainland Europe and classified in the Phlebotomus and Sergentomyia genera (both: Diptera: Psychodidae). The current version of the tool consists of 111 questions and 36 drawings obtained from classical written keys, and 107 photographs for the quick and easy identification of 26 species of the genus Phlebotomus and four species of the genus Sergentomyia. The tool guides users through a decision tree using yes/no questions about the morphological characters of the specimen. The tool was applied by 30 individuals, who then completed study questionnaires. The results of subsequent analyses indicated that the usability (x‾SUSScore=75.4) and users' level of appreciation (86.6%) of the tool were quite high; almost all of the participants considered recommending the tool to others. The tool may also be useful in training new entomologists and maintaining their level of expertise. This is a dynamic tool and can be improved or upgraded according to feedback. The tool is now available online at http://parasitology.ege.edu.tr/sandflyDST/index.php. © 2016 The Royal Entomological Society.Item Leishmaniasis in Turkey: First clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regions(Blackwell Publishing Ltd, 2016) Özbilgin A.; Çulha G.; Uzun S.; Harman M.; Topal S.G.; Okudan F.; Zeyrek F.; Gündüz C.; Östan I.; Karakuş M.; Töz S.; Kurt Ö.; Akyar I.; Erat A.; Güngör D.; Kayabaşi Ç.; Çavuş I.; Bastien P.; Pratlong F.; Kocagöz T.; Özbel Y.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. © 2016 John Wiley & Sons Ltd.Item Molecular detection and identification of Leishmania spp. in naturally infected Phlebotomus tobbi and Sergentomyia dentata in a focus of human and canine leishmaniasis in western Turkey(Elsevier B.V., 2016) Özbel Y.; Karakuş M.; Arserim S.K.; Kalkan Ş.O.; Töz S.Human visceral leishmaniasis (VL) is reported from 38 provinces of Turkey and dogs are accepted as main reservoir hosts. Kuşadasi town, belonging to Aydin province and located in western part of Turkey, is endemic for human and canine visceral leishmaniasis caused by Leishmania infantum MON1 and MON98.In this study, phlebotomine survey was conducted to determine the vector sand fly species and to identify sand fly blood meal sources. In August and September 2012, 1027 sand fly specimens were caught using CDC light traps. Eight Phlebotomus and two Sergentomyia species with the dominancy of Phlebotomus tobbi (61.34%) were detected. A total of 622 female sand flies (571 Phlebotomus; 51 Sergentomyia) were checked for Leishmania infection by direct dissection of the midgut. The half of the midgut content was inoculated into NNN culture for isolation of the parasite. Leishmania species-specific ITS1 real time PCR, conventional PCR assays of ITS1 and hsp70 genes and subsequent sequencing were performed from extracted DNAs. A region of cytochrome b (cyt-b) gene of vertebrates based PCR was used to determine the source of blood meal of sand flies.In microscopical examinations, two female specimens (0.32%) were found naturally infected with high number and different stages of promastigotes. No growth was observed in NNN culture but Leishmania DNA was obtained from both specimens. First positive specimen was identified as P. tobbi and L. infantum DNA was detected. Second specimen was Sergentomyia dentata, but Leishmania DNA could not be identified on species level. A total of 16 blood-fed female P. tobbi specimens were used for blood meal analysis and eight, three and one specimens were positive for human, dog and mouse, respectively.This is the first detection of Leishmania promastigotes using microscopical examination in P. tobbi and S. dentata in human and canine visceral leishmaniasis endemic area in western part of Turkey. Our results indicate that, (i) P. tobbi is the principal vector species and (ii) human and dogs are main blood sources. The detection of Leishmania sp. in Sergentomyia species may be an evidence for natural cycle of Sauro-leishmania agents in the area. © 2016 Elsevier B.V.Item Epidemiological analysis of Leishmania tropica strains and giemsa-stained smears from Syrian and Turkish leishmaniasis patients using multilocus microsatellite typing (MLMT)(Public Library of Science, 2017) Karakuş M.; Nasereddin A.; Onay H.; Karaca E.; Özkeklikçi A.; Jaffe C.L.; Kuhls K.; Özbilgin A.; Ertabaklar H.; Demir S.; Özbel Y.; Töz S.Turkey is located in an important geographical location, in terms of the epidemiology of vector-borne diseases, linking Asia and Europe. Cutaneous leishmaniasis (CL) is one of the endemic diseases in a Turkey and according to the Ministry Health of Turkey, 45% of CL patients originate from Şanlıurfa province located in southeastern Turkey. Herein, the epidemiological status of CL, caused by L. tropica, in Turkey