Genetic Diversity of Toxoplasma gondii by Serological and Molecular Analyzes in Different Sheep and Goat Tissues in Northeastern Iran.

Background: Toxoplasmosis is a parasitic disease caused by compilation protozoan agent Toxoplasma gondii, leading to significant financial and quality-adjusted life-year losses. Overcooked or raw meat consumption has been a considerable transmission route. The present study was conducted to determine the seropositivity rate of T. gondii in sheep and goats by serological and molecular tests and genotyping of obtained isolates in northeast Iran. Methods: Blood and tissue samples (diaphragm, heart) of 296 animals (including 168 sheep and 128 goats) were collected from the slaughterhouse in Quchan Country from august 2016 to April 2017. Modified agglutination test (MAT) and the PCR method performed to detect parasite DNA on tissues.PCR-RFLP method of GRA6 gene was used to determine the genotype of T. gondii. In addition, sequencing analysis was performed to evaluate the Toxoplasma type strains. Results: Serum positive for MAT results were found in 27.4% of sheep and 23.4% of goats. Positive PCR of B1 gene results in diaphragm and heart tissues of sheep and goats was 47.8% and 26.1%, 40% and 23.3%, respectively. PCR of GRA6 gene results were positive in 10 samples that RFLP technique results using MseI enzyme revealed genotype I. Sequencing and phylogenetic analysis revealed DNA of all samples was closely related to Toxoplasma type I. Conclusion: Concerning the high seropositivity rate of toxoplasmosis, undertaking an appropriate preventive program for reducing the prevalence of T. gondii infection by raw or undercooked meat consumption of livestock is recommended. Our study supports the notion that these animals' consumption of raw and undercooked meat can be a probable source of human toxoplasmosis.

Giant Brain Hydatid Cyst in an Adult: A New Case Report

Cystic echinococcosis is an important zoonotic disease that occurs in humans and mammals in general, which causes considerable economic loss and poses health concerns in different parts of the world. The patient involved in this case report was a 28-year old man living in Birjand city who had been suffering from intermittent headache, nausea and vomiting for the past two weeks. The other symptoms presented by the patient were dizziness, blurred vision, seizures and imbalance. The patient only complained of headache in the last two weeks and had no symptoms of visual or speech impairment. He had a history of consuming raw vegetables, but did not have canine contact. In brain computed tomography and magnetic resonance imaging, a large cyst was evident in his brain. The patient was admitted to Razi Hospital in Birjand and followed-up by surgical treatment of the hydatid cyst, with no complications observed.

Biogenic selenium nanoparticles target chronic toxoplasmosis with minimal cytotoxicity in a mouse model.

Introduction. Nanoparticles (NPs) have numerous biological benefits due to their large surface-volume ratio and convenient entry into cells compared to other particles. Previous research has shown the antimicrobial properties of biogenic selenium NPs (SeNps) and their effects on cellular immunomodulatory cytokines that play a key role in controlling infections.Aim. This study aimed to evaluate the therapeutic effects of SeNPs against chronic toxoplasmosis in mice.Methodology. Infected mice with Toxoplasma gondii (Tehran strain) were orally treated with SeNPs at doses of 2.5, 5 and 10 mg kg-1 once a day for 14 days. On the fifthteenth day, the mean number of brain-tissue cysts and the mRNA levels of TNF-α, IL-12, IL-10, IFN-γ and inducible nitric oxide synthase (iNOS) in the mice of each group were recorded. Moreover, serum clinical chemistry factors in the treated mice were examined to determine the safety of SeNPs.Results. The mean number of tissue cysts was significantly (P<0.001) decreased in mice treated with SeNPs in a dose-dependent manner compared with the control group. The mRNA levels of inflammatory cytokines were significantly increased in mice treated with SeNPs at a dose of 10 mg kg-1 compared with the control subgroup (P<0.05). No significant variation (P>0.05) observed in clinical chemistry parameters among the mice in the control subgroup compared with those treated with SeNPs.Conclusion. The findings demonstrated the therapeutic effects of SeNPs with no considerable toxicity against latent toxoplasmosis in the mouse model. Nevertheless, further studies are obligatory to reveal the exact anti-Toxoplasma mechanisms of SeNPs.

Differential expression of TLRs 2, 4, 9, iNOS and TNF-α and arginase activity in peripheral blood monocytes from glucantime unresponsive and responsive patients with anthroponotic cutaneous leishmaniasis caused by Leishmania tropica.

BACKGROUND: Detection of the mechanism of host/parasite interactions in unresponsive forms of anthroponotic cutaneous leishmaniasis (ACL) caused by Leishmania tropica is helpful for immunotherapy and vaccine development. In the present study, the gene expression of toll-like receptors (TLRs), TNF-α, iNOS and also arginase (ARG) activity in monocytes from Glucantime unresponsive in comparison to responsive patients infected with L. tropica was investigated. METHODS: In this case-control study, patients with unresponsive (n = 10) and responsive (n = 10) ACL were recruited. Gene expression of TLR2, TLR4, TLR9, TNF-α and iNOS was analyzed in L. tropica-exposed monocytes. The level of ARG activity in both isolated promastigotes and the lysates of monocytes was also determined. RESULTS: L. tropica-exposed monocytes represented higher expression of all three TLRs and TNF-α and lower expression of iNOS compared to unexposed ones in both groups of patients. Results revealed a significant down-regulation of TLR2 and TNF-α and up-regulation of TLR9 expression in unresponsive isolates in comparison to responsive ones. Besides, ARG level showed a significant increase in L. tropica-stimulated monocytes and cultured promastigotes from unresponsive isolates versus responsive ones. CONCLUSIONS: The decreased TLR2, TLR4, TNF-α and iNOS and the increased level of TLR9 expression in L. tropica-exposed monocytes from unresponsive isolates and also the increment in ARG activity in their promastigotes and monocytes, might possibly be involved in the severity of the disease and leading to Glucantime unresponsiveness.

