Echinococcus granulosus sheep strain (G1) as the predominant genotype in definitive host (dogs) isolates in northeastern Iran.

Echinococcus granulosus sensu lato (E. granulosus s.l.) is a zoonotic parasite, causing cystic echinococcosis in humans. In the present study, prevalence and genotypes of E. granulosus s.l. was assessed in stools collected from 244 dogs including 138 stray and 106 domestic animals using high resolution melting curve (HRM) method. Initially, to detect taeniid eggs in feces, all samples were examined using the formalin-ether techniques. Genomic DNA was extracted from the positive samples and E. granulosus s.l. was differentiated from other Taeniidae parasites using SSU-rDNA gene and E. granulosus s.l. was analyzed for genotyping using HRM based on the cox1 gene. In total, 12.7% (31/244) of the samples were positive for Taeniidae eggs. In addition, among the positive samples, 77.4% (24/31) were positive for E. granulosus s.l.. In details, 11.3% (12/106) of the domestic dogs and 8.7% (12/138) of the stray dogs were positive for E. granulosus s.l.. The results of HRM analysis showed that all E. granulosus s.l. isolates were G1 strain. Findings of the present study indicated a considerable prevalence of E. granulosus G1 among dogs in the northeast of Iran and imply a serious risk of transmitting to humans and livestock.

Iranian Hydatid Disease Registry: Establishment and Implementation of a Neglected Tropical Disease Registry.

BACKGROUND: Cystic echinococcosis (CE) or hydatid disease is a global public health concern which imposes considerable economic costs on the communities in endemic regions. CE surveillance data are not adequately reliable. The present study reports the development and outcomes of a CE registry in Iran. METHODS: Hydatid Registry (HydatidReg) was initially established as a single-center registry in 2014 after the ethical approval of KMU. Following a call from MoHME to promote registry of different diseases and health outcomes, a call for participation was announced and all the Iranian Universities of Medical Sciences were requested to contribute to the registry. Subsequently, a nation-wide registry of hydatid disease was established in 2016. With a global perspective, HydatidReg joined the European Register of Cystic Echinococcosis (ERCE). A data collection form based on minimum dataset was designed and standard operating procedures (SOPs) were prepared to ensure standardized patient enrolment in the registry. A biobank system with two-dimensional barcoding was established along with HydatidReg for management and organization of biological specimens. RESULTS: As of March 2021, a total of 690 patients were enrolled in the registry. HydatidReg registered 362 (17.3%) out of the total 2097 patients enrolled in ERCE. Quality control (QC) of the data demonstrated 91.2% completeness and 80% timeliness. In the biobank, 322 biological specimens from 184 CE patients have been deposited including 70 blood, 96 sera and 156 parasite materials. CONCLUSION: High-quality data in the HydatidReg registry provided opportunities for health professionals to improve quality of care and organize meaningful research.

Oxidative stress associated with long term occupational exposure to extremely low frequency electric and magnetic fields.

BACKGROUND: Occupational exposure to extremely low frequency electromagnetic fields (ELF-EMFs) may have harmful effects on biologic systems and has raised many concerns in the last decades. OBJECTIVE: The aim of this study was to determine the effects of exposure to extremely low frequency electric and magnetic fields on lipid peroxidation and antioxidant enzyme activities. METHODS: This study was conducted on 115 power plant workers as the exposed group and 145 office workers as the non-exposed group. The levels of Malondialdehyde (MDA), superoxide dismutase (SOD), Catalase (Cat), and total antioxidant capacity (TAC) were measured in the serum of all subjects. Exposure to ELF-EMFs was measured based on spot measurements and the IEEE Std C95.3.1 standard. RESULTS: The levels of MDA, SOD, and Cat in the exposed group were significantly higher than in the non-exposed group. However, the level of TAC was not significantly different between the exposed (2.45±1.02) and non-exposed (2.21±1.07) groups. The levels of MDA and SOD were higher among workers with higher exposure to electric fields than workers with low exposure. All oxidative stress indicators increased with increased exposure to magnetic fields, except TAC. CONCLUSIONS: The antioxidant system imbalance among power plant workers may be related to long term occupational exposure to electromagnetic fields.

The Effects of Antioxidant Vitamins on Proinflammatory Cytokines and Some Biochemical Parameters of Power Plant Workers: A Double-Blind Randomized Controlled Clinical Trial.

