Background: Cystic echinococcosis can cause severe disease and probable death in humans. Epitopes of its antigens play a key role in the sensitivity and specificity of immunodiagnostic tests.
Methods: Epitope prediction software programs predict the most antigenic linear B-cell epitopes of AgB (8 kD), Ag5, and Ag95. Six such epitopes were predicted and connected by “Gly-Ser” linker and synthesized. The purity of the concentrated recombinant multi-epitope protein was assessed by 15% SDS-PAGE. Overall, 186 serum samples were collected from the Loghman Hakim Hospital and different laboratories, Tehran, Iran, from July 2016 to February 2017. Patients infected with hepatic hydatid cysts, patients infected by other parasites and viruses, and healthy individuals were used to detect the anti-CE IgG using recombinant multi-epitope protein.
Results: Forty-one samples out of 43 cases of hydatidosis were diagnosed correctly as positive, and two were negative. In addition, six negative cases of healthy individual group were diagnosed as positive and negative with rMEP-ELISA and the commercial kit, respectively. Therefore, these six samples were considered as false positive using our method. In addition, a diagnostic sensitivity of 95.3% (95% CI, 84.19% to 99.43%) and a specificity of 95.0% (95% CI, 89.43% to 98.14%) were obtained using optimum cutoff value (0.20). The sensitivity and specificity of the commercial kit was 100%.
Conclusion: Our findings showed high diagnostic accuracy of the ELISA test using the developed recombinant protein, which encourages the use of this recombinant multi-epitope protein for rapid serological diagnosis of hydatidosis.

Background: KMP-11 (Kinetoplastid membrane protein-Π) exists in all species of kinetoplastid family. It is fully conserved and the protein produced by this gene can induce a very high cellular immune response. We aimed to design a suitable construction for a Leishmania major DNA vaccine and evaluate the protective efficacy of it as a candidate for DNA vaccine against cutaneous leishmaniasis in BALB/c mice.
Methods: This experimental study was conducted in Tehran City, Iran, between April 20, 2015 and May 30, 2016. KMP-11 gene of L. major (MRHO/IR/75/ER, Iranian strain) and NT-GP96 of Xenopus GP96 DNA from a pBluescript-GP96 plasmid were amplified by PCR and the purified PCR products were cloned into the pJET1.2/blunt plasmid vector, then, subcloned into pEGFP-N1 plasmid as an expression vector. Finally, the KMP-11 gene was fused with GP96 and afterward the combination cloned in pEGFP-N1. All the cloned genes confirmed by enzyme digestions. Then, four groups of mice were immunized with PBS, pEGFP-N1, pEGFP-N1-KMP, and pEGFP-N1-fusion. Four weeks after immunization, all animals were challenged with L. major virulent promastigotes.
Results: The constructed fusion potentially showed an ability to elicit Th1 responses that led to cutaneous lesion healing. Interestingly, the group received KMP11-GP96 –GFP showed the highest ratio of IFN- γ /IL-4 and IgG2a/IgG1 compare to other groups. No side effect was observed after using the fusion in the mice.
Conclusion: The constructed fusion could well stimulate both the cellular and humoral immune systems that led to cutaneous lesion healing in mice.

Background: Broad spectrums of pharmacological properties, including antimicrobial activity have been attributed to Zataria multiflora Boiss (Laminaceae). The in vivo efficacy and safety of Z. multiflora essential oil (ZM-EO) were evaluated against acute toxoplasmosis caused by Toxoplasma gondii (Sarcocystidae) in mice.
Methods: Z. multiflora (aerial parts) was obtained from the rural districts of Kerman city (Kerman Province) Southwestern Iran, in May of 2016. Male NMRI mice were orally treated with normal saline (control group) and ZM-EO at the doses of 0.2 and 0.4 mL/kg once a day for 14 d (8 mice in each group). On the 15^th day, the mice were infected with 10⁴ tachyzoites of T. gondii RH strain by intraperitoneal route. The mortality rate and parasite load were determined in the infected mice. Additionally, 24 mice were applied to examine the sub-acute toxicity of ZM-EO at the above doses after treatment during 14 d.
Results: GC/MS analysis displayed that the key constituents were thymol (45.4%), carvacrol (23%) and p-cymene (10.6%), respectively. Overall, 100% mortality was observed on the 8^th and 9^th days in treated mice with the concentrations of 0.2 and 0.4 mL/kg, respectively. The mean number of tachyzoites in the mice treated with 0.2 and 0.4 mL/kg of ZM-EO were 189×10⁴ and 76×10⁴ cell/mL, respectively, meaningfully (P<0.05) reduced compared with the control mice. Results also demonstrated that ZM-EO had no important toxicity on mice.
Conclusion: The results demonstrated the efficacy of ZM-EO against acute toxoplasmosis. Nevertheless, supplementary surveys are mandatory to examine its precise effects, mainly immunomodulatory effect on toxoplasmosis.

