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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. [ABSTRACT FROM AUTHOR]

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This study aimed to evaluate clinical signs related with canine leishmaniosis (CanL) and to determine their relation with serological results in the differential diagnosis of this disease in dog populations. A total of 253 dogs from houses and a dog shelter in KuSadasý, an endemic region for leishmaniosis, were investigated for clinical signs related to CanL. Sera collected from the study group were examined by indirect fluorescence antibody (IFA), whole ELISA and rK39 ELISA tests for the serological diagnosis. Popliteal lymph node aspiration materials were examined for the parasitological diagnosis. Clinical signs were separated into two groups as cutaneous and visceral signs and no clinical signs were observed in 51.7% (131/253) of the dogs, while 48.3% (122/253) of the dogs had at least one sign. A total of 42 dogs [29 (23.7%) in the symptomatic and 13 (9.9%) in the asymptomatic group] were diagnosed as CanL serologically and/or parasitologically. Two strains were isolated and identified as Leishmania infantum MON-1. There was no correlation between IFAT titers and the number of the clinical signs (P > 0.05; R square = 0.002). Weight loss was found to be the most common sign of CanL while epistaxis was the least common but specific sign. The seropositivity ratio of CanL is 16.6% (42/253) among dogs in the region and clinical signs were found to be helpful for suspecting CanL but at least one serological and/or parasitological method should be performed for the accurate and differential diagnosis in the dogs. Our findings also showed that the one-third of the dogs in an endemic area for CanL could be asymptomatic. [ABSTRACT FROM AUTHOR]

Copyright of Turkish Journal of Veterinary & Animal Sciences is the property of Scientific and Technical Research Council of Turkey and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

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., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The acid-fast staining method is widely used in the diagnosis of cryptosporidiosis, a disease causing diarrhea in humans. However in this technique, some of Cryptosporidium spp oocysts can not be stained and seen as formed "ghost-like bodies" and which can only be evaluated by experienced microscopists. In the recent years, PCR technique which is proven to be also highly sensitive in diagnosis and genotyping, is used as an alternative method. In our study we aimed to evaluate the efficacy of PCR in diagnosis of cryptosporodiosis. Thirty-three stool samples, belonging to 22 patients who applied to Ege University Hospital, Parasitology Clinic between August 2001 - August 2003 and microscopically diagnosed as cryptosporodiosis has been included in the study. Twenty-three of these 33 samples were processed immediately, while ten samples were stored in 10% formalin. As the control group, 11 stool samples including 8 specimens with different parasites and 3 negative samples were selected. Nested-PCR is applied to all samples. The sensitivity and specifity of the method for fresh and formalin preserved samples were found to be 100% and 50%, respectively. As a conclusion, PCR technique is found to be useful for diagnosis in cryptosporodiosis patients with especially those including few oocysts in their fresh feces.