In vitro evaluation of albendazole nanocrystals against Echinococcus granulosus protoscolices.

Echinococcus granulosus is a zoonotic parasite causing hydatidosis in humans and animals. This study has been done in order to investigate the effect of albendazole nanocrystals on the viability of E. granulosus protoscolices. The average size and hydrodynamic diameter of albendazole nanocrystals were 976±218 and 1334±502 nm, respectively. Fertile hydatid cysts were isolated from the liver of slaughtered sheep. The isolated cysts were further identified using morphological and molecular techniques. The nucleotide sequence analysis indicated that the genotype of the protoscolices was E. granulosus sensu stricto with 100% similarity. The parasites were examined precisely for susceptibility to albendazole nanocrystals. The results revealed that albendazole nanocrystals are effective in removing protoscolices. It was observed that 1 µg/ml albendazole nanocrystals and albendazole completely inhibited the viability of the protoscolices within 17 and 23 days, respectively. The results suggested that albendazole nanocrystals can be used as an alternative effective treatment for E. granulosus infection.

A novel delivery of curcumin by the efficient nanoliposomal approach against Leishmania major.

Several side effects and drug resistance accompany the current therapies for Leishmaniasis. Nanoliposomal curcumin is applied as a new therapy approach instead of current therapy. In this study, nanoliposomal curcumin was prepared using thin-film hydration method and characterized based on encapsulation efficiency, size, and zeta potential. Curcumin was successfully loaded into nanoliposomes with an encapsulation efficiency of 92%. The surface charge of the nanoparticle was neutral, and the size of nanoparticle was 176.5 nm. Nanoliposomal curcumin is in spherical shape without any agglomeration. Cell viability assay was performed on HFF cell line to show biocompatibility of liposome nanoparticles. Anti-Leishmanial effect of different concentrations of liposomal curcumin (0.05-30 µg mL-1) and amphotericin B (25 µg mL-1) were studied on Leishmania major [MRHO/IR/75/ER] at various hours (24, 48, and 72) using hemocytometer technique. Nanoliposomal curcumin inhibitory concentration (IC50) at hours 24, 48, and 72 were 6.41, 3.8, and 2.33 µg mL-1, respectively. As prepared nanoliposomal curcumin showed a significant antileishmanial effect and induced a better and more tangible effect on the survival of L. major promastigotes and could be suitable candidates for further investigations.

Serum levels of CC chemokine ligands in cutaneous leishmaniasis patients.

The crucial functions of chemokine/receptors in numerous parasitic infections, including leishmaniasis, are well documented. This study aimed to assess the serum levels of CC ligand (CCL) 2, CCL5, and CCL11 in cutaneous leishmaniasis (CL) patients. 64 patients, suffering from CL and 100 healthy people were selected, and their blood serum concentrations of CCL2, CCL5, and CCL11 were measured using enzyme-linked immunosorbent assay. The results demonstrated that while the mean serum levels of CCL5 and CCL11 increased significantly in CL patients, the mean serum levels of CCL2 decreased, compared to the control group. Despite the sufficient production of CCL5 and CCL11 in CL patients, they suffered from CCL2 deficiency, as the defense mechanism against parasitic infection. These findings suggest a mechanism that might partially explain the patients' susceptibility to persistent infection and their inability to clear the parasites.

Isolation and Molecular Identification of Leishmania spp. Agents in Patients with Cutaneous Leishmaniasis in Yazd Province, Endemic Region of Central Iran.

BACKGROUND: Cutaneous Leishmaniasis (CL) is a major health problem in many parts of Iran. Many methods have been introduced for detection and identification of Cutaneous Leishmaniasis. The purpose of this study was isolation and molecular identification of Leishmania spp. agents in patients with CL from endemic region of central Iran. In this study, one of the main loci of central Iran named Yazd will be assessed CL identification using PCR-RFLP. METHODS: For this cross-sectional study, sampling was done from 372 suspicious patients with CL who referred to Health Centers of Yazd Province from 2016 to 2017. After collection samples of patients, DNA extraction was done from samples on slides. Genus detection was done using specific primers by PCR. RFLP analysis was done for species identification. RESULTS: Out of 372 samples, 159 samples were positive using PCR based method. Out of 159 samples, 87 (54.7%) L. major and 72 (45.3%) L. tropica were identified using RFLP analysis. The number of lesions in each patient was different but 119 (74.8%) patients showed the number of 1-3 lesions, and more lesions (more than 10 lesions) was showed in 4 (2.5%) person. CONCLUSION: The CL found in Yazd province resulted from L. major and L. tropica as the agents of rural and urban types, respectively. The prevalence of L. major and L. tropica was almost the same. This indicated that control programs could be designed for treatment and vector and reservoir control.

Prevalence and genotyping identification of Cryptosporidium in adult ruminants in central Iran.

