Browsing by Author "Karakus, M"
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Item Detection of Leishmania RNA virus 2 in Leishmania species from TurkeyNalçaci, M; Karakus, M; Yilmaz, B; Demir, S; Özbilgin, A; Özbel, Y; Töz, SBackground: 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.Item Synthesis, crystal structure and ab initio/DFT calculations of a derivative of dithiophosphonatesKarakus, M; Solak, S; Hökelek, T; Dal, H; Bayrakdar, A; Kart, SÖ; Karabacak, M; Kart, HHThe compound 2 has been synthesized from the reaction of 2,4-Bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfide and (R)-1-[3,5-Bis(trifloromethyl)phenyl]ethanol in toluene. The obtained crude dithiophosphonic acid 1 has been treated with the excess of N(C2H5)(3) to give rise to 2, [(+HN(C2H5)(3)][(O-CH3CH-C6H3(CF3)(2))(CH3OC6H4)PS2-]. The compound 2 has been characterized by using the spectroscopic methods such as IR, H-1, C-13, P-31 NMR and structural analysing method such as X-ray crystallography. It crystallizes in the orthorhombic system, whose space group is P2(1)2(1)2(1). It consists of a dithiophosphonate bridged methoxyphenyl and bis(triflorophenylethyl) groups and a triethylammonium moiety linked by N-H center dot center dot center dot S and C-H center dot center dot center dot F hydrogen bonds. In the crystal structure, the C17H14F6O2PS2 molecule is elongated along the b-axis and stacked along the a-axis. The triethylammonium, N(CH2CH3)(3), molecule fill in the cavities between the C17H14F6O2PS2 molecule. Moreover, ab initio methods based on Hartree-Fock (HF) and Density Functional Theory (DFT) calculations with the basis set of 6-31G(d) are also carried out to determine the molecular structural properties and to calculate FT-IR and NMR spectrum of the compound 2. The experimental results and theoretical calculations have been compared, and they are found to be in good agreement. (C) 2013 Elsevier B.V. All rights reserved.Item sandflyDST: a dynamic web-based decision support tool for the morphological identification of sandflies present in Anatolia and mainland Europe, and user studyKarakülah, G; Karakus, M; Suner, A; Demir, S; Arserim, SK; Töz, S; Özbel, YSpecies 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) over bar (SUS Score) = 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.Item Does vaginal douching affect the risk of vaginal infections in pregnant women?Sakru, N; Inceboz, T; Inceboz, U; Zeren, I; Karakus, M; Kirca, UObjective: To evaluate the relationship between vaginal douching and vaginal infections among women in early pregnancy. Methods: We conducted this study in the Department of Gynecology and Obstetrics, Family Planning Center, Dr. E. Hayri Ustundag Hospital, Izmir, Turkey, between March 2003 and December 2004. We examined the vaginal swabs of 129 women, asking for termination of pregnancy in a family-planning center as both wet-preparations and cultures for vaginal microorganisms, and recorded the informations on women's vaginal douching habit. Results: Among 129 women examined, 80 had at least one type of vaginal microorganisms. Of 67 vaginal douche users, 48 (71.6%) had at least one type of vaginal organisms, whereas of 62 nonusers, only 32 (51.6%) had microorganism, although age, educational status, coital frequency, age at the first intercourse were not statistically different between the vaginal douche-users and non-users. Especially, Group B Streptococcus (GBS), Enterococcus spp. and Candida spp. were found more frequent in vaginal douche-user women. Conclusion: We found that vaginal douching tends pregnant women to genital tract the incidence of vaginal infections, especially those caused by Enterococcus spp. and GBS. As such infections may render such women to high risk in terms of perinatal mortality and morbidity, it would be appropriate to discourage vaginal douching in pregnant women.Item Evaluation of conjunctival swab sampling in the diagnosis of canine leishmaniasis: A two-year follow-up study in Cukurova Plain, TurkeyKarakus, M; Töz, S; Ertabaklar, H; Pasa, S; Atasoy, A; Arserim, SK; Ölgen, MK; Alkan, MZ; Durrant, C; Özbel, YThe 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 Cukurova 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 Cukurova 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. (C) 2015 Elsevier B.V. All rights reserved.Item Vector and reservoir surveillance study in a canine and human leishmaniasis endemic area in most western part of Turkey, KaraburunKarakus, M; Arserim, SK; Kasap, ÖE; Pekagirbas, M; Aküzüm, D; Alten, B; Töz, S; Özbel, YLeishmaniasis 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.Item 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 ProvinceLimoncu, ME; Balcioglu, IC; Töz, S; Demir, S; Kavur, H; Karakus, M; Vardarli, AT; Özbel, YCutaneous (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/syriacusTonnoir 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 tropics 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.Item Detection of Leishmania major and Leishmania tropica in domestic cats in the Ege Region of TurkeyPasa, S; Vardarli, AT; Erol, N; Karakus, M; Töz, S; Atasoy, A; Balcioglu, IC; Tuna, GE; Ermis, ÖV; Ertabaklar, H; Özbel, YLeishmaniosis 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. (C) 