was examined using multilocus microsatellite typing (MLMT) of strains obtained from Turkish and Syrian patients. A total of 38 cryopreserved strains and 20 Giemsa-stained smears were included in the present study. MLMT was performed using 12 highly specific microsatellite markers. Delta K (ΔK) calculation and Bayesian statistics were used to determine the population structure. Three main populations (POP A, B and C) were identified and further examination revealed the presence of three subpopulations for POP B and C. Combined analysis was performed using the data of previously typed L. tropica strains and Mediterranean and Şanlıurfa populations were identified. This finding suggests that the epidemiological status of L. tropica is more complicated than expected when compared to previous studies. A new population, comprised of Syrian L. tropica samples, was reported for the first time in Turkey, and the data presented here will provide new epidemiological information for further studies. © 2017 Karakuş et al.Item Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused by Leishmania donovani in Turkey(Elsevier B.V., 2017) Özbilgin A.; Harman M.; Karakuş M.; Bart A.; Töz S.; Kurt Ö.; Çavuş İ.; Polat E.; Gündüz C.; Van Gool T.; Özbel Y.In Turkey, the main causative agents are Leishmania tropica (L. tropica) and Leishmania infantum (L. infantum) for cutaneous leishmaniasis (CL) and L. infantum for visceral leishmaniasis (VL). In this study, we investigated leishmaniasis cases caused by L. donovani and established animal models for understanding its tropism in in vivo conditions. Clinical samples (lesion aspirations and bone marrow) obtained from CL/VL patients were investigated using parasitological (smear/NNN) and DNA-based techniques. For species identification, a real time ITS1-PCR was performed using isolates and results were confirmed by hsp70 PCR-N/sequencing and cpb gene PCR/sequencing in order to reveal Leishmania donovani and Leishmania infantum discrimination. Clinical materials from CL and VL patients were also inoculated into two experimental groups (Group CL and Group VL) of Balb/C mice intraperitoneally for creating clinical picture of Turkish L. donovani strains. After 45 days, the samples from visible sores of the skin were taken, and spleens and livers were removed. Measurements of the internal organs were done and touch preparations were prepared for checking the presence of amastigotes. The strains were isolated from all patients and amastigotes were seen in all smears of the patients, and then isolates were immediately stored in liquid nitrogen. In real time ITS1-PCR, the melting temperatures of all samples were out of range of L. infantum, L. tropica and L. major. Sequencing of hsp70 PCR-N showed that all isolates highly identical to previously submitted L. donovani sequences in GenBank, and cpb gene sequencing showed five isolates had longer cpbF allele, whereas one isolate contained a mixed sequence of both cpbF and cpbE. All mice in both experimental groups became infected. Compared to controls, the length and width of both liver and spleen were significantly elevated (p < 0.001) in both groups of mice. However, the weight of the liver increased significantly in all mice whereas the weight of spleen increased only in VL group. Amastigotes were also seen in all touch preparations prepared from skin sores, spleen and liver. L. donovani strain was isolated from autocutaneous a VL patient first time in Turkey. Animal models using clinical samples were successfully established and important clinical differences of the isolated strains were observed. © 2017 Elsevier B.V.Item In vitro anti-leishmanial activity of Sarcopoterium spinosum against Leishmania tropica; [Sarcopoterium spinosum’un Leishmania tropica’ya karşı in vitro anti-leishmanial etkisi](Chartered Inst. of Building Services Engineers, 2018) Can H.; Kayalar H.; Bozkurt B.; Can Ş.; Döşkaya M.; Töz S.Complex clinical symptoms such as ulcerative skin lesions, destructive mucosal inflammation, and disseminated visceral infection can reveal in leishmaniasis. The conventional drugs are toxic and expensive. In addition, patients receive a long treatment with these drugs which have adverse effects and unfortunately there are some limitations during the treatment. The aim of this study is to investigate the in vitro anti-leishmanial activities of four different extracts of Sarcopoterium spinosum against Leishmania tropica. Initially, different concentrations of ethanol, methanol, n-hexane, and water extracts of S. spinosum were incubated with L. tropica promastigotes. After 72 hours of incubation, the growth of L. tropica promastigotes was significantly inhibited and the percentage of inhibition ranged between 42.8 and 100 %. Among these