Background: The purpose of this study was to determine the prevalence and genotype of Cryptosporidium spp. in different groups of immunocompromised patients admitted to the referral hospitals in center of Iran during 2015–2016.
Methods: This cross-sectional study was conducted on 346 immunocompromised patients (HIV⁺/AIDS, Lymphoma, Leukemia and organ transplants) in referred hospitals from central parts of Iran including Isfahan, Markazi, Yazd and Chaharmahale Bakhtiari provinces. Stool samples were analyzed for Cryptosporidium species, modified Ziehl–Neelsen staining techniques followed by the semi-nested PCR and DNA sequencing methods.
Results: The total rate of Cryptosporidium spp. was 3.46% (12/346) in the patients, however, the prevalence of the parasite, was 4.6% (4/87) in HIV⁺/AIDS patients, 3.6% (6/168) in patients with blood malignancy and 2.1% (2/91) in organ transplant recipients. The SSU rRNA gene of Cryptosporidium spp. in all microscopic-positive samples was effectively amplified by the semi-nested PCR and DNA sequences, exposed the existence of two Cryptosporidium species, including C. hominis 91.6% (11/12) and C. parvum 8.3% (1/12).
Conclusion: The predominance of C. hominis in the present study may be certifies the importance of anthroponotic transmission of cryptosporidiosis in center of Iran.

Background: The current study was conducted to investigate the antigenic effect of Marshallagia marshalli on the treatment of asthma by measuring the secreted inhibitory cytokine.
Methods: Case patients and controls were selected from clinics in Mashhad, Khorasan Razavi Province, Northeastern Iran in 2017-18. In this experimental study, peripheral blood mononuclear cells (PBMCs) were isolated from 15 patients with asthma and 10 healthy controls and were cultured. PBMCs were then converted to tolerogenic DCs through exposure to GM-CSF, IL-4 and M. marshalli antigen. Then, tolerogenic DCs were exposed to autologous T cells for five days and finally, the level of secreted TGF-β1 was measured.
Results: The mean TGF-β1 level in the control and control groups was 210.2 ± 8.2 and 225.4 ± 6.1 pq/ml, respectively. The results showed that TGF-β1 levels in both groups significantly increased in both groups (P<0.001). In addition, TGF-β1 levels in the case group were significantly higher than the control group (P<0.001).
Conclusion: M. marshalli antigen increase the level of TGF-β1 and can create antigen-bearing dendritic cells and shift T lymphocytes to the regulatory type. This parasite can be used in dendritic cell therapy to control allergic diseases.

Background: Visceral leishmaniasis (VL) is the most severe form of leishmaniasis in Iran with high mortality rates in the case of inaccurate diagnosis and treatment. This study aimed to prepare and evaluate a new rk39 recombinant antigen from an Iranian strain of Leishmania infantum for diagnosis of VL in humans and dogs.
Methods: rK39-based enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for the detection of anti L. infantum antibodies. We screened 84 human sera and 87 dog sera from clinical cases in the endemic area of Meshkin-Shahr, Iran along with 176 sera from healthy controls (collected from 86 humans and 90 dogs) during 2013 -2016.
Results: Using the rK39 ELISA, a sensitivity of 85.7% (95% CI, 95-99%) and a specificity of 86.0% (95% CI, 95%-99%) were detected in human sera at a 1:800 (cut-off) titer when DAT-confirmed cases were compared with healthy controls; a sensitivity of 96.6% (95% CI, 95%-99%) and specificity of 94.4% (95% CI, 95%-99%) were found at a 1:80 (cut-off) titer compared with DAT. Kappa analysis indicated agreement between the rK39 ELISA and DAT (0.718) when using human sera at a 1:800 (cut-off) titer as well as (0.910) at a 1:80 (cut-off) titer when using dog sera (P<0.05).
Conclusion: New rk39 recombinant antigen from an Iranian strain of Leishmania infantum seems to be used for diagnosis of VL in humans and dogs. Further extended field studies are recommended.

