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Mice were infected intraperitoneally with 10 000 tachyzoites of Toxoplasma gondii (RH) strain and, 24 h later, were treated orally for 10 days with atovaquone and azithromycin, either alone or in combination. Evaluation of the efficacy of the drugs was performed by microscopic examination of smears prepared from the organs of the mice, and by subinoculation of visceral and brain suspensions from surviving mice into healthy mice at the end of the experiments. It was found that 58%, 83% and 100% of the mice survived after administration of 75, 150 or 200 mg/kg per day of azithromycin, respectively. Moreover, 8%, 17% and 25% of the mice survived after treatment with atovaquone at 20, 50 or 100 mg/kg per day, respectively. No synergistic or additive effects of combinations of atovaquone and azithromycin were observed. However, azithromycin did not eradicate the parasite from the brain and viscera of the infected mice, whereas atovaquone at 20, 50 and 100 mg/kg per day removed the parasite from viscera and at 100 mg/kg per day eradicated the parasite from the brain of infected mice. The combinations of atovaquone and azithromycin failed to completely eradicate the parasite from the brain and viscera of infected mice.

During intraocular surgery of a 44-year-old woman from Guilan Province in north Iran, a small flat parasite was removed from the anterior chamber angle of the left eye. According to morphological characterisation of the parasite, it was identified as Fasciola hepatica. The route of entry of the parasite was not identified. Meanwhile, stool and serology examinations of the patient for diagnosis of Fasciola infection were negative. In endemic areas of F. hepatica infection, ocular involvement should be considered in cases of uveitis, despite no other systemic involvement.

Toxoplasmosis is one of the most common parasitic diseases worldwide. It is estimated that approximately one-third of the world’s population is latently infected. Infection generally occurs via oral the route and maternal transmission. Damage of the central nervous system is one of the most serious consequences of congenital toxoplasmosis. Moreover, recent investigations proposed that acute and sub-acute congenital toxoplasmosis as well as latent toxoplasmosis during pregnancy; play various roles in the etiology of different neuropsychiatric disorders in mothers and their offspring. This paper reviews new findings about the role of latent toxoplasmosis in the etiology of various neuropsychiatric disorders in mothers and their offspring.

Introduction: Toxoplasma gondii is one of the most important pathogen that has adverse effect on reproductive function. Evidence Acquisition: Recent studies revealed that infection with T. gondii not only affect female reproduction, also cause male reproductive impairment. In clinical studies, high prevalence of toxoplasmosis in sterile men has been reported. In animal models, toxoplasmosis is associated with male reproductive impairment. Moreover, there are some evidences about venereal transmission of T. gondii. Drugs used for treatment of toxoplasmosis may cause adverse effects on male reproductive function. Results: In present article, effect of Toxoplasma infection on male reproductive system of human and animal was reviewed. There are several reports expressing association between Toxoplasmosis and male genital tract impairment in both human and animals. Conclusions: These findings suggest that T. gondii infection can cause temporary impairment on the reproductive parameters of human or animal male as well as impairment of different hormones which may cause insufficient male reproductivity.

Background: Visceral leishmaniasis (Kala-azar) is one of the most serious tropical diseases, and it can lead to death. The kinetoplastid membrane protein-11 (KMP-11) is highly conserved in all stages of the Leishmania life cycle. Objectives: In the present study, the KMP-11 gene was extracted from Leishmania infantum and then, cloned and expressed in an expression vector.The main objective of this study was to provide an introduction to DNA vaccine production against visceral leishmaniasis. Materials and Methods: DNA was extracted from L. infantum (MHOM/TN/80/IPI1), and amplified by PCR and a specific primer. Afterwards the purified PCR products were successfully ligated into the pTZ57R/T plasmid vector. The pcDNA3 plasmid was used as an expression vector and the KMP-11 gene subcloned into this plasmid. The pTKMP-11 plasmid was digested by restriction enzymes to confirm cloning of this gene in the pTZ57R/T plasmid. In the last step, the subcloned gene was expressed in a eukaryotic cell. Results: The KMP-11 gene was successfully subcloned in pcDNA3 as a eukaryotic expression vector. Recombinant KMP-11 protein was confirmed by the reverse transcriptase polymerase chain reaction (RT-PCR) method. Conclusions: The results of the present study will increase our knowledge about molecular cloning and expression of the L. infantum KMP-11 gene, and this may be used as an effective target for controlling visceral leishmaniasis.

