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 MseІ enzyme revealed genotype І. Sequencing and phylogenetic analysis revealed DNA of all samples was closely related to Toxoplasma type І. 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

Scolicidal effects of biogenic selenium nanoparticles against protoscolices of hydatid cysts

Cystic echinococcosis (hydatid cyst, CE) as a zoonotic parasitic infection caused by the larval stage of the dog tapeworm Echinococcus granulosus is still an important economic and public health concern in the world. One of the treatment options for CE is surgical removal of the cysts combined with chemotherapy using albendazole and/or mebendazole before and after surgery. Currently, many scolicidal agents, which have some complications, have been used for inactivation of the cyst contents. Therefore the development of new scolicidal agents with low side effects and more efficacies is an urgent need for surgeons. The present study was aimed to investigate the in vitro scolicidal effect of selenium nanoparticles biosynthesized by a newly isolated marine bacterial strain Bacillus sp. MSh-1 against protoscoleces of E. granulosus. Protoscolices were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations (50e500 mg/ml) of Se NPs (in size range of about 80e220 nm) were used for 10e60 min. Viability of protoscoleces was confirmed by 0.1% eosin staining. The results indicated that biogenic Se NPs at all concentrations have potent scolicidal effects especially at concentrations 500 and 250 mg/ml after 10 and 20 min of application, respectively. In conclusion, the findings of present study proven that Se NPs have potent scolicidal effects, therefore may be used in CE surgery. However, the in vivo efficacy of these NPs remains to be explored

Acetylcholinesterase Inhibitor Improves Learning and Memory Impairment Induced by Toxoplasma gondii Infection

Background: Here, we established the mouse models of chronic toxoplasmosis by T. gondii Tehran strain to provide a good understanding about defining the possible association between T. gondii exposure and learning and memory impairments. Moreover, as secondary objective of the present study, we hypothesized whether administration of an acetylcholinesterase (AChE) inhibitor could reduce learning and memory impairments induced by T. gondii infection. Methods: Twenty-four male BALB/c mice were used to establishment of latent toxoplasmosis. The animal model of Toxoplasma infection was established by the intraperitoneal inoculation of 20-25 tissue cysts from Tehran strain of T. gondii. Donepezil (2 mg/kg) an AChE inhibitor to treat Alzheimer disease was injected intraperitoneally once a day for two weeks starting from post-infection day 90. Morris water maze (MWM) task was used to assay spatial learning and short term spatial memory in all groups. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups.  P<0.05 was considered statistically significant. Results: Toxoplasma infection impaired spatial leaning and short term spatial memory of the infected BALB/c mice, whereas donepezil, an AChE inhibitor, improved impairments induced by Toxoplasma infection. Conclusion: T. gondii infection through increasing AChE reduces the level of Acetylcholine (Ach) and consequently affects learning and memory activity in infected hosts, whereas, donepezil as an AChE inhibitor improves these impairments by restoring ACh levels at synapses of neurons in brain

Scolicidal effects of biogenic selenium nanoparticles against protoscolices of hydatid cysts

AbstractCystic echinococcosis (hydatid cyst, CE) as a zoonotic parasitic infection caused by the larval stage of the dog tapeworm Echinococcus granulosus is still an important economic and public health concern in the world. One of the treatment options for CE is surgical removal of the cysts combined with chemotherapy using albendazole and/or mebendazole before and after surgery. Currently, many scolicidal agents, which have some complications, have been used for inactivation of the cyst contents. Therefore the development of new scolicidal agents with low side effects and more efficacies is an urgent need for surgeons. The present study was aimed to investigate the in vitro scolicidal effect of selenium nanoparticles biosynthesized by a newly isolated marine bacterial strain Bacillus sp. MSh-1 against protoscoleces of E. granulosus. Protoscolices were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations (50–500 μg/ml) of Se NPs (in size range of about 80–220 nm) were used for 10–60 min. Viability of protoscoleces was confirmed by 0.1% eosin staining. The results indicated that biogenic Se NPs at all concentrations have potent scolicidal effects especially at concentrations 500 and 250 μg/ml after 10 and 20 min of application, respectively. In conclusion, the findings of present study proven that Se NPs have potent scolicidal effects, therefore may be used in CE surgery. However, the in vivo efficacy of these NPs remains to be explored

Molecular and Serological Detection of Acute and Latent Toxoplasmosis Using Real-Time PCR and ELISA Techniques in Blood Donors of Rafsanjan City, Iran, 2013

Background: The differentiation between acute and latent forms of the Toxoplasma gondii (T. gondii) infection is still considered as a complicated issue. This study was aimed to elucidate the status of infection in the blood donors and the probable importance of blood transfusion in the transmission of the infection through detecting both immunological and genetic markers of acute and latent infection. Methods: Totally 235 blood samples from blood donors were collected. The levels of anti-T. gondii IgG and IgM antibodies were examined by specific ELISA kits. cDNA were synthesized from total extracted mRNA molecules from the serum samples and SAG1 gene, specific for tachyzoite form, were amplified using Real-Time PCR technique.Demographic information of study subjects including their gender, age, job,and habitat were recorded. Results: Out of 235 serum samples, 80 (34.04%) and 4 (1.71%) were positive regarding anti-T. gondii IgG and IgM antibodies, respectively. Real-Time PCR results showed that 14 out of 200 (6.97%) of blood donor had mRNA molecules of SAG1 gene. The positive results of Real-Time PCR of SAG1 in female gender and housekeepers were significantly higher than those of male gender and other job categories. Conclusion: The prevalence of chronic and acute infection is high in Iranian blood donors. Additionally, evaluation of antibodies could not be reliable, because several donors negative for anti-T. gondii IgM antibodies had detectable SAG1 mRNA molecules.Hence, it seems that molecular diagnostic tests are essential to detect acute infections