Browsing by Author "Tunali, V"
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Item Effects of Efflux Pump Inhibitors and Antileishmanial Drug Combinations on Leishmania tropica and Leishmania infantum IsolatesÖzel, Y; Çavus, I; Tunali, V; Aksoy, T; Ünlü, M; Özbilgin, ADrug resistance, one of the most important public health problems facing humanity, necessitates new strategies and approaches in the development of antileishmanial agents. Although developments regarding efflux pump inhibitors (EPIs) and other candidate agents are promising, the search continues to increase the duration of use and efficacy of existing antileishmanials. The aim of this study was to investigate the antileishmanial effects of three EPIs, namely reserpine, berberine and verapamil. The antileishmanial activities of EPIs, pentostam and miltefosine against Leishmania tropica and Leishmania infantum strains were determined by broth microdilution method. Minimum parasiticidal concentration (MPC) values were determined by inverted microscope and IC50 values were determined by MTT viability assay method. The effects of EPIs with determined antileishmanial activities on miltefosine and pentostam were investigated by checkerboard method. The MPC values of antileishmanial drugs miltefosine and pentostam for L.tropica and L.infantum were determined as 64 and 196 pg/mL at 24 and 48 hours, respectively. The MPC values of EPIs reserpine and berberine were determined as 314 and 64 pg/mL for the same incubation times and for verapamil, they were determined as 80 pg/mL at 24 hours and 40 pg/mL at 48 hours. Among anti-leishmanials, the IC50 values of miltefosine were calculated as 4.91/3.47 and 4.05/2.91 pg/mL for L.tropica and L.infantum at 24 and 48 hours, respectively and that of pentostam were calculated as 34.58/59.86 and 18.48/40.63 pg/mL at the same incubation times, respectively.The IC50 values of EPIs, reserpine, berberine, and verapamil were calculated as 74.05/50.61, 7.27/6.1, and 12.52/4.53 pg/mL for L.tropica at 24/48 hours, respectively and 64.52/51.72, 8.21/8.01, and 11.59/7.69 pg/mL for L.infantum. When miltefosine was combined with reserpine, berberine or verapamil, synergistic interactions were observed at 24 and 48 hours of incubation. When pentostam was combined with reserpine, partial synergy was observed at 24 hours and synergy was observed at 48 hours. When pentostam was combined with berberine or verapamil, synergistic interactions were observed at both incubation conditions. Synergy results were found to be the same in both L.tropica and L.infantum strains. In recent years, the speed of research on the discovery of new antimicrobials has significantly decreased and it has become necessary to investigate new molecules that can affect resistance mechanisms. It is thought that EPIs may be a promising approach that can increase the clinical performance of antileishmanial agents and reduce the level of side effects in the fight against drug resistance.Item Knock, knock, knocking on Europe's door: Threat of leishmaniasis in Europe with a focus on TurkeyTunali, V; Özbilgin, ALeishmaniasis epidemiology is currently undergoing substantial transformations in both Turkey and Europe, signifying potential implications for public health. This review analyzes the evolving patterns within Turkey and their potential ramifications for Europe. Within Turkey, the dynamics of leishmaniasis are undergoing noteworthy alterations, manifesting in a rise in cutaneous leishmaniasis (CL) cases and the emergence of Leishmania major and Leishmania donovani. These transformations are predominantly driven by factors such as the distribution of vectors, human activities, climate fluctuations, and migration. Across Europe, particularly in countries within the Mediterranean basin, leishmaniasis is endemic, primarily attributed to Leishmania infantum. Recent evidence suggests a resurgence of the disease even in previously non-endemic areas, propelled by climate change, urbanization, and migration. The changing landscape of leishmaniasis in Turkey carries direct implications for Europe. The presence and distribution of Leishmania tropica, L. major, and L. donovani raise concerns regarding cross-border transmission. Turkey's strategic position along migration routes further compounds the risk, alongside the facilitative effects of climate change and host mobility. Embracing a One Health approach with public awareness campaigns should be a priority. To ensure the protection of public health in Europe, it is imperative to adopt a proactive approach by establishing robust surveillance mechanisms, implementing preventive measures, and cultivating collaboration with Turkey. The invaluable experience, strategic geographical location, and well-established infrastructure of Turkey make this collaboration crucial in effectively addressing the evolving dynamics of leishmaniasis and its potential impacts on Europe.Item Effectiveness of Tablet