Iranian Journal of Parasitology
Volume:11 Issue: 2, Apr-Jun 2016

Malaria is the most important transfusion-transmitted infection (TTI) in worldwide after viral hepatitis and human immunodeficiency virus (HIV) infection. The main objective of the present study was to review and evaluate the transmission of malaria via blood transfusion in Iran.
A literature search was done without time limitation in the electronic databases as follows: PubMed, Scopus, Google Scholar, Web of Science, Science Direct, scientific information database (SID), Magiran, IranMedex and Irandoc. The searches were limited to the published papers to English and Persian languages.
Six papers were eligible. From 1963 to 1983, 344 cases of Transfusion-transmitted malaria (TTM) had been reported from different provinces of Iran. The most prevalent species of involved Plasmodium in investigated cases of TTM was Plasmodium malariae (79.24%). The screening results of 1,135 blood donors for malaria were negative by microscopic examination of peripheral blood smears and rapid diagnostic test (RDT) methods.
Lack of TTM report from Iran in the last three decades indicates that the screening of blood donors through interviewing (donor selection) may be effective in the prevention of the occurrence of transfusion-transmitted malaria.

The aim of present study was to evaluate the protective efficacy of live recombinant L. tarentolae expressing KMP11-NTGP96-GFP fusion as candidates for live engineered recombinant vaccine against visceral leishmaniasis in BALB/c mice.
KMP-11 and NT-GP96 genes cloned into the pJET1.2/blunt cloning vector and then into pEGFP-N1 expression vector. The KMP-11, NT-GP96 and GFP fused in pEGFP-N1 and subcloned into Leishmanian pLEXSY-neo vector. Finally this construct was transferred to L. tarentolae by electroporation. Tranfection was confirmed by SDS-PAGE, WESTERN blot, flowcytometry and RT-PCR. Protective efficacy of this construct was evaluated as a vaccine candidate against visceral leishmaniasis. Parasite burden, humoral and cellular immune responses were assessed before and at 4 weeks after challenge.
KMP- NT-Gp96-GFP Fusion was cloned successfully into pLEXSY -neo vector and this construct successfully transferred to L. tarentolae. Finding indicated that immunization with L. tarentolae tarentolae-KMP11-NTGP96-GFP provides significant protection against visceral leishmaniasis and was able to induce an increased expression of IFN-γ and IgG2a. Following challenge, a reduced parasite load in the spleen of the KMP11-NTGP96-GFP immunized group was detected.
The present study is the first to use a combination of a Leishmania antigen with an immunologic antigen in live recombinant L. tarentolae and results suggest that L. tarentolae-KMP11-NTGP96-GFP could be considered as a potential tool in vaccination against visceral leishmaniasis and this vaccination strategy could provide a potent rout for future vaccine development.

The effects of extremely low frequency electromagnetic fields (ELF-EMF) on Toxoplasma gondii have not been explained yet. The aim of this study was to assess the possible effects of ELF-EMF on growth, survival time and viability of Toxoplasma gondii. In addition, the life span of Toxoplasma infected animals was investigated.
Sixty adult male BALB/c mice were used for in vivo and in vivo experiments in Laboratory of Biopyhsics and Parasitology of Medical Faculty, Adnan Menderes University, Turkey, in 2010. During in vivo experiments, pulsed and continuous EMFs were applied for 5 d to the infected mice. During in vivo experiments, pulsed and continuous EMF was applied to the tachyzoites within peritoneal exudates for 8 h/d at 4 °C and the tachyzoites were then injected to mice. In both experiments, the number of T. gondii in peritoneal exudates was counted and T. gondii protein bands patterns were investigated with polyacrylamide gel electrophoresis and Western Blotting.
Pulsed and continuous EMF exposure reduced the number of T. gondii tachyzoites in comparison to controls. However, no statistically significant differences were observed at the patterns of protein bands among the samples.
EMF exposure induces a decrease in the number of T. gondii. Further studies are required to understand the mechanism of EMF on intracellular parasites.

