The First Survey of Isolation and Molecular Typing of Toxoplasma gondii by Bioassay and PCR Method in BALB/c Mice in Camels (Camelus dromedarius) from Eastern Iran.

BACKGROUND: We isolated Toxoplasma gondii from camels by bioassay method in mice model and detect parasitic DNA in brain mice by molecular methods. METHODS: One hundred tissue samples including heart (n=50), and diaphragm (n=50) were collected from camels (n=50) slaughtered in abattoirs from Feb to Oct 2015 in three provinces located in eastern Iran. In first, blood sample from 50 camels was assayed for anti-Toxoplasma antibodies by modified agglutination test (MAT) test. Bioassay method was done in positive MAT blood camels in BALB/c mice and Nested PCR performed in seropositive tissue samples to amplify the B1 and GRA6 genes. The existence of polymorphic restriction sites for endonuclease MseI was used with PCRRFLP method and Sequencing analysis to evaluate the prevalence of type strains (I, II and III). RESULTS: Overall, 13 (26%) of camels were positive with titer of 1:20 for toxoplasmosis and 13(26%) tissue samples of camels were found positive for the T. gondii B1 gene, including 7(14%) diaphragm, 6(12%) heart. Moreover, 3(6%) tissue samples of camels were found positive with GRA6 gene for T. gondii. There are three genotypes and mix genotype using MseI enzyme among all positive samples. CONCLUSION: The obtained results from serological and molecular tests demonstrated the infection of T. gondii with previously recognized genotypes in the tissues of camels for first time from Iran. Since consumption of meat camels are raising in Iran, there may be a high risk of toxoplasmosis through consumption of products from these hosts due to their susceptibility to the infection.

Prevalence of Toxoplasma gondii Infection among Healthy Blood Donors in Northeast of Iran.

BACKGROUND: This cross-sectional investigation aimed to evaluate the prevalence of IgM and IgG anti-Toxoplasma gondii antibodies and the associated risk factors among healthy blood donors in Khorasan Razavi Province, northeast of Iran from Nov 2014 to May 2015. METHODS: Overall, 491 serum samples from apparently healthy blood donors referred the six biggest blood centers in Razavi Khorasan, Iran, were screened for IgG and IgM anti-T. gondii antibodies by enzyme-linked immunosorbent assay (ELISA). A structured questionnaire was used to obtain information on risk factors for T. gondii infection. Nested PCR was also used to detect DNA of T. gondii in the IgM-positive samples by using of B1 and RE (Repetitive Element) as marker for amplifying fragment size of 531 bp and 164 bp in PCR method. RESULTS: Totally, 200 (40.7%) samples were seropositive for anti-T. gondii antibodies; 184 (37.5%) donors tested seropositive for only IgG antibody, 8 (1.6%) tested seropositive for both IgM and IgG and 8 (1.6%) were positive for IgM antibody alone. Several risk factors significantly related to T. gondii seropositivity in the univariate analysis at P<0.05 included age (P<0.001), and raw/half-cocked meat consumption (P=0.015). T. gondii DNA was found in all sixteen IgM-positive samples. CONCLUSION: T. gondii infection was present among healthy blood donors in northeast of Iran. Thus, it is suggested to design screening programs for preventing transfusion-transmitted toxoplasmosis.

Seroprevalence of Toxoplasma gondii infection among childbearing age women in Kerman city, southeastern Iran.

This cross-sectional study aims to determine the prevalence of IgM and IgG anti-T. gondii antibodies and the associated risk factors among childbearing age women referring to counseling centers before marriage in Kerman city, southeast of Iran. Totally, 300 serum samples were collected from women referred to Central Laboratory for Marriage Consultation in Kerman city were screened for IgG and IgM anti-T. gondii antibodies by enzyme linked immunosorbent assay (ELISA). Out of the 300 serum samples, 38 (12.6 %) tested seropositive for anti-T. gondii antibodies; 31 (10.3 %) samples tested seropositive for only IgG antibody, 1 (0.33 %) tested seropositive for both IgM and IgG and 6 (2.0 %) were positive for IgM antibody alone. Statistical analyses also indicated that seroprevalence of anti-T. gondii antibodies increased with age (p < 0.05). Moreover, some risk factors such as, living in rural regions, contact with cats, raw/half-cooked meat consumption, and agricultural activities were significantly (p < 0.05) related to T. gondii seropositivity. The findings revealed that more than three-quarters of the childbearing age women studied in the present investigation are susceptible to infection during pregnancy. Thus, by adopting correct and improved practices we can improve their living conditions, and prevent infection and awareness and control of pathogens associated with disease is recommended.

