Genotyping and phylogenetic analysis of fasciola spp. isolated from sheep and cattle using PCR-RFLP in Ardabil province, Northwestern Iran

The aim of this study was to detect the genotype of Fasciola spp. in Meshkin-Shahr, Ardabil Province, northwestern Iran in different hosts using PCR-RFLP. The parasite hosts included cattle, and sheep. Overall, 70 adult flukes from livers of slaughtered animals were collected from the abattoirs of aforementioned area. The included 35 samples from infected sheep and 35 samples from 35 infected cattle. PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer [ITS 1] region from Fasciola species were used to conduct the study. The fragment of approximately 700bp in all of the Fasciola samples was amplified. PCR products of ITS 1 were subjected for digestion by restriction enzyme. RsaI restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species. Amplicons with the sequences of F. hepatica had a pattern of about 360, 100, and 60 bp band size, whereas F. gigantica worms had a profile of 360, 170, and 60 bp in size, respectively. Results based on PCR-RFLP analysis were confirmed by sequence analysis of representative ITS 1 amplicons. No hybrid forms were detected in the present study. All sheep were infected with F. hepatica but cattle were infected with both species. Both species of Fasciola are present in Ardabil. The method described here can be valuable for identification of Fasciola species in endemic parts for fasciolosis, regions with intermediate species and in that overlapping distribution area

Seroprevalence of human fascioliasis using indirect ELISA in Isfahan district, central Iran In 2013

The aim of this study was to detect the seroprevalence of human fascioliasis in Isfahan County, central Iran in 2013. Overall, 471 sera samples were collected from people recalled randomly to 20 health centers in the city of Isfahan and 10 related villages in 2014. Sera were examined using ELISA test. A questionnaire was filled out for each participant. Altogether eight cases [1.7%] were seropositive which had the OD absorbance in ELISA test more than the calculated cutoff of 0.36. All of them were female. One positive subject had a history of consuming Delar [Local dish] and three seropositive cases with history of eating Zeitoon-Parvadeh [Proceeded olive]. Isfahan County might be considered as one area apt for fascioliasis. More studies in terms of veterinary investigation and verifying the risk factors are necessary

Genotyping of echinococcus gtanulosus isolates from human clinical samples based on sequencing of mitochondrial genes in Iran, Tehran

The present study was aimed to investigate molecular diversity of Echinococcus gmnulosus isolates collected from human clinical samples using two mitochondrial genes cox1 and nod1 in Iran. Forty seven human hydatid cysts were collected through surgery from two hospitals in Tehran during 2010-2012. To determine the fertility of protoscoleces, the cyst fluids were subjected to morphological microscopic examinations. Protoscoleces were removed from each cyst and their total genomic DNAs were extracted. PCR was performed to amplify fragments of 450 and 400 base pair [bp] for cox1 and nod1 genes, respectively. Genotype diversity and sequence variation of the strains were studied by bioinformatics software and in comparison with those mtDNA sequences already dposited in GenBank. Sixteen, [53.3%], 13 [43.3%], and 1 [3.3%] samples were related to lung, liver, and spleen, respectively. The remained 17 unfertile samples were excluded from the study. From the 29 isolates, 86.7% [n=26] and 10% [n=3] were related to G1, and G3 genotypes, respectively. The sole isolate with G6 genotype was obtained from lung sample. Analysis of concatenated sequences of cox1+nad1 indicated the presence of 11 haplotypes among our strains that were related to genotypes G1 [n=9], G3 [n=1] and G6 [n=1]. In consistent to other reports from Iran, genotypes G1, G3, and G6 were observed in our human isolates. The rate of G3 genotype was however higher than other studies implying that human can be considered as a new appropriate host for G3 genotype. Further studies with more sample size from different geographic areas of Iran are needed for E. granulosus mapping

