Browsing by Author "Östan, I"
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Item In vitro Efficacy of Quercus infectoria Oliv. and Achillea millefolium L. Extracts Against Blastocystis spp. IsolatesÖzbilgin, A; Durmuskahya, C; Kilimcioglu, AA; Kayalar, H; Kurt, Ö; Ermis, VÖ; Tabak, T; Östan, IBlastocystis is a common intestinal parasite that can inhabit the intestinal tract of humans and many animals. Despite it was firstly described almost 100 years ago; many subjects are still under debate about Blastocystis, including its life-cycle, pathogenic potential and treatment of infected individuals. Historically, local plant species have been used for therapeutic purposes by the local people of Anatolia. Here, hexane and methanol extracts of two local plants, Quercus infectoria (Fagaceae) and Achillea millefolium, which have been used against diarrhea in Anatolia, were examined for their in vitro efficacies against Blastocystis. LC50 and EC50 values of the plant extracts were determined by Brine Shrimp and Graphpad Prism 5 (R) methods, respectively. The results showed that LC50 (500 mu g/ml) and EC50 (198.8 mu g/ml) concentrations of the methanol extract of A. millefolium were lowest compared to other extracts, its anti-Blastocystis activity was found to be comparable to metronidazole and it showed no cytotoxic activity. These initial results suggest that the methanol extract of A. millefolium may be a novel option for the treatment of Blastocystis infections in humans in future, if confirmed by further, larger-scale studies.Item Comparison of culture media in the isolation and diagnosis of cutaneous leishmaniasisÖzbilgin, A; Zeyrek, F; Limoncu, ME; Östan, I; Tabak, T; Asar, K; Türkmen, H; Kilimcioglu, AALeishmaniasis is a common and significant parasitic disease among the Middle East and Mediterranean countries. Cutaneous and visceral forms of leishmaniasis are seen in Turkey. Direct microscopic examination of Giemsa-stained smears and inoculation into a culture medium are gold standards in diagnosis. In the present study, three diffferent culture media (pepton-yeast extract, brain heart and nutrient broth) were used in comparison with Novy-MacNeal-Nicolle and RPMI 1640 as control media to assess the effectiveness of new culture media both in diagnosis in patients and after isolation from the mice. In addition, smears prepared from the infectious material were stained with Giemsa and examined under a light microscope. Samples obtained from the lesions of cutaneous leishmaniasis produced on the sole of mice were inoculated into all culture media; growth were detected in pepton-yeast extract and nutrient broth media. Smears prepared from the lesions of these mice were stained with Giemsa and amastigotes were detected on them. Growth was also seen in control media. Five of 12 cultivations of the human samples suspected of having cutaneous leishmaniasis revealed growth in pepton-yeast extract medium and nutrient broth medium. Microscopic examination of these five patients showed the presence of amastigotes and growth in control media, while none was observed in the remaining seven cases. These results indicated that pepton-yeast extract and nutrient broth media could be preferred in both isolation of the parasites and diagnosis of cutaneous leishmaniasis.Item A mass screening survey of cystic echinococcosis by ultrasonography, Western blotting, and ELISA among university students in Manisa, TurkeyKilimcioglu, AA; Girginkardesler, N; Korkmaz, M; Özkol, M; Düzgün, F; Östan, I; Pabuscu, Y; Dinç, G; Ok, ÜZCystic echinococcosis (CE) is one of the most important zoonotic diseases in a wide geographic area, including Turkey. In the present project, a total of 4275 students from Celal Bayar University, Manisa, Turkey, were screened by ultrasonography (US) and specific antibodies for CE were examined by Western blotting (WB) and ELISA in finger prick blood samples of 2034 of 4275 volunteered students. We aimed to report the apparent prevalence of CE based on different diagnostic procedures and to compare WB and ELISA with US in diagnosis of CE in a mass screening setting. Six new cases were diagnosed as CE by US during the survey. In addition to these cases, three students were also detected to have been previously operated and pathologically confirmed for hepatic CE. US revealed parenchymal changes in these cases in concordance with their operation history; so, the prevalence of CE by US was calculated as 0.21% (9/4275) (95%CI, 0.11-0.39%) among university students in Manisa. Bands were detected at 8, 28, 32, 38, 42, 47, 70 and 90 kDa by WB and the cases were considered to be positive for CE when at least three of the bands were seen together. Apparent prevalence of CE by ELISA and WB were found to be 2.11% (43/2034) (95%CI, 1.57-2.83%) and 0.25% (5/2034) (95%CI, 0.10-0.57%), respectively. Of the six US positive cases, WB was positive