Human fascioliasis has been reported in many countries, including Iran. Various techniques have been evaluated for diagnosis of human fascioliasis using different antigens. We evaluated Fasciola gigantica partially purified fraction antigen (PPF) isolated from sheep's liver fluke for the diagnosis of human fascioliasis.Two hundred sixty-one sera were collected from 104 patients living in an area endemic for human fascioliasis, from 89 non-fascioliasis patients living in a non-endemic area, and from 68 healthy individuals. Micro-ELISA was used in the evaluation of the sensitivity and specificity of Dot-ELISA.With a 1:800 sera dilution as the cut-off titer, the sensitivity of the Dot-ELISA test in diagnosis of human fascioliasis was 94.23% and the specificity was 99.36%.Dot-ELISA using PPF antigen is a sensitive and specific method for diagnosis of human fascioliasis that is also rapid and inexpensive.

Objectives: Drugs used for toxoplasmosis have limited efficacy and also severe side effects. A new drug with good efficacy and limited side effects is need of the hour. We studied the effects of artemether on Toxoplasma gondii in vitro conditions. Materials and Methods: Artemether (methyl-ether-qinghaosu) was tested for tachyzoites, J774, and Vero cell lines infected by T. gondii. For evaluating the effect of drugs on Vero cells infected with T. gondii, we designed two separate experiments; in the first experiment, the Vero cells were infected with tachyzoites and then treated with artemether; while in the second one, the tachyzoites were exposed to artemether and then Vero cells were infected with treated tachyzoites. For evaluating the apoptotic effect of artemether on tachyzoites and infected J774 macrophages cell line with T. gondii , we used flow cytometry method. Inhibitory concentration (IC 50 ) was evaluated by intracellular replication of tachyzoites in Vero cells. Results: IC 50 for infected Vero cells with tachyzoites was determined as 49.13 μg/ml. In pretreated tachyzoites with artemether before entering into Vero cells, IC 50 was calculated as 13.15 μg/ml. In both experiments, artemether showed a higher inhibitory effect than sulfadiazine (positive control). Artemether even at the highest concentrations only showed low cytotoxicity on Vero and J774 cell lines. Apoptosis in tachyzoites rise with an increasing concentration of artemether. Conclusions: Our findings indicate that artemether is effective to control the tachyzoites of T. gondii in vitro and maybe a good alternative drug for toxoplasmosis.

Background: The Thiol-specific antioxidant (TSA) is an antigen of Leishmania major which is believed to be the most promising molecule as a vaccine candidate against leishmaniasis. Objectives: In this study, we investigated the protective efficacy of TSA-based DNA vaccine against L. major infection. Materials and Methods: Recombinant plasmid construction TSA (pcTSA) was prepared and transfected into eukaryotic cells and expression was confirmed with western blot and RT-PCR. The mice were assigned to six different groups and DNA immunization was performed with 100 µg intramuscular recombinant plasmid with a two-week interval. Cytokines and lymphocyte proliferation assay, antibody responses and determination of parasite burden were performed following immunization and the challenging infection with L. major. Results: The antibody and IFN-γ titers were higher in pcTSA + AlPO4 group the immunized mice with pcTSA alone, but there was no statistically significant difference between the two groups. Additionally the IL-4 titer was not statistically different between the groups following immunization and challenge. After infection with L. major promastigotes, the immunized mice with pcTSA and the one immunized with both pcTSA + AlPO4 presented a considerable reduction in diameter of lesion but there was no statistical difference between the two groups. The immunized mice had significantly lower parasite loads. No significant differences were observed between the two vaccinated groups. However the highest reduction in parasite burden was observed in the group immunized with pcDNA + AlPO4. No significant differences were observed in survival rate of the immunized mice after the challenge with L. major. Conclusions: In conclusion, TSA-based DNA vaccine induced Th1 platform immune response and aluminum phosphate could improve the efficacy of these vaccines with induction of humoral and cellular immune responses against L. major infection. There were no significant differences observed between pcTSA and pcTSA + AlPO4 groups.

