Browsing by Author "Çavuş İ."
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Item Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused by Leishmania donovani in Turkey(Elsevier B.V., 2017) Özbilgin A.; Harman M.; Karakuş M.; Bart A.; Töz S.; Kurt Ö.; Çavuş İ.; Polat E.; Gündüz C.; Van Gool T.; Özbel Y.In Turkey, the main causative agents are Leishmania tropica (L. tropica) and Leishmania infantum (L. infantum) for cutaneous leishmaniasis (CL) and L. infantum for visceral leishmaniasis (VL). In this study, we investigated leishmaniasis cases caused by L. donovani and established animal models for understanding its tropism in in vivo conditions. Clinical samples (lesion aspirations and bone marrow) obtained from CL/VL patients were investigated using parasitological (smear/NNN) and DNA-based techniques. For species identification, a real time ITS1-PCR was performed using isolates and results were confirmed by hsp70 PCR-N/sequencing and cpb gene PCR/sequencing in order to reveal Leishmania donovani and Leishmania infantum discrimination. Clinical materials from CL and VL patients were also inoculated into two experimental groups (Group CL and Group VL) of Balb/C mice intraperitoneally for creating clinical picture of Turkish L. donovani strains. After 45 days, the samples from visible sores of the skin were taken, and spleens and livers were removed. Measurements of the internal organs were done and touch preparations were prepared for checking the presence of amastigotes. The strains were isolated from all patients and amastigotes were seen in all smears of the patients, and then isolates were immediately stored in liquid nitrogen. In real time ITS1-PCR, the melting temperatures of all samples were out of range of L. infantum, L. tropica and L. major. Sequencing of hsp70 PCR-N showed that all isolates highly identical to previously submitted L. donovani sequences in GenBank, and cpb gene sequencing showed five isolates had longer cpbF allele, whereas one isolate contained a mixed sequence of both cpbF and cpbE. All mice in both experimental groups became infected. Compared to controls, the length and width of both liver and spleen were significantly elevated (p < 0.001) in both groups of mice. However, the weight of the liver increased significantly in all mice whereas the weight of spleen increased only in VL group. Amastigotes were also seen in all touch preparations prepared from skin sores, spleen and liver. L. donovani strain was isolated from autocutaneous a VL patient first time in Turkey. Animal models using clinical samples were successfully established and important clinical differences of the isolated strains were observed. © 2017 Elsevier B.V.Item In vivo and in vitro Models for Scanning Drug Substances in Malaria: Prestudy(2017) Özbilgin A.; Çavuş İ.; Nuraydın A.; Kaya T.OBJECTIVE: The Wolrd Health Organization (WHO) encourages all countries to investigate antimalarial drug substances derived from herbal sources with the slogan "Hunt of the Next Artemisinin" due to the emergence of resistant strains of Plasmodium species to artemisinin. In the broad and simple sense, it was planned to help guide the young researchers set in-vitro and in-vivo models of malaria in order to be used in drug research and active ingredient studies.; METHODS: In-vitro study, young Plasmodium berghei trophozoites were removed from the liquid nitrogen tank and resuspended in appropriate conditions, followed by incubation with chloroquine and tetracycline at concentrations of 0.1, 0.4, 0.8, 1.6, 6.4, 12.8 μg/mL for 24 hours at +37°C in a shaking incubator. In-vivo studies, Tetracycline group (TG) and Chloroquine group (KG) were administered 50 mg/kg of tetracycline and chloroquine by intragastric lavage and untreated control group (TACG) were administered the same amount of saline via the same route. The suppression of parasitemia in mice was followed for 24 days.; RESULTS: In our in-vitro study it was observed that 0.8 μg/mL of chloroquine and 1.6 μg/mL of tetracycline was enough to suppress parasitemia. In our in-vivo drug study, all of the mice in the TG group died at day 24, and all of the mice in the TAKG group died at day 12, with no parasitemia observed in the mice in the KG group.; CONCLUSION: Our study suggests that if tetracycline therapy is administered when the induction of chloroquine therapy is delayed, the exacerbation of the parasitemia may be prevented and when chloroquine is obtained chloroquine therapy can be commenced thus preventing the loss of the patient.Item Cryopreservation of Leishmania Species in Manisa Province(2017) Çavuş İ.; Ocak F.; Kaya T.; Özbilgin A.OBJECTIVE: It was aimed to assess the success of the cryopreservation process which is carried out in order to preserve the genetic material and the virulence of the Leishmania species that are an important health problem in our region.; METHODS: Leishmania tropica, L. infantum, L. major, and L. donovani strains in Novy-MacNeal-Nicolle (NNN) medium in MCBU were used. Promastigotes cultured in the NNN medium were transferred to RPMI 1640 medium; promastigotes in the logarithmic phase were washed three times with PBS, and 15% dimethylsulfoxide (DMSO) was added. Leishmania species were transferred to 12 separate tubes. The tubes