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Öğe Assessment of diagnostic doses for widely used synthetic pyrethroids (Deltamethrin & Permethrin) in an endemic focus of leishmaniasis in Turkey(Biomed Central Ltd, 2016) Karakus, Mehmet; Sarikaya, Yasemen; Oguz, Gizem; Dogan, Mert; Ergan, Gokhan; Gunay, Filiz; Kasap, Ozge Erisoz; Ozbel, Yusuf; Alten, BulentBackground: Leishmania is a group of parasitic flagellated protozoons, which are transmitted by female sand flies and produces health problems in humans and also in wild and domestic animals. So far, 25 Phlebotomus and 4 Sergentomyia species were recorded in Turkey including proven or possible vectors of Leishmania spp. As no single insecticide susceptibility test was conducted targeting the sand flies in Turkey, we aimed to determine the diagnostic dose against two commonly used synthetic pyrethroids (deltamethrin and permethrin) in a hyperendemic area for leishmaniasis. Methods: Sand flies were collected from villages of Adana in 2-4 September 2013 using Centers for Disease Control and Prevention (CDC) light traps and transferred to the laboratory. The World Health Organisation tube test method was conducted using self-prepared filter papers with different concentrations. In order to determine the diagnostic dose, lethal doses (LD) were calculated by EPA Probit Analysis. Sand flies used in the experiments were dissected, mounted and identified. Results: For the lowest (0.025 %) and highest dose of permethrin (0.5 %), the mortality rate was recorded as 52.6 % and 100 % by the end of 24-h period and the diagnostic dose was recorded as 0.36 %. The mortality rate for lowest (0.0025 %) and highest (0.05 %) doses of deltamethrin was recorded as 54.8 % and 100 %. The diagnostic dose of deltamethrin was determined as 0.9 %. Conclusion: An insecticide susceptibility study was conducted in Turkey for the first time and effective doses were determined by calculating the LDs. According to presented results, the wild population of sand flies collected from a hyper-endemic region of Adana Province is still susceptible to deltamethrin and permethrin.Öğe 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, YusufMyiasis 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.Öğe Could Phlebotomus mascittii play a role as a natural vector for Leishmania infantum? New data(Bmc, 2016) Obwaller, Adelheid G.; Karakus, Mehmet; Poeppl, Wolfgang; Toez, Seray; Oezbel, Yusuf; Aspoeck, Horst; Walochnik, JuliaBackground: The occurrence of phlebotomine sand flies in Central Europe was questioned until they were recorded for the first time in Germany in 1999, and ten years later also in Austria. The aim of this study was to investigate sand flies collected in Austria for their carrier status of Leishmania spp. Findings: From 2012 to 2013 field studies were conducted in eastern Austria. Altogether, 22 individuals of sand flies were found, all morphologically identified as Phlebotomus (Transphlebotomus) mascittii Grassi, 1908. Twelve non-engorged female specimens with no visible remnants of a blood meal in their bodies were individually investigated for Leishmania spp. by ITS-1 real-time PCR. One out of these was positive for Leishmania, identified as Leishmania infantum by DNA sequencing. This finding suggests that L. infantum is not excreted by P. mascittii and possibly can establish an infection within P. mascittii. Interestingly, an asymptomatic dog living on the farm where this sand fly had been caught was also Leishmania-positive. Conclusions: This study provides new data on the suspected vector capacity of P. mascittii, being the northernmost sand fly species in Europe and in most central European regions the only sand fly species found. Proven vector capacity of P. mascittii for Leishmania spp. would be of significant medico-veterinary importance, not only with respect to expanding sand fly populations in Central Europe related to global warming, but also in the light of globalization and increasing movements of humans.Öğe 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, YusufLeishmaniases 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.