Toxoplasma gondii is an obligate intracellular protozoan that is a causative agent of toxoplasmosis, a disease which may result in a spectrum of consequences. It has been shown that DNA vaccine can be effective in partial protection against this parasite. In the present work, a single DNA vaccine containing ROP1 was evaluated against T. gondii infection in Balb/c mice. To enhance the immune response, alum was used as an adjuvant. After intramuscular immunization, cytokine and antibody assays and mortality rate were evaluated. The results showed that mice immunized by pcROP1 with or without alum produced high Th1 immune response compared with control groups. This type of DNA vaccine prolonged slightly the survival time. The current study showed that ROP1 DNA vaccine can induced partial protective response against toxoplasmosis.Keywords: Toxoplasma gondii, DNA vaccine, ROP1gene, immunity, mice

Project : This study investigated the in vitro and in vivo effectiveness of biogenic selenium nanoparticles (Se NPs), biosynthesized by Bacillus sp. MSh-1, against Leishmania major (MRHO/IR/75/ER). Procedure : The 3-(4,5-dimethylthiozol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity effects of the biogenic Se NPs against both promastigote and amastigote forms of L. major . In a separate in vivo experiment, we also determined the preventive and therapeutic effects of biogenic Se NPs in BALB/c mice following subcutaneous infected with L. major . Results : The MTT assays showed that the highest toxicity occurred after 72 h against both promastigote and amastigote forms of L. major . The cytotoxicity of Se NPs was higher at all incubation times (24, 48, and 72 h) against the promastigote than the amastigote form ( p 50 ) of the Se NPs were 1.62 ± 0.6 and 4.4 ± 0.6 μg ml −1 against the promastigote and amastigote forms, respectively, after a 72-h incubation period. Apoptosis assays showed DNA fragmentation in promastigotes treated with Se NPs. In an animal challenge, prophylactic doses of biogenic Se NPs delayed the development of localized cutaneous lesions. Moreover, daily administration of Se NPs (5 or 10 mg kg −1 day −1 ) in similarly infected BALB/c mice that had not received prophylactic doses of Se NPs also abolished the localized lesions after 14 days. Conclusion : Based on these in vitro and in vivo studies, biogenic Se NPs can be considered as a novel therapeutic agent for treatment of the localized lesions typical of cutaneous leishmaniasis.

Visceral leishmaniasis is one of the most important sever diseases in tropical and subtropical countries. In the present study the effects of antimony sulfide nanoparticles onLeishmania infantum in vitrowere evaluated. Antimony sulfide NPs (Sb2S5) were synthesized by biological method fromSerratia marcescensbacteria. Then the cytotoxicity effects of different concentrations (5, 10, 25, 50, and 100 μg/mL) of this nanoparticle were assessed on promastigote and amastigote stages ofL. infantum. MTT method was used for verification results of promastigote assay. Finally, the percentages of apoptotic, necrotic, and viable cells were determined by flow cytometry. The results indicated the positive effectiveness of antimony sulfide NPs on proliferation of promastigote form. The IC50(50% inhibitory concentration) of antimony sulfide NPs on promastigotes was calculated 50 μg/mL. The cytotoxicity effect was dose-dependent means by increasing the concentration of antimony sulfide NPs, the cytotoxicity curve was raised and the viability curve of the parasite dropped simultaneously. Moreover, the IC50of antimony sulfide NPs on amastigote stage was calculated 25 μg/mL. On the other hand, however, antimony sulfide NPs have a low cytotoxicity effect on uninfected macrophages but it can induce apoptosis in promastigote stage at 3 of 4 concentrations.

