Introduction of protocols for mass production of Toxoplasma gondii tachyzoites of the genotype II PRU strain

Background:Few investigations of genotype II of Toxoplasma gondii,the most preva-lent form of the Toxoplasma parasite in humans,have been carried out on due to the rapid conversion of tachyzoites to bradyzoites in its life cycle.The current study aimed to create animal and in vitro models for production of the tachyzoites of the Prugniaud(PRU)genotype II strain.Methods:To develop an immunocompromised model and obtain tachyzoites of the PRU strain,BALB/c mice were orally treated with dexamethasone(10 mg/kg),cyclo-phosphamide(36 mg/kg),and cyclosporine(18 mg/kg)from 5 days prior to inocula-tion.Then,10-15 tissue cysts of PRU strain were inoculated intraperitoneally into the mice.The tachyzoites obtained from mice were then cultivated in a HeLa cell culture.The resulting yield of tachyzoites was cryopreserved in 92%fetal calf serum,8%dimethyl sulfoxide.The infectivity of these tachyzoites was evaluated using in vivo and in vitro examinations.Results:Numerous tachyzoites were observed in the peritoneal fluid of the immuno-suppressed mice within 10-15 days after inoculation,and many tachyzoites were har-vested from the HeLa cell culture.Trypan Blue staining showed 80%viability of the tachyzoites recovered from cryopreservation and this was confirmed by HeLa cell culture.In addition,mice infected intraperitoneally with the recovered tachyzoites presented with cysts in the brain after 2 months.Conclusion:We have developed an animal model for mass production of T.gondii tachyzoites of the PRU strain.This method can provide fresh viable tachyzoites of Toxoplasma gondii for use as and when required in future investigations.

THE ULTRASTRUCTURE OF TOXOPLASMA GONDII TACHYZOITES AND THE EFFECT OF USNIC ACID ON IT

The normal fine structures of toxoplasma gondii tachyzoite and the erfect of usnic acid on the ultrastructure of the tachyzoites were observed by transmission electron microscope (TEM).The studies indicate that the pathological changes or the ultrastructure of the parasite took place under the effect of the 10 mg/L usnic acid for 30 min.These changes can be illustrated as folio'vs; 1)The posterior portion of the rhopties were destroyed.2)The membrane of the daughter cell fractured. 3)The membranate organellae or the organisms were demaged.

Analysis Concentration of <i>Toxoplasma gondii</i>on Anti-Toxoplasma IgG-IgM Antibody Levels, and the Outcomes of Pregnancy in Mice Balb/c

Introduction: Toxoplasma gondii is an obligate intracellular protozoan parasite that can infect any warm blood vertebrae, and if first trimester pregnant woman infected, it may cause abortion. The objective is to prove the effect of the Toxoplasma gondii concentration in anti-toxoplasma IgG-IgM antibody levels, and the outcomes of Balb/c mice pregnancies. Materials and Methods: The study was conducted in Balb/c mice with inclusion criteria, and was conditioned pregnant. The pathogen strains of Toxoplasma gondii tachyzoite injected intraperitoneally. The blood samples were taken serially to be tested for anti-toxoplasma IgG-IgM antibody levels. After the mice were injected with tachyzoite, they are assessed every day to observe their body weight, vaginal bleeding, and labor. Anti-toxoplasma IgG-IgM antibody levels examined using qualitative mouse IgG-IgM antibody ELISA KIT. Results: Anti-toxoplasma IgM antibody levels increased significantly after 24 hours of injection tachyzoites in all dose groups, and remained high through day 21. Anti-toxoplasma antibody IgG levels increased significantly after 72 hours post injection and remained elevated until day 21. The incidence of abortion is 100% in mice which injected tachyzoite levels 1 × 103 and 1 × 104, and the incidence of abortion approximately 2 - 4 days post injection. 100% of mice that were injected with tachyzoites 1 × 101 and 1 × 102 have labor at term. Physical anomaly was found in baby mice from mice that were injected with tachyzoite 1 × 102. Conclusion: There is a significant correlation between the concentrations of Toxoplasma gondii tachyzoite with anti-toxoplasma IgG-IgM antibody levels, and there is a significant relationship between the concentrations of tachyzoite with abortion.