Seroepidemiology of human hydatidosis using AgB-ELISA test in Arak, central Iran

On the continuation of sorting the puzzle of the situation of hydatid disease in Iran and considering that so far no survey was conducted in this context in Arak City, Markazi Province central Iran, the present study was conducted to determine the seroprevalence of human hydatidosis using AgB-ELISA test. Totally 578 serum samples randomly were collected from patients referred to hospitals and different health centers in the city and 3 nearby villages of Arak. All sera were examined by ELISA tests using AgB. Before sampling, a questionnaire was filled out for each case. Data were analyzed using Chi-square test and multivariate logistic regression for risk factors analysis. P<0.05 was considered significant. Cut-off value was calculated 0.32. Twenty cases [3.46%] were seropositive for hydatidosis in the region. This rate for females was 3.99% and for males 2.26%, respectively. There was no significant difference as regards all factors studied except location [P<0.001]. As for job, farmers and ranchmen had the highest rate of infection as 6.67%. The seropositivity rate infection was 4.8% in illiterate people, which showed the highest rate. As regards residency, rural life showed significant difference with urban life [1.5% vs. 7%]. Age group of 40-49 yr old [6.25%] had the highest rate of positivity. The rate of prevalence in this region shows more or less the same range with other cities of Iran. Obtained result might assist the policy makers to take sanitary measures to control the disease

Endoparasites of stray dogs in Mashhad, Khorasan Razavi province, northeast Iran with special reference to zoonotic parasites

To find out different species of helminthes and blood/tissue protozoan parasites of stray dogs and their potential role for transmission of zoonotic species to human in Mashhad, Khorasan Razavi Province, northeast Iran, during 2008-2009. Totally, 100 stray dogs were selected among Mashhad municipal collection from different sites of the city. Internal organs were examined for any parasites. Helminthes were identified based on morphological characteristics. Smears prepared from peripheral blood as well as liver, spleen and any skin lesion were stained by Giemsa and examined microscopically. Samples obtained from spleen were aseptically cultured in three culture media including NNN, Schneider's Drosophila [HIMEDIA] and RPMI1640 [GIBCO] for isolation of Leishmania spp. The titer of anti-Leishmania and anti-Toxoplasma antibodies were measured by direct agglutination test [DAT] and indirect fluorescent antibody test [IFAT], respectively. 84% of dogs were infected at least with one species of intestinal helminthes. The species of parasites and rate of infection were as follows: Taenia hydatigena [61%], Dipylidium caninum [46%], Mesocestoides lineatus [19%], Echinococcus granulosus [10%], Toxascaris leonina [53%] and Toxocara canis [7%]. Anti-Leishmania antibodies were detected by DAT in 8 dogs [8%] at 1:320 titers and higher. Forty seven dogs [47%] showed anti-Toxoplasma titer at 1:10 and 17 [17%] showed titer of >/= 1:100. No blood parasites were found in prepared blood smears. The high rate of parasitic infection and presence of zoonotic species especially E. granulosus and T. canis emphasizes the risk of diseases spread in urban areas by stray dogs

Morphometric differentiation between camel and sheep strains of Echinococcus granulosus using computer image analysis system (CIAS)

OBJECTIVE@#To find importance of morphometric criterion of larval rostellar hook of Echinococcus granulosus (E. granulosus) and the easy and reliable method for distinguish sheep and camel strains in epidemiologic studies.@*METHODS@#Larval rostellar hooks (n=1860) of 31 camel and sheep isolates in Iran, which already had been characterized by PCR, were carefully processed by computerized imagime analysis system (CIAS) and acquired data about rostellar hooks were analyzed using software SPSS.@*RESULTS@#Measurement analysis of rostellar hooks [mean length (24.23±3.12) μ m] indicated that length of the large hook was a remarkable parameter for strain differentiation. Data analysis demonstrated that CIAS could be used as a reliable tool to distinguish camel from sheep strains with high sensitivity (95.2%) and specificity (91.5%).@*CONCLUSIONS@#CIAS as a specific, sensitive, economic, fast, and reliable means might be used for differentiation of E. granulosus strains. Although perimeter and area were measured by digital technology, they were not shown as discriminative criterion as total hook length did.

