---

Aims: Toxoplasmosis is a cosmopolitan zoonotic disease caused by an obligate apicomplexan intracellular parasite known as Toxoplasma gondii (T. gondii). Recently, toxoplasmosis has been suggested as a risk factor for diabetes. Thus, the present study aimed to assess the association between T. gondii infection and two types of diabetes in Tehran, the capital of Iran.
Materials & Methods: In the current cross-sectional study, 98, 95, and 94 blood samples were collected from Type 1 and Type 2 diabetic and nondiabetic individuals, referring to Imam Sajad hospital from February to August 2018, respectively. Anti-T. gondii specific IgG and IgM antibodies were measured using enzyme-linked immunosorbent assay (ELISA). Moreover, a structured demographic questionnaire was completed for each person.
Results: IgG antibody was found to be positive in 16.32 (16 of 98) and 57.89% (55 of 95) of patients with diabetes Type 1 and Type 2 and 17.02% (16 of 94) of nondiabetic individuals as controls, respectively. However, the prevalence of positive IgM antibody in these groups was determined as 2.04 (2 of 98), 6.32 (6 of 95), and 17.02 % (16 of 94), respectively.
Conclusion: This finding revealed that toxoplasmosis could be considered as a possible risk factor for diabetes Type 2, while no statistically significant association was found between T. gondii infection and diabetes Type 1.  More research is required to be conducted in the future in order to better understand this association.

---

Backgrounds: Toxoplasma gondii is a zoonotic parasite of increasing concern to humans and animals. Considering the side effects of drugs used to treat toxoplasmosis, it is essential to find alternative drugs.
Materials & Methods: In this study, colchicine and propranolol at four concentrations (1, 5, 10, and 15 µg/mL) were added to the RPMI medium containing peritoneal macrophages and incubated for 60 min, Then tachyzoites were added to the medium, and the efficacy rates of colchicine and propranolol in inhibiting tachyzoites entry into macrophages were evaluated after 30 and 60 min. For in vivo assay, one group received no drugs, and the second group was treated with colchicine and propranolol at different concentrations for different durations.
Findings: The in vitro experiment showed that treatment with 15 mg/mL of colchicine and propranolol for 60 min following tachyzoites addition was the most efficient method to inhibit tachyzoites penetration, indicating the efficacy rates of 80.20%±1.20 and 89.97%±1.30, respectively (p< .05). Based on the in vivo test, pretreatment with 2 mg/kg of colchicine one hour before tachyzoites injection had the best inhibitory effect (70.32%±4.07). Also, pretreatment with 2 mg/kg of propranolol 90 min before tachyzoites injection (78.54%±1.99) induced the best inhibitory effect (p< .05).
Conclusion: According to the results, colchicine and propranolol could inhibit tachyzoites entrance into nucleated cells in vitro and in vivo. In this study, the most efficient concentrations and times for using these substances were determined.
 

---

Objectives: Toxoplasma gondii is a coccidian protozoon which forms tissue cyst in different organs of infected intermediate host. The present study was conducted to demonstrate the active form of parasite in different tissues of rat, which was experimentally infected with RH strain of Toxoplasma gondii using bioassay method inmice. Materials & Methods: In the experimental assay, 75 rats and 500 mice were used. The rats were infected experimentally with 40-50 thousands of tachyzoites intrapritoneally. Three rats were killed every 24 h then with up to three days 3 days interval for 60 days and their different organs were searched biologically for presence of the parasite. In this regard, the organs were squeezed with mortar in normal saline, diluted and following adding streptomycin and penicillin injected intrapritoneally in three outbreed mice each. Then the mice were followed up for 10 days. Results: Toxoplasma cyst was found in brain, liver and spleen of the rats within 5 to 9 post infection. The parasite was appeared in heart and muscles within 9-13 days after infection but not found in the kidney. The parasite was disappeared in spleen and liver following12 and 28 days of infection respectively but remain active in the brain, heart and muscles within 60 days of the study. Implication: Toxoplasma gondii could remain active in spleen and liver of infected rat for a short time. Meanwhile brain and muscles (heart and skeletal muscles) of the infected animal can keep the parasite for a long time. Kidney is not appropriate tissue for Toxoplasma gondii localization.