were stored at -86°C for one night by placing them in Coolcell boxes. The tubes were transferred into a liquid nitrogen tank. One cryotube per Leishmania strain is thawed monthly and cultured in NNN medium.; RESULTS: For the duration of study it was observed that each Leishmania isolate preserved 60-65% of their viability and entered the logarithmic phase on the 7th day following the inoculation in the NNN medium. Abnormalities in the structures and movements of the promastigotes were not observed in microscopic examinations.; CONCLUSION: The following conclusions were made: cryopreservation is important for studies planned related to leishmaniasis and cryopreservation with DMSO is successful.Item Imported cutaneous leishmaniasis cases detected in Truck drivers in Hatay; [Hatay’da Tır Şoförlerinde saptanan yurt Dışı kaynaklı kutanöz leyşmanyazis olguları](Ankara Microbiology Society, 2018) Çulha G.; Doğramaci A.Ç.; Kaya T.; Çavuş İ.; Gülkan B.; Özbi̇Lgi̇n A.Leishmaniasis, seen in tropical and subtropical regions, is an infectious disease caused by the protozoan parasite Leishmania species. There are three main forms of leishmaniasis: cutaneous, mucocutaneous and visceral leishmaniasis. Cutaneous leishmaniasis (CL) has become an increasing problem as the number of travels around the world increases and people go to work in endemic areas. Turkey has received great numbers of immigrants in recent years, from its neighboring countries like Iraq, Islamic Republic of Iran, Afghanistan, Turkmenistan and the Syrian Arab Republic because of the political instabilities in these countries as well as the job opportunities caused by large-scale development projects undertaken by Turkey. In this report, imported CL cases detected in five truck drivers transporting from Hatay to Turkmenistan, Syria, Saudi Arabia, Iran and Georgia, Uzbekistan and Azarbaijan countries were presented. The patients admitted to Mustafa Kemal University, Faculty of Medicine Dermatology Policlinic, with wound complaints on their bodies were directed to the Department of Parasitology to obtain smear samples from their wounds. The age range of the patients were 38 to 43 years. Patients with wound trail for a period ranging from one month to one year had a number of lesions varying between 2-7 and in all cases, a smear preparation was prepared from the lesions for diagnostic purposes. Clinical material obtained from five patients with pre-diagnosis of CL was firstly examined with Giemsa stain. Samples taken from the patients were inoculated into modified NNN (Novy-MacNeal-Nicolle) medium for the evaluation of the presence of the promastigotes. Promastigotes obtained from the inoculated medium were also genotyped using the ITS1 region. In all of the slides prepared from the clinical material taken from the patients amastigotes were determined. The growth of promastigotes were observed in only three of the clinical specimen inoculated media. The genotyped three species were Leishmania tropica, Leishmania infantum/donovani and Leishmania major. In this study, the importance of support for the diagnosis of different microbiological methods used in the diagnosis of leishmaniasis infection which occurred during the outbreaks of the disease has been put forward. In addition, it was aimed to draw attention to the importance of imported CL cases in our country diagnosed in five truck drivers making transportation from Hatay to Turkmenistan, Syria, Saudi Arabia, Iran, Georgia, Uzbekistan and Azerbaijan. © 2018 Ankara Microbiology Society. All rights reserved.Item Do the rodents have a role in transmission of cutaneous leishmaniasis in Turkey?; [Türkiye’de kutanöz leyşmanyaziste kemiricilerin rolü var mı?](Ankara Microbiology Society, 2018) Özbi̇Lgi̇n A.; Çavuş İ.; Yildirim A.; Gündüz C.Leishmaniasis is a zoonotic/anthroponotic vector borne parasitic infection which is caused by Leishmania species and transmitted by sand flies (Phlebotomus spp.) The reservoirs of Leishmania species in nature are various wild and domestic carnivores, rodents and human. The aim of this study was to investigate whether the rodents in genera Meriones, Mesocricetus, Rattus and Mus which inhabit in the natural habitat of our country could be natural reservoirs of Leishmania tropica, Leishmania infantum, Leishmania major and Leishmania donovani for cutaneous Leishmaniasis (CL)., The rodents Mus musculus (Balb/C mouse), Mesocricetus auratus (hamster), Meriones unguiculatus (gerbil) and Rattus norvegicus (rat) which are part of the natural habitat in Turkey were used in the study. L.tropica, L.infantum, L.major and L.donovani promastigote isolates obtained from CL patients and cultured in enriched media were injected in the footpads of the animals intradermally using the density of 108 promastigote/ml. The scale of the lesions on the footpads of the animals were measured for 12 weeks. At the end of the experiment, the animals were sacrificed and “touch preparations” were prepared using footpad, liver, spleen and testicles of the sacrified animals and were examined using Giemsa stained slides following culturing in enriched NNN medium. Leishmania amastigotes were seen in the slides prepared from the footpads of the all experimental animals and all cultures were positive for