Preparation and evaluation of a glycerol-preserved direct agglutination antigen for long-term preservation: a comparative study of the detection of anti-Leishmania infantum antibodies in human and dog

OBJECTIVE@#To prepare and evaluate a glycerol-preserved antigen from an Iranian strain of Leishmania infantum (L. infantum) for use in glycerol-preserved direct agglutination tests (GP-DAT) as an alternative to freeze dried direct agglutination tests (FD-DAT) that use freeze-dried antigen.@*METHODS@#Glycerol-preserved DAT antigen was prepared and stored at different temperatures. We tested antigen stored at 4 °C, 22-37 °C and 50 °C over a period of 365 days. Seven hundred twenty-nine serum samples were collected from different geographical zones of Iran from 2007-2009, and 80 of these samples were pooled to produce sera. Each pooled serum contained 10 sera. All positive and negative pooled sera were separately tested for anti-L. infantum antibodies with GP-DAT, FD-DAT and formaldehyde-fixed direct agglutination test (FF-DAT) antigens; tests were performed on both human and dog sera over a period of 12 months.@*RESULTS@#There was strong agreement between the results obtained using GP-DAT and FD-DAT antigens stored at 22-37 °C for 12 months for both human (100%) and dog (100%) pooled sera. The direct agglutination test results were highly reproducible (weighted kappa: GP=0.833, FD=0.979 and FF=0.917).@*CONCLUSIONS@#Because GP-DAT antigen is highly stable over a range of temperatures and is easy to transport in the field, this type of antigen may be particularly useful in areas with endemic visceral leishmaniasis.

Evaluation of rabbit antibody response against 8 and 16 kDa recombinant subunits of antigen B from Echinococcus granulosus

OBJECTIVE@#To immunize rabbits with 12 and 16 kDa recombinant subunits of antigen B from Echinococcus granulosus (E. granulosus) and measuring polyclonal antibody and humoral immune response using ELISA and gel diffusion.@*METHODS@#Two mentioned antigens were cloned and expressed in expression vector and purified by affinity chromatography. Four young rabbits were selected and challenged intradermally with yielded recombinant antigens. Rabbits' sera were collected post infection and were tested using ELISA and gel diffusion for polyclonal antibody detection 10 days after last injection.@*RESULTS@#The specific antibody against the recombinant peptides was efficiently produced within 4 weeks post infection.@*CONCLUSIONS@#Produced recombinants proteins could induce the immune response of the rabbits successfully. This process might improve the clarification of diagnosis and vaccination as regards hydatidosis.

comparative analysis of writing scientific references manually and by using endnote bibliographic software

By writing scientific paper, one gradually starts the academic career. Academic staff members, most often have no choice but to engage in scientific writing to fulfill the requirements for selection or promotion in their academic career. Those who do not find themselves competent enough to write, at times indulge in plagiarism. The present study was conducted to compare writing references for scientific paper manually and by using EndNote software, which is quite helpful for the authors. In this study time consumed in inserting twenty references in a paper mannually was compared with putting references using EndNote software. [version 9] In addition, the format of references was changed in different manners to find out the time required for making these changes. Comparison of time spent for inserting one reference or all references in both formats Vancouver and Harvard showed significant difference [P<0.001]. The study showed that persuading the authors to use electronic software for writing references and scientific papers would significantly improve the quality of the manuscripts besides enlarging the scope of medical journalism

Evaluation of a dot-ELISA for the serodiagnosis of human hydatid disease

A dot-ELISA utilizing antigen-B from sheep hydatid fluid, dotted onto nitrocellulose filter discs was developed for the rapid diagnosis of human hydatidosis. 1micro g of antigen per dot, serum dilution of 1:800, dilution conjugate of 1:1000 and 45 min incubation were found optimal. Thirty four patients infected with hydatidosis, 32 cases with other parasitic diseases and 36 healthy subjects were included in the assay and were examined using dot-ELISA to detect antibody against the aforementioned parasite antigen. Sensitivity, specificity and positive and negative predictive values of the assay were calculated as 97.1%, 98.5%, 97.1% and 98.5% respectively. No false positive reaction was observed when 36 sera healthy subjects were assayed. One case of cross-reactions was observed as for a serum infected with fasciolosis. It was concluded that dot-ELISA is rapid, antigen and serum conservative as well as encompasses great importance to confirm clinical diagnosis either in the laboratory or in the filed