BACKGROUND: Apicomplexan parasites of the genus Cryptosporidium infect a wide range of animal species as well as humans. Cryptosporidium spp. can cause life threatening diarrhea especially in young animals, children, immunocompromised patients and malnourished individuals. Asymptomatic cryptosporidial infections in animals can also occur, making these animals potential reservoirs of infection. METHODS: In the present study, a molecular survey of Cryptosporidium spp. in ruminants that were slaughtered for human consumption in Yazd Province, located in central Iran was conducted. Faeces were collected per-rectum from 484 animals including 192 cattle, 192 sheep and 100 goats. DNA was extracted from all samples and screened for Cryptosporidium by PCR amplification of the 18S rRNA gene. Positives were Sanger sequenced and further subtyped by sequence analysis of the 60 kDa glycoprotein (gp60) locus. RESULTS: In total, Cryptosporidium spp. were detected in 22 animals: C. andersoni and C. bovis in seven and two cattle faecal samples, respectively, C. ubiquitum in five sheep, and C. xiaoi in six sheep and two goat samples, respectively. To our knowledge, this study provides for the first time, molecular information concerning Cryptosporidium species infecting goats in Iran, and is also the first report of C. ubiquitum and C. xiaoi from ruminants in Iran. CONCLUSION: The presence of potentially zoonotic species of Cryptosporidium in ruminants in this region may suggest that livestock could potentially contribute to human cryptosporidiosis, in particular among farmers and slaughterhouse workers, in the area. Further molecular studies on local human populations are required to more accurately understand the epidemiology and transmission dynamics of Cryptosporidium spp. in this region.

Molecular typing of Giardia duodenalis in cattle, sheep and goats in an arid area of central Iran.

Giardia duodenalis is one of the most common intestinal parasites in humans as well as livestock and wildlife. It is of both public and veterinary health importance in developing nations. A molecular survey of Giardia duodenalis assemblages in ruminants from Yazd Province, Iran was conducted on 484 animal faecal samples collected per rectum from slaughtered ruminants including 192 cattle, 192 sheep and 100 goats from June to November 2017. Species-specific and assemblage-specific PCRs for assemblages A, B and E at the triose phosphate isomerase (tpi) gene were performed, and samples positive for Giardia were confirmed by sequencing. In total, 25 (5.16%) of examined faecal samples including eight cattle (4.2%), twelve sheep (6.2%) and five goats (5%) were infected with G. duodenalis. Assemblage-specific PCR detected G. duodenalis assemblage E in seven faecal samples (six in sheep and one in a goat). Assemblages A and B were not detected. This study provides the first insight into Giardia infection in slaughtered livestock in Iran. Although the prevalence of infection with Giardia in this hot-arid area of Iran was low, educating people about direct contact with livestock such as farmers and abattoirs workers about this zoonotic infection is important.

Polyurethane sheet impregnated with Arabinogalactan can lead to increase of attachment of promastigotes and Amastigote of Leishmania major (MRHO/IR/75/ER) by GP63 and HSP70 genes.

The aim of this study was to investigate the effect of polyurethane sheet (PUS) and polyurethane sheet impregnated with Arabinogalactan (PUSIAG) on the cell attachment and viability of Promastigotes and Amastigotes of Leishmania major (MRHO/IR/75/ER), and mouse macrophages, and its whole skin cells (WSCs). In a sterile condition, 10 mL of Arabinogalactan 5% w/v was poured into a falcon. Then, a piece of PUS was placed inside it, and incubated at 37 °C for 24 h. Next, it was washed, and cut. Then, one piece of PUS and PUSIAG was separately added to 1 mL of cell suspension (Promastigotes, Amastigote, and WSCs), and then incubated for 1, 2, 3, and 4 days at 37 °C. After incubation times, the quantity of adhered cells was counted, and cell viability was measured by MTT assay. Also, for WSCs and macrophages, the expression of integrin, fibronectin and GAPDH was investigated, and for Promastigotes and Amastigotes, the expression of GP63, Cpb, and 18s rRNA was measured. This study showed that with increase of exposure time, the percentage of attached cells was increased. There was a significant difference between attached cells to PUSIAG and PUS in case of Promastigotes and Amastigotes. It seems that Promastigotes and Amastigotes have higher interest to PUSIAG than WSCs and Macrophages. Also, this study showed with increase of exposure time, the percentage of viable cells was decreased. There were significant differences between cell viability of Promastigotes and Amastigotes when exposed to PUSIAG and PUS, especially in long time incubation. Also, when incubation time was increased the relative expression of integrin and fibronectin in WSCs and macrophages, and GP63 and HSP70 in Promastigotes and Amastigotes were increased.

The Effect of Nonsurgical Periodontal Therapy on Trichomonas Tenax and Entamoeba Gingivalis in Patients with Chronic Periodontitis.