Some epidemiological studies have suggested that exposure to extremely low-frequency magnetic fields (ELF-MFs) can affect the immune system. This study aimed to investigate the effects of antioxidant vitamin consumption on proinflammatory cytokines and biochemical parameters changes. In this randomized, controlled double-blinded trial study, power plant workers exposed to ELF-MFs were enrolled based on inclusion criteria. Ninety-one eligible subjects were randomly divided into four groups: the first group (400 units of vitamin E/day), second group (1,000 mg of vitamin C/day), third group (400 units of vitamin E and 1,000 mg of vitamin C/day), and control group. The intervention was conducted for 3 months. Proinflammatory cytokines interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α (TNF-α), and biochemical parameters (fasting blood sugar, total cholesterol, triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol [HDL-c], total protein, and albumin) were measured among the participants' serums, before and after the intervention. The mean level of IL-6 in all vitamin-receiving groups, the mean level of IL-1ß in vitamin C and E&C groups, and the mean level of TNF-α in the vitamin C group, decreased significantly after intervention. The arithmetic means of the effect sizes for IL-1ß, IL-6, and TNF-α were 0.71, 0.57, and 0.2, respectively. The level of HDL-c in the vitamin E and vitamin E&C groups and the level of triglyceride in the vitamin C group significantly increased after the intervention. Taking antioxidant vitamins can prevent an increase of proinflammatory cytokines induced by prolonged exposure to ELF-MFs. Bioelectromagnetics. 2021;42:18-26. © 2020 Bioelectromagnetics Society.

Efficacy and Safety of Zataria multiflora Boiss Essential Oil against Acute Toxoplasmosis 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 15th 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 8th and 9th 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.

The effect of vitamin E and C on comet assay indices and apoptosis in power plant workers: A double blind randomized controlled clinical trial.

Extremely low frequency electromagnetic fields have been classified as a possible human carcinogen by the International Agency for Research on Cancer and this has raised some concern about its health effects on employees extensively exposed to these fields at thermal power plants. In this study, the effect of using vitamin E and C supplements have been examined on employees working at a thermal power plant. In this randomized controlled, double-blind clinical trial, 81 employees from different parts of the thermal power plant were enrolled between July and November 2017, and divided into four groups: Group 1 received vitamin E (400 units/day), Group 2: vitamin C (1000 mg/day), Group 3: vitamin E + C and Group 4: no intervention. DNA damage was measured in peripheral blood lymphocytes using comet assay and apoptosis, using flow cytometry. Based on the results, tail intensity and tail length in the vitamin E group, and all comet assay indices in the vitamin E + C and vitamin C groups (except DNA damage index) significantly decreased after the intervention, while the comet assay indices did not change significantly in the control group. None of the flow cytometry indices including early apoptosis, late apoptosis and necrosis changed after intervention in either group. The use of antioxidant vitamins such as E and C, can increase the activity of the non-enzymatic antioxidant defense system, and protect DNA from damage caused by exposure to extremely low frequency magnetic fields. But, taking these vitamins has no effect on apoptosis. It seems that consumption of vitamin E affected all investigated comet assay indices and can be probably considered as the best intervention.

Isolated Human and Livestock Echinococcus granulosus Genotypes Using Real-Time PCR of cox1 Gene in Northeast Iran.

PURPOSE: Cystic echinococcosis (CE), caused by the metacestode of Echinococcus granulosus, is highly endemic over large parts of Iran. The purpose of this study was to investigate the prevalence rate of hydatidosis and mitochondrial cox1 real-time PCR with high-resolution melting curve (HRM) analysis of E. granulosus isolated from human and livestock. METHODS: In this cross-sectional study, 61 formalin-fixed paraffin-embedded tissue isolates were collected from human CE cases and 83 hydatid cysts from the liver and lung lesions of the livestock in Khorasan Razavi province, Northeast Iran. DNA was extracted from each isolate and amplified by real-time PCR and analyzed using the HRM method. RESULTS: The HRM analysis using the cox1 gene of 40 E. granulosus human isolates showed that 35 (87.5%), 4 (10%), and 1 (2.5%) of the isolates were categorized as G1, G3, and G6 genotypes, respectively. Out of the total 1342 livestock inspected, 39 (4%) goats and 44(12%) cattle were found harboring hydatid cysts all belonging to E. granulosus sensu stricto. CONCLUSION: The results confirmed that the high prevalence of E. granulosus sensu stricto in intermediate hosts is remarkable in northeast of Iran coupled with the high prevalence of infection in livestock, which reinforced the need for hydatidosis control programs in this region.