---

Background: Multiple sclerosis (MS) is an autoimmune inflammatory disease of the central nervous system. Toxoplasma gondii infection is one of the risk factors of MS. Knowing the correlation between T. gondii infection and MS could lead to a better understanding of the disease incidence. This study aimed to assess the correlation between T. gondii infection and the disease incidence in infected individuals.
Materials & Methods: Serum samples of 38 MS patients referring to the neurology clinic of Ghaem hospital in Mashhad in 2019 were analyzed by Pishtazteb commercial kit for anti-T. gondii IgG and IgM antibodies using enzyme-linked immunosorbent assay (ELISA). The obtained data were analyzed with SPSS software Version 20.
Findings: Among the 79 tested individuals, 22 were positive, and 57 were negative for anti-T. gondii IgG antibodies. Among the negative cases, 29 (50.9%) had MS, and 28 (49.1%) were controls. Among the positive cases, nine (40.9%) patients had MS, and the remaining 13 (59.1%) were controls. The frequency of IgG antibody in the case and control groups was not significantly different (p= .427). Anti-T. gondii IgM antibody was negative in all samples. There was no significant difference between the types of MS regarding the frequency of anti-T. gondii antibodies (p= .402).
Conclusion:  No significant difference was found in the frequency of anti-T. gondii IgG antibodies between the two groups. However, further studies with larger sample sizes are recommended to gain a better understanding of the relationship between anti-T. gondii IgG antibody positivity and MS incidence

---

Objective: Toxoplasmosis, caused by an intracellular protozoan parasite, and the Toxoplasma gondii, is widespread throughout the world. In recent years, significant progress has been made in the identification of vaccine candidates which could induce a protective response. GRA7, an excretory 29 kDa Toxoplasma gondii a dense granular antigen released by infected host cells. In tachyzoite-infected cells, p29 accumulates within the parasitophorous vacuole and co-localizes with its delimiting membrane. Materials and Methods: In the present work, first genomic DNA of Toxoplasma gondii was extracted and used for amplifying of GRA7 gene as a template. Then PCR product was extracted from agarose gel and cloned into TOPO vector. The plasmid containing GRA7 gene was extracted from the transformed bacteria (TOP10 strain) and sequenced. Results: Sequence analysis of GRA7 gene cloned into TOPO vector showed only one base difference when composed with the gene bank sequence for RH strain was only one base. Conclusion: The results indicated that this clone is suitable for subcloning in Prokaryotic and Eukaryotic plasmid.

---

Background: In thyroiditis, the cellular immune response plays a crucial role in triggering the production of autoantibodies. Toxoplasma gondii elicits a strong innate and adaptive immune response within the host organism. This study aims to assess the seroepidemiological prevalence of toxoplasmosis and the levels of interferon-gamma (IFN-γ) in patients with autoimmune thyroiditis (AITD) in Iraq.
Methods: This case-control survey was conducted on 100 patients diagnosed with AITD and 70 healthy individuals (non-AITD) who attended general hospitals in Thi-Qar Province, Iraq, between July and November 2023. The prevalence of anti-Toxoplasma gondii antibodies was evaluated using enzyme-linked immunosorbent assay (ELISA) kits. Serum levels of IFN-γ were measured using ELISA kits, while the expression levels of IFN-γ were assessed using real-time polymerase chain reaction (PCR).
Findings: Among the participants, 33 patients (33.00%) in the AITD group and 9 patients (12.85%) in the non-AITD group tested positive for T. gondii IgG antibodies (p < 0.001). Additionally, 2.00% of AITD patients and 1.40% of non-AITD patients were positive for anti-Toxoplasma IgM antibodies. PCR analysis revealed the presence of T. gondii parasites in 2.00% of AITD patients and 1.4% of non-AITD patients. In AITD patients with T. gondii antibodies, both serum levels and gene expression of IFN-γ were significantly elevated compared to AITD patients without T. gondii antibodies (p < 0.05).
Conclusion: Current findings suggest that individuals infected with T. gondii may experience direct effects on the thyroid gland due to elevated levels of IFN-γ. However, further analyses are necessary to validate these results.