Computer Use in Achievement Of Schedule-Following Skills by Children with Autism Using Graduated GuidanceEliçin, Ö; Tunali, VTablet computers are frequently used in the education of children with autism. Studies show that educational activities involving tablet computers have positive effects on the acquisition of target skills by children with autism. In this research, the effectiveness of the use of tablet computers in teaching children with autism to use schedules was evaluated. Three male students of ages 5 to 7 diagnosed with autism participated in the research. A single-case multiple-probe-design-across-participants was used in the research. Independent variable of the research was the education provided via tablet computers using the graduated guidance technique, and the dependent variable was the acquisition of schedule-following skill. Follow-up and generalization data were collected on the 7th, 14th, and 21st days after the teaching sessions ended. Follow-up sessions were completed with 100% performance for all three subjects. Generalized correct responses were found 90% for one subject, and 100% for the other two. The results indicated that the subjects acquired, maintained, and generalized the schedule-following skills after the teaching sessions completed. In addition, the teachers of the subjects were asked their opinions about the application. These opinions that form the social validity data of the research support the research findings.Item The Importance of the Contribution of Rapid Test, Serological and Molecular Methods in the Diagnosis of Two Imported Malaria Cases with Atypical MicroscopyZorbozan, O; Pullukçu, H; Sahar, EA; Karakavuk, M; Can, H; Tunali, V; Döskaya, M; Turgay, N; Töz, S; Özbilgin, AMalaria is a widespread and life-threatening disease in tropical and subtropical regions. In patients with typical clinical symptoms, malaria is considered as a preliminary diagnosis if there is a travel history to malaria-endemic areas. The basis of the laboratory diagnosis of malaria is the microscopic examination of Giemsa stained smears. On the other hand, the diagnosis and differentiation of Plasmodium species with microscopic examination may have some difficulties. In the first case, adifferent appearance from the classical Plasmodium vivax erythrocytic forms in infected erythrocytes were detected in 1% of all erythrocytes in thin smear blood preparations of a 26-year-old male with complaints of fever and chills and a story of travel to Nigeria. It was observed that parasitic nuclei were not prominent, and were located in the cytoplasm irregularly as chromatin or dye particles, nucleus fragments similar to Schuffner's granules in the form of scattered and granular spots were present in some erythrocytes, the cytoplasm of some Plasmodium erythrocytic forms were irregular and nuclei were not seen. There were no Schuffner's granules in any of the infected erythrocytes. P. vivax was detected by the rapid diagnostic test (OptiMAL, DiaMed GmbH, Switzerland), which searches for the antigens of Plasmodium species, in the peripheral blood sample of the patients. The P. vivax 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibodies against Plasmodium species were searched by using the Pan Malaria Antibody CELISA (CeLLabs Pty Ltd, Brookvale, Australia) kit in the patient's serum sample and the optical density (OD) value of the patient sample was measured five times the OD value of the positive control. In the second case, adifferent appearance from the classical P. falciparum erythrocytic forms in infected erythrocytes were detected in 12% of all erythrocytes in thin smear blood preparations of a 31-year-old male who has been suffering from persistent fever, severe headache, pain in the eyes and was known to be working in Nigeria. It was observed that some Plasmodium trophozoites have 1/3 of the size of erythrocytes such as P. vivax and have non-granular cytoplasm, some erythrocytic forms were round and the nucleus and cytoplasm were hardly distinguished, some of them were seen as crescent and close to the nucleus of the cytoplasm and some erythrocytic forms had characteristically a single nucleus and a scattered cytoplasm, similar to mature trophozoites of P. vivax. Although the Plasmodium young trophozoites were similar to P. vivax in means of magnitude, the forms in which the nuclei adhered to the erythrocyte wall were common. There were no P. falciparum gametocyte forms. P. falciparum like young trophozoite was observedonly in one of the four smears. P. falciparum was detected by the commercial rapid diagnostic test and P. falciparum 18S rRNA gene was also detected by the multiplex real-time polymerase chain reaction. Antibody formation against Plasmodium species was not detected in the ELISA test. In these case reports, the importance of the support of rapid diagnostic tests, serological and molecular methods to microscopic diagnosis and species determination of two imported