extracts, the most efficient growth inhibition (100 %) was obtained with methanol extract (at a dose of 50 µg/ml). In conclusion, S. spinosum may be a potential source for the development of novel therapeutic agents to treat L. tropica infection. © 2018, Chartered Inst. of Building Services Engineers. All rights reserved.Item Vector and reservoir surveillance study in a canine and human leishmaniasis endemic area in most western part of Turkey, Karaburun(Elsevier B.V., 2019) Karakuş M.; Arserim S.K.; Erişöz Kasap Ö.; Pekağırbaş M.; Aküzüm D.; Alten B.; Töz S.; Özbel Y.Leishmaniasis is an arthropod borne disease that is endemic in 102 countries and one and half million new cases are reported each year. Sand flies are the one and only proven vectors of the disease and dogs are the main reservoirs in urban areas. Karaburun peninsula is located in most western part of Turkey and is reported to be an endemic area for human and canine leishmaniasis. The most recent study was undertaken more than 15 years ago in The peninsula and no clear data available for vectors or reservoirs. Thus, we aimed to update the information regarding sand fly diversity, infection status of reservoirs and vectors in the study area. Sand flies were collected using CDC light traps at 13 different sites of Karaburun and species identification was made using previously published keys. Monospecific pools were generated using midguts with blood retention and were screened for the presence of Leishmania spp. DNA by molecular techniques. A non-invasive conjunctival swab sampling was performed to identify the infection status among reservoirs and species typing of the causative agent was also undertaken using ITS1 PCR. Three out of 30 pools were found positive for Leishmania infantum that were generated using guts of Phlebotomus tobbi (n:36). Among all sampled dogs (44) and cats (19), 11 and one of them were found positive for L. infantum, respectively. There was a decrease in the number of P. papatasi during the study period, while increase was observed in the number of P. tobbi. The presence of proven vectors and reservoirs as well as Leishmania DNA in cats was shown in the present study. Sand fly fauna is updated and Leishmania DNA presence in cats was reported in the study area for the first time. © 2018 Elsevier B.V.Item The current clinical and geographical situation of cutaneous leishmaniasis based on species identification in Turkey(Elsevier B.V., 2019) Özbilgin A.; Töz S.; Harman M.; Günaştı Topal S.; Uzun S.; Okudan F.; Güngör D.; Erat A.; Ertabaklar H.; Ertuğ S.; Gündüz C.; Çavuş İ.; Karakuş M.; Östan Ural İ.; Ölgen M.K.; Kayabaşı Ç.; Kurt Ö.; Özbel Y.Leishmaniases are a group of vector-borne diseases caused by the members of Leishmania genus, and there are three main clinical forms of the infection as visceral, cutaneous, and mucocutaneous. Cutaneous leishmaniasis is a growing public health problem in Turkey due to increasing detection of autochthonous cases caused by L. major and L. donovani in some regions in addition to Syrian imported cases. For this reason, we aimed to evaluate the current epidemiological situation of CL in the view of causative agents and their geographical distribution throughout Turkey. The samples were collected from 356 CL patients admitted to different centers in 18 provinces between January 2013 and December 2016. Direct microscopy, culture (regular and enriched NNN) and molecular techniques (real-time ITS1 PCR and hsp70 PCR/sequencing) were performed. By molecular techniques, 299, 28, 19 and 10 isolates/clinical samples were identified as L. tropica, L. major, L. infantum and L. donovani, respectively. Most of the patients (65.73%) had one lesion usually on their face/head. Dry-nodular type lesions (n = 291) were mainly associated with L. tropica while L. major was mainly found related to wet-ulcerative ones. Leishmaniasis recidivans was also detected in 2.52% among 356 patients. L. tropica was detected as most widespread species causing CL in Turkey. L. infantum and L. major was also found in one third of the provinces. Enriched NNN culture was worked well for isolating the parasite and 346 isolates were successfully grown and stored in liquid nitrogen. The comparison of all diagnostic techniques showed that the parasitological positivity rate could increase if the combination of direct microscopy and real-time ITS1 PCR is used. Besides well-known anthroponotic L. tropica cases, the increasing detection of CL cases caused by zoonotic species, L. infantum and L. major, is one of the most important findings in the present study. In our opinion to ensure timely and accurate diagnosis, proper treatment and countrywide effective control of CL in Turkey a systematic approach is needed on the base of