promastigotes prepared from the same clinical material. But not all the experiment groups were positive for the liver, spleen and testicle preparations. According to these results it was concluded that while all rodents in the experiment groups were positive for CL, only a part of the experiment groups were positive for internal organ involvement. Accordingly, (a) All Leishmania strains caused both CL and internal organ involvement in M.unguiculatus and M.musculus, (b) only L.tropica caused CL and internal organ involvement in R.norvegicus, while other Leishmania strains only caused CL in this group, (c) in M.auratus only L.donovani caused CL while other strains caused both CL and internal organ involvement. In our study, it was determined that the rodents Meriones, Mesocricetus, Rattus and Mus genera which are part of our country’s natural habitat could serve as natural reservoirs of L.tropica, L.infantum, L.major and L.donovani, thus having the potential for the spreading of Leishmaniasis in our country and important information were gathered concerning the clinical aspects of the infection caused by Leishmania species in their potential reservoir hosts. © 2018 Ankara Microbiology Society. All rights reserved.Item The diagnostic value of lesional skin smears performed by experienced specialist in cutaneous leishmaniasis and routine microbiology laboratory; [Kutanöz Leishmaniasiste Deneyimli Uzman ve Rutin Mikrobiyoloji Laboratuvarı Tarafından Yapılan Lezyonal Yaymaların Tanısal Değeri](Turkish Society of Hematology, 2019) An İ.; Harman M.; Çavuş İ.; Özbilgin A.Objective: Leishmaniasis is a common vector-borne infection affecting 12 million people in 98 countries. The most frequently used method in diagnosis is the microscopic investigation of the leishmania smears. The diagnostic value of this method varies according to the experience of the evaluator. In this prospective study, it was aimed to emphasize the importance of experience in the evaluation of lesional smears used in the diagnosis of cutaneous leishmaniasis. Methods: In this study, patients who were admitted to Dicle University Medical Faculty Hospital Dermatological and Venereal Diseases Outpatient Clinic between January and December 2016 and who had lesions with suspicious cutaneous leishmaniasis were included. For all the cases, both in the routine microbiology laboratory and in the diagnosis and treatment of cutaneous leishmaniasis, separate smears were performed by an experienced specialist and evaluated independently from each other. Results: In 70 of 98 cases studied, the diagnosis of cutaneous leishmaniasis was confirmed by laboratory evaluations. The rate of positivity was significantly higher in the smears analyzed by experienced specialist in the clinical and diagnosis of cutaneous leishmaniasis (95.7%) than in the smears analyzed by the routine microbiology laboratory (42.9%) (p<0.001). Conclusion: The data in our study showed that smears should be performed and evaluated by experienced specialists in the clinical and diagnosis of cutaneous leishmaniasis. © Telif Hakkı 2019 Türk Dermatoloji Derneği.Item The current clinical and geographical situation of cutaneous leishmaniasis based on species identification in Turkey(Elsevier B.V., 2019) Özbilgin A.; Töz S.; Harman M.; Günaştı Topal S.; Uzun S.; Okudan F.; Güngör D.; Erat A.; Ertabaklar H.; Ertuğ S.; Gündüz C.; Çavuş İ.; Karakuş M.; Östan Ural İ.; Ölgen M.K.; Kayabaşı Ç.; Kurt Ö.; Özbel Y.Leishmaniases are a group of vector-borne diseases caused by the members of Leishmania genus, and there are three main clinical forms of the infection as visceral, cutaneous, and mucocutaneous. Cutaneous leishmaniasis is a growing public health problem in Turkey due to increasing detection of autochthonous cases caused by L. major and L. donovani in some regions in addition to Syrian imported cases. For this reason, we aimed to evaluate the current epidemiological situation of CL in the view of causative agents and their geographical distribution throughout Turkey. The samples were collected from 356 CL patients admitted to different centers in 18 provinces between January 2013 and December 2016. Direct microscopy, culture (regular and enriched NNN) and molecular techniques (real-time ITS1 PCR and hsp70 PCR/sequencing) were performed. By molecular techniques, 299, 28, 19 and 10 isolates/clinical samples were identified as L. tropica, L. major, L. infantum and L. donovani, respectively. Most of the patients (65.73%) had one lesion usually on their face/head. Dry-nodular type lesions (n = 291) were mainly associated with L. tropica while L. major was mainly found related to wet-ulcerative ones. Leishmaniasis recidivans was also detected in 2.52% among 356 patients. L. tropica was detected as most widespread species causing CL in Turkey. L. infantum and L. major was also found in one third of the provinces. Enriched NNN culture was worked well for isolating the parasite and 346 isolates were successfully grown and stored in liquid nitrogen. The comparison of all diagnostic techniques showed that the parasitological positivity rate could increase if the combination of direct microscopy and real-time ITS1 PCR is used. Besides well-known anthroponotic L. tropica cases, the increasing detection