Molecular phylodiagnosis of Echinococcus granulosus sensu lato and Taenia hydatigena determined by mitochondrial Cox1 and SSU-rDNA markers in Iranian dogs: Indicating the first record of pig strain (G7) in definitive host in the Middle East.

Unawareness of canine parasitic diseases among at-risk hosts and an uncontrolled program of stray dog population have caused that zoonotic parasites received great attention in endemic regions of the Middle East. A total of 552 faecal samples were collected between December 2016 to January 2018 from stray (n = 408) and domestic (n = 144) dogs of Iran. All specimens were coproscopically observed following concentration and flotation techniques. Subsequently, the DNAs of taeniid eggs were extracted, amplified, and sequenced by targeting of mitochondrial cytochrome oxidase subunit 1 and small-subunit ribosomal DNA markers. The overall prevalence of canine intestinal parasites found 53.6%. The following parasites and their total frequencies were identified: taeniid (10.5%), Dicrocoelium dendriticum (0.7%), Trichuris vulpis (1.2%), Capillaria spp. (2.3%), Blastocystis spp. (5.2%), Ancylostoma spp. (2%), Eimeria spp. (13.2%), Dipylidium caninum (2.3%), Toxocara canis (3.8%), Giardia spp. (8.5%), and Toxascaris leonina (3.6%). Stray dogs were characterized more likely to be poliparasitized and indicated a higher prevalence of taeniid (10.9%), T. canis (4.4%) Giardia spp. (10.1%) than domestic dogs (P > 0.05). Phylogenetic and sequence analysis of Cox1 and SSU-rDNA indicated a low genetic diversity (Haplotype diversity; 0 to 0.495) in E. granulosus sensu lato G1, G3, G7 genotypes, and Taenia hydatigena. The pairwise sequence distances between G7 isolates showed an intra-diversity of 0.7%-1.5% and identity of 98.5%-100%. The first occurrence of pig strain (G7) from Iranian dogs might have substantial implications in the drug treatment of infected dogs due to the shorter maturation time of G7 compared with G1 genotype. Thus, the preventive strategies should be noticed to determine the risk factors, the importance of applying the hygienic practices, and well adjusting deworming programs for the Iranian dogs and at-risk individuals.

The effects of industrial noise exposure on lipid peroxidation and antioxidant enzymes among workers.

PURPOSE: The aim of this study was to measure individual exposure to noise and its effect on the levels of lipid peroxidation and enzymatic antioxidant defense. METHODS: In this cross-sectional study, 94 male workers working in a food factory in Shahroud, Iran, were selected as the case group and 112 people were also included as the control group. The level of exposure to noise was measured using a dosimeter and the equivalent level was calculated. The levels of malondialdehyde (MDA), catalase (Cat), superoxide dismutase (SOD), and Total Antioxidant Capacity (TAC) were measured in the serum samples of all participants. Independent t test, one-way ANOVA, and multivariate linear regression were used to analyze the data. RESULTS: The levels of MDA, Cat, and TAC were significantly higher in staff exposed to noise than in the controls. The starch and warehouse units' staff were exposed to the highest (99.3 ± 3.23 dB) and lowest (77.1 ± 9.68 dB) mean levels of sound pressure. Based on the linear regression model, noise exposure level was the most important predictor variable for levels of MDA (ß = 0.48, CI 95% = 0.04-0.93), Cat (ß = 0.43, CI 95% = 0.05-0.83), and TAC (ß = 0.11, CI 95% = 0.06-0.16). CONCLUSIONS: Noise exposure increases the production of free radicals. As exposure rate increases, the levels of MDA, Cat, and TAC enzymes increase as well.

In vitro Protoscolicidal Effects of Cinnamomum zeylanicum Essential Oil and Its Toxicity in Mice.

