Genotyping of echinococcus granulosus isolates by sequencing of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene in Aydin [Aydın’da echinococcus granulosus İzolatlarının mitokondriyal sitokrom c oksidaz alt Ünite 1 geni (cox1) dizi analizi ile genotiplendirilmesi]

PubMed ID: 29933737Cyst hydatid (CH) is a zoonotic infection that is characterized by the development of metacestode form of Echinococcus granulosus primarily in liver of humans and ruminants. With a worldwide distribution, the infection is still considered as an important parasitic disease that threatens the public health in Turkey as in the other developing countries. Morphological and biological features of parasite fail to discriminate isolates for typing so molecular methods should be used for this purpose. Recently, a total of eleven genotypes of E.granulosus (G1-10 and lion strain) have been identified and these genotypes are highly correlated with host specificity of the parasite. The aim of this study, was to determine the genotypes of E.granulosus isolates from human samples in Aydın. Cyst fluids from CH operated cases in Adnan Menderes University Faculty of Medicine, Training and Research Hospital were used in the present study. Samples were washed with phosphate buffered saline (PBS) and stored in 70% ethanol at -20ºC. Mitochondrial cytochrome C oxidase subunit 1 (cox1) gene was amplified partially by polymerase chain reaction (PCR). The PCR products were sequenced initially, compared to other database in Genbank and evolutionary distances were estimated with references. The genotypes of E.granulosus isolates were determined according to the closest or exact matches to the references. A total of 20 E.granulosus isolates were genotyped in the present study, most of them (15 isolates, 75%) were identified as Genotype 1 (G1), that is defined as sheep genotype and the remaining isolates were defined as pig/camel genotype G6/7 (five isolates, 25%). A possible explanation to our results May be related to the geographical position of Turkey. The identification of G6/7 in addition to sheep genotype G1 indicated that pigs and camels in this area have role in the transmission and distribution of E.granulosus to humans. There is still limited information about the molecular epidemiology of E.granulosus in Turkey. This study reveals the first data about the genotype distribution of E.granulosus in our city, therefore the findings May help to design control program for the disease with a combination of epidemiological data. © 2018 Ankara Microbiology Society. All rights reserved

Transfection of Leishmania tropica with green fluorescent protein (gfp) gene and investigation of the in vitro drug effect

PubMed 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. © 2019 Ankara Microbiology Society. All rights reserved

Transfection of Leishmania tropica with green fluorescent protein (gfp) gene and investigation of the in vitro drug effect [Leishmania tropica'nin Yeşil Floresan Protein (gfp) Geni ile Transfeksiyonu ve İn Vitro İlaç Etkisinin Araştirilmasi]

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. © 2019 Ankara Microbiology Society. All rights reserved

Epidemic Adenoviral Keratoconjunctivitis Possibly Related to Ophthalmological Procedures in a Neonatal Intensive Care Unit: Lessons from an Outbreak

Purpose: Epidemic adenoviral keratoconjunctivitis can spread rapidly among preterm infants who frequently undergo ophthalmological examination. Here we present our experience on a nosocomial outbreak that affected 8 nursery staff members and 26 premature infants. We focus on the presentation and progress of the outbreak, the diagnosis of the disease and the measures taken for its control