Öğe Detection and identification of cutaneous leishmaniasis isolates by culture, polymerase chain reaction and sequence analyses in Syrian and Central Anatolia patients(Saudi Med J, 2017) Beyhan, Yunus E.; Karakus, Mehmet; Karagoz, Alper; Mungan, Mesut; Ozkan, Aysegul T.; Hokelek, MuratObjectives: To characterize the cutaneous leishmaniasis (CL) isolates of Syrian and Central Anatolia patients at species levels. Methods: Skin scrapings of 3 patients (2 Syrian, 1 Turkish) were taken and examined by direct examination, culture in Novy-MacNeal-Nicole (NNN) medium, internal transcribed spacer polymerase chain reaction and sequence analysis (PCR). Results: According to microscopic examination, culture and PCR methods, 3 samples were detected positive. The sequencing results of all isolates in the study were identified as Leishmania tropica. The same genotypes were detected in the 3 isolates and nucleotide sequence submitted into GenBank with the accession number: KP689599. Conclusion: This finding could give information about the transmission of CL between Turkey and Syria. Because of the Syrian civil war, most of the Syrian citizens circulating in Turkey and different part of Europe, this can be increase the risk of spreading the disease. So, prevention measurements must be taken urgently.Öğe 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, SerayBackground: 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.Öğe Detection of permethrin resistance and phylogenetic clustering of turkish head lice (Pediculus humanus capitis; de Geer, 1767 populations(Elsevier, 2020) Karakus, Mehmet; Atici, Tugce; Karabela, Semsi Nur; Baylan, Orhan; Limoncu, Mehmet Emin; Balcioglu, Ibrahim CuneytHead lice infestation caused by Pediculus humanus capitis de Geer, 1767 is one of the most common public health problems. the relationship between humans and head lice dates back millions of years ago that differentiated into different phylogenetic clades. Treatment of head lice infestation usually based on insecticide-based products, which promotes the resistance in the head lice populations. in the present study, we aimed to screen the presence of permethrin resistance among collected P. h. capitis specimens in Turkey. Three mutation sites (T917I, L920F, and M815I) were screened using real-time PCR and resistance was identified by melt analysis. of the studied specimens, resistance allele frequency (RAF) was found 0.98 for T917I, 0.99 for L920F, and 1.00 for M815I. the phylogenetic study revealed that Clade A and Clade B are present and overlap in Turkey. the present study is first to screen the resistance among Turkish head lice specimens. To not stimulate the pyrethroids resistance in head lice populations, early detection of resistance is crucial and will help the health professionals to choose suitable formula in the treatment. We suggest that the resistance status needs to be screened in randomly selected populations before any treatment application is given.Öğe 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. CuneytVisceral 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.Öğe Development of polymorphic EST microsatellite markers for the sand fly, Phlebotomus papatasi (Diptera: Psychodidae)(Bmc, 2018) Hamarsheh, Omar; Karakus, Mehmet; Azmi, Kifaya; Jaouadi, Kaouther; Yaghoobi-Ershadi, Mohammad Reza; Krueger, Andreas; Amro, Ahmad; Kenawy, Mohamed Amin; Dokhan, Mostafa Ramadhan; Abdeen, Ziad; McDowell, Mary AnnBackground: Phlebotomus papatasi is a widely distributed sand fly species in different tropical and sub-tropical regions including the Middle East and North Africa. It is considered an important vector that transmits Leishmania major parasites, the causative agents of cutaneous leishmaniasis. The development of microsatellite markers for this sand fly vector is of high interest to understand its population structure and to monitor its geographic dispersal. Results: Fourteen polymorphic microsatellite markers were developed with simple di-, tri- and tetra-nucleotide repeats. The F statistics calculated for the 14 markers revealed high genetic diversity; expected heterozygosity (He) ranged from 0.407 to 0.767, while observed heterozygosity (Ho) was lower and ranged from 0.083 to 0.514. The number of alleles sampled fall in the range of 9-29. Three out of 14 markers deviated from Hardy-Weinberg expectations, no significant linkage disequilibrium was detected and high values for inbreeding coefficient (F-IS) were likely due to inbreeding. Conclusions: The development of these functional microsatellites enable a high resolution of P. papatasi populations. It opens a path for researchers to perform multi locus-based population genetic structure analyses, and comparative mapping, a part of the efforts to uncover the population dynamics of this vector, which is an important global strategy for understanding the epidemiology and control of leishmaniasis.