malaria cases were demonstrated.Item Culture-independent assessment of probiotic supplement consistency in commercially available probiotic supplementsGundogdu, A; Nalbatoglu, OU; Sarikaya, I; Karis, G; Erdogan, MN; Hora, M; Tunali, VThe probiotic products market has experienced significant growth in recent years, driven by increasing consumer demand for health benefits. This demand is largely fueled by the widespread promotion of probiotic products, and is compounded by relatively lax regulations regarding these products compared to those for drugs. In this study, we sought to evaluate the consistency of product labels and contents for probiotic supplements in the Turkish and USA markets. A total of 44 commercially available probiotic products were included in the analysis, comprising 40 products from Turkey and 4 from the USA. Shotgun metagenome sequencing was performed to determine the microbiological content and functional characteristics of the products. Additionally, we evaluated the presence of antibiotic-resistance genes and virulence genes in each product. Our findings showed that 38.6% of the products contained microorganisms not indicated on their labels, while 15.9% of products did not contain one or more of the probiotic microorganisms specified on their labels. Moreover, over half of the probiotic products (68.2%) were found to contain at least one resistance gene, and 40.9% of the products carried multiple virulence genes. Streptococcus thermophilus being the primary contributor to the detected virulence genes. This study is the first to examine the label and content discrepancies of probiotic products in the Turkish market and one of the largest to analyze resistance gene profiles of probiotic supplements in the literature. These findings may suggest a potential public health risk and the need for whole genome sequencing analysis of probiotic supplements.Item Infecting Glial Cells with Antimony Resistant Leishmania tropica: A New ex-vivo ModelZorbozan, O; Harman, M; Evren, V; Erdogan, MA; Kilavuz, A; Tunali, V; Çavus, I; Yilmaz, Ö; Özbilgin, A; Turgay, NLeishmaniasis is a vector-borne zoonotic disease that shows different clinical features like cutaneous, mucocutaneous, visceral and viscerotropic forms. The protocols used in the treatment of leishmaniasis are toxic and have many limitations during administration. One of the limitations of treatment is the resistance against the protocols in practice. There is also a need to define new treatment options especially for resistant patients. Ex-vivo models using primary cell cultures may be a good source for evaluating new drug options in patients with antimony resistance, in addition to in-vitro and in-vivo studies. In this study, it was aimed to define a new ex-vivo culture model to evaluate treatment options in patients with cutaneous leishmaniasis who did not respond to treatment. In our experimental model of ex-vivo infection, Leishmania tropica promastigotes isolated from a case previously diagnosed with cutaneous leishmaniasis were used. The primary astroglial cell culture used for the ex-vivo model was prepared from 2-3 days old neonatal Sprague Dawley rat brains under sterile conditions by the modification McCarthy's method. The astroglia cells, which reached sufficient density, were infected with antimony resistant L.tropica promastigotes. After 24 hours of incubation, the supernatant on the cells were collected, the cell culture plate was dried at room temperature, then fixed with methyl alcohol and stained with Giemsa to search for L.tropica amastigotes. Amastigotes were intensely observed in glia cells in primary cell cultures infected with L.tropica promastigotes. No promastigotes were seen on Giemsa stained preparations of the precipitates prepared from the bottom sediment after the centrifugation of the liquid medium removed from the infected plates. In this study, promastigotes from a cutaneous leishmaniasis patient unable to respond to pentavalent antimony therapy were shown to infect rat glia cells and converted to amastigote form. This amastigote glial cell model, as far as we know, is the first model in the literature produced by L.tropica. The occurrence of L.tropica amastigote forms in glia cells may be indicative of the ability of Leishmania species to infect the central nervous system. The central nervous system may be an area for the Leishmania amastigotes to escape from the immune system in cases of leishmaniasis without a treatment response. Our study is important because it is the first study to show the infection of glia cells with L.tropica amastigotes.Item Emerging and Re-Emerging Parasitic Infections of the Central Nervous System (CNS) in EuropeTunali, V; Korkmaz, MIn a rapidly evolving global landscape characterized by increased international travel, migration, and ecological shifts, this study sheds light on the emergence of protozoal and helminthic infections targeting the central nervous