Sarcocystis cameli was first described in one-humped camels (Camelus dromedarius), and it is the only species which have so far reported in camels. Although more than 150 species of Sarcocystis were described in various animals, only a few data on camel Sarcocystis ultrastructure were published, and this report is the first for molecular information (DNA sequence and RLFP digestion pattern). The main objective of the present work is to characterize Sarcocystis isolated from camels by electron microscopy and PCR-RFLP methods. Muscle samples were taken from the fresh esophagus, diaphragm, skeletal muscles, and heart of one-humped camels (C. dromedarius) slaughtered in abattoirs of Tehran and Ghazvin provinces, Iran. The dissection and trypsin digestion techniques were applied for the detection of the cysts. The infected samples were fixed in glutaraldehyde and/or frozen at -20°C until use for ultrastructural and molecular studies, respectively. The ultrastructural and molecular studies were carried out contemporaneously. The 18S rRNA gene of the parasites was amplified by PCR. The PCR products were cloned into a pTZ57R/T and sequenced. In addition, the PCR products were digested separately with each of the four restriction enzymes for RFLP. Our results indicated that only microcysts were observed in muscle samples. The microcysts were white, elongated, spindled, and a few spiral-shaped, with mean size 260 × 75 μm which are identical with S. cameli. The ultrastructure of microcyst wall had many non-branched finger-like protrusions irregularly folded. There was a 600-bp specific band amplified after PCR with specific primers. The molecular data for camel Sarcocystis is reported for the first time in Iran and the world.

Herpes simplex virus type 2 (HSV-2) is highly prevalent and major cause of genital herpes in humans. The life-long nature of infection and the increasing prevalence of genital herpes imply that vaccination is the best strategy for controlling the spread of infection and limiting HSV disease. HSV glycoprotein D (gD) is one of the most important viral immunogen which has an essential role in virus infectivity and induction of immune responses.HSV-2 DNA was extracted and used as template in polymerase chain reactions to amplify gD2 gene. The PCR product was confirmed by restriction enzyme analysis, cloned into a cloning vector and then sequenced. The Bac-to-Bac expression system was used to express HSV-2 gD in insect cells. The expressed protein was used as subunit vaccine to immunize guinea pigs after confirmation.The expressed protein was confirmed with SDS-PAGE and Western-blot analysis. In Western-blot analysis, two major protein bands, with approximate molecular weights of 52-55 and 41-43 kDa corresponding to the glycosylated and non-glycosylated forms of gD2 protein, were observed, respectively. Immunization with the recombinant gD2 could elicit humoral responses in guinea pigs as measured by neutralization test and ELISA, and offered high protection against induced HSV-2 genital disease.The baculovirus expression of heterologous genes permits proper folding, post-translational modification and oligomerization in manners that are often identical to those that occur in mammalian cells. Expression of proteins under the control of the strong polyhedrin promoter, allowing high level protein production, can be used as subunit vaccine.

Cryptosporidiosis is a widespread cause of diarrheal diseases of humans, young calves, and many mammals. Although in recent years molecular investigations on Cryptosporidium have increased, no data are available on Iran in this regard. Two species of Cryptosporidium are known to infect human beings; Cryptosporidium hominis which shows anthroponotic transmission among humans and Cryptosporidium parvum which shows zoonotic transmission between animals and humans. Cryptosporidium oocysts, morphologically identified as C. parvum, were isolated from 24 human and 35 bovine cases in Shahriar (suburb of Tehran, Iran), and genotyped by means of a Nested-polymerase chain reaction/restriction fragment length polymorphism analysis of the 18s rRNA gene. The isolates from bovine samples gave zoonotic or 2 genotype (C. parvum), and DNA profiles of human isolates gave two distinct genotypes, namely an anthroponotic or 1 (C. hominis) and zoonotic genotype or 2 (C. parvum).

