Sequence Analysis of Pvama-1 among Plasmodium Vivax Isolates in Sistan-Baluchistan

BACKGROUND: Apical Membrane antigen 1 (AMA-1) is an important membrane protein that presents in all Plasmodium species and participates in critical phases in the attraction of cells. In human, it is one of the most immunodominant antigens with a protective immune response simulation role Apical Membrane antigen 1 (AMA-1) is an important membrane protein which presents in all Plasmodium species and is located on the surface of merozoite and sporozoites that participates in critical phases in attraction of human red blood cells by merozoites and hepatocytes by sporozoites, so in human, it is one of the most immunodominant antigens with a protective immune response simulation role. Since extra information is necessary to lighten of AMA-1 scope, we equaled genetic variation in P.vivax AMA-1 from 40 Iranian isolates with those reported from the other malarious countries.METHODS: Blood samples were collected from 40 patients’ positive of P.vivax, and genomic DNA was extracted from the blood. The nucleotide sequence for 446 amino acid (AA) residues (42–488 of PvAMA-1) of AMA-1 gene was amplified via PCR and then sequenced.RESULTS: A total of 24 different haplotypes were recognized between samples. No new haplotype was determined in this research that was reported previously in other regions of Iran and the world. We detected 37-point mutations at the nucleotide level in their sequences and showed 43 amino acid variations, at 37 positions in which 6 sites demonstrate trimorphic polymorphism, and the others were dimorphic.CONCLUSION: Sequence analysis of the major haplotype showed 95% similarity with P.vivax Sal-1 AMA-1 gene and high level of allelic diversity at the domain I of PvAMA-1 among P. vivaxisolates of Iran. Because PvAMA-1 is noticeable as vaccinecandidate antigen, these documents provide valuable informationfor the development of malaria vaccine

Identification of Zoonotic Parasites isolated from Stray Dogs in Bojnurd County Located in North-East of Iran

Dog can represent as an important source of zoonotic disease and important health problem for human. They can carry dangerous parasitic diseases such as hydatidosis, toxocariasis and Coenurus cerebralis to humans and animals. This study was performed in order to determine the prevalence and intensity of zoonotic parasites among stray dogs from Bojnurd, the capital city of North Khorasan province in North West of Iran. During a program performing by Bojnurd municipal on the slow killing of stray dogs, 32 dogs from Jun 2013 till March 2015 were selected. At necropsy their alimentary canals were removed and to identify the species of helminthes, the nematodes were cleared in lactophenol and cestodes were stained using carmine acid. Intestinal protozoan parasites were detected with parasitological methods. 28 (87.5%) of 32 stray dogs infected at least with one helminth. Seven species of cestodes were isolated from examined dogs and three species of nematode were detected. Giardia sp. and Cryptosporidium sp. detected from fecal samples. This is the first study of the prevalence of intestinal zoonotic parasites in dogs in this area. It seems control of bearing stray dogs can help human health and reduction economic losses caused by stray dog’s zoonotic parasites

Bionomics and phylo-molecular analysis of Leishmania species isolated from human lesions using ITS1 genes in north-east of Iran

Leishmaniasis is a zoonotic infectious disease caused by Leishmania species. The identification of parasite species and the type of disease is beneficial for treatment and preventive modalities. Leishmania tropica and L. major have been reported as the main etiological agents of cutaneous leishmaniasis (CL) in Iran. The incidence of zoonotic CL has increased and different in distinct loci of Iran. Hence, we perused the Leishmania species and its genetic traits in the North East of Iran. The investigation was conducted on 200 positive smears prepared from patients’ lesions suffering from CL referred to the health care centers of northeastern provinces in Iran from 2013 to 2019. The obtained positive microscopy samples were divided to score the ranges from + 1 to + 6, of them 40 smears exhibited low-parasitemia. Leishmania species analyzed using PCR–RFLP, genetic diversity indices evaluation, phylogenetic analysis, and sequencing comparison with other species in the GeneBank based on ITS1 gene. The isolated L. major strains were similar to other Iranian isolates in this region. Pairwise fixation index (FST) index was statistically significant in different L. major populations and showed the genetic differences in pairwise population of different geographical locations of Iran. The current study confirmed an old pattern endemicity of zoonotic CL in North-east of Iran. Therefore, in order to assess the hybrid formation, more epidemiological, ecological, and gene polymorphism studies are needed to understand the pathogenic role of Leishmania species in Iran.acceptedVersionPeer reviewe