of CL cases caused by zoonotic species, L. infantum and L. major, is one of the most important findings in the present study. In our opinion to ensure timely and accurate diagnosis, proper treatment and countrywide effective control of CL in Turkey a systematic approach is needed on the base of information about characteristics of lesions and patients and epidemiological features of the disease. © 2018 Elsevier B.V.Item Comparison of liver extract medium with novy-macneal-nicolle medium and the molecular method for the diagnosis of leishmaniasis; [Layşmanyaz tanısı için karaciğer ekstreli besiyerinin novy-macneal-nicolle besiyeri ve moleküler yöntemle karşılaştırılması](AVES, 2020) Özbilgin A.; Tünger Ö.; İnanır I.; Çavuş İ.; Perk N.E.; Özel Y.Objective: Diagnosis of Leishmaniasis is based on culture, mi-croscopic examination, serological tests, molecular methods. Novy-MacNeal-Nicolle (NNN) medium is mostly used for culture all over the world. The use of molecular methods for diagnostic purposes led to the questioning of the sensitivity of NNN medium. In this study, it is aimed to compare the performance of a new culture medium with those of NNN medium and the molecular method. Methods: Samples of 22 patients with suspected cutaneous leishmaniasis (CL) and 4 patients with suspected visceral leishmaniasis (VL) were sent to Manisa Celal Bayar University Faculty of Medicine Department of Medical Parasitology for diagnosis or confirmation from Kilis, Osmaniye, Bitlis, Gazian-tep and Manisa provinces. Pre-prepared media were sent to the health centers in these provinces and inoculation of these media with clinical samples was requested. Needle aspiration fluid for CL and bone marrow samples for VL were cultured simultaneously on NNN medium and newly developed liver extracted medium (LEM), and were sent to our institution with their smears. Smears were examined by staining with Giemsa method. In addition, parasite DNA was identified with real-time polymerase chain reaction method in these materials. Results: Leishmania amastigotes were detected in 14 of 26 samples. Promastigote proliferation was observed in 8 and 24 of the samples cultured in NNN and LEM respectively. No proliferation was detected by either method in samples of 2 patients with suspected CL. L. tropica, L. major and L. infantum/donovani were detected among isolates showing dermotropic location, and L. tropica and L. infantum/don-ovani were detected among isolates with viscerotropic location by genotyping. Conclusions: NNN medium, which is widely used in leishmaniasis di-agnosis, is not sensitive enough for the diagnosis of some Leishmania strains in Turkey. Therefore, an enrichment medium such as liver extract medium which we have presented in this study should be used for di-agnosis. However our results should be confirmed with more studies. Klimik Dergisi 2020; 33(2): 137-41. © 2020, AVES. All rights reserved.Item In vitro Cultivation of Plasmodium falciparum; [Plasmodium falciparum’un in vitro Kültürü](Galenos Publishing House, 2020) Özbilgin A.; Çavuş İ.; Haghi M.; Özel Y.Objective: Plasmodium falciparum is a protozoan parasite that causes many deaths worldwide. It’s cultivation in an in vitro culture setting contributes significantly to scientific studies. However, there are no laboratories in Turkey that cultivate P. falciparum in vitro. Hence, the purpose of this study was to cultivate P. falciparum in vitro. Methods: Five P. falciparum strains were used in our study and were kept frozen in liquid nitrogen tanks. These parasite strains were then thawed in a 37 °C water bath and transferred to the Albumax-complete medium that was previously prepared. After that, the petri dishes were placed in the chamber. For 30 seconds, a special gas mixture containing 5% CO2, 5% O2 and 90% N2 was added into the chamber which was placed in a 37 °C oven and left for incubation for 2 days. At the end of the incubation period, thin smear preparations were prepared from the medium, stained with Giemsa and examined using an immersion lens. Results: Examination of the smears revealed that trophozoite and schizont forms of all P. falciparum isolates were present at a rate of 2% in in vitro culture medium. Conclusion: As a result of our study, the in vitro culture of P. falciparum was successfully developed. With this, several projects such as biological and chemical characteristics, pathogenicity, phenotypic and molecular-level drug sensitivities and parasite vaccination studies can be carried out more easily in our country. © 2020 Turkish Society for Parasitology.Item Evaluation of Imported Malaria Cases in Northern Cyprus between 2016 and 2019: First Data Series; [Kuzey Kıbrıs’ta 2016-2019 Yılları Arasındaki İmport Sıtma Olgularının Değerlendirilmesi: İlk Veriler](Galenos Publishing House, 2020) Güler E.; Özbilgin A.; Çavuş İ.; Şanlıdağ T.; Süer K.Objective: In present times, malaria remains an infectious disease with a high mortality rate in some regions of the world. It is predicted to preserve its importance as a disease in the future because of the traveling human populations from malaria-endemic African countries into the regions where malaria has been eradicated. The objective of this study is to evaluate the increasing imported malaria cases in