Treatment of Cutaneous Leishmaniasis (CL) is being faced with serious difficulties in Fars Province, due to emerging of resistance against meglumine antimonite (Glucantime®). In this context, determining some biomarkers for drug sensitivity monitoring seems to be highly essential. Different studies have been carried out to decipher the genes might be involved in antimony resistant phenotype in Leishmania spp. Here, we selected three genes: AQP (as drug transporter), TDR-1-1(as drug activator), and γ-GCS (inducing reduction environment) for comparative expression analysis on clinical resistant and sensitive isolates of L. major.
The clinical isolates of L. major were collected from CL patients referred to Valfajr Health Center, Shiraz from Oct 2011 to Feb 2012. The susceptibility test was performed to confirm drug sensitivity of strains in vitro as well. Then, the gene expression analysis was performed by quantitative real-time PCR using SYBR® Green.
By comparison of expression level between strains, up regulation of γ-GCS gene and down regulation of AQP gene were observed in resistant strains compared to the sensitive isolates; however, down regulation of AQP was not statistically specific. Analysis of TDR-1-1 gene unexpectedly showed a high level of expression in the non-responsive cases.
The γ-GCS, at least, can be considered as a suitable molecular marker for screening antimony sensitivity in clinical isolates, although AQP and TDR-1-1gene seem not to be reliable resistant markers.

Here, we established the mouse models of chronic toxoplasmosis by T. gondii Tehran strain to provide a good understanding about defining the possible association between T. gondii exposure and learning and memory impairments. Moreover, as secondary objective of the present study, we hypothesized whether administration of an acetylcholinesterase (AChE) inhibitor could reduce learning and memory impairments induced by T. gondii infection.
Twenty-four male BALB/c mice were used to establishment of latent toxoplasmosis. The animal model of Toxoplasma infection was established by the intraperitoneal inoculation of 20-25 tissue cysts from Tehran strain of T. gondii. Donepezil (2 mg/kg) an AChE inhibitor to treat Alzheimer disease was injected intraperitoneally once a day for two weeks starting from post-infection day 90. Morris water maze (MWM) task was used to assay spatial learning and short term spatial memory in all groups. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups. P Toxoplasma infection impaired spatial leaning and short term spatial memory of the infected BALB/c mice, whereas donepezil, an AChE inhibitor, improved impairments induced by Toxoplasma infection.
T. gondii infection through increasing AChE reduces the level of Acetylcholine (Ach) and consequently affects learning and memory activity in infected hosts, whereas, donepezil as an AChE inhibitor improves these impairments by restoring ACh levels at synapses of neurons in brain.

The aim of the present study was genotyping of Echinococcus granulosus isolates from dogs and jackals in Mazandaran Province, northern Iran, and using partial sequence of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1).
E. granulosus isolates (n = 15) were collected from 42 stray dogs and 16 jackals found in south of the Caspian Sea in northern Iran. After morphological study, the isolates were genetically characterized using consensus sequences (366bp) of the cox1 gene. Phylogenetic analysis of cox1 nucleotide sequence data was performed using a Bayesian Inference approach.
Four different sequences were observed among the isolates. Two genotypes [G1 (66.7%) and G3 (33.3%)] were identified among the isolates. The G1 sequences indicated three sequence profiles. One profile (Maz1) had 100% homology with reference sequence (AN: KP339045). Two other profiles, designated Maz2 and Maz3, had 99% homology with the G1 genotype (ANs: KP339046 and KP339047). A G3 sequence designated Maz4 showed 100% homology with a G3 reference sequence (AN: KP339048).
The occurrence of the G1 genotype of E. granulosus sensu stricto as a frequent genotype in dogs is emphasized. This study established the first molecular characterization of E. granulosus in the province.

The aim of the present work was to investigate the prevalence and species of intestinal microsporidiosis among animals in Giza, Egypt.
A total of 869 animal fecal samples were collected from domesticated animals (dogs, cats, rabbits, cattle, buffaloes, sheep, goats, donkeys and pigs) living in Giza, Egypt. Spores of microsporidia were concentrated from collected samples by centrifugation and finally stained with modified trichrome (MT) stain to detect microsporidial spores. Microsporidial spores in microscopically-positive samples were molecularly confirmed and identified using species-specific primers.
Spores of microsporidia were microscopically detected in 17.0% of the examined animal fecal samples. The highest and lowest rates of infection with intestinal microsporidia were recorded in dogs (33.3%) and buffaloes (6.9%), respectively. Molecularly, the obtained microsporidial spores were classified as Enterocytozoon bieneusi and E. intestinalis. Dual infection with both identified species was observed in fecal samples from buffalo, rabbit, goat, cat, pig and dog.
Domestic animals may play a role in dissemination of intestinal microsporidiosis in the environment. Examined animals were infected with E. bieneusi in a higher percentage than E. intestinalis.