STATEMENT OF THE PROBLEM: Trichomonas tenax and Entamoeba gingivalis are commensal protozoa which inhabit the human oral cavity. These parasites are found in patients with poor oral hygiene and might be a reason for progressive periodontal diseases. PURPOSE: The aim of this study was to evaluate the effect of nonsurgical periodontal treatment on the frequency of these protozoa in saliva and plaque samples. MATERIALS AND METHOD: In this clinical trial, samples of saliva and dental plaque were collected from 46 patients with moderate to severe chronic periodontitis before and after periodontal therapy. The samples were assessed for the frequency of parasites. RESULTS: The frequency of Entamoeba gingivalis was reduced in saliva (p= 0.007) and plaque (p= 0.027) three weeks after the treatment. Likewise, the frequency of Trichomonas tenax reduced in saliva (p= 0.030); however, the decrease was not significant in plaque (p= 0.913). Trichomonas tenax frequency in dental plaque directly related to the severity of periodontitis (r= 0.565, p≤ 0.000). In contrast, the number of Entamoeba gingivalis in both saliva (r= -0.405, p≤ 0.005) and plaque (r= -0.304, p= 0.040) was inversely related with the severity of the periodontal disease. CONCLUSION: Nonsurgical periodontal treatment could reduce the number of Trichomonas Tenax and Entamoeba gingivalis in the oral environment of patients with chronic periodontitis.

The Effect of Smoking on Mineral and Protein Compositionof Saliva.

INTRODUCTION: To assess the salivary composition of proteins and minerals in smokers compared with non-smokers. MATERIALS AND METHODS: In this study we compared the total protein and Ca, Na, K, Mg, Pb of whole saliva in two groups of men (28 smokers and 31nonsmokers) aged between 29-41years. RESULTS: Fifty-nine participants were evaluated. The mean age was 33.14±5.32 years among smokers and 32.15±5.12 years among non-smokers (P>0.05). The mean concentration of total protein, Ca, Pb, and Zn of whole saliva in smokers was lower than that in non-smokers, but the difference was not statistically significant (P>0.05). The mean concentration of Na, K, Mg in whole saliva was not significantly different between smokers and non-smokers (P>0.05). CONCLUSION: We specified that smoking reduced the value of total protein, Ca and Pb of saliva, however it did not have an impact on Na, K, and Mg of saliva.

Anti Leishmanial Effect of Zinc Sulphate on the Viability of Leishmania tropica and L. major Promastigotes.

BACKGROUND: Cutaneous leishmaniasis (CL) is an endemic disease in developing countries. Although pentamidine orantimonite (Glucantime) has been recommended for cutaneous leishmaniasis treatment by the World Health Organization, there are some concerns too such as high cost, side effects, need for frequent injections, and restricted efficacy. Therefore, different methods have been used for CL treatment so far. OBJECTIVES: This study assessed the sensitivity of two parasite agents of cutaneous leishmaniasis: Leishmania major and L. tropica to zinc sulphate in vitro. In the present study, the zinc sulphate effect on urban and rural strains of cutaneous leishmaniasis, viability of old world, in vitro is under investigation. MATERIALS AND METHODS: The design of the present study was experimental (laboratory-trial) based. Iranian endemic species of L. major and L. tropica were appropriately collected, proliferated, and maintained in the standard culture. Afterward, the proper concentrations of zinc sulphate were provided, sterilized, and added to the cultures containing parasites. In different intervals, parasites were counted by two methods: the slide and cell proliferation ELISA. RESULTS: Both parasite species showed sensitivity to zinc sulphate in vitro and in comparison with the control group, their numbers were reduced. Zinc sulphate (in concentrations of 0.5, 1, 2, and 3 percent) was added to the cultures containing parasites, and the total number of the live parasites was counted through the slide method (Neubauer slide) every day up to the fifth day. The results were analyzed and found statistically significant (P < 0.05). In the second phase, the counting process was repeated with the addition of zinc sulphate compound with different concentrations (3, 4, 5, and 6 percent) and live parasite numbers were counted by ELISA method after 24 hours. The findings revealed that all the cultures containing zinc sulphate showed a slower growth in comparison to the control group. The mentioned difference was statistically significant (P < 0.05). CONCLUSIONS: Considering the safety of zinc sulphate compound in comparison with Glucantime, there is a possibility of using it in the treatment of CL caused by both species of L. major and L. tropica. It is obvious that more researches are mandatory both in vivo and in vitro to figure out its daily dosage, proper concentration, time and duration, and possible side effects.

Rare variants of cutaneous leishmaniasis presenting as eczematous lesions.

Cutaneous Leishmaniasis may present with clinical presentation such as zosteriform, sporotrichoid and erysipeloid. The eczema variant has rarely been reported. We report a 27- year- old patient with atypical cutaneous leishmaniasis resembling eczema on the hand of a man in Yazd province in the central of Iran.