Hatice Ertabaklar

A real-time ITS1-PCR based method in the diagnosis and species identification of Leishmania parasite from human and dog clinical samples in Turkey.

Human visceral leishmaniasis (VL) caused by L. infantum and cutaneous leishmaniasis (CL) caused by L. tropica and L. infantum have been reported in Turkey. L. infantum is also responsible for canine leishmaniasis (CanL) and it is widely common in the country. The main aim of the present study was to design a real-time PCR method based on the internal transcribed spacer 1 (ITS1) region in the diagnosis of all clinical forms of leishmaniasis in Mediterranean, and to identify the species directly from clinical samples. Totally, 315 clinical specimens, human/canine visceral (blood, bone marrow, lymph node) and cutaneous (lesion aspiration) samples, and 51 Turkish Leishmania isolates typed by isoenzymatic method were included in the study. For optimization, DNA samples of the 34 strains were amplified by conventional ITS1-PCR and then sequenced for designing the primers and probes, allowing the species identification. Following the validation with the isolates, the test was applied on clinical samples and melting temperatures were used for genotyping. A group of PCR products were further sequenced for confirmation and assigning the inter- and intraspecies heterogeneity. The diagnosis of leishmaniasis is successfully achieved by the new real-time PCR method, and the test identified 80.43% of human and canine VL samples as L.infantum and 6.52% as L.tropica; 52.46% of CL samples as L. infantum and 26.90% as L. tropica. In 13.04% of visceral and 20.62% of cutaneous samples, two peaks were observed. Hovewer, the higher peak was found to be concordant with the sequencing results in 96.96%, in terms of species identification. The real-time ITS1 PCR assay clearly identified the leishmanial species in 81.58% of all clinical samples. Genotypic variations of Leishmania parasites in Turkey within species and intraspecies were observed, and L. tropica is also found as causative agent of human and canine VL in Turkey.

Epilepsy and seropositivity rates of Toxocara canis and Toxoplasma gondii

PURPOSE Increased seropositivity for Toxoplasma gondii and Toxocara canis have been observed in epileptic patients. Our aim is to determine whether there is any relationship between these agents and epilepsy in our cryptogenic epilepsy group. MATERIAL AND METHODS We studied specific IgG antibodies against T. gondii and T. canis in 100 cryptogenic epileptic patients and 50 healthy volunteers that had no history of epilepsy in their first degree relatives. We studied T. gondii and T. canis-specific IgG antibody serum levels and compared the values of these two groups. RESULT We found similar T. gondii and T. canis serum IgG antibodies in patients with cryptogenic epilepsy and in the control group, even though the control group included more animal owners. CONCLUSION We did not show any relationships between epilepsy and positive T. gondii and T. canis serology in our epileptic patients.

Leishmaniasis in Turkey: molecular characterization of Leishmania from human and canine clinical samples

Human leishmaniasis, both visceral and cutaneous, and canine leishmaniasis have been reported in Turkey for centuries. However, the advent of new diagnostic tools during the last 30 years has led to the recognition that leishmaniasis is an important public health problem throughout the country. In most disease foci both canine and human leishmaniases exist together and identification of parasite species causing these diseases is a pre‐requisite for understanding disease epidemiology. A total of 109 samples obtained from human and canine leishmaniasis cases were examined using internal transcribed spacer 1 PCR followed by restriction fragment length polymorphism analysis. Our results indicate that two species, Leishmania tropica and Leishmania infantum, are primarily responsible for cutaneous and visceral leishmaniasis, respectively, in Turkey. However, a new focus of human cutaneous leishmaniasis caused by L. infantum in Hatay region is described. This finding further stresses the importance of Leishmania species molecular characterization in prescribing appropriate therapy and understanding the disease’s transmission in different endemic foci.

Spatial distribution of phlebotomine sand flies in the Aydin Mountains and surroundings: the main focus of cutaneous leishmaniasis in western Turkey.

ABSTRACT: An entomological survey was conducted to determine the spatial distribution of phlebotomine fauna and understand the effect of environmental factors. The entomological survey was carried out during 2006–2007 in a study area in the rural area of Aydin province, near the Kusadasi town where VL, CL, and canine leishmaniasis (CanL) are endemic. In 2006 and 2007, 132 locations were sampled using sticky traps mainly on embankments. Detailed environmental and meteorological information was also collected for each location. The results of entomological studies indicated that the probable vectors are Phlebotomus tobbi and P. neglectus for VL and CanL, and P. similis for CL in this western leishmaniasis focus. The data revealed a correlation between their presence and spatial variables such as altitude, sampling site location, and humidity. The distribution areas of probable vector species in this study area allowed the identification of risk levels, which may provide useful information to guide the leishmaniasis research in endemic regions.

