Blastocystis: Emerging Protozoan Parasite with High Prevalence in Iran

Background: Blastocystis is a zoonotic protozoan parasite habit in intestinal tract of humans and wide range of animals. Because of the mysterious nature and unknown or less-known aspects of Blastocystis, comprehensive information about epidemiology of this parasite is not available. The objective of this study was to investigate the available parasitology studies during the last decade in Iran and determine the prevalence of Blastocystis and its position among other intestinal parasites. As well as, investigate the effective factors in its prevalence.Materials and Methods: All available studies related to the prevalence of intestinal parasites in Iran during the recent decade were collected using information databases. After determinant the mean prevalence of each parasite, the most common parasites, and effective factors on their prevalence were assessed and analyzed.Results: Different studies showed that the most common intestinal parasite at this period of time was Blastocystis spp. with 14.6% prevalence rate. Additionally, in 44.5% of cases Blastocystis spp. allocated the first and in 100% of cases, the first to third rank of the most common intestinal parasites in Iran. Giardia lamblia and Entamoeba coli were in the next category.Conclusion: To our knowledge, the present study is the first survey in which the Blastocystis spp. introduces as one of the most common intestinal parasites in human. Various factors, including the low sensitivity of routine diagnostic methods, hosts multiplicity, easy transportation and unknown impressive factors are effective in the increased prevalence of this parasite. The results of this study could improve the attitude of teachers and researchers towards Blastocystis spp

Clinical and pharmacological applications of silymarin components at cellular and molecular level: A review

Silymarin, a flavonolignan from ‘milk thistle’ (Silybum marianum) plant is used almost exclusively for hepatoprotection and amounts to 180 million US dollars business in Germany alone. In this review, we discuss about its safety, efficacy and future uses in liver diseases. The use of silymarin may replace the polyherbal formulations and will avoid the major problems of standardization, quality control and contamination with heavy metals or bacterial toxins. Silymarin consists of four flavonolignan isomers namely; silybin, isosilybin, silydianin and silychristin. Among them, silybin being the most active and commonly used. Silymarin is orally absorbed and is excreted mainly through bile as sulphates and conjugates. Silymarin offers good protection in various toxic models of experimental liver diseases in laboratory animals. It acts by antioxidative, anti-lipid peroxidative, antifibrotic, anti-inflammatory, membrane stabilizing, immunomodulatory and liver regenerating mechanisms. Silymarin has clinical applications in alcoholic liver diseases, liver cirrhosis, Amanita mushroom poisoning, viral hepatitis, toxic and drug induced liver diseases and in diabetic patients. Though silymarin does not have antiviral properties against hepatitis virus, it promotes protein synthesis, helps in regenerating liver tissue, controls inflammation, enhances glucuronidation and protects against glutathione depletion. Silymarin may prove to be a useful drug for hepatoprotection in hepatobiliary diseases and in hepatotoxicity due to drugs. The non-traditional use of silymarin may make a breakthrough as a new approach to protect other organs in addition to liver. As it is having a good safety profile, better patient tolerability and an effective drug at an affordable price in near future new derivatives or new combinations of this drug may prove to be useful. © Medwell Journals, 2016

Efficacy of Myrtus communis L. to inactivate the hydatid cyst protoscoleces

Purpose: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. Materials and Methods: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5–100 μl/ml) were used for 5–30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups. Results: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 μl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 μl/ml was 100%. However, lower concentrations (12.5 and 25 μl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. Conclusion: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery

Short and long term evaluation of the efficiency of PermaNet® 2.0 bed net against environmental factors and washing using bioassay tests

The aim of the present study was to examine the resistance of PermaNet® 2.0 bed nets against repeated washing and environmental factors by using bioassay tests. After 5, 15 and 21 washings with detergents and by using bioassay tests, the resistance of 40 PermaNet® 2.0 bed nets was compared with that of 40 bed nets conventionally treated with one K-O tablet. To examine the long-term resistance, 31 PermaNet® 2.0 bed nets were also distributed among villagers, and were re-collected to perform bioassay tests after 1, 2 and 5 years. In the first phase of this study, the insecticidal effect of the conventionally-treated nets significantly decreased due to repeated washings (P < 0.001); however, it was not significant regarding PermaNet® 2.0 bed nets (P = 0.92 in continuous exposure and P = 0.12 in mortality tests). In the long-term phase of this study, the time required for knockdown of PermaNet® 2.0 increased over the first 2 years and then decreased. In addition, the mortality rate decreased over the first 2 years and then increased. In conclusion, it seems that the technique used by the manufacturer for impregnation of PermaNet® 2.0 bed nets has an acceptable efficiency in comparison with conventional techniques