in only one case with two cysts in the liver. All of four cases with liver involvement were positive by ELISA. The high prevalence of CE among university students in Manisa indicated that CE is a major health problem in this area of Turkey. Our results supported that WB is rather difficult and not feasible as a mass screening test and may not be effective for confirmation especially in asymptomatic cases. As a result, we recommend US to be used initially in mass screening surveys for CE followed by confirmation by ELISA for suspected cases. Further examination primarily by chest X-ray followed by computed tomography and/or magnetic resonance imaging, if needed, should be recommended for US negative, ELISA and WB positive individuals who may have non-abdominal cysts. (C) 2013 Elsevier B.V. All rights reserved.Item Health inequitiesÖstan, I; Kilimcioglu, AA; Girginkardesler, N; Özyurt, BC; Limoncu, ME; Ok, ÜZBackground: Intestinal parasitic infections affect child health and development and slow down growth, while reducing adults' productivity and work capacity. The aim of the present study was to determine and compare the incidences of intestinal parasitic infections and the socio-economic status of two near primary school children in Manisa, a western city of Turkey. Methods: A total of 352 children were involved a questionnaire study from a private school (Ulkem Primary School-UPS, 116 children) and a community-based school (Sehzadeler Primary School-SPS, 236 children). Of these, stool samples could be obtained from a total of 294 students; 97 (83.6%) from UPS, and 197 (83.5%) from SPS. The wet mount preparations of the stool samples were examined; samples were also fixed in polyvinyl alcohol and examined with modified formalin ethyl acetate sedimentation and trichrome staining techniques. Data were analyzed using SPSS for Windows version 10.0. The chi-squared test was used for the analytic assessment. Results: The percentages of the students found to be infected with intestinal parasites, were 78 (39.6%) and 13 (13.4%) in SPS and UPS, respectively. Totally 91 (31.0%) of the students from both schools were found to be infected with at least one intestinal parasite. Giardia lamblia was found to be the most common pathogenic intestinal parasite and Blastocystis hominis was prevalent independently from the hygienic conditions. The factors which significantly (p < 0.05) increase the incidence of intestinal parasites were uneducated and unemployed mother, lower social status of father, living in crowded houses with insufficient indoor spaces, using the tap water as drinking water, and living at shanty areas. Conclusion: Intestinal parasitic infections in school children were found to be a public health problem that increased due to lower socio-economic conditions. We conclude that organization of education seminars including the topics such as prevention of the infectious diseases, improving general hygienic conditions, and application of supportive programs for the parents may be suggested not only to reduce intestinal parasitic infections, but also to elevate the socio-cultural levels.Item The value of ultrasonography alone in screening surveys of cystic echinococcosis in children in TurkeyKilimcioglu, AA; Özkol, M; Bayindir, P; Girginkardesler, N; Östan, I; Ok, ÜZA total of 1205 primary school children were examined for cystic echinococcosis in five villages of Manisa, Turkey, to evaluate the efficacy of diagnostic methods of this infection in community-based screening surveys. Six hundred and thirty children from three villages, examined by a portable ultrasound scanner, chest microfilm and serological methods (ELISA, indirect hemagglutination) in our previous study, were designated as Study Group 1; and 575 children, from two adjacent villages, examined by ultrasonography alone in the present study, were designated as Study Group 2. In Study Group 1, hepatic cystic echinococcosis was detected in two cases (0.3%) by ultrasonography, while 43 (8.9%) and 49 (10.1%) cases were found to be positive for cystic echinococcosis by ELISA and indirect hemagglutination, respectively. Three of 575 children (0.5%) were diagnosed with cystic echinococcosis (two hepatic and one renal involvement) by ultrasonography alone in Study Group 2; and lung lesions were later detected in both cases with liver involvement by chest radiography. Our results suggested that serological tests may be beneficial in suspected cases for confirmation and differential diagnosis, but have some drawbacks, such as discrepancy in results and high false seropositivity rates. Chest microfilm is not easy in field studies and exposure to X-ray is undesirable. As a reliable, simple, inexpensive and rapid technique, ultrasonography alone is recommended to be used in community-based screening surveys for cystic echinococcosis with confirmatory tests for suspected cases