the Turkish Republic of Northern Cyprus. Methods: In this study, we investigated 13 patients who were diagnosed with malaria between 2016 and 2019. We clinically evaluated all the cases. More importantly, we made the diagnosis of these patients by Giemsa-stained thin and thick blood smears, rapid malaria antigen tests, and genotyping (only for five patients) by real-time polymerase chain reaction. Additionally, we evaluated patients with malaria in terms of age, gender, and seasons. Results: In the diagnosed malaria cases, 11 (84.4%) of them were male and 2 (15.6%) were female. There was no significance between malaria infection and gender (p=0.358). Plasmodium falciparum, Plasmodium vivax, and Plasmodium ovale infection were detected in ten patients (76.9%), two (15.4%) patients, and one (7.7%) patient, respectively. There was a significant increase (p=0.003) in the malaria cases in 2019 (n=9). The seasonal comparison revealed that malaria infections are most common in autumn (8/13, 61.5%). Conclusion: Despite the eradication of malaria in Turkish Republic of Northern Cyprus, the rising number of recently imported cases increases the risk of emerging local cases. Malaria infection should be immediately suspected, particularly, in foreign patients who travel from the malaria-endemic region and present with symptoms such as fever and shivering if the laboratory findings especially detect thrombocytopenia. © 2020 Turkish Society for Parasitology.Item The Production of Trypanosoma Brucei Rhodesiense, Cause of African Sleeping Sickness, and Trypanosoma Cruzi, Cause of American Chagas Disease, on Different Medias and Testing a New Media; [Afrika Uyku Hastalığı Etkeni Trypanosoma brucei rhodesiense ve Amerika Chagas Hastalığı Etkeni Trypanosoma cruzi’nin Farklı Besiyerlerinde Üretilmesi ve Yeni Besiyerinin Denenmesi](Galenos Publishing House, 2020) Özbilgin A.; Çavuş İ.; Nuraydın A.; Özel Y.Objective: Human African trypanosomiasis, also known as sleeping sickness, is a parasitic disease in which Glossina is transmitted by human intervention and Trypanosoma b. rhodosiense and Trypanosoma b. gambiense are the causative agents Production of parasites in axenic cultures provides great advantage in parasite biochemistry, immunological, physiological and molecular studies. In this study, it is aimed to determine the medium which will produce in vigorous amount of Trypanosoma b. rhodosiense and Trypanasoma cruzi and to establish a new medium. Methods: In this study, Trypanosoma b. rhodosiense and Trypanasoma cruzi strains stored in Manisa Celal Bayar University Parasite Bank will be removed from liquid nitrogen tank under suitable conditions, planted in Medium I, Medium II, Medium III and newly developed medium. Reproductive densities of the media will be statistically analyzed on Thoma lamina depending on the time, using the Sidak’s multiplequality test. Results: As a result of this study, it has been concluded that the best medium, to produce abundantly Trypanosoma b. rhodosiense and Trypanasoma cruzi strains, to be used in diagnosis and active substance screenings, molecular studies, metabolic analyzes and drug studies is the medium IV. Conclusion: This study is one of the first studies related to the production of Trypanosoma species in Turkey and planned to provide a basis for the studies of African sleeping disease, Chagas disease and their agents. © 2020, Galenos Publishing House. All rights reserved.Item Comparison of in vitro Resistance of Wild Leishmania İsolates, Which are Resistant to Pentavalent Antimonial Compounds, Against Drugs Used in the Treatment of Leishmaniasis; [Pentavalent Antimonial Bileşiklere Dirençli Vahşi Leishmania İzolatlarının Leishmaniasis Tedavisinde Kullanılan İlaçlara Karşı in vitro Dirençlerinin Karşılaştırılması](Galenos Publishing House, 2020) Özbilgin A.; Çavuş İ.; Kaya T.; Yıldırım A.; Harman M.Objective: Meglumine antimoniate (Glucantime®) and Sodium stibogluconate (Pentostam®) are used for the treatment of cutaneous leismaniasis in Turkey. There is a reported resistance to these drugs in recent years. The aim of the present study was to compare the in vitro sensitivities of resistant Leishmania isolates against Amphotericin B, Miltefosine, Meglumine Antimoniate, Paromomycin and Sodium Stibogluconate. Methods: Five Leishmania isolates of patients with cutaneous leishmaniasis, who showed no clinical recovery despite two consecutive meglumine antimoniate treatments, which were stored in the Parasite Bank in Manisa Celal Bayar University Medical Faculty were selected. They were genotyped with Real-Time PCR using specific primers and probes to ITS1 region. Drug resistance levels of each Leishmania isolate were analysed for Amphotericin B, Miltefosine, Meglumine Antimoniate, Paromomycin, and Sodium Stibogluconate at concentrations of 500, 250, 125, 50, 25 µg/mL with XTT method and hemocytometer. Results: It was observed that the resistant Leishmania tropica isolates showed no resistance to Amphotericin B, and were sensitive to Miltefosine, Sodium Stibogluconate, Paromomycin and Meglumin Antimonate, respectively. In addition, Leishmania tropica (MHOM/AZ/1974/SAF-K27) isolate of the control group could stay viable in none of the drug concentrations of five agents in the