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    Analysis of morphological variations of three Adlerius (Diptera: Psychodidae) species collected in two cutaneous leishmaniasis endemic foci of Turkey
    (Springer Int Publ Ag, 2022) Kavur, Hakan; Arikan, Huseyin; Toz, Seray; Balcioglu, Ibrahim Cuneyt; Ozbel, Yusuf
    Leishmaniasis, transmitted to human by sand flies, is a neglected vector-borne diseases and has two clinical forms: cutaneous (CL) and visceral leishmaniasis (VL) in Turkey. Subgenus of Adlerius are known as probable vectors of both forms of leishmaniasis in Turkey. Examined three species belonging to Adlerius are difficult to distinguish morphologically and have similar taxonomic characters. The aim of this study was to analyse and identify the possible morphological variations, of wild-caught sand fly specimens belonging to Adlerius subgenus. Six and seven morphological characters for female and male specimens were used, respectively. ANOVA, principal component analysis (PCA) and discriminant function analysis (DFA) test were performed to determine the significant morphometric variables of the three species. Univariate and multivariate analyses of 13 morphometric variables via ANOVA, DFA and PCA showed that eight (six for females, two for males) morphometric variables had an accuracy of 100% for discriminating three Adlerius subgenus specimens. The analyses revealed significant differences on several morphological structures of Adlerius specimens. These results suggested that morphological and morphometrical features can be used to discriminate three Adlerius species, in which one of them is proven vector Leishmania (L.) tropica in two foci of Turkey.
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    The association between Cytomegalovirus co-infection with Pneumocystis pneumonia and mortality in immunocompromised non-HIV patients
    (Wiley, 2018) Ekren, Pervin Korkmaz; Toreyin, Zehra Nur; Nahid, Payam; Doskaya, Mert; Caner, Ayse; Turgay, Nevin; Zeytinoglu, Aysin; Toz, Seray; Bacakoglu, Feza; Guruz, Yuksel; Erensoy, Selda
    Introduction Impact of Cytomegalovirus (CMV) co-infection pneumonia in non-HIV patients with Pneumocystis jirovecii pneumonia (PCP) is unclear. Objectives Methods The aim of our study was to determine whether CMV co-infection is associated with an increased risk of mortality. Our study was conducted at Ege University Hospital, Turkey. We used molecular assays to diagnose Pneumocystis jirovecii in respiratory samples, and CMV in both respiratory and blood samples. We compared morbidity and mortality stratified by CMV co-infection status. Results Conclusion Between 2009 and 2015, 43 patients (mean age: 56.7 +/- 15.3 years) were diagnosed with PCP. Only 3 of 43 patients had received PCP prophylaxis. We microbiologically confirmed CMV co-infection in 28 of 43 (65.1%) patients. Acute respiratory distress syndrome (ARDS) and requirement of mechanical ventilation were more common in the CMV co-infection group (P = .019 and P = .031 respectively), and duration of intensive care unit was also longer (P = .006). In univariate analyses, mortality at 30 days was higher in the CMV co-infection group as compared to the group with PCP alone (78.6% and 46.7% respectively; P = .046). In multivariate analyses, mortality was independently associated only with the presence of ARDS [OR: 6.22 95% CI 1.3-29.32] and the association with CMV co-infection was no longer significant [OR: 2.6 95% CI 0.49-13.72, P = .257]. The risk of mortality appears to be increased in the setting of CMV and PCP co-infection in HIV-uninfected immunocompromised patients. PCP prophylaxis use was lower than expected, suggesting low physician awareness of the risks of PCP in this population.