---

Objective: The entry of Toxoplasma gondii into the bloodstream and other tissues (such as liver, muscle, cardiac muscle, …) of intermediate hosts and its multiplication in nucleated cells may cause changes in plasma levels of various enzymes due to tissue damage. In present study the serum levels of AST, ALT, ALK/P, CPK, LDH, and ACP in rats infected experimentally with Toxoplasma gondii have been investigated. Materials and Methods: Totally, 116 uninfected rats were divided into 87 as case group and 29 as control group. The case group was infected intraperitoneally with 50000 tachyzoites. Blood specimens were taken from cases and its control once every 8 hours in the first three days and then once every three days for a period of 60 days and serum levels were measured for the mentioned enzymes. Results: During the study, the following changes were observed: AST in the first 8 hours, from the 32th hour till the 40th hour and from the 48th hour till the 56th hour; ALT in the first 8 hours and from the 48th hour till the 56th hour; ALK/P from the 24th hour till the 40th hour and from the 48th hour till the 64th hour; CPK and LDH from the 24th hour till the 40th hour and from the 48th hour till 56th hour; ACP from the 16th hour till the 48th hour. But afterward, whole offer mentioned enzyme shifted to normal levels. Conclusion: Alteration in serum enzyme levels of rats during infection with Toxoplasma gondii found is not permanent.

---

Objective: The aim of the present study was to investigate the effect of protein and DNA components of Toxoplasma gondii on maturation of dendritic cells and their efficiency in IL-12 production and proliferation of T cells. Materials and Methods: for DC generation, Bone marrow cells were cultured in the presence of GM-CSF and IL-4 for 5 days. Tumor lysate and protein or DNA components of Toxoplasma gondii were added to the culture media and incubated for another 2 days. LPS was added as control for DC maturation. Proliferation of T cells were determined by MLR and IL-12 production was measured by ELISA kit. Maturation of dendritic cell were determined by flowcytometry. Results: DCs treatment with protein components of Toxoplasma gondii caused a significant increase in IL-12 production and proliferation of T cells (P<0.001). Conclusions: Different compositions of microbial body like protein and DNA components of Toxoplasma gondii can cause augmentation of antigen presentation capacity of DC and their IL-12 production capability. Among these components the protein was more effective as compared to DNA.

---

Objective: Toxoplasma gondii is an obligate intracellular protozoan that causes Toxoplasmosis in human and animal. In recent years, significant progress has been made in the identification of vaccine candidates which can induce protective responses. In this study we used complete Rhoptry protein 2 gene of Toxoplasma gondii as a single DNA vaccine and evaluated its immune responses in comparison with control groups. Materials and Methods: BALB/c mice were immunized intramuscularly with three weaks time interval with pcROP2 (as case group) and pc-DNA3 and PBS (as control groups). After immunization, we evaluated the immune response using cytokine and antibody measurements. Results: The results of cytokine (IFN-γ, IL-4) assays showed that mice immunized with pcROP2, elicited stronger Th1-type cellular immune responses than those immunized with empty plasmid, or PBS (high level of IFN-γ and low-level of IL-4).Also Anti-T. gondii IgG titres (OD) increased markedly in the pcROP2 group, which was significantly higher than those of control groups (P<0.05). When challenged with the highly virulent Toxoplasma gondii RH strain, mice immunized with pcROP2 had siginificantly higher survival rates compared to control groups (P<0.05). Conclusion: This study showed that pc-ROP2 as a single DNA vaccine is effective to prime enhanced and balanced cellular and humeral immunity responses, and relatively improved mice survival time against toxoplasmosis.