The aim of this study was to survey the Iranian Parasitology researchers’ performance, and analyse and visualize the scientific outputs of their co-authorship network.
This study was conducted using scientometric method and social network analysis (SNA). The data extracted from the Web of Science (WoS) databases in July 10th 2014. Totally, 1048documents of all types in research area of Parasitology during 1972-2013 by Iranian researches retrieved. The co-authorship map was drawn utilizing NETDRAW, Coauthor.exe, and UCINET softwares and was analysed based on SNA measures.
The researchers’ co-authorship network consisted of 78 authors and its density degree is 0.57. “Mohebali” ranked top in all of centrality measures.The most of the publications were related to 2012, “Mohebali” with about 9% of all documents was the Iranian most prolific author in Parasitology field. The Iranian researches have published mostly (266 documents) in “Iranian Journal of Parasitology”, and the most of the documents belong to “Tropical Medicine” subject field. The most of Iranian researchers’ scientific cooperation was performed with England and United States.
Bringing forth density degree (is 0.57) showed that this network has an almost medium density. Indeed, the authors have had relations in moderate level with each other in the network. The findings of this study can be identified aspects of scientific collaboration, and help policy makers of Parasitology field research.

Rodents have important role as reservoirs of different parasites. The aim of this study was to determine helminth parasites of abundant rodents in Meshkin-Shahr, Ardabil Province northwest Iran.
From April 2014 to March 2015; 205 rodents including 118 Meriones persicus, 63 Mus musculus and 24 Cricetulus migratorius were collected, using live traps. All rodents were dissected and their different tissues examined for infectivity with helminth parasites.
Overall, 74.2% of rodents were infected with helminth parasites. The rate of infectivity in M. persicus, M. musculus and C. migratorius was 82.2%, 61.9%, 66.7%, respectively. In general, among all 205 rodents, the species and infection rates of helminthes were as follows: Nematoda: Trichuris sp. (46.8 %), Capillaria hepatica (18.1%), Syphacia frederici (14.2%), Aspicularis tetraptera (3.4%), Trichuris rhombomidis (2%), Heligmosomom sp. (2%), Streptopharagus kuntzi (0.5%), Spiruridae gen. sp. (0.5%); Cestoda: Hymenolepis nana fraterna (16.6%) Hymenolepis diminuta (7.3%) tetratiridium of Mesocestoides sp. (1%), Paranoplocephala sp. (0.5%), Cysticercus fasciolaris (0.5%), Taenia endothoracicus larva (0.5%), and Acanthocephala: Moniliformis moniliformis (18.5%).
Variable species of helminthes circulate in the rodents of the study area. Presence of several zoonotic species highlights the potential risk of infections for public health.

This study aimed at investigating the prevalence and risk factors of bovine thelaziosis, performed in the southeast of Iran, an endemic area for Iranian Sistani cattle.
Between September 2012 to October 2014, 1924 cattle, Sistani breed (n= 1235) and Brahman breed (n=689) of all sex and age groups collected from Sistan and Baluchestan Province were examined using visual observation of the eyes by flushing the conjunctival sac and lachrymal duct with sterile saline solution.
The overall prevalence for thelaziosis was 50 of 1924 cows (2. 6%; 95% CI: 1. 9-3. 3%), with significant higher prevalence of infection in Sistani breed than in Brahman breed (3. 15% vs 1. 59%). Sixty adult worms (84. 5% of females and 15. 5% of males) were collected from the conjunctiva of the infected cattle: Thelazia gulosa (50/60, 83. 3%) was the most represented species followed by Thelazia. rhodesi (10/60, 16. 7%). The number of worms collected per cow ranged from one to seven (average ± standard deviation: 2. 08 ± 1. 49). Worms were gathered from cows throughout all months of the year. The difference in the seasonal variations of prevalence and the intensity of infection were significant, however, no significant correlation between prevalence, sex and age of cattle was noted.
Bovine thelaziosis needs special attention by veterinarians in the differential diagnosis of ocular manifestations and considering its impact on cattle production.