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    Blood Meal Analysis and Molecular Detection of Leishmania DNA in Wild-Caught Sand Flies in Leishmaniasis Endemic Areas of Turkey and Northern Cyprus
    (Springer Int Publ Ag, 2022) Yetismis, Kardelen; Mert, Ufuk; Caner, Ayse; Nalcaci, Muhammed; Toz, Seray; Ozbel, Yusuf
    Introduction Phlebotomine sand flies (Diptera: Psychodidae) are known as the vector of diseases such as leishmaniasis, bartonellosis and viral diseases. The aim of this study is to detect the host feeding pattern of sand flies in the endemic areas for leishmaniasis in Turkey (Antalya, Kayseri) and Northern Cyprus (TRNC) as well as the presence of Leishmania DNA in the specimens. Methods One-hundred seventy-six blood-fed sand fly specimens were examined for blood meal analysis. A SYBR Green-PCR assay was performed with specific forward primers for each host and a universal reverse primer. Primers of human and goat were used together in multiplex PCR while goat and cow were studied separately. ITS-1 qPCR assay was also performed on both blood-fed and non-blood-fed females to detect Leishmania parasites. Results Blood sources could be detected in 69 out of 176 blood-fed sand fly specimens. The results of blood meal analysis showed that specimens were fed mostly on cows (22.2%) followed by humans (5.7%), goats (2.8%) and dogs (0.6%). Multiple feeding patterns were also detected as human + cow (3.4%), cow + goat (2.8%) and human + goat (1.7%). Five of the blood-fed specimens were Leishmania spp. positive: P. major s.l. (n = 1), P. tobbi (n = 2) were L. tropica positive from Antalya, P. simici was positive for L. infantum from Kayseri and P. papatasi (n = 1) was positive for L. major from Cyprus. Leishmania infection rates were determined as 3.79%, 1.69% and 2.63% among the blood-fed sand fly specimens in Antalya, Kayseri and TRNC, respectively. Conclusion The SYBR-Green-based multiplex PCR assay is a cost-effective and promising tool for blood meal identification of wild-caught sand flies as well as other blood-sucking arthropods. Feeding patterns of important vector species detected in the present study show the high risk in these endemic areas. As a next step, to identify the blood source in a shorter time and to make the test more sensitive, development of this assay to probe-based and multiplex PCR will be also planned.
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    A Case Report: Subcutaneous Myiasis Caused by Dermatobia Hominis After a Trip to Brazil
    (Galenos Yayincilik, 2017) Balci, Fatma Kamer Varici; Ataman, Saniye; Guler, Erkan; Karakus, Mehmet; Yolasigmaz, Aysegul Unver; Toz, Seray; Turgay, Nevin; Ozbel, Yusuf
    Myiasis is an invasive diseases caused by larvae of various Dipterian flies. Subcutaneus myiasis cases are commonly observed by Dermatobia hominis larvea. A 26-year-old female patient with a history of travel abroad. Brasil diagnosed with subcutaneous myiasis originating from D. hominis. After spending six months in Amazon forests, Brasil, patient observed two small erythematous papules on right lower quadrant abdomen and delicate and itchy in the sacral region. Two larvae removed from the papules were sent to Ege University Medical Faculty Polyclinic of Infectious Diseases and sent to the parasitology polyclinic for the identification of larval species. After the necessary macroscopic and microscopic examinations, D. hominis was obtained as aresult of myiasis effect. Geographically, natural distribution of D. hominis is South American countries. In this case report we want to introduce the mechanism of egg distribution and take attention to the importance of medical history on myiasis cases. Since it is the first case seen in Turkish patients, it is thought to be presented in terms of revealing the importance of anamnesis in myiasis cases.