found during the screening program. (c) 2006 Elsevier Ireland Ltd. All rights reserved.Item In Vitro Cultivation of Plasmodium berghei: Application of Drug Resistance Tests with Chloroquine and ArtesunateÖstan, I; Kurt, Ö; Özbilgin, AThe aim of this study is to assess the drug resistance testing of Plasmodium berghei after short-term culture in vitro. First stage, the life cycle of the malaria parasites was completed in an average of 24 h inside the red blood cells in vitro. Second stage, the inhibition rates of Chloroquine and Artesunate on the infected erythrocytes were determined. The results showed that P. berghei parasites were completed their erythrocytic schizogony in short term in vitro cultivation. In the drug resistance tests with Chloroquine and Artesunate, antiplasmodial resistance did not occur with both drugs and the effect of Artesunate is higher than Chloroquine.Item Two novel serum-free media for the culture of Trichomonas vaginalisLimoncu, ME; Kilimcioglu, AA; Kurt, Ö; Östan, I; Özkütük, N; Özbilgin, ADefinitive diagnosis of Trichomonas vaginalis, one of the common causes of sexually transmitted diseases in Turkey, relies on the microscopic examination of both fresh preparations and culture material of genital secretions and urine. The aim of this study was to compare the diagnostic efficacies of two culture media, serum-free TB1 and TB2, including iron and vitamin B12, respectively, with the well-known medium, TYM. Growth rate reached peak levels 48 h in TYM and 72 h in both TB1 and TB2 after inoculation. The highest amount of viable trophozoites has been obtained from TB1, almost equal to TYM but significantly higher than TB2. Fresh preparations obtained from the vaginal secretions of 119 patients have been examined and vaginal samples have been inoculated in TB1, TB2, and TYM. Viable T. vaginalis trophozoites have been detected in ten (8.4%) of fresh preparations and 11 (9.9%) of each medium. It is concluded that serum-free TB1 medium could be used effectively in both the isolation and maintenance of T. vaginalis culture in vitro.Item In vitro and in vivo activities of Haplophyllum myrtifolium against Leishmania tropicaÖstan, I; Saglam, H; Limoncu, ME; Ertabaklar, H; Toz, SÖ; Özbel, Y; Özbilgin, AThis study aimed to evaluate the in vitro and in vivo leishmanicidal activity of an endemic Turkish plant and compare its efficacy with a reference drug. In addition to the in vitro activities of the ethanol, acidified and alkaloid extracts and furoquinoline alkaloids skimmianine and gamma-fagarine, in vivo antileishmanial activitiy of the acidified extract of Haplophyllum myrtifolium Boiss. (Rutaceae) were investigated against Leishmania tropica (L. tropica), a causative agent of cutaneous leishmaniasis. All the extracts and pure compounds showed in vitro inhibitory activity against the promastigotes of. L. tropica. The in vitro 50% inhibitory concentrations of gamma-fagarine, acidified extract, ethanol extract, skimmianine and alkaloid extract against promastigotes were determined as 8.7, 9.4, 10.9, 25.7 and 25.8 mu g/ml respectively. In vivo results of Haplophyllum myrtifolium acidified extract showed that this plant has a limited effect on decreasing the lesion size of experimental mice infected with Leishmania tropica. To the best of our knowledge, this is the first time both the in vitro and in vivo antileishmanial activity of Haplophyllum mrytifolium have been reported in the same research.Item The prevalence of intestinal parasites in the province of Izmir, TurkeyDagci, H; Kurt, Ö; Demirel, M; Östan, I; Azizi, NR; Mandiracioglu, A; Yurdagül, C; Tanyüksel, M; Eroglu, E; Ak, MStool samples of a total of 2,047 people in Izmir province were examined by wet mount, formalin ethyl acetate concentration, and trichrome staining methods with an aim to reveal the prevalence of intestinal parasites in Izmir together with related personal and environmental risk factors. Geographical mapping showing the density and variation of the species of intestinal parasites in Izmir was done after all maps were scaled, and the coordinates were determined with GeoMedia5.0(R). The prevalence of the intestinal parasites was found to be 25.6% in Izmir, with a variation between the districts. Blastocystis hominis was the leading parasite, and the prevalence of parasites was higher in children compared to adults; however, the difference was statistically insignificant. There was also no significant difference between the parasite prevalence and sex, marital status, education, income, frequent eating outside, and habitual raw meat eating. Significant differences were found between the parasite prevalence and crowded families, early immigrants, individuals with no social security, and people living in close contact with their livestock. The parasites were found to be less common among individuals who had been drinking bottled water and