Background: Dirofilariasis is a globally distributed arthropod-borne parasitic disease of mainly canids and felids. We evaluated to extend the knowledge of morpho-molecular characteristics and outer ultrastructure of Dirofilaria immitis isolated from Northwest of Iran.
Methods: Overall, 67 filarial worms including 41 females and 26 males parasites were collected from the cardiovascular system of the 43 stray dogs in Meshkinshar, Ardebil Province, Northwest of Iran in 2017, and subjected to light and scanning electron microscopy (SEM) as well as carmine alum staining for morpho-molecular and identification. Molecular methods were used for confirmation of morphological findings by sequencing of Cytochrome c oxidase subunit I (cox1) gene.
Results: The partial DNA sequencing of cox1 gene of adult parasites showed considerable homology and close proximity to the previously isolated from Kerman and Meshkinshahr, Iran. The lowest genetic variation and the highest intra-species variability was found in D. immitis and Dirofilaria repens, respectively. No similarity was identified between D. immitis nucleotide sequence and Wolbachia species as its endosymbiont bacteria.
Conclusion: The SEM technique is an excellent tool for differential recognition of the parasite surface morphology and molecular techniques could differentiate and identify Dirofilaria spp.

Background: Several species of streptomycetes, saprophytic bacteria found widely distributed in soil, water and plants, produce bioactive compounds such as intra and extracellular hydrolases including lytic enzymes which reflecting on their importance in the biological control of insects and parasites. This study assessed the in vitro effects of Streptomyces tyrosinase, produced from Streptomyces spp. isolated from Egyptian soil, on animal-parasitic nematode Toxocara vitulorum, in terms of egg development and adult worm’s cuticular structure, and as an alternative strategy to alleviate this infection.
Methods: This study was conducted at the National Research Centre, Egypt in 2018. Five different concentrations of tyrosinase, ranged from 1%-30% were tested against the development of T. vitulorum eggs. The concentration induced the highest inhibitory activity was tested against adult T. vitulorum cuticle, which is essential for the protective and nutritive functions. The results were compared with those observed in the egg development and worm cuticle following incubation in Streptomyces protease (as a reference enzyme).
Results: Compared to Streptomyces protease, higher inhibitory activity on T. vitulorum egg development and extreme cuticular alterations of the treated adult worms had been observed following 24 h exposure to Streptomyces tyrosinase. Once the cuticle had been damaged, the enzyme would be able to penetrate deeper into the internal tissues of the nematode and caused more widespread disruption.
Conclusion: The current study could offer a promising bio-control agent, Streptomyces tyrosinase, against T. vitulorum alternative to the more expensive synthetic anthelmintics.

Background: Echinococcus granulosus parasite causes a zoonotic disease which is important for public and veterinary health. Since pumpkin seeds (Cucurbita sp.) are used as traditional vermifuge in Iran, they may be a potential herbal anthelmintic.
Methods: This study was designed in 2016 to evaluate the in vitro scolicidal effect of Cucurbita moschata seeds form northern part of Iran. Hydroalcoholic and petroleum ether extracts were prepared by maceration and soxhlet respectively. Both extracts with four different concentrations (100, 10, 1, 0.1 mg/ml) were incubated against protoscoleces in 5, 15, 30 and 60 min.
Results: Maximum mortality was 16% with 1% hydroalcoholic extract in 60 min. The highest mortality with organic extract was 4% with 10% concentration in 60 min (P=0.015).
Conclusion: Since highest mortality was 16%, the extract did not reach to LD50 (50% mortality). Therefore, the potency of the total extract is not sufficient as potential scolicidal drug.