Toxoplasma gondii is the most common parasite causing latent cerebral infections in human. It has been shown that some anti-psychotic drugs are able to inhibit the proliferation of the parasite in in vitro study. There is very limited data regarding the inhibitory effect of anti-psychotics on Toxoplasma in in vivo. In this study, we evaluated anti-Toxoplasma activity of fluphenazine and thioridazine drugs on T. gondii in mice.
Mice were divided into six groups: Control, sesame as vehicle, thioridazine 10 mg/kg, thioridazine 20 mg/kg, fluphenazine 0.06 mg/kg and fluphenazine 0.6 mg/kg. They were inoculated intraperitoneally with brain suspension containing tissue cysts of T. gondii Tehran strain. Two months after inoculation, the number of cysts in crushed smears of mice brain were counted microscopically and considered as an indicator of anti-Toxoplasma activity. This work has conducted in Qazvin, central Iran, 2014.
Our study showed that fluphenazine and thioridazine could not significantly inhibit the brain cystogenesis of T. gondii in mice. However, the number of brain cysts was less at higher dose compared to lower doses for both drugs.
Further studies need to clear the mechanism of different structure of anti-psychotic drugs on activity of Toxoplasma.

Intestinal parasitic infections have a worldwide distribution. High prevalence of intestinal parasitic infections in individuals with low socioeconomic status and environmental conditions was found. No study has ever been conducted on the prevalence of these infections in Jiroft. Therefore, in this study prevalence of intestinal parasitic infections was evaluated in Jiroft, Kerman Province, Iran.
A total of 1060 individuals from rural and urban areas of Jiroft were sampled accidentally, during 2013-2014. Fresh stool samples were collected from all individuals and examined by formalin ether concentration and agar plate culture. Direct examination was performed on watery samples.
Out of 1060 individuals, 563 (53.1%) and 497 (46.9%) people were from rural and urban areas, respectively. In general, 297 individuals (28%) were infected with intestinal parasites. The prevalence of infection for protozoa and helminthes infections were 27.4% and 1.8%, respectively. The most prevalent protozoans were Blastocystis hominis (13.7%) and Giardia lamblia (7.8%), and that of helminth was Hymenolepis nana (1.1%).
Intestinal protozoan parasites were more prevalent than helminth parasites. Source of water supply and personal hygiene were important factors in the distribution of parasites in the study area.

Anisakid nematodes are common parasites of fish, mammals, fish-eating birds, and reptiles with a worldwide distribution, causing diseases in human, fish and important economic losses.
A preliminary epidemiological study was carried out on Anisakid nematodes larvae in some commercially important fish species to evaluate the anisakid nematode larvae from greater lizardfish, (Saurida tumbil), Japanese thread fin bream (Nemipterus japonicus), crocodile longtom (Tylosurus crocodilus crocodiles) and longfin trevally (Carangoides armatus) from the Persian Gulf of Iran.
The collected larvae were identified mainly as the third larval stage (L3) of Hysterothylacium larval type A, B and C, Anisakis sp., Raphidascaris sp., Pseudoterranova sp. and Philometra sp. (Nematoda: Philometridae). The prevalence of Anisakid larvae infection of examined fishes was 97.2% in N. japonicus, 90.3% in S. tumbil, 20.5% in crocodile longtom and 5.5% in longfin trevally. Anisakis type III for the first time was different from Anisakis type I and Anisakis type II.
Zoonotic anisakids by high prevalence in edible fish could be a health hazard for people. So health practices should be considered in these areas.

This study was proposed to monitor the situation of asymptomatic malaria among the native population and Afghani and Pakistani immigrants in Kahnooj and Ghale-Ganj districts from Kerman Province, Southeastern Iran.
A number of 180 and 120 individuals from Kahnooj and Ghale-Ganj respectively were registered and considered based on a cross-sectional surveillance method. From 300 registered cases, 200 individuals (66.7%) were selected among Afghani and Pakistani immigrants and the rest (33.3%) were native resident individuals. All samples were processed with employing microscopical examination, Rapid Diagnostic Tests (RDTs) and Semi- nested Multiplex PCR techniques.
None of the samples collected from native residents showed any malaria parasite, but among Afghani immigrants, one asymptomatic vivax malaria was detected in a 12 yr old girl with 280 parasites per microliter of blood. Moreover, one symptomatic vivax malaria was detected from a Pakistani immigrant with 47560 parasites per microliter of blood. All results obtained via microscopical method, confirmed by RDTs and PCR techniques.
To achieve the malaria elimination program different studies are needed that to be performed. Monitoring the asymptomatic malaria in all over the malaria endemic areas especially among the immigrant individuals is the most crucial necessity.