system (CNS) within Europe. Despite being traditionally associated with tropical regions, these infections are progressively becoming more prevalent in non-endemic areas. By scrutinizing the inherent risks, potential outcomes, and attendant challenges, this study underscores the intricate interplay between diagnostic limitations, susceptibility of specific population subsets, and the profound influence of climate fluctuations. The contemporary interconnectedness of societies serves as a conduit for introducing and establishing these infections, warranting comprehensive assessment. This study emphasizes the pivotal role of heightened clinician vigilance, judicious public health interventions, and synergistic research collaborations to mitigate the potential consequences of these infections. Though rare, their profound impact on morbidity and mortality underscores the collective urgency required to safeguard the neurological well-being of the European populace. Through this multifaceted approach, Europe can effectively navigate the complex terrain posed with these emergent infections.Item Response to Dahly and MorrisTunali, V; Ermis, BH; Nalbantoglu, ÖUItem Autochthonous transmission of Leishmania donovani and Leishmania major with all the components of infection cycle at Europe's doorstepÖzbilgin, A; Tunali, V; Akar, SS; Yildirim, A; Sen, S; Çavus, I; Zorbozan, O; Gündüz, C; Turgay, N; Inanir, IObjectives: Leishmaniasis is a vector-borne disease and dogs may act as urban reservoirs. Turkey and most of the Mediterranean basin countries are endemic for leishmaniasis. In this study, it is aimed to report the autoch-thonous leishmaniasis cases, with all the components of the infection cycle (reservoir, vector, and the host) in a region close to Europe. Methods: Nine human and four canine autochthonous leishmaniasis cases were included in the study. Direct microscopy, culture methods, serological, and molecular tests were applied to the samples obtained from the cases. Results: VL and CL patients consisted of 2 L.infantum, 1 L. donovani, 2 L. tropica, and 2 L. tropica,1 L. major,1 L. infantum infected patients respectively. CanL cases were infected with L. infantum, L. donovani, L. tropica, and L. major. Conclusions: All the cases were autochthonous cases located in Manisa province. As Greece and all the Mediterranean basin countries in Europe share competent vectors, it is concluded that the detection of all 4 species of Leishmania parasites in such proximity to Europe poses an important public health threat for Europe. This study reports all four species of Leishmania spp., including L. major and L. donovani in close proximity to continental Europe.Item Efficacy of AI-Assisted Personalized Microbiome Modulation by Diet in Functional Constipation: A Randomized Controlled TrialArslan, NÇ; Gündogdu, A; Tunali, V; Topgül, OH; Beyazgül, D; Nalbantoglu, ÖUBackground: Currently, medications and behavioral modifications have limited success in the treatment of functional constipation (FC). An individualized diet based on microbiome analysis may improve symptoms in FC. In the present study, we aimed to investigate the impacts of microbiome modulation on chronic constipation. Methods: Between December 2020-December 2021, 50 patients fulfilling the Rome IV criteria for functional constipation were randomized into two groups. The control group received sodium picosulfate plus conventional treatments (i.e., laxatives, enemas, increased fiber, and fluid intake). The study group underwent microbiome analysis and received an individualized diet with the assistance of a soft computing system (Enbiosis Biotechnology (R), Sariyer, Istanbul). Differences in patient assessment constipation-quality of life (PAC-QoL) scores and complete bowel movements per week (CBMpW) were compared between groups after 6-weeks of intervention. Results: The mean age of the overall cohort (n = 45) was 31.5 +/- 10.2 years, with 88.9% female predominance. The customized diet developed for subjects in the study arm resulted in a 2.5-fold increase in CBMpW after 6-weeks (1.7 vs. 4.3). The proportion of the study group patients with CBMpW > 3 was 83% at the end of the study, and the satisfaction score was increased 4-fold from the baseline (3.1 to 10.7 points). More than 50% improvement in PAC-QoL scores was observed in 88% of the study cohort compared to 40% in the control group (p = 0.001). Conclusion: The AI-assisted customized diet based on individual microbiome analysis performed significantly better compared to conventional therapy based on patient-reported outcomes in the treatment of functional constipation.Item Artificial intelligence-based personalized diet: A pilot clinical study for irritable bowel syndromeKarakan, T; Gundogdu, A; Alagözlü, H; Ekmen, N; Ozgul, S; Tunali, V; Hora, M; Beyazgul, D; Nalbantoglu, OUWe enrolled consecutive IBS-M patients (n = 25) according to Rome IV criteria. Fecal samples were obtained from