Background: In the current study, we described the epidemiological features, clinical presentation, diagnosis and management of patients with suspicion of fascioliasis in Kohgiluyeh and Boyer-Ahmad Province in southwest of Iran.   
Methods: Overall, 56 patients with suspicion of fascioliasis, based on their clinical signs and symptoms that referred to Clinic of Internal Medicine in Yasuj city, from 2014 to 2016 were enrolled. Demographic data, history of eating aquatic local plants, the chief complains, and laboratory findings were recorded for each patient. Stool samples were obtained from each case for detection of Fasciola eggs. Moreover, blood samples were taken from each patient and evaluated for detection of anti-Fasciola antibodies by an indirect ELISA. Patients who defined as having fascioliasis were treated with triclabendazole and were followed for at least three months for clinical improvement.
Results: Serological test was positive in 5 patients. Of these 5 cases, three cases had a history of ingesting raw aquatic vegetables. The main clinical signs and symptoms in positive cases were; abdominal pain (60%), epigastric pain (40%), anemia (60%), and dermal pruritus (20%). Hypereosinophilia was seen in all of 5 positive cases. No Fasciola egg was found in stool specimens of any of the patients. The fascioliasis cases were treated by triclabendazole and clinical symptoms disappeared in all of 5 cases.
Conclusion: Our observation further confirmed Yasuj district as a human endemic area for fascioliasis in Iran. The study also highlighted the importance of clinical features together with eosinophilia, as key parameters, in the diagnosis of human fascioliasis. Clinicians need to be aware of this disease and should keep in mind fascioliasis when hypereosinophilia present in patients in such endemic areas.

Background: Intestinal parasitic infections (IPIs) are among the most important etiologies of gastrointestinal disorders in developing countries. The present study was performed to determine the prevalence of IPIs in rural inhabitants of Fouman, northern Iran.
Methods: Overall, 31 villages were randomly selected during 2015-2016. Stool samples were collected from 1500 inhabitants aged 2-87. The samples were examined by direct wet smear, formalin ethyl-acetate concentration and agar plate culture. Trichrome staining and modified acid-fast staining were used as confirmatory tests for intestinal amoeba and flagellates and cryptosporidium spp., respectively. Data were analyzed with Chi-Square and Fisher exact tests using SPSS.
Results: 8.06% of participants were positive for at least one intestinal parasite. The prevalence of mixed parasitic infections was 0.87%. The most prevalent IPIs were caused by Trichostrongylus spp. (3.13%), followed by Strongyloides stercoralis (1.5%), Giardia lamblia (1.3%), and Entamoeba coli (1.0%), Blastocystis hominis (0.86%), E. histolytica/dispar (0.53%), Endolimax nana (0.26%), Iodamoeba butschlii (0.13%), Trichuris trichiura (0.07%), Enterobius vermicularis (0.07%), Hook worm (0.07%) and E. hartmani (0.07%). Statistically, the prevalence of IPIs showed significant differences regarding the age groups, education status, occupation (P<0.001), and the habit of eating raw vegetables (P<0.007), whereas, the differences were insignificant with regard to sex (P=0.924) and water supply (P=0.088).
Conclusion: The prevalence of IPIs, especially soil-transmitted helminthes (STHs) has sharply decreased in northern Iran. Excluding Trichostrongylus spp. and S. stercoralis, other intestinal parasites only produce a marginal and unnoticeable health problem in this area, today.