Detection of Toxoplasma gondii in Acute and Chronic Phases of Infection in Immunocompromised Patients and Pregnant Women with Real-time PCR Assay Using TaqMan Fluorescent Probe

Background: Toxoplasma gondii, cause severe medical complications in infants and immune-compromised individuals. As using early, sensitive and rapid technique has major in diagnosis of toxoplasmosis, the present study was aimed to detect parasite by using from repetitive element (RE) and B1genes, in blood samples of seropositive immunocompromised patients and pregnant women. Methods: A total of 110 peripheral blood samples were collected from seropositive cases with anti-T. gondii antibodies, including immunocompromised patients and pregnant women. DNA was extracted by a commercial kit and subjected to TaqMan probe-based real-time PCR assay by using primers and probes specific for RE and B1 genes, separately. The data were analyzed by Kappa test and SPSS-22 software. Results: In the pregnant women, 17 (68%) and 14 (56%) samples from 25 IgM+/ IgG+ cases and, 7 (25%) and 6 (21.4%) samples from 28 IgG+/IgM- cases were positive by RE and B1 real time PCR, respectively. Likewise, in immunocompromised group, 20 (66.6%) and 17 (56.6%) samples from 30 IgM+/ IgG+ cases and 2 (7.4%) and 2 (7.4%) samples from 27 IgG+/ IgM- cases were positive by RE and B1 real time PCR, respectively. Conclusion: Probe-based real time PCR assay is a quantitative approach for early diagnosis of T. gondii infection in clinical samples. Moreover, this method can be more appropriate in diagnosis of acute and reactivated toxoplasmosis. In addition our results indicated that RE gene is more sensitive than B1 gene

Serological and Molecular Diagnosis of Toxoplasma gondii Infections in Thalassemia Patients.

BACKGROUND: This study aimed to the serological and molecular diagnosis of Toxoplasma gondii infections and related risk factors in patients with thalassemia major and healthy controls. METHODS: This case-control study was performed in Shahrekord University of Medical Sciences, Shahrekord, west of Iran from Jan 2014 to Jan 2015. Overall, 235 patients with thalassemia major and 235 healthy controls were enrolled. Assessment of anti-Toxoplasma antibodies in sera samples was performed using commercial ELISA kits. In order to the molecular investigate of T. gondii in blood samples, a relatively new molecular assay, LAMP technique based on Toxoplasma SAG1 gene was conducted for the first time. The specificity of LAMP outer primers for the T. gondii detection was confirmed by sequencing the purified PCR product. RESULTS: 51.9% of thalassemia patients and 34.8% of healthy controls were positive for anti-Toxoplasma IgG antibodies, which the difference was statistically significant (P<0.01). In terms of anti-Toxoplasma IgM antibody, 3.4% of thalassemia patients and 2.1% of healthy individuals were positive, which the difference was not statistically significant (P=1). Based on SAG1-LAMP, 9.78% of the thalassemia patients and 5.95% of healthy controls were positive for T. gondii DNA, which the difference was not statistically significant (P≤0.230). CONCLUSION: Thalassemia patients, probably due to repeated blood transfusion and consequently, immune deficiency, are at risk of transmitting Toxoplasma infection more than healthy people. Therefore, screening of Toxoplasma infection in blood transfusion centers may be effective in the prevention of toxoplasmosis in these patients. KEYWORDS: Loop-mediated isothermal amplification; Serology; Thalassemia major; Toxoplasma gondi

Serological and Molecular Diagnosis of Toxoplasma gondii Infections in Thalassemia Patients

Background: This study aimed to the serological and molecular diagnosis of Toxoplasma gondii infections and related risk factors in patients with thalassemia major and healthy controls. Methods: This case-control study was performed in Shahrekord University of Medical Sciences, Shahrekord, west of Iran from Jan 2014 to Jan 2015. Overall, 235 patients with thalassemia major and 235 healthy controls were enrolled. Assessment of anti-Toxoplasma antibodies in sera samples was performed using commercial ELISA kits. In order to the molecular investigate of T. gondii in blood samples, a relatively new molecular assay, LAMP technique based on Toxoplasma SAG1 gene was conducted for the first time. The specificity of LAMP outer primers for the T. gondii detection was confirmed by sequencing the purified PCR product. Results: 51.9% of thalassemia patients and 34.8% of healthy controls were positive for anti-Toxoplasma IgG antibodies, which the difference was statistically significant (P<0.01). In terms of anti-Toxoplasma IgM antibody, 3.4% of thalassemia patients and 2.1% of healthy individuals were positive, which the difference was not statistically significant (P=1). Based on SAG1-LAMP, 9.78% of the thalassemia patients and 5.95% of healthy controls were positive for T. gondii DNA, which the difference was not statistically significant (P≤0.230). Conclusion: Thalassemia patients, probably due to repeated blood transfusion and consequently, immune deficiency, are at risk of transmitting Toxoplasma infection more than healthy people. Therefore, screening of Toxoplasma infection in blood transfusion centers may be effective in the prevention of toxoplasmosis in these patients

