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

Morphological Characteristics and Molecular Markers in Identification of Anopheles gambiae Complex and Anopheles stephensi Members as Main Malaria Vectors in Africa and Asia

Introduction and purpose: Human malaria is one of the widespread vector-borne diseases worldwide. Lots of efforts have been made to control and eliminate the vectors of the disease. Anopheles gambiae and Anopheles stephensi (A. stephensi) are the main vectors of malaria in Africa and Asia. The members of Anopheles gambiae complex contain the vector and non-vector species. A. stephensi has three biological forms with different vector  capacities. The phenotypic and genotypic diversity has made it more difficult to identify vector populations and consequently the surveillance and control of malaria. The present study aimed to compare the morphological and molecular diagnostic characteristics of the two important malaria vectors in the two world continents. Methods: After searching, studying, and organizing published papers related to the subject, Anopheles gambiae and A. stephensi species were compared based on the morphological and molecular characteristics. Different morphological keys for the two species, the specimens from the insectarium of Urmia University of Medical Sciences, as well as the present species of national insectarium in Pasteur Institute of Iran, were used for morphological comparison. In addition, all the present sequences of five molecular markers, including COI, COII, D3, ITS2, and OBP1, were extracted from GenBank and analyzed using bioinformatics software. Results: Based on the obtained results of the present study, the number of ridges on  Anopheles gambiae eggs was more than that on A. stephensi eggs. However, unlike A. stephensi in Anopheles gambiae, the number of ridges was not a diagnostic characteristic for the identification of Anopheles gambiae complex. There were four diagnostic characteristics in the larvae comparison of the two species and there were three different characteristics in adults. All the molecular markers were capable of separating the two species; however, ITS2 and D3, as well as COI, COII, and OBP1 markers were preferred for inter-species and intra-species comparisons, respectively. Conclusion: The combination of traditional diagnostic and new molecular methods can be simultaneously used in inter-species separation. As a result, in order to eliminate malaria in Iran and the countries covered by The World Health Organization Regional Office for the Eastern Mediterranean, it is recommended that the combination of morphological, molecular, and field epidemiological data can provide practical solutions for vector control programs. Obviously, none of these data exclusively respond to the needs of the mentioned programs. Furthermore, the coordination, design, implementation, and evaluation of applied projects, as well as executive actions, are necessary for the success of these programs regarding the elimination and eradication of malaria and other vector-borne diseases

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

Three cases of brain hydatidosis in North Khorasan, Iran

Abstract Cystic hydatidosis is a serious public health problem in Iran. Although cysts can develop in almost all organs and the brain cysts are very rare. Here, we present 3 confirmed cases of brain hydatidosis and the patients who underwent successful surgery. Pathological examinations demonstrated the presence of cystic hydatidosis

Scolicidal Effect of Some Herbs on Echinococcus granulosus Protoscoleces: a Systematic Literature Review

Surgery is the best choice of treatment for human hydatidosis. Secondary infection is one of the end points of surgery in treating the hydatidosis which results from the spillage of protoscolices into the peritoneal cavity. Many chemical scolicidal agents have been used for inactivation of the cyst’s content, but most of them are associated with adverse side effects. Some studies have reported that traditional plants might have a potential application in prevention of post-surgery infections. We searched Medline, PubMed and Google scholar to identify the potentially relevant studies on implication of traditional plants against Echinococcus granulosus protoscoleces. In this study, we have reviewed scolicidal effects of some plant species (such as Zataria multiflora, Berberis vulgaris, Allium spp. Mentha spp. and etc.) and their products (essential oils, methanolic extract, aqueous extract and etc.) on Echinococcus granulosus protoccoleces. The scolicidal activity of these herbs could be helpful in hydatid cyst surgery. However, the mechanisms by which plant extracts killed protoscolices and also their safety for human cells are unclear and needed to be more investigated

Molecular characterization of Fasciola hepatica and phylogenetic analysis based on mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I and cytochrome oxidase subunit I) genes from the North-East of Iran