2015 Elsevier B.V. All rights reserved.Item The current clinical and geographical situation of cutaneous leishmaniasis based on species identification in TurkeyÖzbilgin, A; Töz, S; Harman, M; Topal, SG; Uzun, S; Okudan, F; Güngör, D; Erat, A; Ertabaklar, H; Ertug, S; Gündüz, C; Çavus, I; Karakus, M; Ural, IÖ; Ölgen, MK; Kayabasi, Ç; Kurt, Ö; Özbel, YLeishmaniases are a group of vector-borne diseases caused by the members of Leishrnania 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. infant= 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.Item Leishmaniasis in Turkey: first clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regionsÖzbilgin, A; Çulha, G; Uzun, S; Harman, M; Topal, SG; Okudan, F; Zeyrek, F; Gündüz, C; Östan, I; Karakus, M; Töz, S; Kurt, Ö; Akyar, I; Erat, A; Güngör, D; Kayabasi, Ç; Çavus, I; Bastien, P; Pratlong, F; Kocagöz, T; Özbel, YObjectiveTo report isolation of Leishmaniamajor strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L.major between 2011 and 2014. MethodsInitial 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. ResultsMelting 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 3weeks, which even progressed to extremity amputation. ConclusionCutaneous 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. ObjectifRapporter sur l'isolement de souches de Leishmania major obtenues a partir de 18 cas de leishmaniose cutanee (LC) de patients turcs autochtones infectes par L. major entre 2011 et 2014. MethodesLe diagnostic initial a porte sur la microscopie et la culture sur un milieu enrichi, prepare en ajoutant des quantites specifiques d'extrait de foie, de proteines et de sources de lipides au milieu NNN. Les promastigotes ont ensuite ete transferes dans le milieu RPMI contenant 10% de serum fOEtal de veau pour la culture de masse. La PCR en temps reel specifique de l'espece et ciblant la region ITS1 de Leishmania spp. a ete realisee en utilisant a la fois les echantillons d'aspiration de la lesion et de promastigotes cultives. Deux des 18 isolats ont ete identifies par analyse des isoenzymes dans le Centre de reference de la leishmaniose a Montpellier, en France. Chaque isolat a ete inocule dans les coussinets plantaires de six souris pour observer la pathogenicite de L. major. Les lesions en developpement ont ete observees et l'epaississement des coussinets plantaires ont ete mesures chaque semaine. ResultatsLes analyses de courbe de fusion des 18 isolats ont montre un pic concordant avec L. major et deux d'entre eux ont ete confirmes par des analyses d'isoenzyme comme L. major de zymodeme MON103. Dans le modele murin, des lesions aigues observees au jour 21 ont ete acceptees comme une indication de forte infection. Des deficiences severes ont ete observees sur tous les coussinets plantaires des souris pendant plus de trois semaines, qui ont meme progresse jusqu'a l'amputation de l'extremite. ConclusionL. major causant la LC a ete recemment identifie dans la province d'Adana dans le sud de la Turquie par la PCR. Notre etude montre que de tels cas de LC ne sont pas limites a Adana, mais sont actuellement presents dans l'ouest et dans le sud-est de l'Anatolie, et le long de la cote mediterraneenne. Le role des petits mammiferes, principaux reservoirs de L. major en Anatolie, devrait etre elucide, de meme que les facteurs sous-jacents qui causent les manifestations cliniques severes dans les infections a L. major en Turquie, contrairement aux infections dans les pays voisins. ObjetivoReportar el aislamiento de cepas de L. major obtenidas de 18 pacientes turcos con leishmaniosis cutanea (LC) autoctona, infectados con Leishmania major entre 2011 y 2014. MetodosEl diagnostico inicial se realizo mediante microscopia y cultivo en medio enriquecido, preparado mediante la adicion de cantidades especificas de extracto de higado, proteina y fuentes de lipido al medio NNN. Los promastigotes fueron transferidos al medio RPMI con un 10% de suero fetal para su cultivo masivo. Se realizo PCR en tiempo real, especie-especifica, que detecta la region ITS1 de Leishmania spp., utilizando tanto muestras aspiradas de las lesiones como promastigotes de cultivo. Dos de los 18 aislados se identificaron mediante analisis isoenzimatico en el Centro de Referencia de la Leishmaniosis en Montpellier, Francia. Cada aislado fue inoculado en las almohadillas de las patas de seis ratones para observar la patogenicidad de L. major. Se observo el desarrollo de las lesiones y se midio semanalmente el engrosamiento de las almohadillas. ResultadosEl analisis de la curva de fusion de los 18 aislados mostro un pico de concordancia con Leishmania major, y dos de ellos fueron confirmados mediante un analisis isoenzimatico como L. major zymodeme MON103. En el modelo de raton, las lesiones agudas observadas en el dia 21 se aceptaron como indicativas de una infeccion masiva. Se observaron danos graves en todas las almohadillas de los ratones a lo largo de tres semanas, que progresaron hasta la amputacion de las extremidades. ConclusionRecientemente se ha identificado, mediante PCR, LC causada por L. major en la provincia de Adana al sur de Turquia. Nuestro estudio muestra que estos casos de LC no estan limitados a Adana y que actualmente existen desde el oeste al sudeste de Anatolia y a lo largo de la costa