Öğe Effects of environmental factors and storage conditions on the performance of Olyset(A (R)) Plus against sand flies in WHO cone bioassays(Oxford Univ Press, 2016) Karakus, Mehmet; Kasap, Ozge Erisoz; Gunay, Filiz; Oguz, Gizem; Demir, Samiye; Suner, Asli; Alten, Bulent; Ozbel, YusufLeishmaniasis, visceral and cutaneous, is seen in Turkey and has both public and veterinary importance. So far, four Leishmania species and their vectors have been detected in Turkey. Vector control is essential in endemic areas and several personal protection methods are available including long-lasting insecticidal nets (LLINs). In this study, we aimed to measure the effects of usage and storage conditions on LLINs in a village-scale study. Olyset(A (R)) Plus bed nets were set up in different climatic conditions (rain, exposed to sunlight and humidity) and collected after 6 months. The effectiveness of bed nets were tested by WHO's cone test method using wild-caught sand flies. Bed nets, which were placed directly exposed to sunlight (A1, A2) showed lower (17.2%) knock down effect compared to bed nets placed indoors (A3, B1). Twenty-four hour mortality was 100% for the five study groups (A2, A3, B1, C1, C2) whereas group A1 was found to have a lower mortality rate (44.4%). Bed nets need to avoid direct exposure to sunlight. When used and stored in appropriate conditions (cool, well-ventilated place away from sunlight) they can be used as an effective vector control tool in endemic areas.Öğe 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, YusufCutaneous (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.Öğe 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, SerayTurkey 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.Öğe 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, YusufThe 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.Öğe Evaluation of the efficacy of Olyset (R) Plus in a village-based cohort study in the Cukurova Plain, Turkey, in an area of hyperendemic cutaneous leishmaniasis(Soc Vector Ecology, 2014) Gunay, Filiz; Karakus, Mehmet; Oguz, Gizem; Dogan, Mert; Karakaya, Yasemen; Ergan, Gokhan; Kaynas, Sinan; Kasap, Ozge Erisoz; Ozbel, Yusuf; Alten, BulentThe aim of this study was to measure the protective efficacy of Olyset (R) Plus, a new long-lasting factory-treated insecticidal net (LLIN) incorporated with 2% permethrin and 1% of the synergist piperonyl butoxide (PBO), against cutaneous leishmaniasis (CL) transmission under field conditions. A village-scale trial, promoting the use of LLIN by the local inhabitants of the study area was conducted as a pilot study in a new hyperendemic focus of CL caused by a Leishmania infantum/L. donovani hybrid parasite transmitted by proven vector species Phlebotomus tobbi in Cukurova Plain, Adana, Turkey, between May, 2013 and May, 2014. The study area comprised eight villages; two of them were selected as an intervention village with Olyset (R) Plus net (Kizillar) and a control village without net application (Malihidirli). Six villages with surrounding allopatric barriers were utilized as a buffer zone cluster between intervention and control villages. Monthly entomological surveys were performed in the intervention and control villages and Damyeri, representing the other six villages, to collect adults of Phlebotomus tobbi. Results showed a significant reduction in cutaneous leishmaniasis incidence in the intervention village from 4.78% to 0.37%. The protective efficacy rate of LLIN was 92.2%. In contrast, incidence rates increased in the control village from 3.67% to 4.69%. We also evaluated residual insecticide levels of used nets after six and 12 months of usage. It was determined that the nets had retained full insecticidal strength. These results highlight the value of real-world data on bed net effectiveness and longevity to guide decisions regarding sand fly control strategies. To the best of our knowledge, this is the first field study to evaluate Olyset (R) Plus efficacy in a hyperendemic cutaneous leishmaniasis area.Öğe 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, SerayLeishmaniasis 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.Öğe First time identification of Acanthamoeba genotypes in the cornea samples of wild birds; Is Acanthamoeba keratitis making the predatory birds a target?