Introduction and objective: Dicrocoelium dendriticum is a worldwide spread parasite of liver, bile ducts and gallbladder of especially ruminants and humans as well. Identification of specific antigens is useful for early diagnosis of the infection. The goal of this study was the isolation and identification of excretory-secretory and somatic antigens from D. dendriticum by sodium dodecyl sulphate (SDS)-PAGE and evaluation of humoral immune response against these antigens.

  Methods: The parasites were collected and washed by phosphate buffered saline (PBS) and supplemented by antibiotic for several times. For preparing somatic antigens, parasites were sonicated and centrifuged prior to collect supernatant. For preparing excretory-secretory antigens the viable parasites were transferred to the sterile medium. The samples were centrifuged and supernatants were collected. The sera of infected sheep with different infection degrees were collected too. Somatic and excretory-secretory proteins were isolated with SDS PAGE and stained with coomassie blue. Immunogenicity properties of the resulting proteins were determined using western blot analysis.

  Results: The total extract of somatic antigens analyzed by SDS-PAGE revealed 21 proteins. In mild infection, bands of 130 KDa were immune dominant. In moderate infections 48, 80 and 130 KDa and in heavy infections 48, 60, 80, 130 KDa were detected as immune dominant bands. In excretory- secretory antigens seven bands of protein were detected. In mild infection 130 KDa, in moderate infection 100, 120 and 130 KDa and in heavy infection 45, 80, 85, 100, 120 and 130 KDa were immune dominant bands.

  Conclusion: Probably the most immunogenic protein band during different degrees of infection was 130KDa that can be used for vaccination and inducing immunity.

  Background & objectives: The leishmaniases are considered among the major infectious diseases affecting public health in several regions. There are many chemical agents which are effective in treatment of visceral leishmaniasis. But, overall treatment of visceral leishmaniasis is often difficult. Thus, identification of new chemotherapeutic agents is important for treatment of disease. Since targeting of the ergosterol synthesis pathway of Leishmania may be useful therapeutically, the aim of this study was to investigate the effect of alone or in combination of amiodarone and ketoconazole on Leishmania infantum.

  Methods : To obtain logarithmic promastigotes of L. infantum, the parasites were cultured in BHI medium with FCS 10% together with antibiotics of penicillin and streptomycin and incubated at 24° C. Amastigote forms were obtained in BHI medium supplemented with 20% FCS at pH of 5.5 which incubated in 37° C. L.infantum susceptibility to amiodarone and ketoconazole was evaluated by proliferation of parasites in the absence or presence of these drugs with MTT assay. For evaluation of antiproliferative synergism against promastigotes and axenic amastigotes, fractional inhibitory concentrations (FIC) were calculated. An isobologram curve was constructed too.

  Results: Amiodarone produced a marked reduction in the viability of L.infantum promastigotes and axenic amastigotes. On the other hand ketoconazole induced a dose dependent effect on the parasites proliferation for promastigotes and axenic amastigotes. When the drugs were used in combination, the results indicated clear synergistic as shown by a concave isobologram and FIC value.

  Conclusion: The present study represents the evidence that the combination of amiodarone plus ketoconazole acts synergistically in controlling L. infantume in vitro. It is possible that amiodarone could be used in combination with ketoconazole to combat infection at low doses, thus reducing its side effects such as cardiotoxicity, thyroid dysfunction and pulmonary fibrosis.

Background & objectives: One of the most important zoonotic parasitic diseases, hydatidosis, is caused by the larval stage of Echinococcous granulosus. Investigations have shown that plants secondary metabolites, such as essential oils have anti parasitic properties. Based on previous reports on antiparasitic properties of Lepidium sativum, in this study we investigated the scolicidal effects of the essential oil (EO) extracted from this plant.

Methods: Lepidium EO was obtained by hydrodistillation method. Gas chromatography-mass spectrometry (GC-MS) was employed to determine the chemical composition of the EO. Protoscolices were exposed to various concentrations of EO (1, 3, 5, 10 and 15 mg/ml) for 10, 20, 30 and 60 min. Viability of protoscolices was confirmed by 0.1% eosin staining.

Results: A total of 19 compounds representing 95.5% of the total oil, were identified. α-Thujene (88.86%), Myrcene (2.9%) and P-cymene (1.67%) were found to be the major EO constituents. Based on the results, protoscolices mortality rates at 1, 3 and 5 mg/ml of EO didn’t have a significant relationship with the control group. While, the difference in mortality rate at a concentration of 10 mg/ml of EO in 30 and 60 min was significant. Also, the concentration of 15 mg/ml of EO at all times of incubation had significantly higher protoscolicidal effect. In the present study there was a significant relation between the amount of protoscolicidal activity of different EO concentrations and different incubation times. In other words mortality rates enhanced with increasing concentrations and incubation times.

Conclusion: The results of present study revealed that the EO of Lepidium is rich in α-Thujene and has a high scolicidal power. This plant may be used as a natural scolicidal agent

Background & objectives: Polyamines such as putrescine, spermidine, and spermine are ubiquitous in all eukaryotic cells and play an essential role in cell division and differentiation. One way of polyamine biosynthesis is done by ornithine decarboxylase (ODC) which catalyzes the transformation of ornithine to putrescine. The aim of the present study was to evaluate the level of putrescine, spermidine and spermine in protoscolices, hydatid fluid and germinal layer and also to evaluate ODC activity.

Methods: In the present study putrescine, spermidine and spermine levels were investigated in germinal layers, hydatid fluids and protoscolices. To evaluate the activity of ODC, protoscolices were incubated with ornithine and changes in polyamines level were assayed. The samples were homogenized and liquid chromatography (HPLC) was used for polyamines measurement.

Results: Based on the results, putrescine was the lowest polyamine and since its level was not increased in protoscolices incubated with ornithine, ODC activity was not detected. Spermidine was the highest polyamine and the results showed that germinal layer contained the highest level of polyamines.

Conclusion: Overall, the results showed that ODC activity was not detected in hydatid cyst and level of polyamines in germinal layers  which contained rapidly proliferating cells was higher than other parts.