Background: Natural polysaccharides such as chitosan (CS) are widely used as antimicrobial agents. In recent years, and considering that CS has a strong antimicrobial potential, interest has been focused on antimicrobial activity of chitosan nanoparticles (CS NPs). The main factors affecting the antibacterial activity of chitosan include molecular weight (MW) and concentration. In this regard, the aim of this study was to produce various MWs and concentrations of CS NPs, through the ionic gelation method, and investigate their potential anti-parasitic activity against tachyzoites of Toxoplasma gondii RH strain. Materials and methods: The MWs and degree of deacetylation of the CS were characterized using viscometric and acid–base titration methods, respectively. The efficacy of various MWs and concentrations of NPs was assessed by performing in vitro experiments for tachyzoites of T. gondii RH strain, such as MTT assay, scanning electron microscopy, bioassay in mice and PCR. In vivo experiment was carried out in BALB/c mice which were inoculated with tachyzoites of T. gondii RH strain and treated with various MWs of CS NPs. Results: The results of in vitro and in vivo experiments revealed that anti-Toxoplasma activity strengthened as the CS NPs concentration increased and the MW decreased. In vitro experiment showed 100% mortality of tachyzoites at 500 and 1,000 ppm concentrations of low molecular weight (LMW) CS NPs after 180 min and at 2,000 ppm after 120 min. Furthermore, a 100% mortality of tachyzoites was observed at 1,000 and 2,000 ppm concentrations of medium molecular weight (MMW) CS NPs and at 2,000 ppm concentration of high molecular weight (HMW) CS NPs after 180 min. Growth inhibition rates of tachyzoites in peritoneal exudates of mice receiving low, medium and high MWs of CS NPs were found to be 86%, 84% and 79% respectively, compared to those of mice in sulfadiazine treatment group (positive control). Conclusion: Various MWs of CS NPs exhibited great anti-Toxoplasma efficiency against tachyzoites of RH strain, with the greatest efficacy shown by LMW CS NPs in both experiments. It seems that CS NPs can be used as an alternative natural medicine in the treatment of toxoplasmosis.

Amplification of internal transcript spacer 1 of ribosomal RNA (ITS1-RNA) gene followed by RFLP analysis and sequencing was used to identify the causing agents of cutaneous and visceral leishmaniasis (CL and VL) in humans and animal reservoir hosts from various geographical areas in Iran. We also used random amplified polymorphic DNA (RAPD-PCR) to obtain polymorphisms among isolates ofLeishmaniaspp. Totally, 362 suspected human and animal cases including 173 CL, 49 VL, 60 rodents, and 80 domestic dogs were examined forLeishmaniainfection. From 112 culture-positive samples prepared from CL cases, 75 (67%) were infected withL. majorand 37 (33%) withL. tropica. Of the 60 rodents examined, 25 (41.6%) harbored theLeishmaniainfection; 21 were infected withL. majorand 4 withL. turanica. From 49 suspected VL, 29 were positive by direct agglutination test (DAT), whereas microscopy detected parasite in bone marrow of 25 and culture in 28 of the patients. Two VL patients were infected withL. tropicaand 26 withL. infantum. Of the 80 domestic dogs, 56 showed anti-Leishmaniaantibodies with DAT. Of these, 55 were positive by both microscopy and culture. Molecular identity, obtained only for 47 samples, revealedL. infantumin 43 andL. tropicain 4 dogs. The polymorphisms amongL. tropicaandL. majorisolates were 3.6% and 7.3%; the rate among human and canine VL isolates was 2.8% and 9.8%, respectively. Our results showed that at least four differentLeishmaniaspecies with various polymorphisms circulate among humans and animal hosts in Iran.

Background: The aim of this study was to prepare curcumin nanoemulsion (CR-NE) to solve the problems associated with poor water solubility and low bioavailability of CR and to test its efficiency in the treatment of acute and chronic toxoplasmosis in mouse models. Materials and methods: CR-NE 1% was prepared using spontaneous emulsification by soybean as oil phase; a mixture of Tween 80 and Tween 85 as surfactant; ethanol as cosurfactant and distilled water. Particle size and zeta potential of NE were assessed using Nano-ZS90 dynamic light scattering. Stability testing of NE was assessed after storage for 2 months at room temperature. In vivo experiments were carried out using 50 BALB/c mice inoculated with virulent RH strain (type I) and 50 BALB/c mice inoculated with avirulent Tehran strain (type II) of Toxoplasma gondii and treated with CR-NE (1% w/v), CR suspension (CR-S, 1% w/v), and NE without CR (NE-no CR). Results: The mean particle size and zeta potential of CR-NE included 215.66±16.8 nm and -29.46±2.65 mV, respectively, and were stable in particle size after a three freeze–thaw cycle. In acute phase experiment, the survival time of mice infected with RH strain of T. gondii and treated with CR-NE extended from 8 to 10 days postinoculation. The differences were statistically significant between the survival time of mice in CR-NE-treated group compared with negative control group (P<0.001). Furthermore, CR-NE significantly decreased the mean counts of peritoneum tachyzoites from 5,962.5±666 in negative control group to 627.5±73 in CR-NE-treated mice (P<0.001). Growth inhibition rates of tachyzoites in peritoneum of mice receiving CR-NE, CR-S, and NE-no CR included 90%, 21%, and 11%, respectively, compared with negative control group. In chronic phase experiment, the average number and size of tissue cysts significantly decreased to 17.2±15.6 and 31.5±6.26 µm, respectively, in mice inoculated with bradyzoites of T. gondii Tehran strain and treated with CR-NE compared with that in negative control group (P<0.001). Decrease of cyst numbers was verified by downregulation of BAG1 in treatment groups compared with negative control group with a minimum relative expression in CR-NE (1.12±0.28), CR-S (11.76±0.87), and NE-no CR (14.67±0.77), respectively, (P<0.001).
Conclusion: Results from the current study showed the potential of CR-S and CR-NE in treatment of acute and chronic toxoplasmosis in mouse models for the first time. However, CR-NE was more efficient than CR-S, and it seems that CR-NE has a potential formula for the treatment of acute and chronic toxoplasmosis, especially in those with latent bradyzoites in brain.

Toxoplasma gondii is a protozoan parasite with worldwide distribution, infecting a broad-range of humans and warm-blooded animals. In the current study, role of this parasite on secondary sex ratio and risk of miscarriage was investigated. In this cross-sectional study, 850 cord blood samples were collected in Tehran, Iran, 2014–2015. Enzyme-linked immunosorbent assay (ELISA) was used to assess anti-Toxoplasma IgG in samples. Information such as sex of the neonates and age, number of previous pregnancies and history of miscarriage of the mothers were recorded in questionnaires. Logistic regression analysis was used to assess the possible relationship between the latent toxoplasmosis and the highlighted parameters. Logistic regression analysis showed that the odds of having a male neonate in seropositive women is nearly 64% higher than that in seronegative women (OR = 1.64, CI95 = 1.16–2.33, P = 0.005). The odds ratio of having male neonate increased to 2.10 (CI95 = 1.24–3.57, P = 0.006) in high-titer seropositive women, compared to that in seronegative control group. The odds of having a miscarriage history was approximately two and a half times greater in seropositive women than in seronegative ones (OR = 2.45, CI95 = 1.56–3.87, P < 0.001). The odds ratio of having miscarriage increased to 2.76 (CI95 = 1.61–4.73, P < < .001) in low-titer seropositive women, compared to that in seronegative control group. Results of the current study have shown that T. gondii infection affects secondary sex ratio in human offspring and can be addressed as one of the major miscarriage causes in women.