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    The comparison of microscopy and real time polymerase chain reaction methods for the diagnosis of Pneumocystis Jirovecii pneumonia: evaluation of clinical parameters
    (Turkish Assoc Tuberculosis & Thorax, 2017) Toz, Seray; Gunduz, Cumhur; Tetik, Asli; Tasbakan, Meltem; Pullukcu, Husnu; Bacakoglu, Feza; Tasbakan, Mehmet Sezai; Gulen, Figen; Unver, Aysegul; Turgay, Nevin
    Introduction: Pneumocystis jirovecii pneumonia (PCP) causes serious infections, especially in patients with immunosuppressive diseases. In this study, it was aimed to evaluate the results of samples obtained from PCP suspected patients using two different methods together with clinical data. Materials and Methods: Microscopy and real time polymerase chain reaction (real time PCR) methods were performed with bronchoalveolar lavage (BAL) samples sended to Ege University Medical Faculty Direct Parasitology Diagnostic Laboratory between March 2009 and June 2010. Demographic characteristics, clinical and laboratory data were also recorded retrospectively. The data were evaluated using the SPSS 16.0 program. Results: A total of 42 BAL samples collected from patients (24 males, mean age: 31.49 +/- 26.14) were included. There were totally 16 P. jirovecii positives either one of the tests. Sixteen and three samples were detected positive by real time PCR and microscopy, respectively. Trimethoprim-sulfamethoxazole was prescribed in 11 PCP diagnosed cases and 6 of them died. Conclusion: Today, despite the growing opportunities in diagnosis and treatment, PCP pneumonia is associated with high mortality. Careful examination of clinical data and immune status of the patients are important. Multidisciplinary approach is required for early PCP diagnosis.
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    Comparison of Polymerase Chain Reaction Using Kinetoplast DNA Specific Primers and Other Parasitological Methods in the Diagnosis of Clinical Samples of Suspected Patients with Cutaneous Leishmaniasis in Sanliurfa
    (Ankara Microbiology Soc, 2017) Yildiz Zeyrek, Fadile; Toz, Seray; Yuksel, Fehmi; Turgay, Nevin; Ozbel, Yusuf
    Visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are seen endemically in Turkey and CL caused by Leishmania tropica is an important public health problem in southeastern as well as other regions of Turkey. The diagnosis has been usually made by clinical view of lesion and/or parasitologically using lesion aspiration smears. Histological examination does not, always reveal the parasite in the skin biopsy, particularly in chronic lesions. Besides this, due to CL infections caused by different species in endemic areas, diagnostic methods enabling species identification are in great need. Species identification, in the time of diagnosis, is an important procedure for helping the clinicians in the planning of treatment as well as control measures. Polymerase chain reaction (PCR) is a specific and sensitive diagnostic tool that can also identify the parasite at species level. Kinetoplast DNA (kDNA) is one of the genetic regions that can be used for the detection of Leishmania parasites in clinical specimens, kDNA PCR is reported as one of the most sensitive methods related to species-specific variable regions in mini-circle long time ago. It has been considered as one of the most ideal targets for the diagnosis of leishmaniasis. The aim of the study was to perform PCR targeting kDNA by using the primers of Uni21/Lmj4 in clinical samples and compare the results with other parasitological methods like smear and culture, for the diagnosis of CL. The kDNA PCR, parasite culture and microscopical evaluation of stained smears of 62 specimens from suspected CL cases who have referred to Cutaneous Leishmaniasis Diagnosis and Treatment Center in Sanliurfa, Turkey were included in the study. The kDNA PCR showed the highest sensitivity 100% of the samples (35/35) among all diagnostic assays, followed by the microscopy (25/35 positive, 71.4% sensitivity) and culture (19/35 positive, 54.3% sensitivity). The sensitivity of combination of culture and microscopy was 88.6% (31/35 positive). These results suggested that performing kDNA PCR in addition to conventional techniques is important for improving the true diagnosis of CL to the species level and also important for establishing treatment regimens and designing appropriate precautions in highly endemic area like the southeastern region of Turkey.
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    The current clinical and geographical situation of cutaneous leishmaniasis based on species identification in Turkey
    (Elsevier Science Bv, 2019) Ozbilgin, Ahmet; Toz, Seray; Harman, Mehmet; Topal, Suhan Gunasti; Uzun, Soner; Okudan, Fulya; Gungor, Dilek; Erat, Aysegul; Ertabaklar, Hatice; Ertug, Sema; Gunduz, Cumhur; Cavus, Ibrahim; Karakus, Mehmet; Ural, Ipek Ostan; Olgen, M. Kirami; Kayabasi, Cagla; Kurt, Ozgur; Ozbel, Yusuf
    Leishmaniases 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.