Genomic DNA was isolated from Parabronema skrjabini. rRNA region was amplified and sequenced.
The RNA secondary structure was predicted using mfold software (http://mfold.rit.albany.edu). The secondary structure with bulge, hairpins, helices, interior, external and multi loops was predicted for 5.8srDNA of our sequence of P. skrjabini and a sequence of P. skrjabini and two species of Habronema (H. microstoma and H. muscae) in GenBank. RNA motifs were predicted by MEME program version 4.10.2.
The length of 5.8S rRNA sequence for P. skrjabini#1, P. skrjabini#2, H. microstoma and H. muscae was 158, 156, 127 and 127bp, and the DG required for the formation of the secondary structure was -70.50, -56.40, -41.50 and -41.40 kcal/Mol, respectively. Common structural elements were initially recognized with the help of mfold by screening for thermodynamically optimal and suboptimal secondary structures (default settings, with T = 37 °C). The energy levels of the presumptive secondary structures were then calculated with mfold at the DNA level. Both motifs and the sequence of P. skrjabini#1 were completely different from the other analyzed samples. This difference might be due to the differences in host and geographical area.
This is the first molecular study of P. skrjabini in sheep, which could be further used in the structure modeling across Habronematidae.

The aim of this study was to determine the prevalence of the Nosema ceranae and Nosema apis among apiaries using both spore counts and multiplex PCR and the replacement of N. apis by N. ceranae in some regions of Turkey.
A hundred honey bee samples were collected from 99 apiaries in 11 different locations in 2011-2012 in Turkey. Nosema infection degree from collected samples was determined using light microscope and molecular detection of Nosema spp. (N. ceranae and N. apis) was performed using specific primers by multiplex PCR.
N. ceranae was only found spores in sampling areas using molecular diagnosis. N. apis was not detected in whole sampling areas using both techniques. There are no Nosema spores detected in Konya one location using two techniques. The nu­cleotide sequences from amplification products of the Nosema infested honeybee samples were (98%) identical with the sequence of N. ceranae for many countries deposited in the GenBank database in this study.
The present study illustrated that N. ceranae is the only spores for sampled areas in 2011-2012. The study could also indicate that N. ceranae has been replaced instead of N. apis in Turkey. In addition, the prevalence of N. ceranae and two microsporodia spores effects on honey bee colonies in Turkey were needed to determine with intensive sampling, periodically.

Toxocariasis is the clinical terms applied to infection of human with Ascarid nematodes in the order Ascaridida, named Toxocara canis and T. cati. Because in recent years in Iran many people desire to keep pets (cats and dogs), and lacking of seroepidemiological study of toxocariasis in Mashhad, we decided to determine the seroprevalence of toxocariasis among people who own cats and dogs in comparison with control group.
A serological study for detection antibodies to Toxocara in two groups (93 cat and dog owners and 93 healthy people as control group) was conducted from Feb 2013 to Dec 2013. An ELISA method was employed using determination of IgG antibodies against Toxocara. The serum samples were evaluated for anti-Toxocara antibody, using ELISA technique at Parasitology and Immunology Lab of Imam Reza Hospital of Mashhad. Using a questionnaire, epidemiological factors associated with infection were examined. Data were analyzed using Chi-square test.
The seroprevalence of Toxocara antibodies in the pet owners and control group was respectively 20.43% and 1.07%. 47.3% of pet owners were female.
Presented data showed the significant difference between seroprevalence of toxocariasis among pet owners and control group. Education of society and in particular pet owners consisting of preventing contamination of the environment with Toxocara eggs is advised.