information about characteristics of lesions and patients and epidemiological features of the disease. © 2018 Elsevier B.V.Item Detection of Leishmania RNA virus 2 in Leishmania species from Turkey(Oxford University Press, 2019) Nalçacı M.; Karakuş M.; Yılmaz B.; Demir S.; Özbilgin A.; Özbel Y.; Töz S.Background: Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus infecting some Leishmania strains and triggering a destructive hyperinflammatory response in mammalian hosts in the New World. There is limited knowledge of the presence of this virus in Old World Leishmania species and its role in the outcome of the disease. We aimed to investigate the presence of LRV in Leishmania species/strains from Turkey. Methods: Twenty-nine previously identified Leishmania isolates (24 L. tropica, 2 L. infantum, 3 L. major) were examined for LRV positivity using dsRNA visualization in agarose gel after total nucleic acid extraction and RQ-deoxyribonuclease treatment and amplification of a 526 bp fragment of the LRV2-specific RNA-dependent RNA polymerase gene by reverse transcription polymerase chain reaction. Results: Ten (7 L. tropica [24.13%], 3 L. major [10.34%]) of the 29 Leishmania strains gave positive results for LRV. Basic Local Alignment Search Tool analysis showed that all these viruses are LRV2-1. LRV2 was detected for the first time in L. tropica strains in the present study. Conclusions: The clinical manifestation and resistance status of the disease can be different depending on the host and parasite species/strains. The presence of LRV2 may be one of the factors contributing the course of disease. Further studies are needed to elucidate the specific role of LRV2, as it may be a potential target for effective treatment strategies. © The Author(s) 2019. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved.Item The initial detection of Toscana virus in phlebotomine sandflies from Turkey(Blackwell Publishing Ltd, 2020) Özbel Y.; Oğuz G.; Arserim S.K.; Erişöz Kasap Ö.; Karaoglu B.; Yilmaz A.; Emanet N.; Günay F.; Hacioğlu S.; Demirok M.C.; Töz S.; Alten B.; Nalçaci M.; Özkul A.; Ergünay K.Toscana virus (TOSV) is a prominent arthropod-borne viral agent of human central nervous system infections occurring in the Mediterranean region. The main transmission route to susceptible individuals involves sandflies as vectors. Despite several reports revealing widespread TOSV activity in Turkey, vectors remained unidentified. A sandfly field survey was carried out in five provinces in Central, Southeast and Mediterranean Anatolia in 2017 to identify TOSV and related sandfly-borne phleboviruses and Leishmania parasites, with evidence for circulation in the region. A total of 7136 sandfly specimens, collected via standard methods, were evaluated in 163 pools. TOSV was detected in 11 pools (6.7%), comprising Phlebotomus major sensu lato, Sergentomyia dentata and Phlebotomus papatasi species. TOSV partial L and S segment sequences were characterized, that phylogenetically clustered with local and global genotype A strains. An amino acid substitution outside the conserved motifs of the viral polymerase, also present in previous TOSV sequences in endemic regions, was observed. Leishmania tropica was detected in a single pool of Ph. sergentii (0.6%). This is the first report of TOSV in sandflies from Turkey, and this study further provides evidence for additional sandfly species with the potential to transmit TOSV. © 2020 The Royal Entomological SocietyItem Entomological survey for the detection of sand fly fauna and vector species in the cutaneous leishmaniasis endemic area in east mediterranean region of Turkey, Mersin province(Oxford University Press, 2020) Limoncu M.E.; Balcioǧlu I.C.; Töz S.; Demir S.; Kavur H.; Karakuş M.; Vardarli A.T.; Özbel Y.Cutaneous (CL) and visceral (VL) forms of leishmaniasis, transmitted by sand flies, are seen in all countries located in Mediterranean Basin including Turkey. In this study, we aimed to conduct an entomological survey for the detection of sand fly fauna and vector species in Mersin province, one of the important endemic areas for CL in Turkey. In total, 912 sand fly specimens were collected in 2010 and 2011 using CDC light traps. Nine Phlebotomus (Diptera: Psychodidae) and three Sergentomyia (Diptera: Psychodidae) species were detected. Of the collected Phlebotomus sand flies, P. sergenti Parrot, 1917 (30.1%) was the most dominant followed by P. alexandri Sinton, 1928 (18.2%), P. neglectus/syriacus Tonnoir Adler (12.0%), P. tobbi Adler & Theodor, 1930 (11.7%), and P. papatasi Scopoli, 1786 (10.2%), while S. minuta Rondani, 1843 (11.3%) was the dominant species among Sergentomyia. During the field work in 2011, female specimens (n = 81) were screened for the presence of