Adaptive immune response in symptomatic and asymptomatic enteric protozoal infection: evidence for a determining role of parasite genetic heterogeneity in host immunity to human giardiasis.

The genetic basis of the ultimate clinical outcomes of human giardiasis has been the subject of numerous investigations. We previously demonstrated roles for both host and parasite factors in determining the outcome of enteric infection in a murine model of Giardia duodenalis infection. In the current study, fecal and serum specimens from healthy controls and human subjects infected with the intestinal parasite G. duodenalis were assessed. Using a semi-nested PCR method, clinical isolates were genetically characterized based on the gdh and tpi loci, and the phylogenetic trees were constructed. Using a sandwich ELISA method, the serum levels of representative TH1 and TH2 cytokines were measured in infected human subjects and healthy controls. Here we showed that symptomatic human giardiasis was characterized by significantly elevated serum levels of the TH1 cytokine IFN-γ compared to healthy controls, whereas asymptomatic human subjects and healthy controls had comparable levels of serum IFN-γ. Further analyses showed that human subjects infected with G. duodenalis genotype AI had significantly elevated levels of serum IFN-γ and IL-10, but not IL-5, whereas human subjects infected with AII had similar levels of those cytokines compared to healthy controls. These data demonstrate roles for both host and parasite factors in the determination of the outcome of enteric infections and may further broaden our understanding of host-parasite interaction during enteric protozoal infections.

Efficacy of the Bunium persicum (Boiss) Essential Oil against Acute Toxoplasmosis in Mice Model.

BACKGROUND: We evaluated the in vivo activity of Bunium persicum (Boiss) essential oil on infected mice with acute toxoplasmosis. METHODS: To evaluate prophylactic effects, male NMRI mice received B. persicum essential oil at the concentrations of 0.05 and 0.1 mL/kg for 14 days. After 24 h mice were infected intraperitonealy with 1×10(4) tachyzoites of T. gondii, RH strain. In order to investigate therapeutic effects, mice were infected and then received B. persicum oil at the concentrations of 0.05 and 0.1 ml/kg two times a day for 5 days. The time/mean time of death in all infected mice and the number of tachyzoites from infected mice were recorded. RESULTS: The time/mean time of death of infected mice was 8 and 9 days after oral administration of B. persicum oil at the concentration of 0.05 and 0.1 mL/kg, respectively (P<0.05). In contrast, the time/mean time of death control group was 5 days. In addition, B. persicum significantly reduced the mean number of tachyzoites compared with control group. The time/mean time of death of infected mice was 6 and 7 days after oral administration of B. persicum essential oil at the concentration of 0.05 and 0.1 mL/kg, respectively. In contrast, the time/mean time of death control group was 5 days. B. persicum especially at the concentration of 0.1 ml/kg significantly reduced the mean number of tachyzoites compared with control group. CONCLUSION: The results showed the potential of B. persicum essential oil as a natural source for the production of new prophylactic agent for use in toxoplasmosis.

In Vitro Inhibitory Effect of Berberis vulgaris (Berberidaceae) and Its Main Component, Berberine against Different Leishmania Species.

BACKGROUND: Leishmaniasis has been identified as a major public health problem in tropical and sub-tropical countries. The present study was aimed to investigate antileishmanial effects of various extracts of Berberis vulgaris also its active compoenent, berberine against Leishmania tropica and L. infantum species on in vitro experiments. METHODS: In this study in vitro antileishmanial activity of various extracts of B. vulgaris also its active compoenent, berberine against promastigote and amastigote stages of L. tropica and L. infantum was evaluated, using MTT assay and in a macrophage model, respectively. Furthermore, infectivity rate and cytotoxicity effects of B. vulgaris and berberine in murine macrophage cells were investigated. RESULTS: The findings of optical density (OD) and IC50 indicated that B. vulgaris particulary berberine significantly (P<0.05) inhibited the growth rate of promastigote stage of L.tropica and L.infantum in comparison to meglumine antimoniate (MA). In addition, B. vulgaris and berberine significantly (P<0.05) decreased the mean number of amastigotes in each macrophage as compared with positive control. In the evaluation of cytotoxicity effects, it could be observed that berberine as compared with B. vulgaris exhibited more cytotoxicity against murine macrophages. Results also showed that when parasites were pre-incubated with B. vulgaris their ability to infect murine macrophages was significantly decreased. CONCLUSION: B.vulgaris particularly berberine exhibited potent in vitro leishmanicidal effects against L. tropica and L.infantum. Further works are required to evaluate the antileishmanial effects of B.vulgaris on Leishmania species using clinical settings.