Multilocus Microsatellite Typing (MLMT) of Strains from Turkey and Cyprus Reveals a Novel Monophyletic L. donovani Sensu Lato Group

In eastern Mediterranean, leishmaniasis represents a major public health problem with considerable impact on morbidity and potential to spread. Cutaneous leishmaniasis (CL) caused by L. major or L. tropica accounts for most cases in this region although visceral leishmaniasis (VL) caused by L. infantum is also common. New foci of human CL caused by L. donovani complex strains were recently described in Cyprus and Turkey. Herein we analyzed Turkish strains from human CL foci in Çukurova region (north of Cyprus) and a human VL case in Kuşadasi. These were compared to Cypriot strains that were previously typed by Multilocus Enzyme Electrophoresis (MLEE) as L. donovani MON-37. Nevertheless, they were found genetically distinct from MON-37 strains of other regions and therefore their origin remained enigmatic. A population study was performed by Multilocus Microsatellite Typing (MLMT) and the profile of the Turkish strains was compared to previously analyzed L. donovani complex strains. Our results revealed close genetic relationship between Turkish and Cypriot strains, which form a genetically distinct L. infantum monophyletic group, suggesting that Cypriot strains may originate from Turkey. Our analysis indicates that the epidemiology of leishmaniasis in this region is more complicated than originally thought

MIKROBIYOLOJI BULTENI

Cutaneous leishmaniasis (CL) is a parasitic disease transmitted by vector sand flies Phlebotomus and Lutzomyia. This disease is characterized by long time non-healing skin lesions, and caused by Leishmania species. CL is the most common infection in Eastern and Southeastern Anatolia in Turkey and L. tropica is known as the main agent of the disease. Number of cases is increasing in our country in time because of malnutrition, migration, travel, low socioeconomic level and ecological changes. For the treatment, the pentavalent antimonials are often used as intralesionally for many years, and it was reported that resistant cases have increased in recent years. New treatment methods and anti-Leishmanial activity of new agents have been investigated because of side effects, resistance development and toxic reactions of the present drugs. These studies are first carried out in vitro and afterwards with in vivo experimental animal models. Reporter gene technology has been used to investigate a variety of purposes like biological events in microorganisms and the efficacy and resistance of drugs in recent years. The major areas that green fluorescent protein (gfp) used are that they can be incorporated into different genes to determine the amount of expression of these genes in different organisms and can be used as markers in living cells. Especially gfp gene, which encodes the green fluorescent protein, is widely used nowadays. Gene-based assays have several advantages like being easy to follow-up, inexpensive and have improved biosecurity. The aim of the present study was to perform the transfection of L. tropica with enhanced gfp (egfp) and in vitro usefulness of gfp-transfectants as a drug screening model in comparison to the conventional methods. Promastigotes of L. tropica were transfected with p6.5/egfp by electroporation and selected for tunicamycin-resistance as previously described. L. tropica promastigotes transfected with gfp and in vitro effect of meglumine animoniate was assessed using different methods such as fluorescence microscopy, fluorometer and XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxyanilide) assay. The use of gfp-transfected Leishmania strains was found more rapid and more sensitive by fluorescent microscopy and fluorometry than conventional assays for the evaluation of potential anti-leishmanial agents. Consequently, stable gfp-transfected Leishmania species will be used in vitro and in vivo for screening of anti-leishmanial drugs and vaccine development as well as for understanding the biology of the host-parasite interactions at the cellular level. As a result ot this study, gfp transfected model using a Turkish L. tropica isolate was established to be used in further studies

ACTA TROPICA

In the present study, we aimed to carry out an epidemiological and entomological survey on a visceral leishmaniasis (VL) focus located on the northern central part of Anatolia, Turkey. Five villages of Corum province, where five confirmed cases of human visceral leishmaniasis (HVL) (one patient/village) were reported between June 1998 and August 2001 were included in the study. A total of 625 children and 131 dogs were sampled and the physical examination was carried out by authorized physicians and veterinarians. An indirect fluorescent antibody test (IFAT) was performed by standard procedures for human and dog sera, while the direct agglutination test (DAT) was only performed for dog sera. Sand fly collection was performed in three villages by CDC miniature light traps. Hepatosplenornegaly and hepatomegaly were detected in two and eight children, respectively. The seropositivity rate among children was found to be 0.16% (1/625) in the region. The seroprevalence of canine infection in these five villages ranged between 0.0% and 28.26%. In two villages, named Ahlatcik and Asagifindikli, no seropositive dogs were found. A total of 1218 sand flies were collected throughout the study. Six species of Phlebotonius were identified: P. transcaucasicus, P. neglectus, P. halepensis, P tobbi, P. papatasi, and P. jacusieli. P. transcaucasicus was found to be the predominant species in Cevizli (47.44%; 343/723) and Ucoluk (79.95%; 351/439) villages, while P. tobbi was abundant in Kucukerikli (42.85%; 24/56). (c) 2005 Elsevier B.V. All rights reserved

Interleukin (IL) 5 levels and eosinophilia in patients with intestinal parasitic diseases