Leishmania promastigotes by midgut dissection, and all were found negative. The rest of the collected female specimens (n = 334) were pooled according to species (P. alexandri, P. neglectus/syriacus, P. papatasi, P. sergenti, P. simici, and P. tobbi) and location (Mut, Silifke, and Anamur). In total, 29 pools were generated and real-time ITS1 PCR assay was performed to detect and identify natural Leishmania Ross, 1903 (Kinetoplastida: Trypanosomatida) infection. Two pools, both from Mut town, containing P. sergenti specimens were found positive and Leishmania tropica Ross, 1903 was identified as an infectious agent for both pools. In conclusion, the sand fly fauna was determined in an endemic area for CL. The detection of L. tropica DNA in P. sergenti specimens showed the possible vectorial role of this species in Mersin province. © 2020 The Author(s).Item Determination of sand fly fauna and molecular detection of Leishmania in sand flies in Antalya Province, Southern Turkey(Springer Science and Business Media Deutschland GmbH, 2021) Arserim S.K.; Çetin H.; Karakuş M.; Demir S.; Ser Ö.; Töz S.; Balcioğlu I.C.; Ölgen M.K.; Yilmaz B.; Özbel Y.Visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are diseases transmitted by infected female sand flies. Since the eradication of malaria in Turkey, CL is the main vector-borne disease in the country, with more than 2000 cases per year, making it a significant public health problem. The aims of this study were to carry out an entomological survey in Antalya Province, an endemic area for CL in the Mediterranean Region of Turkey, to identify sand fly fauna and to screen female specimens for the presence of Leishmania parasites (Leishmania infantum, L. tropica, L. major, and L. donovani) using molecular analysis. Sand flies were collected in 42 localities of seven districts in Antalya Province using CDC miniature light traps in two different periods, June 2012 and September 2013. The specimens were kept in 96% ethanol until the dissection was done. The head and genitalia of the specimens were cut for preparing individual slides for species identification. The rest of the body of female specimens was kept separately. The specimens were identified at the species level, and 27 pools were generated according to the locations and species for screening the presence of Leishmania. A commercial kit was used for DNA extractions. Real-time and conventional polymerase chain reaction (PCR) targeting the internal transcribed spacer region (ITS1) were then performed. In total, 1306 specimens comprising nine species belonging to the Phlebotomus genus were collected in the study region, with Phlebotomus neglectus/syriacus (38.82%) the most abundant, followed by P. alexandri (21.67%) and P. tobbi (20.44%). In the 27 pools, Leishmania infantum DNA was detected in four pools containing P. neglectus/syriacus and one pool containing P. tobbi. In conclusion, the sand fly fauna in the Antalya Province is diverse. The probable vector sand fly species are P. neglectus/syriacus and P. tobbi with high dominance (59.26%), which indicates a high risk of CL transmission. The data presented here may help to shed more light on the transmission cycles of the Leishmania parasite in this CL endemic area. © 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.Item Diversity of leishmania strains isolated from cutaneous leishmaniasis patients in Turkey and its reflection to clinics in mice model; [Türkiye’de kutanöz leyşmanyazis hastalarından elde edilen leishmania İzolatlarındaki farklılıklar ve bunların fare modeline klinik yansıması](Ankara Microbiology Society, 2021) Özbilgin A.; Çulha G.; Güray M.Z.; Zeyrek F.Y.; Akyar I.; Töz S.; Östan Ural İ.; Kurt Ö.; Kocagöz T.; Çavuş I.; Gündüz C.Although asexual reproduction has been attributed to Leishmania species, genetic exchange has recently been demonstrated, which helped emerging of hybrid isolates. Situated on the crossroads between three continents, Leishmania hybrids may be present in Turkey. In Turkey, visceral leishmaniasis caused by Leishmania infantum is less common, while cutaneous leishmaniasis (CL) caused by Leishmania tropica and L.infantum could reach 2500 reported cases a year. Our aim was to investigate genetic variability of local Leishmania species and presence of hybrid Leishmania strains in Turkey. Twenty CL patients from Sanliurfa and Hatay, where only L.tropica and both L.tropica and L.infantum cause CL, respectively, were registered equally. All isolates were assessed with real-time polymerase chain reaction (Rt-PCR), isoenzyme analysis, gene sequencing, two-dimensional gel electrophoresis (2D-PAGE) and MALDI-TOF/TOFMS followed by in vivo analyses on mouse model. Identification of differentially expressed proteins was