Seroprevalence of Toxoplasma Gondii in Sheep, Cattle and Horses in Urmia North-West of Iran

Background: Toxoplasma gondii is a zoonotic protozoan parasite found worldwide and responsi­ble for major economic losses in most classes of livestock. This study was aimed to deter­mine the prevalence of T. gondii infection in sheep, cattle and horses in Urmia, north-west of Iran, using MAT.Methods: Blood samples of 276 livestock and 26 horses were collected from July 2009 till April 2010. The data were analyzed by the Chi-square, Fisher's Exact and Cochran's and Mantel-Haen­s­zel Tests. The level of significance was set at P < 0.05.Results: Thirty-three (21.1%) sheep, 2 (1.6%) cattle and 3 (11.5%) horses were seropositive to T. gondii. Analysis showed that sheep were 15 times more likely to be seropositive comparing to cattle also 2 times more likely to be seropositive than horses.Conclusion: This study showed seroprevalence of equine T. gondii infection with a considerable rate in sheep in Urmia, northwest of Iran. More comprehensive studies on livestock toxoplasmo­sis are required for further analysis of the parasite reservoir for human infection

Using Human Umbilical Cord Matrix Cells (HUCM) for Culturing Toxoplasma gondii for Serological and Molecular Assays

Introduction & Objective: Toxoplasma gondii is an intracellular parasite and one of the most common parasites between human and animals. Nowadays the molecular methods are being used for the diagnosis. Many molecular and drug assays have been performed on this parasite. Many serological and molecular methods are available for detection of this parasite. The aim of this study was to use human umbilical cord matrix cells for propagation of Toxoplasma gondii for serological and molecular assays. Materials & Methods: In this experimental study, performed in Kerman Medical University, RH strain of tachyzoite of Toxoplasma gondii was isolated from mice and cultured in HUMC, A549 and Hep2 media. The growth and generation conditions of cultured parasite in different times were studied. DMEM medium was used to compare the viability of parasite. The obtained data were analyzed by variance analysis using SPSS software. Results: Parasite entry to cell line was observed in the fist 48 hours of culturing in all media. In day 3-6, propagation of parasites in HUMC cell line was better than other cultures. In the late 6th day volume of cultivated parasites in virulence was acceptable. Conclusion: Use of human umbilical cord matrix cells is a suitable and inexpensive method for proliferation of Toxoplasma gondii. Using these cell lines are useful in providing live parasite for research purposes, drug assays or making laboratory kits

Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: Molecular and phylogenetic studies

Fasciolosis is a zoonotic disease caused by Fasciola hepatica and Fasciola gigantica. Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our knowledge of the distribution of haplotypes and gene flow of this parasite is not comprehensive yet. The purpose of this study was to investigate this gap in the epidemiology of F. hepatica in different provinces of Iran between 2015 and 2017. Isolated Fasciola were collected from abattoirs in 9 provinces. The partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene was used for the identification and molecular analysis of F. hepatica isolates. The amplified PCR products were purified and subjected to direct sequencing for subsequent construction of phylogenetic tree and network analysis. In the 130 subjects analyzed, 37 ND1 haplotypes were detected. This is the first study in Iran which investigates F. hepatica population and its genetic structure, based on mitochondrial ND1 marker in different geographical regions of Iran. © 2019 Elsevier B.V

Detection of species and molecular typing of Leishmania in suspected patients by targeting cytochrome b gene in Zahedan, southeast of Iran

Aim: Cutaneous leishmaniasis (CL) is one of the most important health problems that are capable of involving both tropical and subtropical areas, especially in Iran. This cross-sectional study aimed to differentiate the species that are able to cause CL in Zahedan city by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Materials and Methods: It was conducted on 145 suspected CL patients in Zahedan city between 2014 and 2016. The smears were initially prepared, air-dried, fixed with absolute methanol, and stained with 10% Giemsa. Then, we examined the stained samples by a light microscope under 1000× magnifications. PCR assay targeted cytochrome b (cyt b) gene using LCBF1 and LCBR2 primers and the products digested by Ssp1 enzymes. Results: From 145 suspected CL patients, 76 (52.4%) were positive in microscopic examination. In addition, we detected gene of interest (cyt b) in 98 (67.5%). The results of PCR-RFLP indicated that 53/98 (54%) cases were Leishmania major and 45/98 (46%) were Leishmania tropica, and the main species in these areas was L. major. Conclusion: We concluded that the microscopic examination is not sensitive enough and is not able to distinguish between different Leishmania species. Instead, molecular methods like PCR-RFLP can be appropriately used with promising results