Efficacy of Myrtus communis L. to Inactivate the Hydatid Cyst Protoscoleces

Purpose: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. Materials and Methods: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5–100 μl/ml) were used for 5–30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups. Results: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 μl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 μl/ml was 100%. However, lower concentrations (12.5 and 25 μl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. Conclusion: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery

CHEMICAL COMPOSITION AND PROPHYLACTIC EFFECTS OF SATURJA KHUZESTANICA ESSENTIAL OIL ON ACUTE TOXOPLASMOSIS IN MICE

Background: Toxoplasma gondii is a widespread zoonotic protozoan that infects approximately one third of the global human population and all other warm-blooded animals. The present study aims to evaluate the prophylactic effects of Satureja khuzestanica essential oil (SKEO) on infected mice with acute toxoplasmosis. Materials and Methods: The components of the SKEO were identified by gas chromatography/mass spectroscopy (GC/MS). To evaluate the prophylactic effects of SKEO, mice were divided into four groups. (i) non-treated group, (ii) mice treated with olive oil once a day for two weeks, (iii) mice treated with SKEO at the dose of 0.2ml/kg once a day for two weeks, (iv) and mice orally treated with SKEO at the dose of 0.3 ml/kg once a day for two weeks. After 24 h (fifteenth day) mice in the groups of two-four were infected intraperitonealy with 10-4 tachyzoite of T. gondii, RH strain. The mortality rate in all infected mice and the number of tachyzoites from infected mice were recorded. Results: The main components of SKEO were carvacrol (78.8%), thymol (7.5%), and beta-Bisabolene (1.2%). Findings of prophylactic effects revealed that mortality rate of infected mice was 8 days after oral administration of SKEO at the concentration of 0.2 and 0.3ml/kg (

Utility of blood as the clinical specimen for the diagnosis of ocular toxoplasmosis using uracil DNA glycosylase-supplemented loop-mediated isothermal amplification and real-time polymerase chain reaction assays based on REP-529 sequence and B1 gene

Background: Ocular infection with Toxoplasma gondii is a major preventable cause of blindness, especially in young people. The aim of the present study was to assess detection rate of T. gondii DNA in blood samples of clinically diagnosed of ocular toxoplasmosis using uracil DNA glycosylase-supplemented loop-mediated isothermal amplification (UDG-LAMP) and real-time quantitative PCR (qPCR) based on REP-529 and B1. Methods: One hundred and seventeen patients with clinically diagnosed ocular toxoplasmosis (OT) were participated in the study as well as 200 control patients. Peripheral blood samples were assessed using UDG-LAMP and qPCR techniques targeting REP-529 and B1. Results: Detection limits of qPCR using REP-529 and B1 were estimated as 0.1 and 1 fg of T. gondii genomic DNA, respectively. The limits of detection for UDG-LAMP using REP-529 and B1 were 1 and 100 fg, respectively. In this study, 18 and 16 patients were positive in qPCR using REP-529 and B1, respectively. Based on the results of UDG-LAMP, 15 and 14 patients were positive using REP-529 and B1, respectively. Results of the study on patients with active ocular lesion showed that sensitivity of REP-529 and BI targets included 64 and 63%, respectively using qPCR. Sensitivity of 62 and 61%, were concluded from UDG-LAMP using REP-529 and B1 in the blood cases of active ocular lesion. qPCR was more sensitive than UDG-LAMP for the detection of Toxoplasma gondii DNA in peripheral blood samples of patients with clinically diagnosed toxoplasmic chorioretinitis. Furthermore, the REP-529 included a better detection rate for the diagnosis of ocular toxoplasmosis in blood samples, compared to that the B1 gene did. Moreover, the qPCR and UDG-LAMP specificity assessments have demonstrated no amplifications of DNAs extracted from other microorganisms based on REP-529 and B1. Conclusions: Data from the current study suggest that qPCR and UDG-LAMP based on the REP-529 are promising diagnostic methods for the diagnosis of ocular toxoplasmosis in blood samples of patients with active chorioretinal lesions