PubMed ID: 15534922Aim: Intestinal parasitic diseases are commonly accompanied with diarrhoeal symptoms and allergic reactions. Eosinophilia occurs as a result of IL-5 synthesized from Th2 cells during allergic reactions. IL-5 acts as a factor activating eosinophils. The aim of this study was to compare the IL-5 cytokine measurements in serum samples and cell cultures. And also to compare eosinophilia observed in helminth infections and protozoon infections accompanied with allergy. Methods: Twenty-three patients who presented with diarrhoeal symptoms and allergic complaints were tested positive for intestinal parasites, as well as 21 controls with allergic complaints who did not have any intestinal parasites were included in this study. TL-5 production in in vitro cell cultures prepared by using phytohemaglutinin (PHA) to stimulate peripheral blood mononuclear cells (PBMC) obtained from the blood samples taken from these patients were compared with the IL-5 level in serum. Furthermore, the IL-5 production in protozoon and helminth infections was also compared. Absolute eosinophil values in 1 mm3 of blood were calculated by means of peripheral smear in both groups within the scope of the study. Results: Parasites such as helminth detected in 15 (65.2%) and protozoon in 8 (34.8%) of the patients were included in this study. As regards the values of the sera in both patients with parasite infection and controls, the IL-5 production was found to be higher in the cell culture supernatant (P<0.001 and P<0.05). When the IL-5 level of the patients with helminth parasites was compared with that of those with protozoon, it was determined that the IL-5 level in serum was more significant in the patients with protozoon than in those with helminth (P<0.05). In the study group, the patients were found to have parasites, the percentage of eosinophil was 7.0% compared to 6.5% in the control group. Thus, there was no significant difference between the eosinophil values (P>0.05). Conclusion: It was found that IL-5 cytokine levels in serum samples from the patients with helminth and protozoon displayed more measurable values as compared to the IL-5 levels after stimulation with mitogen. It is concluded that IL-5 acts as a triggering factor in the toxiallergic complaints commonly seen in helminth and protozoon in infections. Copyright © 2004 by The WJG Press

Seroprevalence of canine visceral leishmaniasis around the Aegean cost of Turkey [Ki{dotless}yi{dotless} ege bölgesindeki köpeklerde visseral leishmaniasis'in seroprevalansi{dotless}]

Visceral leishmaniasis (VL) is an infectious and zoonotic disease of people, wild and domestic mammals residing in tropical and subtropical climate zone. VL is a potential risk for human health and regarding the economical loss as a result of the disease, endemic areas should be recognized the effective implementation of control measures, and detailed epidemiological surveys should be performed in an attempt to obtain information about the incidence and prevalence of the disease in locations where the disease is diagnosed. Besides, in our country, most of the traveling has been drifted to areas dominating Mediterranean climate increasing the importance of investigations on visceral leishmaniais in Aegean region. In this study, it was aimed to determine the seroprevalence of the infection from Selçuk/İzmir, Turgutlu/Manisa, Aydi{dotless}n city province, Kuşadasi{dotless}/Aydi{dotless}n, Marmaris/Mugla, Bodrum/Mugla in stray dogs that are regarded as the main reservoirs for Visceral Leishmaniasis. For this purpose, anti-leishmania antibodies were determined with immunofluorence antibody diagnostic prosedure in serum samples obtained from untreated and unprotected 300 stray dogs located in these settlements. According to the results of the study 27 (9%) out of a total 300 dogs were infected with canine visceral leishmaniasis (CanL). It was considered that the result of this study may be used as a reference for further studies regarding human and dogs residing in Turkey

Prevalence of Leishmania infantum and Dirofilaria immitis infection in dogs in Aydin province and the town of Selçuk, İzmir, Turkey [Aydin'in bazi i·lçe ve köyleri ile İzmir'in Selçuk i·lçesindeki köpeklerde Leishmaniosis ve Dirofilariasis'in prevalansi]

The aim of this study was to determine the prevalence of Leishmania infantum and Dirofilaria immitis infection in dogs housed outdoors in Aydin province and Selçuk in western Turkey. Anti-Leishmania antibodies were evaluated in sera samples by IFA and ELISA techniques. The presence of D. immitis microfilariae was determined in blood samples by use of the modified Knott's test. Using the IFA and ELISA on 158 dogs surveyed, 5 (3.2%) were positive for anti-Leishmania antibodies while 22 (13.9%) were found infected with D. immitis. Only 2 out of 5 seropositive dogs exhibited major signs of canine visceral leishmaniosis, and amastigotes were detected in parasitological examinations of popliteal lymph node aspirates. It was noted that there were dogs infected with Leishmania in dog shelters in the towns of Kuşadasi and Selçuk, and the infection rate of dogs for D. immitis was high in 3 villages near the city of Aydin. © TÜBİTAK

Effectiveness of peptone-yeast extract (P-Y) medium in the cultivation and isolation of Entamoeba histolytica/Entamoeba dispar in Turkish patients

WOS: 000181429700006PubMed ID: 12614984Amebiasis is a common protozoan infection worldwide, causing serious health problems in both children and adults. Today, almost 10% of the world population is infected with Entamoeba histolytica/Entamoeba dispar. The aims of this study were both the comparison of the reproduction rates,and densities of E. histolytica/E. dispar in Robinson, Dobell-Laidlaw and P-Y culture media and isolation of E. histolytica/E. dispar from stool samples in Peptone-Yeast (P-Y) medium. Trophozoites and cysts of E. histolytica/E. dispar, maintained in Robinson medium, and stool samples of patients with amebiasis were inoculated into P-Y, Robinson and Dobell-Laidlaw culture media. Reproduction rates reached their peak levels 48 h after the inoculation in all culture media. Reproduction rates in P-Y and Robinson media were found similar; however, they were higher than the reproduction rate in Dobell-Laidlaw medium (p 0.05). Twelve isolates from 12 patients were cultivated in P-Y medium and checked for reproduction everyday for 7 days. Twelve of the 12 (100%) isolates were cultivated in P-Y medium, indicating that the P-Y was an effective medium for the isolation of E. histolytica/E. dispar in stool samples. According to these results, P-Y medium could be preferred in immunologic, serologic and molecular studies and, thus the definitive diagnosis of amebiasis due to its low cost and simple formula. (C) 2003 Elsevier Science Inc. All rights reserved