(Academic Press Inc Elsevier Science, 2017) Karakavuk, Muhammet; Aykur, Mehmet; Sahar, Esra Atalay; Karakus, Mehmet; Aldemir, Duygu; Donduren, Omer; Ozdemir, Huseyin Gokhan; Can, Huseyin; Guruz, Adnan Yuksel; Dagci, Hande; Doskaya, MertAcanthamoeba is a free-living amoeba which can be isolated from environment and among others well known as an opportunist protozoan parasite causing infections in humans and animals. Eyes are extremely important for the wild birds and losing sight ability due to Acanthamoeba can be dangerous. The studies on Acanthamoeba infection in wild birds is very few in world and Turkey therefore we aimed to screen deceased wild birds found in Izmir and Manisa provinces located in western Turkey using PCR and non-nutrition agar (NNA) plate method. Cornea samples were obtained from 18 deceased wild birds. During the external examination, signs of keratitis were observed in two Eurasian sparrowhawks (Accipiter nisus). All of the corneal samples were analyzed by two PCR methods and NNA plate. According to results, the Acanthamoeba positivity in corneal samples was 16.6% and 5.5% by PCR and plate method, respectively. According to sequencing data, two of isolates belonged to genotype T5 and one was genotype T4. In conclusion, Acanthamoeba infection was detected in wild bird cornea samples with/without keratitis for the first time in the world. The result of this study also show that Acanthamoeba can be a cause of keratitis in wild birds of Turkey and thus these predator birds can be a target of other wild animals due to loss of sight ability. In terms of public health, these results show the importance of wild birds as a source of Acanthamoeba infection in nature. (C) 2017 Elsevier Inc. All rights reserved.Öğe Insecticide Susceptibility Status of Wild-Caught Sand Fly Populations Collected from Two Leishmaniasis Endemic Areas in Western Turkey(Iranian Scientific Society Medical Entomology, 2017) Karakus, Mehmet; Gocmen, Bayram; Ozbel, YusufBackground: In Turkey, vector control programs are mainly based on indoor residual spraying with pyrethroids against mosquitoes. No special control program is available for sand flies. Most insecticide susceptibility tests were done for mosquitoes but not for sand flies. We therefore aimed to determine the insecticide susceptibility against two commonly used insecticides; deltamethrin and permethrin, on wild-caught sand fly populations collected in two geographically separated leishmaniasis endemic areas. Methods: Insecticide susceptibility of wild-caught sand flies to deltamethrin (0.05%) and permethrin (0.75%) using ready-to use impregnated insecticide papers of WHO was investigated in 2010 based on knockdown time using standard WHO tube-test kit and procedures. Sand flies used in this study were collected from villages of Aydin (Bascayir) and Mugla (Tepecik). Results: The resistance and early resistance were detected on the sand fly population from Mugla province against deltamethrin and permethrin, respectively. However, populations from Aydin Province were sensitive to both insecticides. Conclusion: The resistance against deltamethrin and permethrin was detected on sand fly population in Mugla Province where both insecticides have been applied for long time while no resistance was found in the insecticide free area, Aydin Province. These findings can be an indicator for showing the ability for developing the insecticide resistance in sand flies. Because of the presence and dominancy of vector sand fly species of Leishmania infantum (Phlebotomus neglectus, P. tobbi) in both study areas, the systematic monitoring for resistance of sand fly populations and more attention are needed by the authorities involved in control programs for sand fly-borne diseases.Öğe Investigation of Phlebotominae (Diptera: Psychodidae) Fauna, Seasonal Dynamics, and Natural Leishmania spp. Infection in Mug?la, Southwest of Turkey(Elsevier, 2021) Pekagirbas, Metin; Karakus, Mehmet; Kasap, Ozge Erisoz; Demir, Samiye; Nalcaci, Muhammed; Toz, Seray; Ozbel, YusufDue to its geographical location, Mug?la province is one of the most frequently used places by refugees. Although leishmaniasis have been previously reported in this region, there is a lack of information on the etiological agent and possible vectors. The main objectives of this study were; i) to investigate the sand fly fauna, ii) to reveal the natural Leishmania spp. infection in wild caught sand flies using molecular tools, and iii) to determine the annual seasonal dynamics of the sand flies in Mug?la region. Totally, 2093 specimens belonging to 15 species [12 Phlebotomus, three Sergentomyia; 51 unidentified] were collected during the one-year (June 2016-June 2017) period. of the collected sand flies, 1928 (92.12%) were caught by the Centers for Disease Control (CDC) light traps, while 165 (7.88%) of them were caught by sticky traps. Phlebotomus major sensu lato (s.l.), the potential vector of visceral leishmaniasis (VL) and canine leishmaniasis (CanL) in the Mediterranean and Aegean region, was detected in all sampling locations and found as the dominant taxon (n=1035; 49.45%) of the study area and followed by Phlebotomus tobbi (n=371; 17.72%). During the sampling period, sand fly activity was started in March and