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    The current epidemiology of leishmaniasis in Turkey, Azerbaijan and Georgia and implications for disease emergence in European countries
    (Wiley, 2022) Ozbel, Yusuf; Toz, Seray; Munoz, Clara; Ortuno, Maria; Jumakanova, Zarima; Perez-Cutillas, Pedro; Maia, Carla
    Leishmania spp. are sand fly-borne protozoan parasites causing leishmaniasis in humans and animals. The aim of the study was to analyse the epidemiology of leishmaniasis in Turkey, Azerbaijan and Georgia from 2005 to 2020 and evaluate the associated risk for disease emergence in European countries. It is based on an analysis of WHO and OIE reported cases between 2005 and 2020, a review of scientific articles published in SCOPUS between 2009 and 2020 and a questionnaire survey to public health and veterinary authorities in these countries. Endemic Leishmania spp. include L. infantum in the three countries, L. major in Azerbaijan and Turkey and L. tropica and L. donovani in Turkey. Leishmaniasis is reported in humans, animals and sand flies and incidence is spatially and temporarily variable. In the southern Caucasus and particularly in Georgia, reported incidence of human visceral leishmaniasis by L. infantum remains high. However, whilst Georgia experienced a gradual decrease from >4.0 cases per 100,000 population in 2005-09 to 1.13 cases per 100,000 population in 2020, the period with highest incidence in Azerbaijan, which ranged between 0.40 and 0.61 cases per 100,000 population, was 2016-2019, and no cases have so far been reported for 2020. Visceral leishmaniasis in the Southern Caucasus affects mostly young children from deprived urban areas and its closely associated to canine leishmaniasis. Turkey reported cases of visceral leishmaniasis between 2005 and 2012 and in 2016 only, and incidence ranged between 0.02 and 0.05 per 100,000 population. In contrast, the reported annual incidence of cutaneous leishmaniasis in Turkey was much greater and peaked at 7.02 cases per 100,000 population in 2013, associated to imported cases from cutaneous leishmaniasis endemic Syria. Leishmaniasis by L. infantum in Azerbaijan and Georgia represents a regional public and animal health challenge that requires support to improve diagnosis, treatment and control. The unprecedented rise of cutaneous leishmaniasis and the spread of L. tropica and L. donovani in Turkey is an important risk factor for their emergence in Europe, especially in Mediterranean countries where competent vectors are widespread.
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    Cutaneous Leishmaniasis Cases Caused by Leishmania infantum in Sanliurfa Province, Turkey
    (Ankara Microbiology Soc, 2020) Zeyrek, Fadile Yildiz; Toz, Seray; Uluca, Nermin; Doni, Nebiye; Toprak, Sahin; Ozbel, Yusuf
    Leishmaniases are a group of vector-borne diseases, and two clinical forms, visceral (VL) and cutaneous leishmaniasis (CL, Oriental sore), are seen in Turkey. While VL cases are recorded as 20-25 per year, CL cases are reported around 2000 per year, and nearly half of CL cases were recorded in anliurfa province. Therefore, by knowing the epidemiology of the disease in anliurfa province, it is possible to develop control measures and reduce the total number of cases across the country. Although Leishmania tropica is known as the main causative agent in anliurfa, other Leishmania species have also been identified as a result of mass human movements in the last 10 years. in this study, we aimed to present the first CL cases caused by Leishmania infantum in $anllurfa. A total of 14 cases, which were admitted with the suspicion of CL and diagnosed as positive by direct microscopy and/or real-time ITS1-PCR using lesion aspiration samples are included in the study. Two or more smears were prepared from the samples taken from the lesions of the patients by fine needle aspiration. One of the smears was stained with Giemsa stain after fixation with methyl alcohol and examined under the light microscope at x1000 magnification for the presence of Leishmania amastigotes. DNA isolation was made from the other unstained preparations with a commercial kit (Qiagen DNeasy, Germany) according to the recommendations of the manufacturer. the real-time ITS1-PCR method was performed by using the Old World species-specific primers and probes. As a result, by the identification of the species with real-time ITS1-PCR, it was determined that the causative agent was L.infantum in five cases, Lmajor in one case and L.tropica in eight cases. It was learned that four of the cases in which L.infantum was detected as the causative agent were local, one was Syrian and they lived in the city center. Also two of the eight cases, which were identified as L.tropica, were Syrian and six of them were domestic cases and all of them lived in the city center. While all 14 patients included in the study were positive with real-time ITS1-PCR, amastigotes were detected in 10 cases only. the cases of CL presented in this study are the first cases caused by L.infantum reported from anliurfa, and are important in terms of concretely demonstrating the effect of mass human mobility and migration on the epidemiology of the infection.
