The Distribution of Blastocystis Subtypes among School-aged Children in Mugla, Turkey

Background: Blastocystis is a common protozoon that inhabits human intestinal tract and has a worldwide distribution. This study aimed to determine subtype (ST) distribution of Blastocystis among school-aged children in a western city of Turkey between Mar and Jun 2014. Methods: This cross-sectional study was conducted among primary school children in Mugla between Mar and Jun 2014. Overall, 468 stool samples from children were examined by direct microscopy and inoculated into Jones medium. Blastocystis partial small subunit ribosomal RNA gene (SSU-rDNA) was amplified and sequenced from culture positive isolates. Subtypes were determined according to closest or exact match at GenBank and Blastocystis ST (18S) database. Results: The positive rate of Blastocystis was 7.4% (n=35) with xenic in-vitro culture (XIVC). The subtypes could be identified for 33 (94.2%) isolates; 12 (34.2%) were ST3, 11 (31.4%) were ST1, 9 (25.7%) were ST2, one was (2.8%) ST7. No relationship was found between Blastocystis infected and non-infected cases in terms of gastrointestinal symptoms. Additionally, none of the possible risk factors was related to Blastocystis infection. Conclusion: Subtypes in children was similar to those reported in most of the studies that found ST3 as the most common subtype.

Transfection of Leishmania tropica with Green Fluorescent Protein (gfp) Gene and Investigation of the In Vitro Drug Effect

WOS: 000465377600009PubMed ID: 31130125Cutaneous 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