peaked in August. Sand fly population size reduced dramatically between mid-September and early October. The number of collected specimens was peaked in August, while there is only one sample collected both in November and March. The majority of the sand flies (78.66%) were collected at an altitude range of 200-400 m. Seventy-two monospecific pools were screened for the presence of Leishmania DNA by real time ITS1 PCR and 24 (nine P. major s.l., eight P. tobbi, two P. papatasi, two S. minuta, one P. alexandri, one P. similis, and one Phlebotomus (Transphlebotomus spp.) of them (33.8%) were found positive (L. infantum, L. tropica, and L. major). To the best of our knowledge, the presence of fifteen sand fly species and their distribution, seasonal dynamics, molecular detection of Leishmania parasites in Mug?la province was reported for the first time. The presence of vector species in the study area, appropriate temperature and humidity conditions, long sand fly activity season, and presence of Leishmania parasite suggests that there is a serious risk in the transmission of leishmaniasis in Mug?la.Öğe Leishmaniasis in Turkey: first clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regions(Wiley, 2016) Ozbilgin, Ahmet; Culha, Guelnaz; Uzun, Soner; Harman, Mehmet; Topal, Suhan Gunasti; Okudan, Fulya; Zeyrek, Fadile; Gunduz, Cumhur; Ostan, Ipek; Karakus, Mehmet; Toz, Seray; Kurt, Ozgur; Akyar, Isin; Erat, Aysegul; Gungor, Dilek; Kayabasi, Cagla; Cavus, Ibrahim; Bastien, Patrick; Pratlong, Francine; Kocagoz, Tanil; Ozbel, YusufObjectiveTo report isolation of Leishmaniamajor strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L.major between 2011 and 2014. MethodsInitial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L.major. Developing lesions were observed, and the thickening of footpads was measured weekly. ResultsMelting curve analyses of 18 isolates showed a peak concordant with L.major, and two of them were confirmed by isoenzyme analyses as L.major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3weeks, which even progressed to extremity amputation. ConclusionCutaneous leishmaniasis-causing L.major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L.major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L.major infections in Turkey, contrary to the infections in neighbouring countries. ObjectifRapporter sur l'isolement de souches de Leishmania major obtenues a partir de 18 cas de leishmaniose cutanee (LC) de patients turcs autochtones infectes par L. major entre 2011 et 2014. MethodesLe diagnostic initial a porte sur la microscopie et la culture sur un milieu enrichi, prepare en ajoutant des quantites specifiques d'extrait de foie, de proteines et de sources de lipides au milieu NNN. Les promastigotes ont ensuite ete transferes dans le milieu RPMI contenant 10% de serum fOEtal de veau pour la culture de masse. La PCR en temps reel specifique de l'espece et ciblant la region ITS1 de Leishmania spp. a ete realisee en utilisant a la fois les echantillons d'aspiration de la lesion et de promastigotes cultives. Deux des 18 isolats ont ete identifies par analyse des isoenzymes dans le Centre de reference de la leishmaniose a Montpellier, en France. Chaque isolat a ete inocule dans les coussinets plantaires de six souris pour observer la pathogenicite de L. major. Les lesions en developpement ont ete observees et l'epaississement des coussinets plantaires ont ete mesures chaque semaine. ResultatsLes analyses de courbe de fusion des 18 isolats ont montre un pic concordant avec L. major et deux d'entre eux ont ete confirmes par des analyses d'isoenzyme comme L. major de zymodeme MON103. Dans le modele murin, des lesions aigues observees au jour 21 ont ete acceptees comme une indication de forte infection. Des deficiences severes ont ete observees sur tous les coussinets plantaires des souris pendant plus de trois semaines, qui ont meme progresse jusqu'a l'amputation de l'extremite. ConclusionL. major causant la LC a ete recemment identifie dans la province d'Adana dans le sud de la Turquie par la PCR. Notre etude montre que de tels cas de LC ne sont pas limites a Adana, mais sont actuellement presents dans l'ouest et dans le sud-est de l'Anatolie, et le long de la cote mediterraneenne. Le role des petits mammiferes, principaux reservoirs de L. major en Anatolie, devrait etre elucide, de meme que les facteurs sous-jacents qui causent les manifestations cliniques severes dans les infections a L. major en Turquie, contrairement aux infections dans les pays voisins. ObjetivoReportar el aislamiento de cepas de L. major obtenidas de 18 pacientes turcos con leishmaniosis cutanea (LC) autoctona, infectados con Leishmania major entre 2011 y 2014. MetodosEl diagnostico inicial se realizo mediante microscopia y cultivo en medio enriquecido, preparado mediante la adicion de cantidades especificas de extracto de higado, proteina y fuentes de lipido al medio NNN. Los promastigotes fueron transferidos al medio RPMI con un 10% de suero fetal para su cultivo masivo. Se realizo PCR en tiempo real, especie-especifica, que detecta la region ITS1 de Leishmania spp., utilizando tanto muestras aspiradas de las lesiones como promastigotes de cultivo. Dos de los 18 aislados se identificaron mediante analisis isoenzimatico en el Centro de Referencia de la Leishmaniosis en Montpellier, Francia. Cada aislado fue inoculado en las almohadillas de las patas de seis ratones para observar la patogenicidad de L. major. Se observo el desarrollo de las lesiones y se midio semanalmente el engrosamiento de las almohadillas. ResultadosEl analisis de la curva de fusion de los 18 aislados mostro un pico de concordancia con Leishmania major, y dos de ellos fueron confirmados mediante un analisis isoenzimatico como L. major zymodeme MON103. En el modelo de raton, las lesiones agudas observadas en el dia 21 se aceptaron como indicativas de una infeccion masiva. Se observaron danos graves en todas las almohadillas de los ratones a lo largo de tres semanas, que progresaron hasta la amputacion de las extremidades. ConclusionRecientemente se ha identificado, mediante PCR, LC causada por L. major en la provincia de Adana al sur de Turquia. Nuestro estudio muestra que estos casos de LC no estan limitados a Adana y que actualmente existen desde el oeste al sudeste de Anatolia y a lo largo de la costa Mediterranea. Es necesario aclarar el papel que en Anatolia juegan los pequenos mamiferos, principales reservorios de L. major, al igual que el de los factores que hay detras de las manifestaciones clinicas severas en infecciones por L. major en Turquia, al contrario del de las infecciones presentes en paises vecinos.Öğe Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused by Leishmania donovani in Turkey(Elsevier Science Bv, 2017) Ozbilgin, Ahmet; Harman, Mehmet; Karakus, Mehmet; Bart, Aldert; Toz, Seray; Kurt, Ozgur; Cavus, Ibrahim; Polat, Erdal; Gunduz, Cumhur; Van Gool, Tom; Ozbel, YusufIn Turkey, the main causative agents are Leishmania tropica (L. tropica) and Leishmania infantum (L. infant:tun) for cutaneous leishmaniasis (CL) and L. infantum for visceral leishmaniasis (VL). In this study, we investigated leishmaniasis cases caused by L. donovani and established animal models for understanding its tropism in in vivo conditions. Clinical samples (lesion aspirations and bone marrow) obtained from CL/VL patients were investigated using parasitological (smear/NNN) and DNA-based techniques. For species identification, a real time ITS1-PCR was performed using isolates and results were confirmed by hsp70 PCR-N/sequencing and cpb gene PCR/sequencing in order to reveal Leishmania donovani and Leishmania infantum discrimination. Clinical materials from CL and VL patients were also inoculated into two experimental groups (Group CL and Group VL) of Balb/C mice intraperitoneally for creating clinical picture of Turkish L. donovani strains. After 45 days, the samples from visible sores of the skin were taken, and spleens and livers were removed. Measurements of the internal organs were done and touch preparations were prepared for checking the presence of amastigotes. The strains were isolated from all patients and amastigotes were seen in all smears of the patients, and then isolates were immediately stored in liquid nitrogen. In real time ITS1-PCR, the melting temperatures of all samples were out of range of L. infcrnuttn, L. tropica and L. major. Sequencing of hsp70 PCR-N showed that all isolates highly identical to previously submitted L. donovani sequences in GenBank, and cpb gene sequencing showed five isolates had longer cpbF allele, whereas one isolate contained a mixed sequence of both cpbF and cpbE. All mice in both experimental groups became infected. Compared to controls, the length and width of both liver and spleen were significantly elevated (p < 0.001) in both groups of mice. However, the weight of the liver increased significantly in all mice whereas the weight of spleen increased only in VL group. Amastigotes were also seen in all touch preparations prepared from skin sores, spleen and liver. L. donovani strain was isolated from autocutaneous a VL patient first time in Turkey. Animal models using clinical samples were successfully established and important clinical differences of the isolated strains were observed.
