Effect of Foeniculum Vulgare Aqueous and Alcoholic Seed Extract against Zoonotic Cutaneous Leishmaniasis

BACKGROUND፡ Cutaneous leishmaniasis is considered one of the major neglected tropical diseases. Drug resistance, limitary efficacy, and severe side effects remain a challenge for treatment. Foeniculum vulgare is known as a medicinal plant belonging to the Apiaceae, and anti-microbial properties of this plant have already been confirmed.METHOD: The F.vulgare sterile aqueous and alcoholic extracts were prepared. In vitro has used RAW 264.7 cell line and L. major parasite (MRHO/IR/75/ER). Cytotoxicity assay on macrophages (CC50), cytotoxicity assay on promastigotes (IC50), and cytotoxicity assay on infected macrophages (EC50) were accomplished with both extracts by MTT and light microscopy methods. Four in vivo were allocated in four groups and five BALB/c mice each group. Stationary phase promastigotes were inoculated into the base of mice tails subcutaneously (SC).Measurement of the body weight, lesion size, parasite burden of the lesion, and spleen after 4 weeks for evaluation effects of the alcoholic extract on CL was done.RESULTS: The results of in vitro revealed that the optimal concentrations of both extracts reducing the promastigotes and amastigotes growth. Alcoholic extract no harmful side effects for the host macrophages, while were indicated has a potent action against L. major. In vivo results after 4 weeks did not show any variation in lesion size and body weight. Also, lesion size and spleen parasite burden decreased in comparison to no treatment group.CONCLUSION: The alcoholic extract could be a new alternative treatment for cutaneous leishmaniasis. However this extract needs more investigation for novel herbal drugs against CL.&nbsp

Fatty acid and retinol-binding protein: A novel antigen for immunodiagnosis of human strongyloidiasis

The tenacious human parasitic helminth Strongyloides stercoralis is a significant health problem worldwide. The current lack of a definitive diagnostic laboratory test to rule out this infection necessitates designing more specific diagnostic methods. Fatty acid and retinol-binding protein (FAR) plays a crucial role in the development and reproduction of nematodes. We generated a recombinant form of this protein and determined its applicability for immunodiagnosis of S. stercoralis. The L3 form of S. stercoralis was harvested and used for RNA extraction and cDNA synthesis. The coding sequence of S. stercoralis FAR (SsFAR) was cloned into pET28a(+) vector, expressed in E. coli BL21 and purified. ELISA and immunoblotting were employed to determine the specificity and sensitivity of rSsFAR using a set of defined sera. In addition, we analyzed the phylogenetic relationship of SsFAR with different FAR sequences from other nematodes. The cloned SsFAR had an open reading frame of 447 bp encoding 147 amino acids, with a deduced molecular mass of 19 kD. The SsFAR amino acid sequence was 93% identical to FAR of S. ratti. For differential immunodiagnosis of strongyloidiasis, rSsFAR exhibited 100% sensitivity and 97% specificity. However, cross-reactivity with FAR proteins of other parasites, namely Toxocara canis and Echinococcus granulosus, was noted. Our results provide a novel approach for immunodiagnosis of S. stercoralis infections using rSsFAR with reliable sensitivity and specificity

Subtypes Distribution and Frequency of Blastocystis sp. Isolated from Diarrheic and Non-diarrheic Patients

Background: Blastocystis is one of the most common parasites, reported from both human and animals. This parasite is more prevalent in regions with low levels of hygiene, close contact with animal and unsuitable disposal systems. The aim of the study was to subtype Blastocystis sp., isolated from diarrheic and non-diarrheic patients using sequencing of 18S ribosomal DNA. Methods: Totally, 300 stool samples were collected from diarrheic and non-diarrheic patients referred to Imam Reza Hospital, Tehran from Apr to Aug 2015. All samples were concentrated using conventional Formalin – ether technique and recognized under light microscope. The fresh stool samples were also cultivated in clotted fetal bovine medium and examined for growing of Blastocystis every 48 h with direct smear slides for 10 d.DNA extraction was performed on all positive samples. Amplified DNA fragment of 18S rDNA was sequenced and compared with reference genes, previously deposited in Genbank database. Results: The number of diarrheic and non-diarrheic patients participated in the study was 134 (44.66%) and 166 (55.34%), respectively. Three subtypes 1, 2, 3 were identified from positive samples. Subtype 2 was the most prevalent (36.5%) followed by subtype 1 (33.3%) and subtype 3 (30.2%). There were no mixed subtypes. Furthermore, the most prevalent subtypes in diarrheic and non-diarrheic patients were subtype 2 (39.28%) and subtype 1 (37.14%), respectively. Conclusion: Blastocystis sp., is one of the most prevalent unicellular parasites among diarrheic and non-diarrheic patients. Indeed, ST2 was the most prevalent subtype particularly in those samples collected from diarrheic patients

Fatty acid and retinol-binding protein: A novel antigen for immunodiagnosis of human strongyloidiasis.

The tenacious human parasitic helminth Strongyloides stercoralis is a significant health problem worldwide. The current lack of a definitive diagnostic laboratory test to rule out this infection necessitates designing more specific diagnostic methods. Fatty acid and retinol-binding protein (FAR) plays a crucial role in the development and reproduction of nematodes. We generated a recombinant form of this protein and determined its applicability for immunodiagnosis of S. stercoralis. The L3 form of S. stercoralis was harvested and used for RNA extraction and cDNA synthesis. The coding sequence of S. stercoralis FAR (SsFAR) was cloned into pET28a(+) vector, expressed in E. coli BL21 and purified. ELISA and immunoblotting were employed to determine the specificity and sensitivity of rSsFAR using a set of defined sera. In addition, we analyzed the phylogenetic relationship of SsFAR with different FAR sequences from other nematodes. The cloned SsFAR had an open reading frame of 447 bp encoding 147 amino acids, with a deduced molecular mass of 19 kD. The SsFAR amino acid sequence was 93% identical to FAR of S. ratti. For differential immunodiagnosis of strongyloidiasis, rSsFAR exhibited 100% sensitivity and 97% specificity. However, cross-reactivity with FAR proteins of other parasites, namely Toxocara canis and Echinococcus granulosus, was noted. Our results provide a novel approach for immunodiagnosis of S. stercoralis infections using rSsFAR with reliable sensitivity and specificity