BACKGROUND: This study investigates the scolicidal effects of Cinnamomum zeylanicum essential oil against the protoscoleces of hydatid cysts and its toxicity in the mice model. MATERIALS AND METHODS: Gas chromatography/mass spectroscopy analyses were used to identify the constituents of essential oil. Protoscoleces were treated with different concentrations of the essential oil (6.25-100 µL/mL) in each test tube for 5-30 min. The viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Forty-eight male NMRI mice were also used to determine the toxicity of C. zeylanicum essential oil (0.5-4 mL/kg). RESULTS: The main components were found to be cinnamaldehyde (91.8%), ρ metoxicinamate (1.57%), and α pinene (1.25%). Findings indicate that C. zeylanicum essential oil with the concentrations of 100 and 50 µL/mL killed 100% of protoscoleces after 5 min of exposure. Also, the lower concentrations of C. zeylanicum essential oil motivated a late protoscolicidal effect. The LD50 value of intraperitoneal injection of C. zeylanicum essential oil was 2.07 mL/kg body weight after 48 h, and the maximum nonfatal dose was 1.52 mL/kg body weight. The results also showed that there was no significant toxicity following oral administration of C. zeylanicum essential oil for 2 weeks. CONCLUSION: The results exhibited the favorable scolicidal activity of C. zeylanicum, which could be applied as a natural scolicidal agent in hydatid cyst surgery. SUMMARY: We evaluated the efficacy of Cinnamomum zeylanicum essential oil against hydatid cyst protoscolecesThe viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining)Forty-eight male NMRI mice were also used to determine the toxicity of C. zeylanicum essential oilC. zeylanicum with potent scolicidal activity could be applied as a natural scolicidal agent in surgery. Abbreviations used: GC/MS: Gas chromatography/mass spectrometry analysis; CE: Cystic echinococcosis; LD50: Lethal dose 50%; I.p: Intraperitoneally.

Toxoplasma gondii Infection Potentiates Cognitive Impairments of Alzheimer's Disease in the BALB/c Mice.

This study tests the hypothesis that in chronic Toxoplasma gondii infection communication among immune cells promotes neuroinflammation through cytokine networks and potentiate cognitive impairments in BALB/c mice with Alzheimer's disease (AD). The animal model of Toxoplasma infection was established by the intraperitoneal inoculation of 20-25 tissue cysts from the Tehran strain of T. gondii . We injected amyloid-beta 1-42 peptide (Aß1-42, 1 and 2 µl) into the hippocampus of BALB/c mice to establish an animal model of AD. The behavioral experiments such as spatial learning and memory were performed using the Morris water maze test. The mRNA levels of TNF-α, IL-1ß, IFN-γ, and inducible nitric oxide synthase (iNOS) were examined by real-time PCR. We found that T. gondii infection caused AD-like symptoms and impaired learning and memory functions of the infected BALB/c mice. We also found that in Toxoplasma infection + Aß1-42 (1 µl) group, T. gondii infection could potentiate AD in infected mice receiving subdoses of Aß1-42 (1 µl) and caused considerable impairment in learning and memory functions similar to AD group. Comparison of the results demonstrated that mRNA levels of IL-1ß, TNF-α, IFN-γ, and iNOS significantly (P < 0.001) increased in T. gondii + Aß1-42 (1 µl) in comparison with the other tested groups. The obtained results showed that chronic T. gondii infection communication among immune cells promotes neuroinflammation through cytokine networks and induces pathological progression of AD in the mice brain, whereas the presence of neuroanatomical Toxoplasma tissue cysts in the brain could also affect the behavioral functions in T. gondii -infected mice.

Efficacy and Safety of Bunium Persicum (Boiss) to Inactivate Protoscoleces during Hydatid Cyst Operations.

BACKGROUND: Current scolicidal agents, which have been used for inactivation of protoscoleces during surgical procedures, are associated with adverse side effects including sclerosing cholangitis. This investigation aimed to evaluate the scolicidal effects of Bunium persicum (Boiss) essential oil against protoscoleces of hydatid cysts and also its toxicity in a mice model. METHODS: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (3.125-50 mcL/mL) were used for 5-30 min. The viability of protoscoleces was confirmed using the eosin exclusion test (0.1% eosin staining). Further, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of B. persicum essential oil. RESULTS: The obtained results revealed that the B. persicum essential oil at the concentrations of 25 and 50 mcL/mL after 5 min of exposure killed 100% protoscoleces. The mean mortality rate of protoscoleces after 10 min of exposure to the concentration of 12.5 mcL/mL was 100%. Lower concentrations (6.25 and 3.125 mcL/mL) of B. persicum essential oil, however, indicated a delayed protoscolicidal effects. The LD50 value of intra-peritoneal injection of the B. persicum essential oil was 1.96 mL/kg body wt. No significant difference (p > 0.05) was observed in the clinical chemistry and hematologic parameters after oral administrations of B. persicum essential oil at the doses 0.05, 0.1, 0.2, and 4 mL/kg for 14 d. CONCLUSION: Our findings demonstrated the potent scolicidal activity of B. persicum with no significant toxicity; it might be used as a natural scolicidal agent in hydatid cyst surgery.