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    The Designing of a Gel Formulation with Chitosan Polymer Using Liposomes as Nanocarriers of Amphotericin B for a Non-invasive Treatment Model of Cutaneous Leishmaniasis
    (Springer Int Publ Ag, 2022) Colak, Nergiz Gurbuz; Uyanikgil, Emel Oyku Cetin; Ozbel, Yusuf; Toz, Seray
    Purpose Leishmaniasis is a disease caused by different Leishmania spp., which are transmitted to humans by a bite of infected female sand flies. Cutaneous leishmaniasis (CL, oriental sore), visceral leishmaniasis (VL), and mucocutaneous leishmaniasis (MCL) are three main clinical forms, however, only CL and VL are seen in Turkey. Cutaneous leishmaniasis is characterized by skin lesion(s) and is one of the most important vector-borne diseases in Turkey with over 2000 cases reported annually in 40 out of 81 provinces. The treatment is usually made invasively and painfully by intralesional injection of pentavalent antimony compounds. Non-invasive and innovative treatment methods are needed as aimed in this study. Methods In the present study, one of the classical antileishmanial drugs, amphotericin B (AmB), encapsulated in liposomes was evaluated using non-invasive design based on chitosan, which is a nontoxic, biocompatible and biodegradable polymer. To avoid the invasive effect of conventional intralesional needle application, the drug was encapsulated in liposomes and incorporated into a chitosan gel for applying topically on the skin lesion. The efficacy of encapsulation of amphotericin B into liposomes and the drug release from liposomes were studied. The chitosan gel was evaluated for viscosity, flowability, appearance and pH. The efficacy of the drug embedded into chitosan gel, liposomal AmB alone and chitosan gel alone in four different concentrations was also tested using Leishmania spp. promastigotes in vitro. Results The findings have shown that AmB was encapsulated into the liposomes with high efficiency (86.6%) and long-term physical and chemical stability. Therefore, designed liposomal formulation was suitable for sustained release. The appearance of the drug-embedded chitosan gel was transparent and appropriate. Chitosan gels showed non- Newtonian behavior and plastic flow. The liposomal AmB also showed higher efficacy with no parasites in all concentrations while drug embedded into chitosan gel and chitosan gel alone were effective in two higher concentrations. The lower efficacy of the drug-embedded chitosan gel in 24 h in in-vitro study was probably due to slow release of the drug. Conclusion The gel design created in this study will provide ease of use for the lesions of CL patients that do not have a specific number, size, and shape. Follow-up studies by the ex-vivo macrophage infection model with Leishmania intracellular amastigote forms and Leishmania-infected animal models are needed to understand the present design's efficacy better.
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    Detection of Leishmania RNA virus 2 in Leishmania species from Turkey
    (Oxford Univ Press, 2019) Nalcaci, Muhammed; Karakus, Mehmet; Yilmaz, Bahtiyar; Demir, Samiye; Ozbilgin, Ahmet; Ozbel, Yusuf; Toz, Seray
    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.