all patients twice (pre-and post-intervention) and high-throughput 16S rRNA sequencing was performed. Six weeks of personalized nutrition diet (n = 14) for group 1 and a standard IBS diet (n = 11) for group 2 were followed. AI-based diet was designed based on optimizing a personalized nutritional strategy by an algorithm regarding individual gut microbiome features. The IBS-SSS evaluation for pre- and post-intervention exhibited significant improvement (p < .02 and p < .001 for the standard IBS diet and personalized nutrition groups, respectively). While the IBS-SSS evaluation changed to moderate from severe in 78% (11 out of 14) of the personalized nutrition group, no such change was observed in the standard IBS diet group. A statistically significant increase in the Faecalibacterium genus was observed in the personalized nutrition group (p = .04). Bacteroides and putatively probiotic genus Propionibacterium were increased in the personalized nutrition group. The change (delta) values in IBS-SSS scores (before-after) in personalized nutrition and standard IBS diet groups are significantly higher in the personalized nutrition group. AI-based personalized microbiome modulation through diet significantly improves IBS-related symptoms in patients with IBS-M. Further large-scale, randomized placebo-controlled trials with long-term follow-up (durability) are needed.Item A Multicenter Randomized Controlled Trial of Microbiome-Based Artificial Intelligence-Assisted Personalized Diet vs Low-Fermentable Oligosaccharides, Disaccharides, Monosaccharides, and Polyols Diet: A Novel Approach for the Management of Irritable Bowel SyndromeTunali, V; Arslan, NC; Ermis, BH; Hakim, GD; Gündogdu, A; Hora, M; Nalbantoglu, OUINTRODUCTION: Personalized management strategies are pivotal in addressing irritable bowel syndrome (IBS). This multicenter randomized controlled trial focuses on comparing the efficacy of a microbiome-based artificial intelligence-assisted personalized diet (PD) with a low-fermentable oligosaccharides, disaccharides, monosaccharides, and polyols diet (FODMAP) for IBS management. METHODS: One hundred twenty-one patients participated, with 70 assigned to the PD group and 51 to the FODMAP diet group. IBS subtypes, demographics, symptom severity (IBS-SSS), anxiety, depression, and quality of life (IBS-QOL) were evaluated. Both interventions spanned 6 weeks. The trial's primary outcome was the within-individual difference in IBS-SSS compared between intervention groups. RESULTS: For the primary outcome, there was a change in IBS-SSS of -112.7 for those in the PD group vs -99.9 for those in the FODMAP diet group (P = 0.29). Significant improvement occurred in IBS-SSS scores (P < 0.001), frequency (P < 0.001), abdominal distension (P < 0.001), and life interference (P < 0.001) in both groups. In addition, there were significant improvements in anxiety levels and IBS-QOL scores for both groups (P < 0.001). Importantly, PD was effective in reducing IBS SSS scores across all IBS subtypes IBS-Constipation (IBS-C; P < 0.001), IBS-Diarrhea (IBS-D; P = 0.01), and IBS-Mixed (IBS-M; P < 0.001) while FODMAP diet exhibited comparable improvements in IBS-C (P = 0.004) and IBS-M (P < 0.001). PD intervention significantly improved IBS-QOL scores for all subtypes (IBS-C [P < 0.001], IBS-D [P < 0.001], and IBS-M [P = 0.008]) while the FODMAP diet did so for the IBS-C (P = 0.004) and IBS-D (P = 0.022). Notably, PD intervention led to significant microbiome diversity shifts (P < 0.05) and taxa alterations compared with FODMAP diet. DISCUSSION: The artificial intelligence-assisted PD emerges as a promising approach for comprehensive IBS management. With its ability to address individual variation, the PD approach demonstrates significant symptom relief, enhanced QOL, and notable diversity shifts in the gut microbiome, making it a valuable strategy in the evolving landscape of IBS care.Item Overcoming the Challenge;In VivoEfficacy of Miltefosine for Chronic Cutaneous LeishmaniasisTunali, V; Harman, M; Çavus, I; Gündüz, C; Özbilgin, A; Turgay, NBackground Cutaneous Leishmaniasis (CL) is the most common form of leishmaniasis. CL can be divided into two major groups: acute CL (ACL) and chronic CL (CCL). The aim of this study is to compare the efficacy of miltefosin and pentavalent antimony compoundsin vivowith the CCL patient samples. Materials Three study groups were formed, each consisting of five male Mus musculus (Balb/C) mice. In this model, promastigotes from the culture of a CCL patient were utilized. 