Acanthamoeba keratitis due to Acanthamoeba genotype T4 in a non-contact-lens wearer in Turkey

An otherwise healthy 22-year-old man presented with Acanthamoeba keratitis (AK) in the right eye. He was not a contact lens wearer and had no history of corneal trauma. The Acanthamoeba strain isolated from a corneal scraping was identified as morphological group II and genotype T4. Three more Acanthamoeba strains isolated from sites of possible human contact with acanthamoebae in the same geographical region, including a lens storage case, tap water and soil, were subjected to morphological and molecular biological identification. Whereas the strain from tap water also exhibited genotype T4, the two other isolates were identified as morphological group I and genotype T9. To the best of our knowledge, this is the first study identifying an AK-causing Acanthamoeba strain in Turkey and the first isolation of genotype T9 in this country.

Prevalence of Demodex folliculorum in young adults: relation with sociodemographic/hygienic factors and acne vulgaris.

474 JEADV 2006, 20, 461–488

Detection of Leishmania major and Leishmania tropica in domestic cats in the Ege Region of Turkey.

Leishmaniosis is a group of diseases caused by different species of Leishmania parasites in mammalian species. The aim of the present study was to investigate the presence of Leishmania spp. DNA in cats using real time polymerase chain reaction (RT-PCR) assays targeting internal transcribed spacer (ITS1) and heat-shock protein 70 gene (Hsp70) regions with Leishmania species-specific primers and probes. Blood samples were collected from 147 cats (73 female; 74 male) in the endemic regions for zoonotic visceral leishmaniasis in the western provinces of Turkey and analyzed using two RT-PCR assays. Additionally, Hsp70 RT-PCR products were sequenced. ELISA assays for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) were also carried out for 145 of the 147 samples. Overall, 13/147 (8.84%) cats were positive for Leishmania by RT-PCR (4 L. major and 9 L. tropica). FIV and FeLV antibody and/or antigen was detected in 4 and 5 cats among Leishmania DNA positives, respectively. To the best of our knowledge, this study is the first to investigate and report the presence of L. major and L. tropica infections in a large group of domestic cats in Turkey. The results obtained indicate that species identification of Leishmania is essential for epidemiological understanding and that clinical signs alone are not indicative for leishmaniosis in cats, as it is in dogs. This study suggests that extensive research should be carried out in cat populations in order to fully understand the role of cats in the epidemiology of the disease.

Blastocystis in ulcerative colitis patients: Genetic diversity and analysis of laboratory findings.

OBJECTIVE To determine Blastocystis frequency and subtypes (ST) in ulcerative colitis (UC) patients and analyse some laboratory findings between Blastocystis positive and negative cases. METHODS Faecal samples from 150 UC patients in Adnan Menderes University, Training and Research Hospital were examined by direct microscopy and cultivated in Jones medium. Blastocystis positive cultures were subjected to DNA isolation and subtypes were identified by sequencing of barcode region. A retrospective analysis was conducted on C reactive protein (CRP), leucocyte counts (WBC), neutrophil counts, and sedimentation rates. RESULTS The overall positive rate of Blastocystis was 8% (12 patients) and the most abundant subtype was ST3 (eight isolates, 66.7%), followed by ST1, ST2 and ST7. Laboratory findings between Blastocystis infected and non-infected UC patients were not significantly different. Blastocystis frequency was 3.8% among the patients in active stage, while it was 11.8% among the patients in remission stage. CONCLUSIONS The present study confirms previous findings that have indicated the predominance of Blastocystis ST3 in humans and contributes additional evidence that suggests the low colonisation of Blastocystis infection in ulcerative colitis patients during active stage.

Evaluation of conjunctival swab sampling in the diagnosis of canine leishmaniasis: A two-year follow-up study in Çukurova Plain, Turkey

The 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 Çukurova 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 Çukurova 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.

[Research to investigate human cystic echinococcosis with ultrasound and serologic methods and educational studies in different provinces of Aydın/Turkey].

OBJECTIVE Cystic echinococcosis (CE) is particularly prevalent in rural areas where domestic animal breeding is common. We planned to investigate the CE among the 7-88 years old inhabitants of (n=209) four villages in the Aydın district, by a portable ultrasound scanner (US) and with serologic methods. METHODS Participants were assessed by ultrasound and sera samples were tested for anti-E. granulosus antibodies by serological tests (ELISA and IHA). A questionnaire was given to the participants in order to assess their knowledge about the disease. RESULTS Hepatic CE was detected in one case (0.47%) by US, while 24 (11.48%) and 28 (13.39%) cases were found to be seropositive for CE by ELISA and IHA, respectively. With both tests, in 9 participants (4.3%), parasite-specific antibodies were detected. According to questionnaire results, 84.22% of the participants were found to have no knowledge about the disease. Only 15.21% of the dog owners were giving anti-parasite drugs to their dogs. CONCLUSION This study is one of the small number of community-based studies about CE performed in Turkey, in which US was applied. Our results suggest that we need national control programs including a multi-sectorial collaboration to eradicate hydatidosis and effective actions to control cystic echinococcosis.