study. Conclusion: It was determined that none of the selected resistant L. tropica isolates showed resistance to Amphotericin B and that was also shown statistically (p<0.05). The results of this study are important in guiding clinicians and researchers who conduct studies on drugs and search for new drug molecules. © 2020 Turkish Society for Parasitology.Item Refugees at the Crossroads of Continents: A Molecular Approach for Cutaneous Leishmaniasis Among Refugees in Turkey(Springer, 2020) Özbilgin A.; Gencoglan G.; Tunali V.; Çavuş İ.; Yıldırım A.; Gündüz C.; Harman M.Purpose: Due to mass population movements driven by internal conflicts and wars, cutaneous leishmaniasis (CL) is becoming increasingly important in Turkey. This study is aimed at determining the clinical aspects, diagnosis and genotyping of CL patients coming to Turkey from abroad. Methods: In our study, the clinical materials obtained from the patients or sent for diagnostic purposes from other centers to our laboratory between years 2012 and 2016 were assessed retrospectively. In total, there were 38 patients from Syria, Iraq, Afghanistan, Iran, and Turkmenistan. Results: 29 (76%), 28 (73%) and 33 (87%) samples were positive by light microscopy, Novy-McNeal-Nicolle(NNN), and enriched medium, respectively. By ITS-1 gene region PCR, 31 (81%) of the cases were positive. 35 of the patients were tested positive by at least one of the diagnostic methods. By genotyping, 21 Leishmania tropica, 8 Leishmania major, 3 Leismania infantum, 2 Leishmania donovani, and 1 Leishmania aethopica were detected. Conclusion: This study is aimed at informing the clinicians working in the field for the import CL cases and recording the changing epidemiological features of CL in the region as well as discussing the possible focus for L. aethiopica infection which has not been shown in the region before. © 2019, Witold Stefański Institute of Parasitology, Polish Academy of Sciences.Item Determination of antimony resistance mechanism of leishmania tropica causing cutaneous leishmaniasis in Turkey; [Türkiye’de kutanöz leyşmanyazis etkeni leishmania tropica’da antimon direnç mekanizmasının belirlenmesi](Ankara Microbiology Society, 2021) Özbilgin A.; Zeyrek F.Y.; Güray M.Z.; Çulha G.; Akyar I.; Harman M.; Özbel Y.; Ertabaklar H.; Çavuş İ.; Gündüz C.World Health Organization reported that approximately one billion people are at risk in endemic areas, one million cases of cutaneous leishmaniasis (CL) and approximately 300,000 cases of visceral leishmaniasis (VL) were reported per year in the last five years. The number of deaths due to VL is reported to be approximately 20,000 per year. Approximately 2500 cases/year have been reported as CL, caused by Leishmania tropica and Leishmania infantum, in Turkey. The significant increase observed in many cities mainly in the provinces of Mediterranean and Aegean regions in cases and foci in recent years, suggests that there may be an increase in this infections in the following years as well. In Turkey, the causative agent of CL is L.tropica and meglumine antimoniate is used in the treatment of CL. We aimed to determine antimony resistance genes specific for L.tropica by comparing the gene and protein expressions of antimony-resistant and non-resistant L.tropica strains. L.tropica isolates obtained from 3 CL patients without antimonate resistance from Aegean, Mediterranean and Southeastern regions of Turkey were provided to transform into 3 resistant isolates against meglumine antimony in the laboratory conditions. Gene expression alterations by microarray method; protein profiles by two-dimensional gel electrophoresis (2D-PAGE) and relevant proteins by MALDI-TOF/TOF MS of these isolates were accomplished and compared. L.tropica isolates from 10 CL patients who did not respond to antimony therapy were analyzed for resistance to antimonial compounds and quantitative real-time polymerase chain reaction was performed to detect the expression of genes responsible for resistance development. Moreover, differences in protein expression levels in isolates with and without antimony resistance were determined by comparing protein profiles and identification of proteins with different expression levels was carried out. Enolase, elongation factor-2, heat shock protein 70, tripanthione reductase, protein kinase C and metallo-peptidase proteins have been shown to play roles in L.tropica isolates developing resistance to antimonial compounds and similar expression changes have also been demonstrated in naturally resistant isolates from patients. In conclusion, it was revealed that L.tropica strains in our country may gain resistance to meglumine antimoniate in a short time. It is foreseen that if the patients living in our country or entering the country are treated inadequately and incompletely, there may be new, resistant leishmaniasis foci that may increase the number of resistant strains and cases rapidly. © 2020 Ankara Microbiology Society. All rights reserved.Item Overcoming the Challenge; In Vivo Efficacy of Miltefosine for Chronic Cutaneous Leishmaniasis(Springer Science and Business Media Deutschland GmbH, 2021) Tunalı V.; Harman M.; Çavuş İ.; Gündüz C.; Özbilgin A.; Turgay N.Background: 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 