Toxoplasmosis is a protozoal infection caused by Toxoplasma gondii. Toxoplasmosis produce severe damage in patients who are immunosuppressed. In those who are immunosuppressed, latent infection can be reactivated resulting in acute disseminating disease. Betamethasone is a synthetic glycocorticoid, used as an anti-inflammatory and immunosuppressant in a wide variety of disorders. The aim of this study was evaluation of betamethasone as an immunosuppressor drug on infected cells by Toxoplasma gondii. In this study, at first HeLa cells were grown in 24 well culture plates in culture medium . When confluent monolayer was obtained, we compared 6 groups to evaluate the effect of betamethasone as a corticosteroid drug (two concentrations 4 and 40 micro g/ml) and the effect of IFN-gamma (100 IU/ml ) on growth, replication and Nitric Oxide (NO) production. The results showed, that high number of plaques were seen in group with 40 mug/ml of betamethasone and the lowest number of plaques were seen in group with 100 IU of IFN-gamma. The difference between plaque number in control and groups treated with IFN-gamma and betamethasone was significant (P0.05). The groups with betamethasone or IFN-gamma without tachyzoites did not show any effect on cell structures. Replication rates in the wells treated with IFN-gamma were decreased significantly 72 h post inoculation in comparison with control group (P0.05). There was no significant difference among different groups in NO production. The results indicated that betamethasone increase the invasion of tachyzoites to host cells in vitro.

Human infection especially with helminth parasites is an emerging health issue, as the human environment is increasingly shared with infected animals, either pets or wild life. In this survey, the intestinal content of 83 stray dogs, 22 red foxes and 10 golden Jackals collected from the West Azarbaijan, Kordestan and Kermanshah provinces in the west of Iran, were studied for the presence of helminth parasites. The percentage of different species recovered from these animals is listed as follows: From stray dogs: Toxocara canis (6.02%), Toxascaris leonina (32.53%), Ancylostoma caninum (3.61%), Oxynema sp. (1.35%), Rictularia affinis (12.05%), Taenia hydatigena (53.01%), Taenia ovis (7.23%), Taenia multiceps (4.82%), Echinococcus granulosus (13.25%), Dipylidium caninum (38.55%), Mesocestoides lineatus (26.50%) and Macracanthorhynchus hirudinaceus (4.82%). From red foxes: T. canis (4.54%), T. leonina (31.82%), A. caninum (4.54%), Uncinaria stenocephala (13.64%), Oxynema sp. (9.09%), R. affinis (54.54%), Strongyloides sp. (4.54%), Physaloptera sp. (4.54%), T. hydatigena (9.09%), E. granulosus (4.54%), D. caninum (9.09%), M. lineatus (81.82%), Joyeuxiella pasqalei (27.27%), Diplopylidium nolleri (4.54%), M. hirudinaceus (22.72%) and Macracanthorhynchus sp. (9.09%). From golden jackals: T. canis (10%), T. leonina (30%), R. affinis (50%), T. hydatigena (10%), D. caninum (20%), M. lineatus (70%), J. pasqalei (30%.), Alaria canis (10%), M. hirudinaceus (30%) and Macracanthomynchus sp. (10%).

In the present study, Echinoccocus granulosus isolates collected from human, sheep and camel samples in Iran were characterized based on rostellar hook morphology of protoscoleces as well as PCR-RFLP. Morphological study on human and animal isolates showed the presence of two distinct strains of the parasite, one in sheep and the other one in camels. In this regard, rostellar hook of sheep isolates were significantly different from those of camel origin, meanwhile human isolates were found to be similar to those isolated from sheep. Molecular analysis of the ITS1 region of rDNA derived from human, sheep and camel isolates were in agreement with the morphological findings. Based on the PCR-RFLP method, the sheep and human isolates appeared to pertain to the same genotype and the camel isolates were appeared to pertain to a different genotype.