Mediterranea. Es necesario aclarar el papel que en Anatolia juegan los pequenos mamiferos, principales reservorios de L. major, al igual que el de los factores que hay detras de las manifestaciones clinicas severas en infecciones por L. major en Turquia, al contrario del de las infecciones presentes en paises vecinos.Item Epidemiological analysis of Leishmania tropica strains and giemsa-stained smears from Syrian and Turkish leishmaniasis patients using multilocus microsatellite typing (MLMT)Karakus, M; Nasereddin, A; Onay, H; Karaca, E; Özkeklikçi, A; Jaffe, CL; Kuhls, K; Özbilgin, A; Ertabaklar, H; Demir, S; Özbel, Y; Töz, STurkey 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 Sanliurfa 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 (Delta K) 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 Sanliurfa 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.Item Determination of sand fly fauna and molecular detection of Leishmania in sand flies in Antalya Province, Southern TurkeyArserim, SK; Çetin, H; Karakus, M; Demir, S; Ser, Ö; Töz, S; Balcioglu, IC; Ölgen, MK; Yilmaz, B; Özbel, YVisceral 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.Item Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused by Leishmania donovani in TurkeyÖzbilgin, A; Harman, M; Karakus, M; Bart, A; Töz, S; Kurt, Ö; Çavus, I; Polat, E; Gündüz, C; Van Gool, T; Özbel, YIn Turkey, the main causative agents are Leishmania tropica (L. tropica) and Leishmania infantum (L. infant:tun) 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. infcrnuttn, 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.Item Detection of permethrin resistance and phylogenetic clustering of turkish head lice (Pediculus humanus capitis; De Geer, 1767 populationsKarakus, M; Atici, T; Karabela, SN; Baylan, O; Limoncu, ME; Balcioglu, ICHead lice infestation caused by Pediculus humanus capitis De Geer, 1767 is one of the most common public health problems. The relationship between humans and head lice dates back millions of years ago that differentiated into different phylogenetic clades. Treatment of head lice infestation usually based on insecticide-based products, which promotes the resistance in the head lice populations. In the present study, we aimed to screen the presence of permethrin resistance among collected P. h. capitis specimens in Turkey. Three mutation sites (T917I, L920F, and M815I) were screened using real-time PCR and resistance was identified by melt analysis. Of the studied specimens, resistance allele frequency (RAF) was found 0.98 for T917I, 0.99 for L920F, and 1.00 for M815I. The phylogenetic study revealed that Clade A and Clade B are present and overlap in Turkey. The present study is first to screen the resistance among Turkish head lice specimens. To not stimulate the pyrethroids resistance in head lice populations, early detection of resistance is crucial and will help the health professionals to choose suitable formula in the treatment. We suggest that the resistance status needs to be screened in randomly selected populations before any treatment application is given.Item Treatment of head lice (Pediculus humanus capitis) infestation: Is regular combing alone with a special detection comb effective at all levels?Kurt, Ö; Balcioglu, IC; Limoncu, ME; Girginkardesler, N; Arserim, SK; Görgün, S; Oyur, T; Karakus, M; Düzyol, D; Gökmen, AA; Kitapçioglu, G; Özbel, YHead lice infestation (HLI) caused by Pediculus humanus capitis has been a public health problem worldwide. Specially designed combs are used to identify head lice, while anti-lice products are applied on the scalp for treatment. In the present study, we aimed to test whether combing only by precision detection comb (PDC) or metal pin comb (MPC) could be effective alternatives to the use of anti-lice products in children. A total of 560 children from two rural schools in Turkey were screened. In the PDC trial, children were combed every second day for 14 days, while in the MPC trial, combing was performed once in every four days for 15 days. Children were divided into two groups (dry combing and wet combing) for both trials and results were compared. The results showed no significant differences between dry and wet combing strategies for both combs for the removal of head lice (p>0.05). The number of adult head lice declined significantly on each subsequent combing day in both approaches, except on day 15 in the MPC trial. In the end, no louse was found in 54.1 and 48.9% of children in the PDC and MPC trials, respectively. Since family members of infested children were not available, they were not checked for HLI. Four times combing within 2 weeks with MPC combs was found effective for both treatment of low HLI and prevention of heavy HLI. In conclusion, regular combing by special combs decreases HLI level in children and is safely applicable as long-term treatment.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 TurkeyÖzbel, Y; Karakus, M; Arserim, SK; Kalkan, SO; Töz, SHuman visceral leishmaniasis (VL) is reported from 38 provinces of Turkey and dogs are accepted as main reservoir hosts. Kusadasi 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 ofPhlebotomus 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. (C) 2016 Elsevier B.V. All rights reserved.