living in a house with a sewage system. The results demonstrated a correlation between the intestinal parasites and environmental conditions in our study group. We further plan to expand the study group to cover all regions of Turkey.Item Leishmaniasis in Turkey: first clinical isolation of Leishmania major from 18 autochthonous cases of cutaneous leishmaniasis in four geographical regionsÖzbilgin, A; Çulha, G; Uzun, S; Harman, M; Topal, SG; Okudan, F; Zeyrek, F; Gündüz, C; Östan, I; Karakus, M; Töz, S; Kurt, Ö; Akyar, I; Erat, A; Güngör, D; Kayabasi, Ç; Çavus, I; Bastien, P; Pratlong, F; Kocagöz, T; Özbel, YObjectiveTo report isolation of Leishmaniamajor strains obtained from 18 Turkish autochthonous cutaneous leishmaniasis (CL) patients infected with L.major between 2011 and 2014. MethodsInitial diagnosis relied on microscopy and culture in enriched medium, prepared by adding specific amounts of liver extract, protein and lipid sources to NNN medium. Promastigotes were then transferred to RPMI medium including 10% of foetal calf serum for mass culture. Species-specific real-time PCR targeting ITS1 region of Leishmania spp. was performed using both lesion aspiration samples and cultured promastigotes. Two of 18 isolates were identified by isoenzyme analysis in the Leishmaniasis Reference Center in Montpellier, France. Each isolate was inoculated into the footpads of six mice to observe the pathogenicity of L.major. Developing lesions were observed, and the thickening of footpads was measured weekly. ResultsMelting curve analyses of 18 isolates showed a peak concordant with L.major, and two of them were confirmed by isoenzyme analyses as L.major zymodeme MON103. In the mouse model, acute lesions seen on day 21 were accepted as an indication of heavy infection. Severe impairments were observed on all mouse footpads over 3weeks, which even progressed to extremity amputation. ConclusionCutaneous leishmaniasis-causing L.major was recently identified in Adana province in southern Turkey, with PCR. Our study shows that such CL cases are not limited to Adana but currently present from western to Southeastern Anatolia, and along the Mediterranean coast. The role of small mammals, the main reservoirs of L.major in Anatolia, needs to be elucidated, as do the underlying factors that cause severe clinical manifestations in L.major infections in Turkey, contrary to the infections in neighbouring countries. ObjectifRapporter sur l'isolement de souches de Leishmania major obtenues a partir de 18 cas de leishmaniose cutanee (LC) de patients turcs autochtones infectes par L. major entre 2011 et 2014. MethodesLe diagnostic initial a porte sur la microscopie et la culture sur un milieu enrichi, prepare en ajoutant des quantites specifiques d'extrait de foie, de proteines et de sources de lipides au milieu NNN. Les promastigotes ont ensuite ete transferes dans le milieu RPMI contenant 10% de serum fOEtal de veau pour la culture de masse. La PCR en temps reel specifique de l'espece et ciblant la region ITS1 de Leishmania spp. a ete realisee en utilisant a la fois les echantillons d'aspiration de la lesion et de promastigotes cultives. Deux des 18 isolats ont ete identifies par analyse des isoenzymes dans le Centre de reference de la leishmaniose a Montpellier, en France. Chaque isolat a ete inocule dans les coussinets plantaires de six souris pour observer la pathogenicite de L. major. Les lesions en developpement ont ete observees et l'epaississement des coussinets plantaires ont ete mesures chaque semaine. ResultatsLes analyses de courbe de fusion des 18 isolats ont montre un pic concordant avec L. major et deux d'entre eux ont ete confirmes par des analyses d'isoenzyme comme L. major de zymodeme MON103. Dans le modele murin, des lesions aigues observees au jour 21 ont ete acceptees comme une indication de forte infection. Des deficiences severes ont ete observees sur tous les coussinets plantaires des souris pendant plus de trois semaines, qui ont meme progresse jusqu'a l'amputation de l'extremite. ConclusionL. major causant la LC a ete recemment identifie dans la province d'Adana dans le sud de la Turquie par la PCR. Notre etude montre que de tels cas de LC ne sont pas limites a Adana, mais sont actuellement presents dans l'ouest et dans le sud-est de l'Anatolie, et le long de la cote mediterraneenne. Le role des petits mammiferes, principaux reservoirs de L. major en Anatolie, devrait etre elucide, de meme que les facteurs sous-jacents qui causent les manifestations cliniques severes dans les infections a L. major en Turquie, contrairement aux infections dans les pays voisins. ObjetivoReportar el aislamiento de cepas de L. major obtenidas de 18 pacientes turcos con leishmaniosis cutanea (LC) autoctona, infectados con Leishmania major entre 2011 y 2014. MetodosEl diagnostico inicial se realizo mediante microscopia y cultivo en medio