compounds in vivo with 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 μL L. tropica promastigote suspension with a density of 108 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 and L. tropica consistent 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. © 2020, Witold Stefański Institute of Parasitology, Polish Academy of Sciences.Item In vitro Anti-Leishmanial Activity of Essential Oils Extracted from Plants Growing in Northern Cyprus Against Leishmania tropica; [Kuzey Kıbrıs’ta Yetişen Bitkilerden Elde Edilen Uçucu Yağlarda Leishmania tropica’ya Karşı in vitro Anti-leishmanial Etkinlik Araştırılması](Galenos Publishing House, 2021) Güler E.; Özbilgin A.; Çavuş İ.; Baddal B.; Etikan İ.; Hüsnü Can Başer K.; Şanlıdağ T.Objective: Natural plant products are considered as a source of novel and effective compounds for the treatment of leishmaniasis. In this study, the in vitro activities of essential oils obtained from Origanum dubium (OD), Origanum majorana (OM), Salvia fruticosa (SF) and Laurus nobilis (LN) plants in Northern Cyprus were investigated against Leishmania tropica. Methods: Leishmania tropica strain (MHOM/TR/2012/CBCL-LT) was obtained. RPMI-1640 was added to 96-well plates in 100 µL aliquots, 100 µg/mL essential oil was added to the first well of each row and serial 2-fold dilutions were performed. A promastigote suspension was pipetted into all wells, and the plates were incubated. The promastigotes were enumerated using a haemocytometer. Results: OD essential oil was effective at killing all promastigotes at a minimum inhibitor height (MIC)=0.2 μg/mL and had high activity at the lowest concentrations. Both SF and LN oils had MIC=1.56 μg/mL and LD50 =0.78 μg/mL. SF was observed to impair promastigote morphology at the lowest concentrations, while LN did not exert any effect at concentrations <0.2 μg/mL. OM essential oil was found to have a MIC=3.13 μg/mL and a LD50 =1.56 μg/mL. Conclusion: All tested essential oils inhibited promastigotes of Leishmania tropica. OD essential oil demonstrated the highest anti-leishmanial activity. © 2021 Turkish Society for Parasitology.Item Investigation of in vitro Efficacy of Miltefosine on Chronic Cutaneous Leishmaniasis; [Kronik Kutanöz Leishmaniasis’te Miltefosin Etkinliğinin in vitro Olarak Araştırılması](Galenos Publishing House, 2022) Tunalı V.; Harman M.; Çavuş İ.; Zorbozan O.; Özbilgin A.; Turgay N.Objective: Leishmaniasis is the second deadliest parasitic disease in the World Health Organisation’s list of neglected diseases, following malaria. Cutaneous leishmaniasis (CL) is the most common form of the disease and it is one of the few communicable diseases with increasing incidence rates owing to factors like armed conflicts and climate change. CL can be divided into two major groups: Acute CL (ACL) and chronic CL (CCL). The aim of this study was to compare the in vitro efficacy of miltefosine and pentavalent antimony compounds in the CCL patient samples. Methods: Five isolates previously isolated from 5 CCL patients were included in this study. Genotyping is performed using internal transcribed spacer 1 (ITS 1) gene region real-time PCR. In vitro drug efficacy tests were applied to determine their activity against meglumine antimoniate (MA) and miltefosine. Serial dilutions (512, 256, 128, 64, 32, 16, 8 and 4 µg/mL) prepared from MA and miltefosine were prepared in 96-well flat-bottom cell culture plates and incubated at 24 °C for 48 hours. The efficacy of the drug on Leishmania spp. promastigotes after 24 and 48 hours was evaluated by hemocytometer slide and XTT cell viability test. Results: All of the samples were genotyped as L. tropica. Evaluation of 24 and 48 hours showed, 128 µg/mL and 256 µg/mL and 32 µg/mL and 64 µg/ mL concentrations of miltefosine and MA were enough to kill all the promastigotes respectively. The results of the hemocytometer slide and XTT were consistent. Conclusion: There are no studies investigating the in vitro efficacy of miltefosine with the CCL patient group. To overcome the treatment challenges experienced in this special patient group, more studies are needed. According to our results, it is concluded that miltefosine is efficient for the treatment of CCL and further clinical studies with miltefosine will reveal valuable data. © 2022 Turkish Society for Parasitology.Item Could RPMI-1640 Medium be Used in the Diagnosis and Isolation of Cutaneous Leishmaniasis Lacking NNN Medium?; [Kutanöz Leishmaniasis Tanı ve İzolasyonunda NNN Besiyeri Olmadığı Durumda RPMI-1640 Besiyeri Kullanılabilir mi?](Galenos Publishing House, 2022) Çavuş İ.; Aksoy T.; Yıldırım A.; Şahin M.T.; Özbilgin A.Laboratory diagnosis of leishmaniasis is based on culture, microscopic examination, serological and molecular methods. The gold standard method is to see amastigotes in microscopic examination and to grow promastigotes in Novy, MacNeal, Nicolle (NNN) medium. NNN medium is frequently used for culture all over the world. In our study, it was aimed to investigate whether the use of RPMI-1640 medium is an appropriate method in cases where the gold standard NNN medium is not available for the diagnosis of cutaneous leishmaniasis (CL). Smears were prepared from the needle aspiration fluid sample from the patient