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    Detection of Pneumocystis jirovecii by PCR in patients with lung cancer: A preliminary study
    (Masson Editeur, 2023) Bagci, Ozlem Ulusan; Guldaval, Filiz; Muftuoglu, Can; Mert, Ufuk; Toz, Seray; Unat, Damla Serce; Unat, Omer Selim
    Introduction: Infection complications in lung cancer (LC), one of the most common cancers in the world, are still among the most important causes of death. Of them, P. jirovecii, which is as an opportunistic infection, causes a life-threatening type of pneumonia in cancer patients. This preliminary study aimed to determine the incidence and clinical status of P. jirovecii by PCR in lung cancer patients compared to the conventional method. Material and methods: Sixty-nine lung cancer patients and fSorty healthy individuals were included in the study. After sociodemographical and clinical features were recorded, sputum samples were collected from attenders. Firstly, microscopic examination was made with Gomori's methenamine silver stain and then PCR was performed.Results: P. jirovecii was detected in three of 69 lung cancer patients by PCR (4.3%), but not by microscopy. However, healthy individuals were negative for P. jirovecii by both methods. Based on clinical and radiologi-cal findings, P. jirovecii was evaluated as probable infection in one patient and colonization in the other two patients. Although PCR is more sensitive than conventional staining methods, it cannot distinguish probable and proven infections from pulmonary colonization.Discussion: It is important to evaluate the decision of infection together with laboratory, clinical and radiolog-ical findings. Moreover, PCR may enable to know the colonization and to take precautions such as prophy-laxis, due to the risk of colonization turning into an infection in immunocompromised patient groups. Further studies involving larger populations and evaluating the colonization-infection relationship in patients with solid tumors are needed.(c) 2023 SFMM. Published by Elsevier Masson SAS. All rights reserved.
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    Determination of sand fly fauna and molecular detection of Leishmania in sand flies in Antalya Province, Southern Turkey
    (Springer, 2021) Arserim, Suha Kenan; Cetin, Huseyin; Karakus, Mehmet; Demir, Samiye; Ser, Onder; Toz, Seray; Balcioglu, I. Cuneyt
    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.
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    Diversity of Leishmania Strains Isolated from Cutaneous Leishmaniasis Patients in Turkey and its Reflection to Clinics in Mice Model
    (Ankara Microbiology Soc, 2020) Ozbilgin, Ahmet; Culha, Gulnaz; Guray, Melda Zeynep; Zeyrek, Fadile Yildiz; Akyar, Isin; Toz, Seray; Gunduz, Cumhur
    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/TOF-MS 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 Linfantum/tropica were also shown to be present.
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    The Effect of BTK Inhibitor Ibrutinib on Leishmania infantum Infection In Vitro
    (Springer Int Publ Ag, 2022) Mert, Ufuk; Muftuoglu, Can; Erdem, Sevgi; Sadiqova, Aygul; Toz, Seray; Ozbel, Yusuf; Caner, Ayse
    Purpose Leishmaniasis is a neglected infectious disease affecting millions of people worldwide. Visceral leishmaniasis (VL), caused by Leishmania infantum and Leishmania donovani, is one of the main clinical forms of the disease and fatal if not treated promptly and properly. Despite being available for the last 70 years, current drugs used in the treatment of leishmaniasis have serious problems as they have high toxicity, require long-term administration and cause serious side-effects, leading to the emergence of resistant and relapse cases. Therefore, there is an urgent need for the discovery of novel antileishmanial molecules and the development of new treatment regimens. The drug used for chemotherapy of B-cell malignancies, Ibrutinib, an inhibitor of Bruton's Tyrosine Kinase (BTK), can offer a new therapeutic perspective due to the functions of BTK on intracellular signaling mechanism of macrophages, which are the primary resident cell for Leishmania. Hence, the study aimed to evaluate ibrutinib as a potential anti-Leishmanial drug. Method In this study, we evaluated the antileishmanial effect of Ibrutinib by in vitroL. infantum infection model using macrophages, with cell viability assay, parasite rescue assay, real-time qPCR. Results We showed that Ibrutinib was significantly more effective than the Glucantime against L. infantum. In addition, our data revealed that Ibrutinib inhibited parasite growth and load without impairing macrophage viability. Conclusions Consequently, due to its efficacy and safety, Ibrutinib may be a promising candidate for the treatment of VL caused by L. infantum as a host-targeted drug.