enriquecido, preparado mediante la adicion de cantidades especificas de extracto de higado, proteina y fuentes de lipido al medio NNN. Los promastigotes fueron transferidos al medio RPMI con un 10% de suero fetal para su cultivo masivo. Se realizo PCR en tiempo real, especie-especifica, que detecta la region ITS1 de Leishmania spp., utilizando tanto muestras aspiradas de las lesiones como promastigotes de cultivo. Dos de los 18 aislados se identificaron mediante analisis isoenzimatico en el Centro de Referencia de la Leishmaniosis en Montpellier, Francia. Cada aislado fue inoculado en las almohadillas de las patas de seis ratones para observar la patogenicidad de L. major. Se observo el desarrollo de las lesiones y se midio semanalmente el engrosamiento de las almohadillas. ResultadosEl analisis de la curva de fusion de los 18 aislados mostro un pico de concordancia con Leishmania major, y dos de ellos fueron confirmados mediante un analisis isoenzimatico como L. major zymodeme MON103. En el modelo de raton, las lesiones agudas observadas en el dia 21 se aceptaron como indicativas de una infeccion masiva. Se observaron danos graves en todas las almohadillas de los ratones a lo largo de tres semanas, que progresaron hasta la amputacion de las extremidades. ConclusionRecientemente se ha identificado, mediante PCR, LC causada por L. major en la provincia de Adana al sur de Turquia. Nuestro estudio muestra que estos casos de LC no estan limitados a Adana y que actualmente existen desde el oeste al sudeste de Anatolia y a lo largo de la costa Mediterranea. Es necesario aclarar el papel que en Anatolia juegan los pequenos mamiferos, principales reservorios de L. major, al igual que el de los factores que hay detras de las manifestaciones clinicas severas en infecciones por L. major en Turquia, al contrario del de las infecciones presentes en paises vecinos.Item Leishmaniasis in Turkey: Determination of Leishmania Species by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS)Culha, G; Akyar, I; Zeyrek, FY; Kurt, Ö; Gündüz, C; Töz, SÖ; Östan, I; Cavus, I; Gülkan, B; Kocagöz, T; Özbel, Y; Özbilgin, ABackground: Cutaneous leishmaniasis (CL) is endemic in Southeastern Anatolia, mainly in Sanliurfa and Hatay provinces, and the causative agents are mostly Leishmania tropica and less frequently L. infantum. Here, we report the first MALDI-TOF analyses of Leishmania promastigotes obtained from the cultures of two CL cases from Osmaniye and Hatay provinces who were initially diagnosed by microscopy, culture and identified as L. infantum with Real-Time PCR (RT-PCR). Methods: Samples obtained from the skin lesions of patients were initially stained with Giemsa and cultivated in NNN medium. Examination of the smears and cultures revealed Leishmania amastigotes and promastigotes, respectively. The promastigotes (MHOM/TR/2012/CBU15 and MHOM/TR/2012/MK05) obtained from the cultures of both patients were used for RT-PCR targeting the ITS-1 region in the SSU of rRNA. The reference strains of four Leishmania species (L. infantum, L. donovani, L. tropica and L. major) were initially assessed with MALDI-TOF and their data were added to MALDI-TOF Biotyper Library. Results: Both RT-PCR and MALDI-TOF analyses indicated that the causative agent in both patient samples was L. infantum. Conclusion: Despite disadvantages such as requirement of culture fluid with nothing but promastigotes and high cost, MALDI-TOF analysis may be a fast, sensitive and specific diagnostic tool in especially large-scale research studies, where the cost declines, relatively.Item A province-based study using sampling method to investigate the prevalence of cystic echinococcosis among primary school children in Manisa, TurkeyOk, UZ; Özkol, M; Kilimcioglu, AA; Dinç, G; Bayindir, P; Östan, I; Pabusçu, Y; Özcan, C; Korkmaz, M; Coskun, S; Yüksel, H; Girginkardesler, NA province-based field study using a portable ultrasound scanner (US) was performed for the first time using sampling method to investigate the prevalence of cystic echinococcosis (CE) in primary school children in Manisa, Turkey. A total of 6093 children from 37 primary schools was selected as the representative sample of the total number of 166,766 primary school children, and examined by the US. Prevalence was found to be 0.15%, as nine children were diagnosed with CE, seven new and two previously operated. A questionnaire applied to the children revealed no significant relationship between the risk factors and the infection (P > 0.05). In conclusion, it would be advisable to repeat the study at the same schools in eight years' time, in order to evaluate the efficacy of the control programs, since nearly all students involved in this study will have graduated by then. Besides, it is recommended to choose a sampling group to find the prevalence of an infection in a defined region. (c) 2007 Elsevier B.V. All rights reserved.