who applied to Manisa Celal Bayar University Faculty of Medicine and had lesions suspicious of CL, and were stained with Giemsa for the presence of amastigotes. The samples taken were directly inoculated into RPMI-1640 broth and incubated at 26 °C for the presence of promastigotes. On consecutive days after incubation, it was checked for promastigote growth. Genotyping of the grown isolate was performed with primers and probes specific to the internal transcribed spacer-1 (ITS-1) gene region with the help of real-time polymerase chain reaction. The amastigote form was observed in the microscopic examination of the needle aspiration fluid sample smear preparations taken from the patient. On the other hand, promastigote growth was observed in RPMI-1640 broth from the 3rd day. In addition, the isolate obtained from the CL patient was determined to be Leishmania tropica as a result of the species determination made by genotyping. It is thought that this study is important in terms of suggesting an alternative medium for the diagnosis of leishmaniasis in laboratories where the gold standard NNN medium is easily accessible. RPMI-1640 medium, which is easily obtained and prepared in parasitology laboratories, can help in the diagnosis of the disease and treatment follow-up, Leishmania spp. isolation and drug resistance studies. © 2022, Galenos Publishing House. All rights reserved.Item Visceral Leishmaniasis Caused by Leishmania Tropica(Springer Science and Business Media Deutschland GmbH, 2023) Özbilgin A.; Tunalı V.; Çavuş İ.; Tetik A.V.; Dinç M.; Yalçın T.; Gündüz C.; Beyaz M.; Köse Ş.Purpose: In 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. Methods: This study includes seven cases of VL caused by L. tropica.Five patients were male (71%) and four were adults (57%). Results: All 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%). Conclusions: In 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. © 2023, The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.Item Autochthonous Case of Malaria Prediagnosed as Leukemia; [Lösemi Ön Tanılı Yerli Sıtma Olgusu](Ankara Microbiology Society, 2023) Gülen H.; Türedi̇ Yildirim A.; Çavuş İ.; Türkmen H.; Özbilgin A.Malaria is a parasitic disease transmitted by infected female Anopheles mosquitoes. There are five species of Plasmodium species that can infect humans. Of these species, especially P.falciparum and P.vivax pose the greatest threat to human health. In the 2014 report of the World Health Organization, it was reported that there were no locally acquired cases of malaria in 16 countries including Türkiye. Malaria cases originating from outside the country and imported due to migration, travel and working abroad are reported as import cases. In this report, a case of non-imported malaria followed with a preliminary diagnosis of leukemia was presented. A 14-year-old female patient who was admitted to a health institution with complaints of high fever, headache, chills, nausea-vomiting, and diarrhea that had been going on for two weeks, was pre-diagnosed as leukemia and was referred to Manisa Celal Bayar University Faculty of Medicine, Hafsa Sultan Hospital, Department of Pediatric Hematology and after pancytopenia was detected in the complete blood count. The anamnesis of the patient revealed that she had no history of international travel and that she had been prescribed medications such as paracetamol, amoxicillin, and metoclopramide for flu-like complaints while working in the Southeastern Anatolia, Aegean, and Mediterranean Regions of Türkiye. Bone marrow aspiration was performed for the etiological examination of pancytopenia. Giemsa-stained blood smears, rapid diagnostics, and real-time quantative polymerase chain reaction (qRt-PCR) analyses were performed in the medical parasitology laboratory and malaria was suspected in both bone marrow and peripheral blood smears. P.vivax erythrocytic forms and gametocytes were present in abundance in smear preparations stained with Giemsa, and rapid diagnosis kit was positive for P.vivax. The strain was genotyped as P.vivax by qRt-PCR analysis. For the treatment of the patient, airalam (artemether + lumefantrine) tablets were provided with 2 x 4 daily posology for three days after the diagnosis, and primaquine was provided after one week of the diagnosis as 1 x 2 tablets (1 x 15 mg) for 14 days, and the patient was discharged without complications following the treatment regimen. The fight against malaria continues uninterruptedly since the establishment of the Republic of Türkiye. Tropical diseases, especially malaria, is of great importance for Türkiye due to numerous reasons such as its location in the subtropical region where Anopheles mosquitoes are capable of malaria transmission, it is situated at the crossroads on the migration routes between continents where human traffic is busy, there are many people who go abroad for work and most importantly rising temperatures due to climate change. For this reason, this case report is important to emphasize the importance of malaria for the country and to increase the awareness of clinicians and laboratories about malaria and the possibility of autochthonous malaria transmission in Türkiye. © 2023 Ankara Microbiology Society. All rights reserved.