Background: Echinococcus granulosus is a worldwide zoonotic cestode that lives mainly in the intestine of dog as definitive host. Its larval stage infects intermediate hosts and forms hydatid cysts mainly in the liver and lungs tissues and less other organs such as brain, eye and bone. In the experimental models, inoculation of protoscoleces into the peritoneum, thoracic cavity, subcutaneous and cerebrum produces hydatid cysts. Experimental echinococcosis in the animal models provides a good opportunity for study of the parasite-host relationship, different transmission ways of infection in the intermediate hosts and effect of new drugs.
Methods: The present study was conducted in the Veterinary School, Shahid Bahonar University of Kerman, Kerman, Iran in 2018. In this study, cerebral hydatidosis was investigated in 6 female Wistar rats weighing (200±20 gr). For this purpose, protoscoleces were collected from hydatid cysts of infected sheep liver. Overall, 300 protoscolices were injected directly in the lateral ventricle by an insulin syringe through the implanted cannula.
Results: After 4 months of inoculation, multiple thin-walled, transparent hydatid cysts were observed in the rat skull. All cysts were infertile. The cysts were localized prominently on the cerebral cortex and lesser in the parenchyma and ventricles. The cyst walls consisted of three layers consist of the outer layer (fibrous capsule), two parasitic layers and the endocyst layer (germinal layer). The cyst was surrounded by the inflammatory cells consist of lymphocytes, plasma cells and macrophages.
Conclusion: To the best of our knowledge, this research is the first experimental cerebral hydatidosis arisen from larval stage of Echinococcus granulosus in the animal model.

Background: The ancient Chehrabad Salt mine, a well-known archaeological site in Iran, has recently received increasing interest from Iranian and international archeologists. Also, the biological remains from this site have provided valuable sources for studying the pathogenic agents of ancient times. This study aimed to identify the parasitic helminth eggs preserved in the herbivores coprolites.
Methods: From 2011 to 2015, we received three coprolites belonging to herbivorous animals recovered during excavations in Chehrabad Salt mine of Zanjan, Iran. The coprolites were dated back to the Sassanid era (224-651 AD) by using radiocarbon accelerator mass spectrometry (AMS) and archeological stratigraphy methods. Following rehydration of the specimens in a 0.5% trisodium phosphate solution, the suspensions were mounted in glycerin jelly on glass slides and examined by a light microscope with 100x and 400x magnifications.
Results: Two coprolites belonged to donkeys and one to an unknown herbivore species. The recovered eggs belonged to members of two helminths families, Strongylidae, and Anoplocephalidae. Also, within the two coprolites, some mites, presumably of the order Oribatida, were observed.
Conclusion: The presence of two different nematodes in the equids coprolites provide clues of the burden of helminths infection on working animal at the Sassanid time and demonstrates the appropriate preservation condition of biological remains in the ancient salt mine of Chehrabad as well.

Background: Chagas disease caused by the parasite Trypanosoma cruzi is considered a neglected disease in several countries. One of the main problems about this disease is the lack of an effective treatment and the absence of adverse effects. T. cruzi, like most pathogenic fungi and yeasts, require specific sterols to maintain viability and proliferative capacity during their life cycle. However, the oxidation of this molecule to ergosterol peroxide has shown several biological effects, including its trypanocidal activity.
Methods: We have synthesized MOFs nanoparticles as carrier system coupled to ergosterol peroxide (MOFs-EP) and we have studied its effect on the circulating forms of the T. cruzi parasite.
Results: MOFs-EP possess an efficient trypanocidal activity at much lower inhibitory concentrations (ng/mL) that the concentrations shown by ergosterol peroxide (mg/mL) when administered unconjugated form.
Conclusion: Our results open a new possibility in the biomedical application of MOFs and ergosterol peroxide in the search for new options for the treatment of Chagas disease.

Background: Intestinal schistosomiasis is a disease caused by infection with one of the blood flukes called Schistosoma mansoni. The distribution of Schistosoma mansoni infection is high in Sub-Saharan Africa due to water source, sanitation and hygiene problems. This study aimed to determine the effect of water source, sanitation and hygiene on the prevalence of schistosomiasis among school-age children in Northwest Ethiopia.
Methods: A cross-sectional study was conducted from Apr 2016 to Aug 2016. Children were selected by systematic random sampling and Formol Ether Concentration Technique (FECT) was used to identify Schistosoma mansoni infection. Statistical analysis was done using descriptive statistics and strength of association of schistosomiasis with determinant factors was calculated by bivariate analysis.
Results: Among 333 children, 7% were infected with Schistosoma mansoni. Using surface water for drinking, poor hand wash habit and latrine utilization were significantly associated (P<0.05) with Schistosoma mansoni infection.
Conclusion: Absence of safe water for bathing, washing and swimming, poor sanitation and hygiene practices were major risk factors for schistosomiasis. Therefore, health education should be given on the transmission of S. mansoni infection, pure water, sanitation and hygiene in S. mansoni endemic areas.