---

Objective: Toxoplasmosis infection has spread throughout the world, it is created by the intracellular protozoan Toxoplasma gondii. Toxoplasma cause changes in the behavior of the rodents. Rodents play an important role in the life cycle of the Toxoplasma gondii, they are the main infection reservoir of the domestic cats. The purpose of this study was to investigation Toxoplasma infection in the Tehran's Rats. Materials and Methods: In this study, 150 mice were collected from all 5 regions of Tehran (North, South, East, West, Center) by live traps over 8-months period. For infection detection, anti-rat conjugate was used in ELISA method. Results: The results showed that 36/7 percent of rats in Tehran have Toxoplasma infection. Maximum number of infected rats were found in the South and Central parts of Tehran with 11.7 percent and with minimum of 1.47 percent in the West of Tehran. Conclusion: The extent of infection indicates the importance of these wild rats in the persistence and life cycle this parasite.

---

Objective: Toxoplasmosis can lead to severe pathological effects in both infected humans and animals. The various DNA vaccines against Toxoplasma compose of single or cocktail antigens have been investigated but they have partial protective against disease. In this study, we used pcROP1 as a DNA vaccine and aluminium phosphate and aluminium hydroxide to compare their efficacy as mineral adjuvants. Materials and Methods: BALB/c mice immunized with pcROP1 alone or with co-administration of Alpo4 or Alum and the effectiveness of these two adjuvants were compared using lymphocyte proliferation assay, cytokine and antibody assay and survival time. Results: The group co-administered alum elicited stronger humoral and Th1-type cellular immune responses than the group co-administered Alpo4, while immune response in group administered with pcROP1 alone is higher than them. When challenged with Toxoplasma gondii RH strain, mice immunized with or without alum had significantly higher survival rates, whereas there was no notable enhancement of survival rate in Alpo4 group (P≤0.05). Conclusions: Our result suggest that pcROP1 plus alum and aluminium phosphate not strongly potentiate the efficacy of this DNA.

---

Objective: Evaluation of the effects of Toxoplasma gondii infection on spermatogenesis in male rats. Materials and Methods: The RH strain of T. gondii tachyzoites were injected interaperitoneally in an infected group of 35 rats, while 21 rats were used as controls. Each ten days from 10- 70 days of post-infection (PI), 5 rats from infected group and 3 rats from control group were scarified. The percentage of body weight to testis weight ratio (BTR) as well as sperm parameters and fructose levels in seminal vesicles and coagulating glands (SVCG) were investigated. An IgG ELISA kit was designed for serologic diagnosis of infection in the rats. Results: All rats injected with T. gondii tachyzoites were infected from 10-70 PI. Sperm motility from 10-70 PI, sperm viability from 10-60 PI and sperm concentration from 20-60 PI were significantly decreased in the infected group (P<0.05); sperm abnormality was significantly increased in the infected group on days 30, 40 and 50 PI (P < 0.05). BTR in the infected group was not significantly changed compared to control group (P>0.05). Fructose level in SVCG in the infected group was significantly decreased on days 10-50 PI (P < 0.05) compared to control. Conclusion: According to the results, toxoplasmosis can cause impermanent impairment on the spermatogenesis in the male rats.

---

Objective: Toxoplasmosis is a worldwide disease, for which different detection methods have been used. The nucleic acid sequence-based amplification (NASBA) method is shown to be highly efficient for diagnosis of live microorganisms. The present research evaluates the molecular isothermal method of NASBA to identify live Toxoplasma gondii (T. gondii) in rat. Methods: Tachyzoites of T. gondii were inoculated in the peritoneal cavities of mice (Mus musculus) and their RNA was extracted. The NASBA method was then used to amplify the tachyzoite B1 rRNA gene. Next, we examined blood samples from 30 experimentally infected case and control rats (Rattus norvegicus) by NASBA. Finally, the resultant band was investigated on an agarose gel. Results: The B1 genes extracted from both the tachyzoites and blood samples were successfully amplified by the NASBA method. This amplified gene yielded an amplicon of approximately 116 bp on gel agarose. Conclusion: NASBA is highly efficient for the identification of live T. gondii. This method can be applied for early diagnosis of active toxoplasmosis in both newborns and immunocompromised individuals.