High-resolution melting analysis (HRM) for differentiation of four major Taeniidae species in dogs Taenia hydatigena, Taenia multiceps, Taenia ovis, and Echinococcus granulosus sensu stricto.

Tapeworms of the genus Taenia include several species of important parasites with considerable medical and veterinary significance. Accurate identification of these species in dogs is the prerequisite of any prevention and control program. Here, we have applied an efficient method for differentiating four major Taeniid species in dogs, i.e., Taenia hydatigena, T. multiceps, T. ovis, and Echinococcus granulosus sensu stricto. High-resolution melting (HRM) analysis is simpler, less expensive, and faster technique than conventional DNA-based assays and enables us to detect PCR amplicons in a closed system. Metacestode samples were collected from local abattoirs from sheep. All the isolates had already been identified by PCR-sequencing, and their sequence data were deposited in the GenBank. Real-time PCR coupled with HRM analysis targeting mitochondrial cox1 and ITS1 genes was used to differentiate taeniid species. Distinct melting curves were obtained from ITS1 region enabling accurate differentiation of three Taenia species and E. granulosus in dogs. The HRM curves of Taenia species and E .granulosus were clearly separated at Tm of 85 to 87 °C. In addition, double-pick melting curves were produced in mixed infections. Cox1 melting curves were not decisive enough to distinguish four taeniids. In this work, the efficiency of HRM analysis to differentiate four major taeniid species in dogs has been demonstrated using ITS1 gene.

Efficacy of Pistacia khinjuk Fruits on Viability of Hydatid Cyst Protoscoleces and Its Acute Toxicity in Mice Model.

BACKGROUND: This investigation aimed to evaluate the in vitro scolicidal effects of Pistacia khinjuk methanolic extract against protoscoleces of hydatid cysts and its acute toxicity in mice NMRI model. METHODS: Protoscoleces were aseptically extracted from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5-100 mg/mL) were used for 10 to 60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Twenty-four male NMRI mice were used to assess the acute toxicity of P. khinjuk. RESULTS: P. khinjuk extract at the concentrations of 100 mg/mL after 10 min of exposure killed 100% of protoscoleces. Similarly, the mean of mortality rate of protoscoleces after 20 min of exposure to the concentration of 50 mg/mL was 100%. The LD50 of the intraperitoneal injection of the P. khinjuk methanolic extract was 2.8 g/kg and the maximum non-fatal dose was 1.7 g/kg. CONCLUSION: The findings demonstrated effective scolicidal effects of P. khinjuk extract with no considerable toxicity that might be a natural source for the producing of new scolicidal agent.

Report: Evaluation of the scolicidal effects of Nectaroscordum tripedale extract and its acute toxicity in mice model.

Current scolicidal agents, which have been used for inactivation of protoscoleces during hydatid cyst surgery are associated with adverse side effects. This study aims to evaluate the in vitro scolicidal effects of Nectaroscordum tripedale L. leave extract against protoscoleces of hydatid cysts and its acute toxicity in mice model. Various concentrations of the extract (12.5-100 mg/mL) were used for 5 to 30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). In addition, the acute toxicity of N. tripedale extract was determined for 2 days in mice model. The results showed that the N. tripedale extract at the concentration of 100 mg/mL after 5 min of exposure killed 100% protoscoleces. Similarly, the mean of mortality rate of protoscoleces after 10 min of exposure to concentration of 50 mg/mL was 100%. The LD50 values of intraperitoneal injection of the N. tripedale extract was 3.36 g/kg body wt. and the maximum nonfatal doses were 2.98 g/kg body wt. The results showed the potential of N. tripedale extract as a natural source for the production of new scolicidal agent for use in hydatid cyst surgery.