A total of 100 jackals, 35 stray dogs, 13 cats, two badgers, one fox and one wild cat were collected from two provinces in northern Iran. On post mortem examination 41 species of helminths were recovered, at least 12 of which have been reported infecting humans in various countries. Hookworms were the most commonly found parasites, and Echinococcus granulosus showed the highest intensity of infection. Seven helminth species from jackals, three species from dogs, four species from cats and four species from badgers are reported for the first time in Iran.

Summary Objectives The main objective of the present study was to detect point mutations at positions 86, 184, 1034, 1042, and 1246 of the Plasmodium falciparum multidrug resistance gene ( pfmdr1 ) in blood samples collected from malaria patients in Chabahar, a harbor city located in Southeast Iran. Methods Twenty-six blood samples from patients infected with P. falciparum , who had a chloroquine (CQ) response failure, were collected pre-treatment. Following treatment with CQ, drug susceptibility was assessed using an in vivo test. Molecular detection of single nucleotide polymorphisms (SNPs) was carried out using the LightCycler hybridization probe assay. Results The pfmdr1 N86Y mutation was found in six isolates (23.1%). Mutations at the four other positions were not observed in any isolates. Conclusion The present study showed no mutation at codon positions 184, 1034, 1042, and 1246 of pfmdr1 in any of the Iranian P. falciparum isolates; thus these alleles cannot serve as markers for CQ resistance in Iran.

Objective: To investigate the protective effect of IL-22 and IL-12 on cutaneous leishmaniasisin BALB/c mice. Methods: The protective effect of IL-22 and IL-12 on cutaneous leishmanias in BALB/c mice was evaluated by measurement of IL-4, INF-毭, total IgG, IgG1 and IgG2a after challenge with Leishamania major. Clinical evaluations were performed by measurement of lesion diameter, and survival rate of the mice. Results: In week 27 post infection, the mortality rates for control groups were 100%. While the survival rates for the IL-12, IL-12 + IL-22, and IL-22(5 ng/ g) groups were 100%. The size of lesions decreased in the presence IL-22 (5 ng/g) of mice weight, which was statistically significant in comparison with other groups (P

Cutaneous leishmaniasis is a common parasitic disease that is endemic in some parts of Iran. The drugs of choice used for leishmaniasis therapy are associated with a risk of recurrence and serious adverse effects. Therefore, finding a safe and effective treatment is of great importance. In the present study, the effect of aloe-emodin on the growth of Leishmania major amastigotes was evaluated under in vitro conditions. In addition, the efficacy of a topical of aloe-emodin ointment was investigated in BALB/c mice with cutaneous leishmanial ulcers. Different concentrations (40 μg/mL, 80 μg/mL, 120 μg/mL, and 160 μg/mL) of aloe-emodin were tested on Leishmania amastigotes twice: 24 hours and 48 hours. The induced apoptosis and necrotic effects of two concentrations (40 μg/mL and 120 μg/mL) of aloe-emodin on promastigotes were investigated by flow cytometry. Under the in vivo condition, aloe-emodin ointment efficacy was evaluated at two concentrations (i.e., 0.1% and 1%). Serum indicator factors of the test and control groups were tested to evaluate the toxic effects of this compound on the liver and kidney. Results showed that aloe-emodin inhibited the growth of Leishmania amastigotes and induced apoptosis in promastigotes. Topical application of aloe-emodin ointment likewise reduced the ulcer size. No significant differences in biochemical analysis were observed between the control and treated groups. In conclusion, aloe-emodin showed antileishmanial effects under in vitro and in vivo conditions and may be used in clinical trials.