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    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 Univ Press Inc, 2020) Limoncu, M. Emin; Balcioglu, I. Cuneyt; Toz, Seray; Demir, Samiye; Kavur, Hakan; Karakus, Mehmet; Ozbel, Yusuf
    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/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.
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    Epidemiological analysis of Leishmania tropica strains and giemsa-stained smears from Syrian and Turkish leishmaniasis patients using multilocus microsatellite typing (MLMT)
    (Public Library Science, 2017) Karakus, Mehmet; Nasereddin, Abed; Onay, Huseyin; Karaca, Emin; Ozkeklikci, Ahmet; Jaffe, Charles L.; Kuhls, Katrin; Ozbilgin, Ahmet; Ertabaklar, Hatice; Demir, Samiye; Ozbel, Yusuf; Toz, Seray
    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 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.
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    Evaluation of conjunctival swab sampling in the diagnosis of canine leishmaniasis: A two-year follow-up study in Cukurova Plain, Turkey
    (Elsevier Science Bv, 2015) Karakus, Mehmet; Toz, Seray; Ertabaklar, Hatice; Pasa, Serdar; Atasoy, Abidin; Arserim, Suha K.; Olgen, M. Kirami; Alkan, M. Ziya; Durrant, Caroline; Ozbel, Yusuf
    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 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.
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    First molecular detection and identification ofLeishmaniaspecies in small wild rodents from Turkey
    (Cambridge Univ Press, 2020) Karakus, Mehmet; oktem, Mehmet Ali; Sozen, Mustafa; Matur, Ferhat; Colak, Faruk; Nalcaci, Muhammed; Toz, Seray
    Leishmaniasis is a parasitic disease infecting animals and humans. Two clinical forms (Visceral and cutaneous leishmaniasis) and four species are reported to be present in Turkey. Several studies have investigated canine and human leishmaniasis in Turkey but no study was performed to screen the infection among wild rodents, so far. the present study aims to investigate the role of small wild rodents as reservoir animals forLeishmaniaspp. in different regions of Turkey. Formalin-preserved tissue samples (spleen, liver, lung) of 712 rodents from 30 provinces were screened for the presence ofLeishmaniaspp. DNA. Before DNA extraction, tissues were dried, rehydrated, and homogenated.Leishmaniascreening in rodent tissues and species determination was performed with a combination of real-time kDNA and ITS1 polymerase chain reaction protocols. Eight (1.12%) out of 712 animals were found to be positive forLeishmaniaspp. DNA and species typing revealed fiveL. infantum, twoL. tropicaand oneL. majoramong positives.Leishmania majorandL. infantumDNA were detected inApodemusspp. from Zonguldak province located in the Western Black Sea Region, whileL. tropicaDNA was found inMerionessp. andGerbillus dasyurusfrom Adana and Hatay provinces located in Eastern Mediterranean Region of Turkey. the present study is first to report natural infection ofL. infantum, L. majorandL. tropicain small wild rodents in Turkey, suggesting their possible roles as reservoirs. Further studies are needed for planning epidemiological studies and also for developing rodent control measures in risky endemic areas to break the transmission cycle.
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    Host-Parasite Interactions: Regulation of Leishmania Infection in Sand Fly
    (Springer Int Publ Ag, 2022) Omondi, Zeph Nelson; Arserim, Suha Kenan; Toz, Seray; Ozbel, Yusuf
    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.