Background: Cystic echinococcosis (CE) is a worldwide zoonotic helminthic disease caused by the larval stage of Echinococcus granulosus. The infection is particularly important in terms of economic and medico-veterinary aspects in endemic areas including Iran. Considering the possibility of organ-tropism in E. granulosus strains, the present study was aimed to identify the genotypes of E. granulosus in different organs involved in patients, undergone surgery in Baqiyatallah Hospital, Tehran, Iran from 2005-2015.
Methods: Overall, 29 formalin-fixed paraffin-embedded tissues (FFPT) from patients with histologically confirmed CE including liver (N: 14) lungs (N: 6) abdomen (N: 2), pancreas (N: 2) and each of spleen, gallbladder and, muscles (N: 1) plus unknown organs (N: 2) were used and genetically characterized using polymerase chain reaction, followed by partial sequencing of mitochondrial cytochrome c oxidase gene subunit 1(cox1) and analyzed.
Results: Nineteen out of 29 isolates including liver (N: 6) lungs (N: 4) abdomen (N: 2), pancreas (N: 2) and each of spleen, gallbladder, and muscle (N: 1), unknown organs (N: 2) obtained from paraffin-embedded blocks of human CE created an acceptable sequence in two directions. All 19 isolates regardless of the organ involved were recognized as E. granulosus sensu stricto (G1).
Conclusion: The sequence alignments of the isolates displayed two profiles. All sequenced samples showed E. granulosus sensu stricto (G1) with no organ-related genotype.

There is no direct evidence to support the existence of regional lymph node metastatic routes in hepatic alveolar echinococcosis, and only a few literature have been reported. There was a case of hepatic alveolar echinococcosis suspected of metastasis of regional lymph node in Clinical Center for Hydatidosis, Qinghai Province, China. The patient was a 24-yr-old male from pastoral area of Seda County, Sichuan Province, China in 2018. He was admitted to the hospital for physical examination and had no special discomfort. Preoperative examination showed liver occupancy and regional lymph node enlargement in the space between liver and stomach. Hepatectomy and lymph node resections were performed. Postoperative pathological results showed that both primary and metastatic lesion were of alveolar echinococcosis. Recovery of patient was good without complications and recurrence. In this case, metastasis was considered because the liver lesion was not directly connected to the lymph node. However, the case was still suspected due to the lack of pathological examination of other lymph nodes in the lymphatic return pathway. Regional lymph node metastasis may be one of the metastatic ways of alveolar echinococcosis.

The pericardial cysts (PC) are rare congenital anomaly. They are usually asymptomatic or incidentally found during surgery or by an imaging modality. We report a 35-yr-old man referred to Imam Ali Hospital, Kermanshah, western Iran in 2017, with palpitation, chest pain and dyspnea and physical exam revealed sign and symptoms of right atrial compression and tamponade.

Dirofilaria repens is the causative agent of human subcutaneous or, less often, ocular dirofilariasis. The work presents a rare case of ocular dirofilariasis manifested by previous subcutaneous migration accompanied by severe headache symptoms. In February 2017, a 58-yr-old man from Trnava region, western Slovakia, noticed red and itchy stripes on his left leg. Inflamed but painless stripes disappeared and showed up again every 5–7 days, migrating gradually towards the head. Approximately one month after the first skin´s alterations, strong pain in the left temple, with the swelling of the left face and the enlargement of mandibular lymph nodes appeared. Several days later, the patient felt excruciating pain of the right eyeball accompanied by strong nausea and subsequent vomiting. Ocular examination revealed the presence of a live worm in the subconjunctival space and morphological and molecular analyses of extracted helminth confirmed D. repens as etiological agent of the infection. According to clinical manifestation of the infection, it could be supposed that ocular form of the disease was the result of the migration of a parasite through the subcutaneous tissues. Moreover, a rare phenomenon of lymphadenitis of underlying lymph nodes and the swelling of left face accompanied the migration.

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