Aim: Fascioliasis is one of the most zoonotic diseases with global extension. As the epidemiological distribution of Fasciola may lead to various genetic patterns of the parasite, the aim of this study is to identify Fasciola hepatica based on spermatogenesis, and phylogenetic analysis using mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I [ND1] and cytochrome oxidase subunit I) gene marker. Materials and Methods: In this study, 90 F. hepatica collected from 30 cattle at slaughterhouse located in three different geographical locations in the North-East of Iran were evaluated based on spermatogenetic ability and internal transcribed spacer 1 gene restriction fragment length polymorphism pattern. Genetic diversity and phylogenetic relationship using mtDNA gene marker for the isolates from the North-East of Iran, and other countries were then analyzed. Results: Partial sequences of mtDNA showed eight haplotypes in both genes. The phylogenic analysis using neighbor joining as well as maximum likelihood methods showed similar topologies of trees. Pairwise fixation index between different F. hepatica populations calculated from the nucleotide data set of ND1 gene are statistically significant and show the genetic difference. Conclusion: F. hepatica found in this region of Iran has different genetic structures through the other Fasciola populations in the world

Canine Visceral Leishmaniasis in Wild Canines (Fox, Jackal, and Wolf) in Northeastern Iran Using Parasitological, Serological, and Molecular Methods

Background: Although many studies had been conducted on various aspects of canine visceral leishmaniasis (CVL) in domestic dogs in the endemic areas of Iran, investigations on CVL in wild canines are rare. Methods: This is a cross-sectional study was conducted from December 2012 to 2013 in northeast of Iran where human VL is endemic. Wild canines were trapped around the areas where human VL cases had been previously identified. Wild canines were collected and examined both clinically and serologically using direct agglutination test (DAT). Microscopically examinations were performed in all the seropositive wild canines for the presence of the amastigote form of Leishmania spp. Some Leishmania sp. which had been isolated from the spleens of wild canines, were examined analyzed by conventional PCR and sequencing techniques using α-tubulin and GAPDH genes. Results: Altogether, 84 wild canines including foxes (Vulpes vulpes, n=21), Jackals (Canis aureus, n=60) and wolves (Canis lupus, n=3) were collected. Four foxes and seven jackals showed anti-Leishmania infantum antibodies with titers of 1:320–1:20480 in DAT. Furthermore, one fox and one jackal were parasitologically (microscopy and culture) positive and L. infantum was confirmed by sequence analysis. Conclusion: The present study showed that sylvatic cycle of L. infantum had been established in the studied endemic areas of VL in northeastern Iran

Iranian Hydatid Disease Registry: Establishment and Implementation of a Neglected Tropical Disease Registry

Background: Cystic echinococcosis (CE) or hydatid disease is a global public health concern which imposes considerable economic costs on the communities in endemic regions. CE surveillance data are not adequately reliable. The present study reports the development and outcomes of a CE registry in Iran. Methods: Hydatid Registry (HydatidReg) was initially established as a single-center registry in 2014 after the ethical approval of KMU. Following a call from MoHME to promote registry of different diseases and health outcomes, a call for participation was announced and all the Iranian Universities of Medical Sciences were requested to contribute to the registry. Subsequently, a nation-wide registry of hydatid disease was established in 2016. With a global perspective, HydatidReg joined the European Register of Cystic Echinococcosis (ERCE). A data collection form based on minimum dataset was designed and standard operating procedures (SOPs) were prepared to ensure standardized patient enrolment in the registry. A biobank system with two-dimensional barcoding was established along with HydatidReg for management and organization of biological specimens. Results: As of March 2021, a total of 690 patients were enrolled in the registry. HydatidReg registered 362 (17.3%) out of the total 2097 patients enrolled in ERCE. Quality control (QC) of the data demonstrated 91.2% completeness and 80% timeliness. In the biobank, 322 biological specimens from 184 CE patients have been deposited including 70 blood, 96 sera and 156 parasite materials. Conclusion: High-quality data in the HydatidReg registry provided opportunities for health professionals to improve quality of care and organize meaningful research