100 mu LL. tropicapromastigote suspension with a density of 10(8)promastigotes/ml were injected into the hint-right footpad of each experimental animal intradermally. Footpads of the mice were measured every two weeks until 24th week. From the 13th week, miltefosin 50 mg/kg/day was administered orally using gavage for 21 days, Meglumin antimoniate (MA) was administered by intramuscular (IM) injection daily for 21 days at 50 mg/kg/day and saline was administered IM for 21 days for the miltefosine, MA and control group, respectively. Results The footpad measurements of the miltefosine group were lower than the control group statistically. Between the MA group and the miltefosine group and MA group and the control group, there was no statistically significant difference. Giemsa stained slides revealed amastigotes in one, two and all of the slides for the miltefosine, MA and control group, respectively. Molecular tests were performed with the Rotor-Gene device andL. tropicaconsistent peaks were obtained in one of the miltefosine group, four in the MA group and all mice in the control group. Conclusions Demonstration of both clinical and laboratory improvement in four of the five experimental animals provides strong evidence that miltefosine is an effective drug in the treatment of CCL. In the literature, no clinical or laboratory studies using miltefosine have been performed with CCL patients only.Item Visceral Leishmaniasis Caused by Leishmania TropicaÖzbilgin, A; Tunali, V; Çavus, I; Tetik, AV; Dinç, M; Yalçin, T; Gündüz, C; Beyaz, M; Köse, SPurposeIn Turkey, the main causative agent of visceral leishmaniasis (VL) is Leishmania. infantum and the main causative agent of cutaneous leishmaniasis (CL) is Leishmania tropica. In this study, we aimed to discuss the possible mechanisms, clinical aspects, and threat of visceralizing L. tropica.MethodsThis study includes seven cases of VL caused by L. tropica.Five patients were male (71%) and four were adults (57%).ResultsAll the VL patients complained of fever and splenomegaly. Fatigue, pancytopenia, and hepatomegaly were present in six patients each (86%), while weight loss and gastrointestinal system (GIS) symptoms were present in 5 patients (71%).ConclusionsIn this study, we have evaluated seven cases of visceralized L. tropica (VLT) in the context of the changing leishmaniasis epidemiology in Turkey. We have evaluated the possible mechanisms of visceralization; inter- and intraspecies genetic exchange with all the old world leishmaniasis agents present in the region, stress induced by inappropriate use of drugs, and possible ongoing adaptation mechanisms of Leishmania spp. The threat posed by VLT is significant as L. tropica is the most widespread and most common cause of leishmaniasis in Turkey. We do not know the vectorial capacity of the sand flies for the transmission of VLT strains or if these strains are in circulation in Turkey. Future studies should be carried out to investigate these issues as the transition of L. tropica from a mild disease-causing agent to a mortal one poses a significant public health concern for Turkey and Europe.Item First Presentation of the Complete Infection Cycle of Visceralizing Leishmania infantum/ donovani Hybrid Strain in Turkey: The Host, Reservoir and VectorÖzbilgin, A; Arserim, SK; Tünger, Ö; Çavus, I; Yildirim, A; Tunali, V; Gündüz, CObjective: Visceral Leishmaniasis (VL) is a widespread infectious disease of the reticuloendothelial system which is mostly caused by Leishmania infantum and L.donovani. VL is a zoonotic infection and dogs act as the main reservoirs of the disease while the vectors are the female Phlebotomus, that is to say, sandflies. Although Leishmania spp. are attributed to reproduce asexually, the genetic exchange between different strains has recently been demonstrated and causes interspecific hybrids. We aimed to present the complete infection cycle of a hybrid strain of Leishmania infantum/donovani with the host, reservoir, and vector.Materials and Methods: A patient presenting with VL proved to have a dog with canine leishmaniasis. Bone marrow aspirate from the patient and lymph node aspirates from the dog were inspected for Leishmania spp. amastigotes under light microscopy. Samples were cultured in enriched NNN medium. Serological examinations were made using IFAT for the patient serum and rK39 test for both human and dog samples. Light traps were placed to collect Phlebotomus. Leishmania molecular identification was executed by primers and probes specific to ITS-1 and cytochrome B gene region of Leishmania parasites.Results: Leishmania spp. amastigotes were detected in the slides while promastigotes were present in the enriched NNN medium. For patient serum, IFAT IgG was 1/256 positive, and rK39 was positive for both dog and human samples. Fifty-two Phlebotomus were caught (24 male, 28 female): 35 of them were P. similis, ten were P. neglectus/synacus, six were P .papatasi, and one was P. tobbi. Molecular analysis showed that the Leishmania spp. isolates from the patient, dog, and P.tobbi were identical. Gel image revealed two bands in all three samples, thus leading to the conclusion that the infecting strain was a hybrid of L. infantum and L. donovani. Conclusion: We found identical isolates that are putative hybrids of L. donovani and L. infantum in human, dog reservoir and sandfly..