Dirofilaria immitis is a common nematode of the cardiovascular system, which infects carnivores all over the world. The infection is prevalent in dogs, but in cats and human is rare. Dirofilariosis is transmitted by mosquitoes. Cats are accidental hosts and are naturally resistant to the infection, compared to the dogs. Mild infection can cause severe illness in cats and may lead to death, so it is clinically important to diagnose the disease.
In the present study, 103 stray cats were collected from Meshkin Shahr district, Aradabil Province, northwestern Iran that is an endemic area for canine dirofilariosis. Blood samples were prepared from the saphenous vein of each cat and were examined for the presence of microfilariae by the modified Knott test.
A 2 yr old male cat (0.97%) was microfilaremic. The cat was subjected to necropsy and 4 adult D. immitis (2 male and 2 female worms) were found in the right ventricle of the heart. PCR was then carried out and D. immitis was confirmed.
Cats and other carnivores in Meshkin Shahr district are at risk of infection due to the high activity of vectors in this climate and it is important to follow up the infection in this area.

Hydatid disease is a rare parasitic disease, which mainly involves liver then lung tissues. Cardiac involvement is very rare, especially when there is not hepatic involvement. We describe a 47-year-old woman with a history of a lung hydatid cyst who was referred to Rajaei Heart Center, Tehran, Iran in 2012. Her chest computed tomographic scan showed a cardiac mass. Echocardiographic examination illustrated a large, well-defined heterogeneous mass (4.5 × 2.5 cm) in the roof of the right atrium with attachment to the crista terminalis without compressive effect on the inferior and superior venae cavae. The patient was candidate for open-heart surgery via median sternotomy. A cystic mass was observed in the lateral aspect of the right atrial wall. After an injection of hypertonic normal saline into the cystic lesion, the mass was excised totally. The right atrial defect was reconstructed with autologous pericardium. The patient was discharged from the hospital in good condition. Histological examination confirmed the diagnosis of the hydatid cyst.

Strongyloides stercoralis is a soil-transmitted helminth (STH) widespread in various part of the world. A 78-yr-old peasant diabetic female from Mazandaran Province northern Iran, was admitted to Infection Department of the Razi Hospital in city of Qaemshahr, north of Iran complaining about abdominal skin rash, pruritus, itching, anorexia, nausea, vomiting, dysuria and cough. This patient had cutaneous migration effects of S. stercoralis larvae in her abdominal skin (larva currents and urticaria). Lung CT without contrast demonstrates bilateral diffuse ground glass opacity draws attention. Upper gastrointestinal endoscopy revealed gastro esophageal reflux with antral gastritis. Duodenal endoscopy showed unusual mucosa and a biopsy from it sent to the pathology laboratory. Histopathology of duodenal bulb and duodenum biopsy showed mild villous atrophy and S. stercoralis infection. The patient was treated with albendazole and clinical sings improved completely after treatment. Strongyloidiasis should be care­fully considered by clinicians who practice in endemic areas. Clinicians must keep a high level of skepticism for patients from endemic area.

A 74 yr old woman from Gonabad, southern part of Khorasan Razavi Province of Iran was admitted to a Hospital of Gonabad, because of respiratory distress, exertional dyspnea and fever. Close contact with domes­tic animals, history of chronic obstructive pulmonary disease (COPD), and completely resolved pulmonary tuberculosis (TB) in remote past, were nota­ble parts of her past medical history. Due to clinical, paraclinical and radio­graphic findings and because of recent hospitalization, she was admitted to internal medicine ward with the diagnosis of health care associated pneumo­nia (HCAP). Despite the application of broad-spectrum antibiotics and ap­propriate supportive care, she had a poor response to the treatment. During the daily visit in Intensive Care Unit (ICU), numerous white larvae were de­tected in both nostrils. Further investigation of oropharynx and tracheal tube aspiration, showed no more larvae in mentioned parts. An hour later, nasal spontaneous bleeding occurred. Otorhinolaryngology consultation was per­formed and led to surgical procedure. In ENT examination, there were nu­merous larvae and massive clot formation in both inferior meatuses and distal nasal septum perforation. Thirty-seven extracted larvae were transferred to Medical Entomology lab by vial 70% ethanol and 5 live larvae for rearing. After pre­cise investigation by aid of light microscopy, the larvae were identified as Chrysomya bezziana. Due to discovered 2nd larvae stage and duration of hospitalization, this infestation was identified as nasal myiasis.