performed. These proteins were confirmed by sequence analysis. All isolates from Sanliurfa were found to be L.tropica which caused cutaneous infection in mice. However, one of 10 isolates from Hatay was found as Leishmania major which caused cutaneous infection. Five isolates were found as L.tropica with Rt-PCR and gene sequencing, one of which had one different protein from the reference L.tropica strain and caused cutaneous infection. Four of the five isolates had five different proteins compared to reference strain and caused both cutaneous and visceral infections. Remaining four isolates showed double melting curves in Rt-PCR, which were concordant with L.tropica and L.infantum. Their sequencing and isoenzyme analyses indicated them as L.infantum. They had six different proteins compared to reference L.infantum strain and caused cutaneous and visceral infections. It is concluded that the isolates with different proteins were hybrid Leishmania species. In the present study, outcomes of the proteomics, genomics, clinical manifestations and tissue tropism on animal models were evaluated together for the first time. In addition to L.tropica and L.infantum, L.major was identified as a causative agent for CL and hybrids of L.infantum/tropica were also shown to be present. © 2020 Ankara Microbiology Society. All rights reserved.Item Host–Parasite Interactions: Regulation of Leishmania Infection in Sand Fly(Springer Science and Business Media Deutschland GmbH, 2022) Omondi Z.N.; Arserim S.K.; Töz S.; Özbel Y.Purpose: Sand flies are the only proven vectors of leishmaniases, a tropical neglected disease endemic in at least 92 countries. Vector–parasite interactions play a significant role in vector-borne disease transmission. There are various bottlenecks to Leishmania colonization of the sand fly midgut. Such bottlenecks include the production of innate immune-related molecules, digestive proteases, parasite impermeable peritrophic membrane, and resident gut microbiota. These barriers determine the parasite load transmitted and, consequently, the disease outcome in mammalian host. Therefore, it is important to understand the molecular responses of both sand fly and Leishmania during infection. Method: Here, we reviewed the published literature on sand fly–Leishmania interactions bringing together earlier and current findings to highlight new developments and research gaps in the field. Conclusion: Recent research studies on sand fly–Leishmania interaction have revealed contrasting observations to past studies. However, how Leishmania parasites evade the sand fly immune response still needs further research. Sand fly response to Leishmania infection can be best understood by analyzing its tissue transcriptome. Better characterization of the role of midgut components could be a game changer in development of transmission-blocking strategies for leishmaniasis. © 2022, The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.Item Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis; [Leishmania RNA Virüs 2 (LRV2) Saptanmasında cDNA Sentezindeki Bir Modifikasyon ile Hassasiyetin Artırılması](Galenos Publishing House, 2022) Nalçacı M.; Karakuş M.; Özbel Y.; Özbilgin A.; Töz S.Objective: Leishmania RNA virus was detected the first time in the New World Leishmania species. Recent studies were also showed the presence of Leishmania RNA virus 2 (LRV2) in Old Word Leishmania species including Turkish L. major and L. tropica isolates. This study aimed to increase the sensitivity of qPCR with a modification in the denaturation step of cDNA preparation protocol. Methods: In this study, LRV2+ three L. major, two L. tropica strains and L. major control strain (MHOM/SU/73/5-ASKH) were included. Total RNA isolation was done using different numbers of Leishmania promastigotes (108, 105 and 103). Before cDNA synthesis, samples were denatured at 95 °C for 2 min, as a modification of the kit procedure. qPCR was undertaken using 0.5 mM primers (LRV F-HR/LRV R-HR) diluted in SYBR Green Master mix. Results: We observed lower Ct values in amplicons with the modified version than with the classical kit protocol for cDNA synthesis, in all of the strains used in the study. The addition of pre-denaturation step at 95 °C showed lower Ct values meaning the sensitivity increased. Different parasite dilutions showed similar results. Conclusion: It is important to increase the sensitivity especially with the aim for detecting LRV in clinical samples obtained from patients probably have less number of parasites. The presence and burden of the virus can help to understand the relationship between the clinical findings and the pathogenicity of the parasite which may lead to changes in the course of treatment. © 2022 Turkish Society for Parasitology.