Detection of methicillin-resistant Staphylococcus aureus (MRSA) in clinical samples of patients with external ocular infection.

BACKGROUND AND OBJECTIVES: Staphylococcus aureus is the main Gram-positive bacteria isolated from patients with ocular infections. Herein, we describe the pattern of antibiotic resistance, presence of resistance genes including ermA, ermB, ermC, msrA, mecA and the pvl cytotoxin gene in S. aureus isolates collected from patients with external ocular infection. MATERIALS AND METHODS: In this study, 8 S. aureus isolates were collected from 81 patients that suffered from eye damage. Antibacterial susceptibility of isolates was determined using the Kirby-Bauer disk diffusion method. Resistance genes including ermA, ermB, ermC, msrA, mecA and the pvl virulence gene were detected by PCR method. Staphylococcal cassette chromosome mec (SCCmec) in MRSA isolates were detected by the multiplex-PCR method. RESULTS: Three isolates were resistant to cefoxitin which is considered MRSA. The mecA gene was identified in MRSA isolates. SCCmec type IV and the pvl gene were detected in one of the MRSA isolates that was recovered from a diabetic patient. CONCLUSION: The emergence of S. aureus isolates belonging to SCCmec type IV and pvl gene among patients with ocular infection is very serious; therefore, identify genetic characterization of MRSA isolates for empirical therapy and infection control is very important.

Scorpion composition and scorpionism in a high-risk area, the southwest of Iran.

BACKGROUND: Scorpions are usually defined as predatory arachnids having venomous and painful stings. Scorpionism is considered as one of the health and medical problems in undeveloped tropical and subtropical countries in the world that endangers thousands of people annually. OBJECTIVE: This study was done for identification of scorpion fauna and scorpionism location in a high-risk area in southwest of Iran. METHODS: This study was carried out in Andimeshk district located in Khuzestan province which has the highest frequency of scorpionism in Iran. In total, 4 urban and 15 rural areas with various climates and topography were selected for monthly scorpion collection in 2016. Additionally, clinical data was obtained from questionnaires over a 5-year period (2012 to 2016) which were completed by physicians and health staff in hospitals and health centers. The data were analyzed by IBM-SPSS Version 20. RESULTS: A total of 850 scorpion samples with the most frequency in rural areas were collected. Eight species belonging to three families; Buthidae, Scorpionidae and Hemiscorpiidae were identified. A. crassicauda was the dominant species followed by H. lepturus. The largest collection due to scorpion's activity was recorded in summer. The five-year prevalence of scorpion sting were 9.1/1000 inhabitants. The highest frequency of scorpionism was observed in the male age group of 25-34 years. Most cases had been treated with polyvalent scorpion antivenom within 6-12 hours of the sting. CONCLUSION: Based on the findings, six species of medically important scorpions that were previously reported from Iran were also collected and caught in this study area. The findings of this study can be useful for policymakers at the Andimeshk Healthcare Center to prevent scorpionism and provide appropriate treatment for scorpionism cases.

Unresponsiveness to meglumine antimoniate in anthroponotic cutaneous leishmaniasis field isolates: analysis of resistance biomarkers by gene expression profiling.

BACKGROUND: Resistance to antimonials is a fundamental determinant of treatment failure in anthroponotic cutaneous leishmaniasis (ACL). Detection of reliable molecular markers to distinguish unresponsive and responsive parasites is critical for consolidating strategies to monitor drug efficacy. METHODS: We analysed the expression of five major antimony resistance-associated genes that is aquaglyceroporin1 (AQP1), γ-glutamylcysteine synthetase (γ-GCS), multidrug resistance protein A (MRPA), trypanothione reductase (TR) and thiol-dependent reductase 1 (TDR1), in unresponsive and responsive Leishmania tropica field isolates by quantitative real-time PCR in comparison with sensitive and resistant reference strains. RESULTS: Gene expression analysis showed the down-regulation of AQP1, γ-GCS and TDR1 by a factor of 1.9, 1.7 and 3.55, respectively, in unresponsive isolates vs. responsive ones. The average RNA expression level of MRPA increased by a factor of 1.9 in the unresponsive group. Isolates exhibited a strong positive linear correlation between gene expression of AQP1 and γ-GCS. A negative correlation between the AQP1 and γ-GCS expression level and lesion duration in responsive patients indicated the potential role in diagnosing drug-unresponsive parasites in endemic areas of ACL. CONCLUSION: In cases of inconclusive outcomes of resistance tests in clinical isolates, expression analysis of a set of influential genes can be beneficial to identify distinctive biomarkers between antimony-unresponsive and responsive parasites.

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.